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Journal articles on the topic 'Fibroblasts'
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1
Plaisancié, Pascale, Charline Buisson, Edwin Fouché, Pierre Martin, Céline Noirot, Claire Maslo, Jacques Dupuy, Françoise Guéraud, and Fabrice Pierre. "Study of the colonic epithelial-mesenchymal dialogue through establishment of two activated or not mesenchymal cell lines: Activated and resting ones differentially modulate colonocytes in co-culture." PLOS ONE 17, no. 8 (August 30, 2022): e0273858. http://dx.doi.org/10.1371/journal.pone.0273858.
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Continuous and rapid renewal of the colonic epithelium is crucial to resist the plethora of luminal deleterious agents. Subepithelial fibroblasts contribute to this turnover by regulating epithelial proliferation and differentiation. However, when intestinal homeostasis is disturbed, fibroblasts can acquire an activated phenotype and play a major role in the progression of intestinal pathologies. To evaluate the involvement of fibroblasts in the regulation of colonocytes under homeostatic or pathological conditions, we established resting and activated conditionally immortalized fibroblast cell lines (nF and mF) from mouse colonic mucosa. We then studied the epithelial-mesenchymal interactions between activated or resting fibroblasts and the normal mouse colonocytes (Co) using a co-culture model. Both fibroblastic cell lines were characterized by RT-qPCR, western blot and immunofluorescence assay. Our results showed that nF and mF cells were positive for fibroblastic markers such as vimentin and collagen 1, and negative for cytokeratin 18 and E-cadherin, attesting to their fibroblastic type. They also expressed proteins characteristic of the epithelial stem cell niche such as Grem1, CD90 or Wnt5a. Only rare nF fibroblasts were positive for α-SMA, whereas all mF fibroblasts strongly expressed this marker, supporting that mF cells were activated fibroblasts/myofibroblasts. In coculture, nF fibroblasts and Co cells strongly interacted via paracrine exchanges resulting in BMP4 production in nF fibroblasts, activation of BMP signaling in Co colonocytes, and decreased growth of colonocytes. Activated-type mF fibroblasts did not exert the same effects on Co cells, allowing colonocytes free to proliferate. In conclusion, these two colonic fibroblast lines, associated with Co cells in coculture, should allow to better understand the role of mesenchymal cells in the preservation of homeostasis and the development of intestinal pathologies.
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Caporarello, Nunzia, Jeffrey A. Meridew, Dakota L. Jones, Qi Tan, Andrew J. Haak, Kyoung M. Choi, Logan J. Manlove, Y. S. Prakash, Daniel J. Tschumperlin, and Giovanni Ligresti. "PGC1α repression in IPF fibroblasts drives a pathologic metabolic, secretory and fibrogenic state." Thorax 74, no. 8 (June 10, 2019): 749–60. http://dx.doi.org/10.1136/thoraxjnl-2019-213064.
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Idiopathic pulmonary fibrosis (IPF) is a fatal ageing-related disease linked to mitochondrial dysfunction. The present study aimed to determine whether peroxisome proliferator activated receptor gamma co-activator 1-alpha (PPARGC1A, encoding PGC1α), a master regulator of mitochondrial biogenesis, is diminished in IPF and controls pathologic fibroblast activation. Primary human IPF, control lung fibroblasts and fibroblasts sorted from bleomycin-injured mice were used to evaluate the expression and function of PGC1α. In vitro PGC1α manipulation was performed by small interfering RNA knockdown or overexpression. Fibroblast activation was assessed by quantitative PCR, Western blotting, matrix deposition, secreted cytokine array, immunofluorescence and traction force microscopy. Mitochondrial function was assessed by Seahorse analyzer and mitochondria mass and number by flow cytometry, mitochondrial DNA quantification and transmission electron microscopy (TEM). We found that PGC1α levels are stably repressed in IPF fibroblasts. After bleomycin injury in young mice, PGC1α expression drops transiently but then increases prior to fibrosis resolution. In contrast, PGC1α expression fails to recover in aged mice with persistent fibrosis. PGC1α knockdown alone in normal human lung fibroblasts reduces mitochondrial mass and function while enhancing contractile and matrix synthetic fibroblast activation, senescence-related gene expression and soluble profibrotic and prosenescence signalling. Re-expression of PGC1α in IPF fibroblasts ameliorates all of these pathological cellular functions. Pharmacological treatment of IPF fibroblasts with rosiglitazone, but not thyroid hormone, elevated PGC1α expression and attenuated fibroblast activation. The sustained repression of PGC1α and beneficial effects of its rescue in IPF fibroblasts identifies PGC1α as an important regulator of the fibroblast’s pathological state in IPF.
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Roy, Bibhas, Luezhen Yuan, Yaelim Lee, Aradhana Bharti, Aninda Mitra, and G. V. Shivashankar. "Fibroblast rejuvenation by mechanical reprogramming and redifferentiation." Proceedings of the National Academy of Sciences 117, no. 19 (April 29, 2020): 10131–41. http://dx.doi.org/10.1073/pnas.1911497117.
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Over the course of the aging process, fibroblasts lose contractility, leading to reduced connective-tissue stiffness. A promising therapeutic avenue for functional rejuvenation of connective tissue is reprogrammed fibroblast replacement, although major hurdles still remain. Toward this, we recently demonstrated that the laterally confined growth of fibroblasts on micropatterned substrates induces stem-cell-like spheroids. In this study, we embedded these partially reprogrammed spheroids in collagen-I matrices of varying densities, mimicking different three-dimensional (3D) tissue constraints. In response to such matrix constraints, these spheroids regained their fibroblastic properties and sprouted to form 3D connective-tissue networks. Interestingly, we found that these differentiated fibroblasts exhibit reduced DNA damage, enhanced cytoskeletal gene expression, and actomyosin contractility. In addition, the rejuvenated fibroblasts show increased matrix protein (fibronectin and laminin) deposition and collagen remodeling compared to the parental fibroblast tissue network. Furthermore, we show that the partially reprogrammed cells have comparatively open chromatin compaction states and may be more poised to redifferentiate into contractile fibroblasts in 3D-collagen matrix. Collectively, our results highlight efficient fibroblast rejuvenation through laterally confined reprogramming, which has important implications in regenerative medicine.
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McDonald, Lindsay T., Meenal Mehrotra, and Amanda C. LaRue. "Hematopoietic Origin of Murine Lung Fibroblasts." Stem Cells International 2015 (2015): 1–11. http://dx.doi.org/10.1155/2015/159713.
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Multiple origins, including the bone marrow, have been suggested to contribute to fibroblast populations in the lung. Using bone marrow reconstitution strategies, the present study tested the hypothesis that the bone marrow hematopoietic stem cell (HSC) gives rise to lung tissue fibroblastsin vivo. Data demonstrate that the nonadherent bone marrow fraction is enriched for CD45+HSC-derived cells and was able to reconstitute hematopoiesis in lethally irradiated animals. Analysis of peripheral blood and lung tissues from engrafted mice demonstrated the ability of this population to give rise to CD45+/Discoidin-Domain Receptor-2+(DDR2) circulating fibroblast precursors (CFPs) in blood and fibroblast populations in lung. An HSC origin for lung fibroblasts was confirmed using a novel clonal cell transplantation method in which the bone marrow is reconstituted by a clonal population derived from a single HSC. Together, these findings provide evidence for an HSC contribution to lung fibroblasts and demonstrate a circulating intermediate through the CD45+/DDR2+HSC-derived CFP.
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Strutz, F., H. Okada, C. W. Lo, T. Danoff, R. L. Carone, J. E. Tomaszewski, and E. G. Neilson. "Identification and characterization of a fibroblast marker: FSP1." Journal of Cell Biology 130, no. 2 (July 15, 1995): 393–405. http://dx.doi.org/10.1083/jcb.130.2.393.
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We performed subtractive and differential hybridization for transcript comparison between murine fibroblasts and isogenic epithelium, and observed only a few novel intracellular genes which were relatively specific for fibroblasts. One such gene encodes a filament-associated, calcium-binding protein, fibroblast-specific protein 1 (FSP1). The promoter/enhancer region driving this gene is active in fibroblasts but not in epithelium, mesangial cells or embryonic endoderm. During development, FSP1 is first detected by in situ hybridization after day 8.5 as a postgastrulation event, and is associated with cells of mesenchymal origin or of fibroblastic phenotype. Polyclonal antiserum raised to recombinant FSP1 protein stained the cytoplasm of fibroblasts, but not epithelium. Only occasional cells stain with specific anti-FSP1 antibodies in normal parenchymal tissue. However, in kidneys fibrosing from persistent inflammation, many fibroblasts could be identified in interstitial sites of collagen deposition and also in tubular epithelium adjacent to the inflammatory process. This pattern of anti-FSP1 staining during tissue fibrosis suggests, as a hypothesis, that fibroblasts in some cases arise, as needed, from the local conversion of epithelium. Consistent with this notion that FSP1 may be involved in the transition from epithelium to fibroblasts are experiments in which the in vitro overexpression of FSP1 cDNA in tubular epithelium is accompanied by conversion to a mesenchymal phenotype, as characterized by a more stellate and elongated fibroblast-like appearance, a reduction in cytokeratin, and new expression of vimentin. Similarly, tubular epithelium submerged in type I collagen gels exhibited the conversion to a fibroblast phenotype which includes de novo expression of FSP1 and vimentin. Use of the FSP1 marker, therefore, should further facilitate both the in vivo studies of fibrogenesis and the mapping of cell fate among fibroblasts.
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Lauritano, Dorina, Alberta Lucchese, Dario Di Stasio, Fedora Della Vella, Francesca Cura, Annalisa Palmieri, and Francesco Carinci. "Molecular Aspects of Drug-Induced Gingival Overgrowth: An In Vitro Study on Amlodipine and Gingival Fibroblasts." International Journal of Molecular Sciences 20, no. 8 (April 25, 2019): 2047. http://dx.doi.org/10.3390/ijms20082047.
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Gingival overgrowth is a serious side effect that accompanies the use of amlodipine. Several conflicting theories have been proposed to explain the fibroblast’s function in gingival overgrowth. To determine whether amlodipine alters the fibrotic response, we investigated its effects on treated gingival fibroblast gene expression as compared with untreated cells. Materials and Methods: Fibroblasts from ATCC® Cell Lines were incubated with amlodipine. The gene expression levels of 12 genes belonging to the “Extracellular Matrix and Adhesion Molecules” pathway was investigated in treated fibroblasts cell culture, as compared with untreated cells, by real time PCR. Results: Most of the significant genes were up-regulated. (CTNND2, COL4A1, ITGA2, ITGA7, MMP10, MMP11, MMP12, MMP26) except for COL7A1, LAMB1, MMP8, and MMP16, which were down-regulated. Conclusion: These results seem to demonstrate that amlodipine has an effect on the extracellular matrix of gingival fibroblast. In the future, it would be interesting to understand the possible effect of the drug on fibroblasts of patients with amlodipine-induced gingival hyperplasia.
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Osorio, José Henry. "Producción de metabolitos en fibroblastos incubados con acido oleico deuterado./Metabolite production of fibroblasts incubated with deuterated oleic acid." Archivos de Medicina (Manizales) 13, no. 2 (December 15, 2013): 202–7. http://dx.doi.org/10.30554/archmed.13.2.24.2013.
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Objetivo: los estudios in vitro, que buscan intermediarios de la degradación mitocondrial de ácidos grasos, facilitan el diagnóstico de alteraciones hereditarias o adquiridas en esa ruta metabólica, el presente estudio analiza la produccion de metabolitos en fibroblastos incubados con acido oleico deuterado. Materiales y métodos: fibroblastos de personas sin antecedentes de alteraciones metabólicas, fueron incubados en presencia de ácido oléico deuterado, y sus metabolitos fueron analizados mediante cromatografía de gases acoplada a espectrometría de masas (GC-MS) Resultados: Se encontró un perfil característico luego de la incubación de este sustrato por fibroblastos. Conclusiones: Este sustrato podría ser usado para realizar estudios in vitro de algunas deficiencias de la β-oxidació mitocondrial de ácidos grasos, mediante la comparación de fibroblastos normales vs fibroblastos de pacientes que presenten deficiciencias de esa vía metabólica. Objective: In vitro studies for searching intermediates of mitochondrial fatty acid degradation,are a tool for diagnosis of hereditary or adquired alterations of the above mentionedmetabolic pathway. The present work analizes the metabolite production in fibroblastsincubated with deuterated oleic acid. Materials and methods: fibroblasts of personswithout antecedents of metabolic alterations, were incubated with deuterated oleic acidand its metabolites were analyzed using gas chromatograpy-mass spectrometry (GCMS).Results. It was found a characteristic profile after incubation of fibroblats with thissubstrate. Conclusion: this substrate could be used to perform in vitro studies of somemitochondrial fatty acid β-oxidación deficiencies, by comparison of normal fibroblastsvs fibroblats of patients who present defiencies of the above named metabolic pathway
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Kurniawati, Yuli, Sudigdo Adi, Achadiyani Achadiyani, Oki Suwarsa, Dimas Erlangga, and Tenny Putri. "KULTUR PRIMER FIBROBLAS: PENELITIAN PENDAHULUAN." Majalah Kedokteran Andalas 38, no. 1 (May 28, 2015): 33. http://dx.doi.org/10.22338/mka.v38.i1.p33-40.2015.
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AbstrakKultur sel fibroblas banyak digunakan untuk penelitian proses penyembuhan luka dan penuaankulit. Metode ini digunakan untuk melihat perkembangan sel, proliferasi kinetik seluler, sertabiosintesis komponen matriks ekstraseluler. Penelitian pendahuluan ini dilakukan untuk optimasiteknik laboratorium serta berbagai kendala yang didapatkan saat kultur fibroblas. Kultur primerfibroblas dibagi menjadi 2 jenis sampel yaitu sampel yang berasal dari embrio mencit usia 7,5–9,5 hari, dan kulit pasien keloid. Sampel dari embrio mencit dilakukan kultur primer denganmetode dissociated fibroblast. Sampel jaringan keloid dan kulit normal dikultur dengan metodeskin explant. Fibroblas yang berasal dari kultur primer embrio mencit tumbuh baik sehinggadapat dilakukan subkultur dan disimpan di dalam nitrogen cair suhu -198°C. Fibroblas yangberasal dari sampel keloid pertama tumbuh sesuai pola pertumbuhan fibroblas, namun padasampel kedua terdapat kontaminasi Paecilomyces sp. yang merupakan salah satu jenis jamurkontaminan. Sel fibroblas mudah untuk dikultur karena memiliki kemampuan tumbuh danmelekat yang tinggi serta regenerasi cepat, namun penelitian lebih lanjut untuk optimasi teknikkultur dan pencegahan kontaminasi masih dibutuhkan sehingga sel dapat tumbuh baik.AbstractFibroblast cell culture method has been used for wound healing and skin aging studies. Thismethod was used for cell development imaging study, celullar kinetic proliferation andextracelullar matrix component biosynthesis. This preeliminary study was done for laboratoricaltechnic optimation as well as problems appeared in fibroblast culture. Fibroblasts primary culturewas divided into 2 type of samples, from 7.5-9.5-day-mice embryo and keloid-patient skin.Primary culture with dissociated fibroblast method was done for mice embryo sample. Keloidtissue sample and normal skin were cultured with skin explant method. Fibroblasts that weretaken from mice embryo primary culture grew well therefore subculture can be done and kept in -198°C liquid nitrogen storage. Fibroblasts that were taken from first keloid sample grewaccording to fibroblast growth pattern, but, there was contamination with Paecilomyces sp. whichwas one of the contaminating fungi. Fibroblast cells are easy to be cultured as they have growthability and high adhesion capability as well as rapid regeneration, but, further study for culturedtechnical optimation and contamination prevention are still neededthereforethe cells can growwell.
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Cai, Zhou, Hua Guo, Jing Qian, Wei Liu, Yuanyuan Li, Liang Yuan, You Zhou, et al. "Effects of bone morphogenetic protein 4 on TGF-β1-induced cell proliferation, apoptosis, activation and differentiation in mouse lung fibroblasts via ERK/p38 MAPK signaling pathway." PeerJ 10 (July 27, 2022): e13775. http://dx.doi.org/10.7717/peerj.13775.
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Fibroblasts, in particular myofibroblasts, are the critical effector cells in idiopathic pulmonary fibrosis (IPF), a deadly lung disease characterized by abnormal lung remodeling and the formation of “fibroblastic foci”. Aberrant activation of TGF-β1 is frequently encountered and promotes fibroblast proliferation, activation, and differentiation in pulmonary fibrosis. Hence, the inhibition of TGF-β1-induced lung fibroblast activation holds promise as a therapeutic strategy for IPF. The present study aimed to investigate the potential effect and underlying mechanisms of bone morphogenetic protein 4 (BMP4) on TGF-β1-induced proliferation, apoptosis, activation and myofibroblast differentiation of adult lung fibroblasts. Here, we demonstrated that BMP4 expression was significantly decreased in TGF-β1-stimulated mouse primary lung fibroblasts (PLFs). BMP4 inhibited proliferation and apoptosis resistance of TGF-β1-stimulated mouse PLFs. BMP4 suppressed TGF-β1-induced fibroblast activation and differentiation in mouse PLFs. We also found that BMP4 inhibited TGF-β1-induced ERK and p38 MAPK phosphorylation. Our findings indicate that BMP4 exerts its anti-fibrotic effects by regulating fibroblast proliferation, apoptosis, activation and differentiation via the inhibition of the ERK/p38 MAPK signaling pathway, and thus has a potential for the treatment of pulmonary fibrosis.
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Jara, Paul, Jazmin Calyeca, Yair Romero, Luis Plácido, Guoying Yu, Naftali Kaminski, Vilma Maldonado, José Cisneros, Moisés Selman, and Annie Pardo. "Matrix metalloproteinase (MMP)-19-deficient fibroblasts display a profibrotic phenotype." American Journal of Physiology-Lung Cellular and Molecular Physiology 308, no. 6 (March 15, 2015): L511—L522. http://dx.doi.org/10.1152/ajplung.00043.2014.
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Idiopathic pulmonary fibrosis (IPF) is a progressive and usually lethal interstitial lung disease of unknown etiology characterized by aberrant activation of epithelial cells that induce the migration, proliferation and activation of fibroblasts. The resulting distinctive fibroblastic/myofibroblastic foci are responsible for the excessive extracellular matrix (ECM) production and abnormal lung remodeling. We have recently found that matrix metalloproteinase 19 (MMP-19)-deficient ( Mmp19−/−) mice develop an exaggerated bleomycin-induced lung fibrosis, but the mechanisms are unclear. In this study, we explored the effect of MMP-19 deficiency on fibroblast gene expression and cell behavior. Microarray analysis of Mmp19−/− lung fibroblasts revealed the dysregulation of several profibrotic pathways, including ECM formation, migration, proliferation, and autophagy. Functional studies confirmed these findings. Compared with wild-type mice, Mmp19−/− lung fibroblasts showed increased α1 (I) collagen gene and collagen protein production at baseline and after transforming growth factor-β treatment and increased smooth muscle-α actin expression ( P < 0.05). Likewise, Mmp19-deficient lung fibroblasts showed a significant increase in proliferation ( P < 0.01) and in transmigration and locomotion over Boyden chambers coated with type I collagen or with Matrigel ( P < 0.05). These findings suggest that, in lung fibroblasts, MMP-19 has strong regulatory effects on the synthesis of key ECM components, on fibroblast to myofibroblast differentiation, and in migration and proliferation.
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Nho, Richard Seonghun, Jintaek Im, Yen-Yi Ho, and Polla Hergert. "MicroRNA-96 inhibits FoxO3a function in IPF fibroblasts on type I collagen matrix." American Journal of Physiology-Lung Cellular and Molecular Physiology 307, no. 8 (October 15, 2014): L632—L642. http://dx.doi.org/10.1152/ajplung.00127.2014.
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Idiopathic pulmonary fibrosis (IPF) is a lethal and progressive lung disease characterized by persistent (myo)fibroblasts and the relentless accumulation of collagen matrix. Unlike normal lung fibroblasts, IPF lung fibroblasts have suppressed forkhead box O3a (FoxO3a) activity, which allows them to expand in this diseased environment. microRNA-96 (miR-96) has recently been found to directly bind to the 3′-untranslated region of FoxO3a mRNA, which subsequently inhibits its function. We examined whether aberrantly low FoxO3a expression is in part due to increased miR-96 levels in IPF fibroblasts on polymerized collagen, thereby causing IPF fibroblasts to maintain their pathological properties. miR-96 expression was upregulated in IPF fibroblasts compared with control fibroblasts when cultured on collagen. In contrast, FoxO3a mRNA levels were reduced in most IPF fibroblasts. However, when miR-96 function was inhibited, FoxO3a mRNA and protein expression were increased, suppressing IPF fibroblast proliferation and promoting their cell death in a dose-dependent fashion. Likewise, FoxO3a and its target proteins p21, p27, and Bim expression was also increased in the presence of a miR-96 inhibitor in IPF fibroblasts. However, when control fibroblasts were treated with miR-96 mimic, FoxO3a, p27, p21, and Bim mRNA and protein levels were decreased. In situ hybridization analysis further revealed the presence of enhanced miR-96 expression in cells within the fibroblastic foci of IPF lung tissue. Our results suggest that when IPF fibroblasts interact with collagen-rich matrix, pathologically altered miR-96 expression inhibits FoxO3a function, causing IPF fibroblasts to maintain their pathological phenotype, which may contribute to the progression of IPF.
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Fontenay-Roupie, M., E. Dupuy, E. Berrou, G. Tobelem, and M. Bryckaert. "Increased proliferation of bone marrow-derived fibroblasts in primitive hypertrophic osteoarthropathy with severe myelofibrosis." Blood 85, no. 11 (June 1, 1995): 3229–38. http://dx.doi.org/10.1182/blood.v85.11.3229.bloodjournal85113229.
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Pachydermoperiostosis or primary hypertrophic osteoarthropathy (HOA) is a rare congenital growth disorder of connective tissue. We report a case of severe myelofibrosis in a patient with HOA. When cultured in vitro, patient bone marrow-derived fibroblasts displayed a high proliferative potential with a shortened doubling time (24 hours v 36 to 48 hours for normal fibroblasts). The role of platelet-derived growth factor (PDGF), previously implicated in the pathogenesis of secondary acquired myelofibrosis, was studied. HOA fibroblasts expressed an increased number of PDGF-BB binding sites (300,000 sites/cell v 200,000 sites/cell for normal fibroblasts) without any modification of affinity. The increased expression of PDGF-R beta appeared to result from an accelerated rate of PDGF-R beta resynthesis with normal kinetics of endocytosis. As a consequence, a several-fold increase of PDGF-R beta tyrosine kinase activity was observed. No autocrine mechanism of growth was suspected as neither spontaneous PDGF-R beta autophosphorylation nor mitogenic activity in HOA fibroblast-conditioned medium was detected. Patient serum and platelet lysate were less potent than controls in inducing [3H]thymidine incorporation into HOA fibroblasts. This was inconsistent with a paracrine mechanism of growth. In vitro, human serum or PDGF-BB were not more mitogenic for HOA than normal fibroblasts. High levels of cyclin D1, a putative oncogene, were detected in serum-deprived HOA fibroblasts. Cyclin D1 overexpression could be implicated in the accelerated growth of these cells. Our results suggest that the mechanism of fibroblastic proliferation observed in this case of myelofibrosis might differ from those reported in other acquired myeloproliferative syndromes and could be associated with an intrinsic abnormality of HOA fibroblast growth.
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Cunningham, Michael F., Neil G. Docherty, John P. Burke, and P. Ronan O'Connell. "S100A4 expression is increased in stricture fibroblasts from patients with fibrostenosing Crohn's disease and promotes intestinal fibroblast migration." American Journal of Physiology-Gastrointestinal and Liver Physiology 299, no. 2 (August 2010): G457—G466. http://dx.doi.org/10.1152/ajpgi.00351.2009.
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Fibroblasts represent the key cell type in fibrostenosing Crohn's disease (FCD) pathogenesis. S100A4 is an EF-hand calcium-binding protein family member, implicated in epithelial-mesenchymal transition and as a marker of activated T lymphocytes and fibroblasts in chronic tissue remodeling. The aim of this study was to examine the expression profile of S100A4 in the resected ileum of patients with FCD. Mucosa, seromuscular explants, and transmural biopsies were harvested from diseased and proximal, macroscopically normal margins of ileocecal resections from patients with FCD. Samples were processed for histochemistry, immunohistochemistry, real-time RT-PCR, Western blotting, and transmission electron microscopy. Primary explant cultures of seromuscular fibroblasts were exposed to transforming growth factor (TGF)-β1 (1 ng/ml), and S100A4 expression and scratch wound-healing activity were assessed at 24 h. CCD-18Co fibroblasts were transfected with S100A4 small interfering RNA, treated with TGF-β1 (1 ng/ml) for 30 min or 24 h, and then assessed for S100A4 and Smad3 expression and scratch wound-healing activity. S100A4 expression was increased in stricture mucosa, in the lamina propria, and in CD3-positive intraepithelial CD3-positive T lymphocytes. Fibroblastic S100A4 staining was observed in seromuscular scar tissue. Stricture fibroblast explant culture showed significant upregulation of S100A4 expression. TGF-β1 increased S100A4 expression in cultured ileal fibroblasts. In CCD-18Co fibroblasts, S100A4 small interfering RNA inhibited scratch wound healing and modestly inhibited Smad3 activation. S100A4 expression is increased in fibroblasts, as well as immune cells, in Crohn's disease stricture and induced by TGF-β1. Results from knockdown experiments indicate a potential role for S100A4 in mediating intestinal fibroblast migration.
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Ivey, Malina J., Jill T. Kuwabara, Kara L. Riggsbee, and Michelle D. Tallquist. "Platelet-derived growth factor receptor-α is essential for cardiac fibroblast survival." American Journal of Physiology-Heart and Circulatory Physiology 317, no. 2 (August 1, 2019): H330—H344. http://dx.doi.org/10.1152/ajpheart.00054.2019.
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Platelet-derived growth factor receptor α (PDGFRα), a receptor tyrosine kinase required for cardiac fibroblast development, is uniquely expressed by fibroblasts in the adult heart. Despite the consensus that PDGFRα is expressed in adult cardiac fibroblasts, we know little about its function when these cells are at rest. Here, we demonstrate that loss of PDGFRα in cardiac fibroblasts resulted in a rapid reduction of resident fibroblasts. Furthermore, we observe that phosphatidylinositol 3-kinase signaling was required for PDGFRα-dependent fibroblast maintenance. Interestingly, this reduced number of fibroblasts was maintained long-term, suggesting that there is no homeostatic mechanism to monitor fibroblast numbers and restore hearts to wild-type levels. Although we did not observe any systolic functional changes in hearts with depleted fibroblasts, the basement membrane and microvasculature of these hearts were perturbed. Through in vitro analyses, we showed that PDGFRα signaling inhibition resulted in an increase in fibroblast cell death, and PDGFRα stimulation led to increased levels of the cell survival factor activating transcription factor 3. Our data reveal a unique role for PDGFRα signaling in fibroblast maintenance and illustrate that a 50% loss in cardiac fibroblasts does not result in lethality. NEW & NOTEWORTHY Platelet-derived growth factor receptor α (PDGFRα) is required in developing cardiac fibroblasts, but a functional role in adult, quiescent fibroblasts has not been identified. Here, we demonstrate that PDGFRα signaling is essential for cardiac fibroblast maintenance and that there are no homeostatic mechanisms to regulate fibroblast numbers in the heart. PDGFR signaling is generally considered mitogenic in fibroblasts, but these data suggest that this receptor may direct different cellular processes depending on the cell’s maturation and activation status.
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Douillet, Camille, Marc Nicodeme, Loïc Hermant, Vanessa Bergeron, Fabien Guillemot, Jean-Christophe Fricain, Hugo Oliveira, and Mikael Garcia. "From local to global matrix organization by fibroblasts: a 4D laser-assisted bioprinting approach." Biofabrication 14, no. 2 (January 24, 2022): 025006. http://dx.doi.org/10.1088/1758-5090/ac40ed.
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Abstract Fibroblasts and myofibroblasts play a central role in skin homeostasis through dermal organization and maintenance. Nonetheless, the dynamic interactions between (myo)fibroblasts and the extracellular matrix (ECM) remain poorly exploited in skin repair strategies. Indeed, there is still an unmet need for soft tissue models allowing to study the spatial-temporal remodeling properties of (myo)fibroblasts. In vivo, wound healing studies in animals are limited by species specificity. In vitro, most models rely on collagen gels reorganized by randomly distributed fibroblasts. But biofabrication technologies have significantly evolved over the past ten years. High-resolution bioprinting now allows to investigate various cellular micropatterns and the emergent tissue organizations over time. In order to harness the full dynamic properties of cells and active biomaterials, it is essential to consider ‘time’ as the 4th dimension in soft tissue design. Following this 4D bioprinting approach, we aimed to develop a novel model that could replicate fibroblast dynamic remodeling in vitro. For this purpose, (myo)fibroblasts were patterned on collagen gels with laser-assisted bioprinting (LAB) to study the generated matrix deformations and reorganizations. First, distinct populations, mainly composed of fibroblasts or myofibroblasts, were established in vitro to account for the variety of fibroblastic remodeling properties. Then, LAB was used to organize both populations on collagen gels in even isotropic patterns with high resolution, high density and high viability. With maturation, bioprinted patterns of fibroblasts and myofibroblasts reorganized into dispersed or aggregated cells, respectively. Stress-release contraction assays revealed that these phenotype-specific pattern maturations were associated with distinct lattice tension states. The two populations were then patterned in anisotropic rows in order to direct the cell-generated deformations and to orient global matrix remodeling. Only maturation of anisotropic fibroblast patterns, but not myofibroblasts, resulted in collagen anisotropic reorganizations both at tissue-scale, with lattice contraction, and at microscale, with embedded microbead displacements. Following a 4D bioprinting approach, LAB patterning enabled to elicit and orient the dynamic matrix remodeling mechanisms of distinct fibroblastic populations and organizations on collagen. For future studies, this method provides a new versatile tool to investigate in vitro dermal organizations and properties, processes of remodeling in healing, and new treatment opportunities.
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Tverdokhlib, I. V., and Yu V. Silkina. "Dermal fibroblasts: the terminology, heterogeneity of subpopulations and common properties." Morphologia 14, no. 4 (September 25, 2021): 108–14. http://dx.doi.org/10.26641/1997-9665.2020.4.108-114.
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Dermal fibroblasts are a dynamic and diverse population of cells whose functions in skin in many respects remain unknown. Normal adult human skin contains at least three distinct subpopulations of fibroblasts, which occupy unique niches in the dermis. Fibroblasts from each of these niches exhibit distinctive differences when cultured separately. Specific differences in fibroblast histophysiology are evident in papillary dermal fibroblasts, which reside in the superficial dermis, and reticular fibroblasts, which reside in the deep dermis. Both of these subpopulations of fibroblasts differ from the fibroblasts that are associated with hair follicles. Fibroblasts engage in fibroblast-epidermal interactions during hair development and in interfollicular regions of skin. They also play an important role in cutaneous structural transformations.
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Sun, Xiaoyan, Huimin Zhu, Wenjuan Li, Li Zhao, Wenhua Li, Xiaoyong Li, and Zhenwei Xie. "Small extracellular vesicles secreted by vaginal fibroblasts exert inhibitory effect in female stress urinary incontinence through regulating the function of fibroblasts." PLOS ONE 16, no. 4 (April 9, 2021): e0249977. http://dx.doi.org/10.1371/journal.pone.0249977.
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Stress urinary incontinence (SUI) is a common condition in women and associated with extra-cellular matrix (ECM) reconstruction, which is mainly regulated by fibroblasts. However, the underlying mechanism remains obscure. Small extracellular vesicles (sEVs) play fundamental biological roles in various cellular functions. Some studies suggested that the sEVs were involved in the metabolism of ECM and the function of fibroblasts. The purpose of our study was to investigate the effect of sEVs secreted by vaginal fibroblasts on the pathogenesis of SUI. We showed that the fibroblasts of female anterior vaginal wall secreted sEVs. Moreover, fibroblasts of females with SUI had significantly elevated secretion of sEVs. The collagen contents, proliferation and migration capacity of fibroblasts were decreased when fibroblasts were co-cultured with fibroblasts-derived sEVs (fibroblast-sEVs) from SUI patients. Proteomic analysis revealed that fibroblast-sEVs contained various differentially expressed proteins including TIMP2, TGF-β and ABCC4, which were involved in signaling pathways of fibroblasts regulation. Therefore, we suggested that fibroblast-sEVs contributed to the pathogenesis of SUI through various proteins including TIMP2, TGF-β and ABCC4.
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Lebel, Mégane, Dominic O. Cliche, Martine Charbonneau, Damien Adam, Emmanuelle Brochiero, Claire M. Dubois, and André M. Cantin. "Invadosome Formation by Lung Fibroblasts in Idiopathic Pulmonary Fibrosis." International Journal of Molecular Sciences 24, no. 1 (December 28, 2022): 499. http://dx.doi.org/10.3390/ijms24010499.
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Idiopathic pulmonary fibrosis (IPF) is characterized by abnormal fibroblast accumulation in the lung leading to extracellular matrix deposition and remodeling that compromise lung function. However, the mechanisms of interstitial invasion and remodeling by lung fibroblasts remain poorly understood. The invadosomes, initially described in cancer cells, consist of actin-based adhesive structures that coordinate with numerous other proteins to form a membrane protrusion capable of degrading the extracellular matrix to promote their invasive phenotype. In this regard, we hypothesized that invadosome formation may be increased in lung fibroblasts from patients with IPF. Public RNAseq datasets from control and IPF lung tissues were used to identify differentially expressed genes associated with invadosomes. Lung fibroblasts isolated from bleomycin-exposed mice and IPF patients were seeded with and without the two approved drugs for treating IPF, nintedanib or pirfenidone on fluorescent gelatin-coated coverslips for invadosome assays. Several matrix and invadosome-associated genes were increased in IPF tissues and in IPF fibroblastic foci. Invadosome formation was significantly increased in lung fibroblasts isolated from bleomycin-exposed mice and IPF patients. The degree of lung fibrosis found in IPF tissues correlated strongly with invadosome production by neighboring cells. Nintedanib suppressed IPF and PDGF-activated lung fibroblast invadosome formation, an event associated with inhibition of the PDGFR/PI3K/Akt pathway and TKS5 expression. Fibroblasts derived from IPF lung tissues express a pro-invadosomal phenotype, which correlates with the severity of fibrosis and is responsive to antifibrotic treatment.
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Melboucy-Belkhir, Sara, Pauline Pradère, Sara Tadbiri, Stéfanie Habib, Antoine Bacrot, Stéphanie Brayer, Bernard Mari, et al. "Forkhead Box F1 represses cell growth and inhibits COL1 and ARPC2 expression in lung fibroblasts in vitro." American Journal of Physiology-Lung Cellular and Molecular Physiology 307, no. 11 (December 1, 2014): L838—L847. http://dx.doi.org/10.1152/ajplung.00012.2014.
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Aberrant expression of master phenotype regulators or alterations in their downstream pathways in lung fibroblasts may play a central role in idiopathic pulmonary fibrosis (IPF). Interrogating IPF fibroblast transcriptome datasets, we identified Forkhead Box F1 (FOXF1), a DNA-binding protein required for lung development, as a candidate actor in IPF. Thus we determined FOXF1 expression levels in fibroblasts cultured from normal or IPF lungs in vitro, and explored FOXF1 functions in these cells using transient and stable loss-of-function and gain-of-function models. FOXF1 mRNA and protein were expressed at higher levels in IPF fibroblasts compared with normal fibroblasts (mRNA: +44%, protein: +77%). Immunohistochemistry showed FOXF1 expression in nuclei of bronchial smooth muscle cells, endothelial cells, and lung fibroblasts including fibroblastic foci of IPF lungs. In normal lung fibroblasts, FOXF1 repressed cell growth and expression of collagen-1 (COL1) and actin-related protein 2/3 complex, subunit 2 (ARPC2). ARPC2 knockdown inhibited cell growth and COL1 expression, consistent with FOXF1 acting in part through ARPC2 repression. In IPF fibroblasts, COL1 and ARPC2 repression by FOXF1 was blunted, and FOXF1 did not repress growth. FOXF1 expression was induced by the antifibrotic mediator prostaglandin E2 and repressed by the profibrotic cytokine transforming growth factor-β1 in both normal and IPF lung fibroblasts. Ex vivo, FOXF1 knockdown conferred CCL-210 lung fibroblasts the ability to implant in uninjured mouse lungs. In conclusion, FOXF1 functions and regulation were consistent with participation in antifibrotic pathways. Alterations of pathways downstream of FOXF1 may participate to fibrogenesis in IPF fibroblasts.
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Zhang, Yihe, Bingjie Jiang, and Meng Huee Lee. "A Novel 3D Model for Visualization and Tracking of Fibroblast-Guided Directional Cancer Cell Migration." Biology 9, no. 10 (October 8, 2020): 328. http://dx.doi.org/10.3390/biology9100328.
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Stromal fibroblasts surrounding cancer cells are a major and important constituent of the tumor microenvironment not least because they contain cancer-associated fibroblasts, a unique fibroblastic cell type that promotes tumorigenicity through extracellular matrix remodeling and secretion of soluble factors that stimulate cell differentiation and invasion. Despite much progress made in understanding the molecular mechanisms that underpin fibroblast–tumor cross-talk, relatively little is known about the way the two cell types interact from a physical contact perspective. In this study, we report a novel three-dimensional dumbbell model that would allow the physical interaction between the fibroblasts and cancer cells to be visualized and monitored by microscopy. To achieve the effect, the fibroblasts and cancer cells in 50% Matrigel suspension were seeded as independent droplets in separation from each other. To allow for cell migration and interaction, a narrow passage of Matrigel causeway was constructed in between the droplets, effectively molding the gel into the shape of a dumbbell. Under time-lapse microscopy, we were able to visualize and image the entire process of fibroblast-guided cancer cell migration event, from initial vessel-like structure formation by the fibroblasts to their subsequent invasion across the causeway, attracting and trapping the cancer cells in the process. Upon prolonged culture, the entire population of fibroblasts eventually infiltrated across the passage and condensed into a spheroid-like cell mass, encapsulating the bulk of the cancer cell population within. Suitable for almost every cell type, our model has the potential for a wider application as it can be adapted for use in drug screening and the study of cellular factors involved in cell–cell attraction.
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Cheng, Maye F., Faizah S. Abdullah, and Matthew B. Buechler. "Essential growth factor receptors for fibroblast homeostasis and activation." F1000Research 13 (February 19, 2024): 120. http://dx.doi.org/10.12688/f1000research.143514.1.
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Fibroblasts are cells of mesenchymal origin that are found throughout the body. While these cells have several functions, their integral roles include maintaining tissue architecture through the production of key extracellular matrix components, and participation in wound healing after injury. Fibroblasts are also key mediators in disease progression during fibrosis, cancer, and other inflammatory diseases. Under these perturbed states, fibroblasts can activate into inflammatory fibroblasts or contractile myofibroblasts. Fibroblasts require various growth factors and mitogenic molecules for survival, proliferation, and differentiation. While the activity of mitogenic growth factors on fibroblasts in vitro was characterized as early as the 1970s, the proliferation and differentiation effects of growth factors on these cells in vivo are unclear. Moreover, recent work exploring the heterogeneity of fibroblasts raises questions as to whether all fibroblast cell states exhibit the same growth factor requirements. Here, we will examine and review existing growth factors known to influence fibroblast homeostasis to begin unpacking the potential growth factors that may influence in vivo fibroblast cell states.
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Elganainy, Dalia, Andressa Dias Costa, Alexander Jordan, Suryun Kim, Sara A. Väyrynen, Hannah L. Williams, Chen Yuan, et al. "Abstract A035: Stromal composition, fibroblast heterogeneity and spatial organization in pancreatic adenocarcinoma." Cancer Research 84, no. 2_Supplement (January 16, 2024): A035. http://dx.doi.org/10.1158/1538-7445.panca2023-a035.
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Abstract Background: The pancreatic ductal adenocarcinoma (PDAC) microenvironment is characterized by highly abundant stromal fibroblasts that are hypothesized to influence tumor behavior. While recent studies have identified multiple populations of stromal fibroblasts with divergent roles in PDAC biology, stromal composition remains incompletely characterized in human tumors, including the landscape of fibroblast subtypes and their spatial organization. Design: We designed multiplex immunofluorescence assays to quantify tumor cells (cytokeratin), immune cells (CD45), fibroblasts (αSMA, CD74, FAP, LaminA), other cells (CD31, CD56, CALD1, NG2), and collagen I in fixed tissue sections. Combinatorial fibroblast marker expression identified myofibroblasts (myCAF), inflammatory fibroblasts (iCAF), antigen-presenting fibroblasts (apCAF), and other hybrid fibroblast subtypes. We characterized fibroblast populations in up-front resected and neoadjuvant-treated cohorts of patients with localized PDAC and in patients with biopsies of metastatic lesions. Using digital imaging, supervised machine learning, and spatial proximity metrics, we quantified cell phenotypes, fibroblast subtype composition and spatial organization at the single-cell level and in the context of tumor clinicopathologic features and patient outcomes. Results: Across 301 up-front resected tumors, we cumulatively analyzed 1.7 million cells, including 611,000 fibroblasts. Stromal fibroblast density ranged from 632 to 4150 cells/mm2. Five main fibroblast subtypes constituted more than 85% of the fibroblast population in these tumors: αSMA+ (myCAF), FAP+αSMA+ (FAP+ myCAF), LaminA+ (iCAF), αSMA+ LaminA+ (myCAF/iCAF), and FAP+ αSMA+ LaminA+ (FAP+ myCAF/iCAF). Unexpectedly, 39% of fibroblasts showed co-expression of markers previously considered distinct for major fibroblast subtypes. By unsupervised clustering of fibroblast subtype proportions, we identified 3 fibroblast-defined tumor subgroups, including tumors rich in FAP+ fibroblasts, hybrid fibroblasts, and single positive fibroblasts (myCAFs, iCAFs, and apCAFs), which were associated with patient survival. Furthermore, fibroblast subtypes were spatially organized, with preferential localization of FAP+ myCAFs near tumor cells and iCAFs more distant from tumor cells yet closer to immune cells. FAP+ myCAFs were more abundant in basal-like tumors than in classical tumors, and FAP+ myCAFs were located closer to basal-like tumor cells than classical tumor cells within individual tumors. Compared to up-front resected primary tumors, neoadjuvant FOLFIRINOX treated tumors and metastatic lesions harbored distinct fibroblast profiles. Conclusion: Fibroblasts in the PDAC microenvironment are heterogeneous both in abundance and subtype composition, exhibit distinct spatial organization and are associated with patient outcomes. Treatment with FOLFIRINOX alters the composition of fibroblast subtypes in primary PDAC, and metastatic PDAC lesions have a different fibroblast composition compared to primary tumors. Citation Format: Dalia Elganainy, Andressa Dias Costa, Alexander Jordan, Suryun Kim, Sara A. Väyrynen, Hannah L. Williams, Chen Yuan, Douglas A. Rubinson, Kimberly Perez, Harshabad Singh, Richard F. Dunne, Thomas E. Clancy, David C. Linehan, Daniel T. Chang, Aram F. Hezel, Albert C. Koong, Andrew Aguirre, Brian M. Wolpin, Jonathan A. Nowak. Stromal composition, fibroblast heterogeneity and spatial organization in pancreatic adenocarcinoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr A035.
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Arlein, Wes J., Jeffry D. Shearer, and Michael D. Caldwell. "Continuity between wound macrophage and fibroblast phenotype: analysis of wound fibroblast phagocytosis." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 275, no. 4 (October 1, 1998): R1041—R1048. http://dx.doi.org/10.1152/ajpregu.1998.275.4.r1041.
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Analysis of phagocytic activity in wound fibroblasts was chosen as a means to assess the possible continuity between macrophage and fibroblast phenotypes. Fibroblast phagocytosis of uncoated, IgG-coated, or collagen-coated fluorescent beads was analyzed by flow cytometry in vivo and in vitro. Phagocytosis of fluorescent beads by procollagen I-positive cells (fibroblasts) was evaluated in vivo by injecting beads into subcutaneously implanted sponge wounds in anesthetized Fisher rats. Phagocytic activity of a purified population of wound fibroblasts was measured in vitro and correlated with oxidation state using hydroethidium. In the wound environment, 50–60% of the cells that engulfed uncoated, IgG-coated, or collagen-coated beads were procollagen I-positive cells (i.e., fibroblasts). Procollagen I-positive cells engulfed uncoated and IgG-coated beads in preference to collagen-coated beads in vivo. Cultured wound fibroblasts engulfed uncoated, IgG-coated, and collagen-coated particles. The majority of fibroblasts that engulfed beads were in an elevated oxidation state. We conclude that substantial fibroblast phagocytosis occurs in the wound, but scavenger receptor-mediated fibroblast phagocytosis is different from that of macrophages. Additional markers will be helpful in defining the macrophage fibroblast continuum.
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Fabisiak, J. P., M. Absher, J. N. Evans, and J. Kelley. "Spontaneous production of PDGF A-chain homodimer by rat lung fibroblasts in vitro." American Journal of Physiology-Lung Cellular and Molecular Physiology 263, no. 2 (August 1, 1992): L185—L193. http://dx.doi.org/10.1152/ajplung.1992.263.2.l185.
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Platelet-derived growth factor (PDGF) is considered a decisive mediator of fibroblast growth and phenotype within the lung. The cellular sources of PDGF within the lung remain undefined. The ability of lung fibroblasts themselves to produce PDGF in vitro was therefore investigated. Northern and Western blot analyses revealed the expression of PDGF-A mRNA and secretion of A-chain containing proteins by fibroblasts derived from adult and fetal rat lung. PDGF-A gene or protein expression were below the limits of detection in two human lung fibroblast lines examined in a similar manner. PDGF-B transcripts or proteins were not detected in any lung fibroblast line examined. Conditioned medium (CM) was collected from these same lung fibroblast lines and tested for its ability to promote cell growth using human fetal lung fibroblasts as targets. Both adult and fetal rat lung fibroblasts were found to produce a potent and efficacious stimulus for cell growth. Growth-promoting activity in rat fibroblast-derived CM functioned as a "competence" factor and was partially inhibited by anti-PDGF antibody. Thus rat lung fibroblasts in vitro produce potent growth factors of which at least one appears to be PDGF-AA. Differences in the expression of PDGF-AA between rat and human lung fibroblasts exist. Growth factor-producing fibroblasts may play a role in lung repair and remodeling through production PDGF-AA in vivo.
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Clark, K., A. Cole, X. Shiwen, V. Ong, C. D. Buckley, and C. P. Denton. "POS0481 DEFINING DISTINCT RESIDENT AND MIGRATORY FIBROBLAST POPULATIONS FROM SKIN BIOPSIES IN SYSTEMIC SCLEROSIS." Annals of the Rheumatic Diseases 81, Suppl 1 (May 23, 2022): 495.2–496. http://dx.doi.org/10.1136/annrheumdis-2022-eular.2678.
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BackgroundRecent studies using single cell RNA sequencing have delineated distinct subpopulations of fibroblasts in skin and other organs. To improve understanding of functional differences between fibroblast subpopulations, we have developed a novel technique to selectively isolate “migratory” fibroblasts, and non-migratory “resident” fibroblasts from heathy control (HC) and SSc skin.ObjectivesTo compare migratory and resident populations of dermal fibroblasts in systemic sclerosis by their differential gene or protein expression, and through functional assays of fibrotic potential.MethodsForearm skin punch biopsies were collected from dcSSc(n=3), and healthy control skin(n=3).Migratory fibroblasts were first isolated by standard explant culture, then the residual biopsy fragments underwent collagenase digestion to yield a population of resident fibroblasts that retained in the biopsy fragments. These populations were further expanded for use at passage 3-6.Functional characterisation included 3-D collagen gel contraction, and migratory scratch-wound assays. Expression of pro-collagen I (Col1), CTGF and αSMA was compared by western blot. Bulk RNAseq on each fibroblast population was performed. Statistical analysis was carried out using Rsoftware “tidyverse”. Criteria for significant differences in gene expression were a fold change of ≥1.5, and adjusted p-value (FDR) of <0.05ResultsCompared with HC, all SSc fibroblast populations showed a hallmark fibrotic phenotype with increased gel contraction, faster migration, and overexpression of Col1, CTGF and αSMA compared with HC. However, SSc resident fibroblasts showed attenuated contraction, migration, and reduced levels of αSMA compared to migratory SSc fibroblasts. Bulk RNAseq performed on each fibroblast population confirmed 5579 significantly differentially expressed genes between SSc resident and SSc migratory fibroblasts, whereas no genes were significantly differentially expressed between HC resident and migratory fibroblasts.SSc and HC migratory fibroblasts and resident fibroblasts were then compared, to understand disease-related differences between the two fibroblast populations. 739 genes were significantly overexpressed in the migratory fibroblast population (including ASPM, TRIP3), whereas 745 genes were significantly upregulated in the resident fibroblast population. The genes upregulated in resident fibroblasts included CCL2, CXCL8, and ICAM1.Many genes typically associated with SSc (such as SERPINE1, COMP), were not significantly different between the SSc fibroblast subpopulations, but were significantly elevated in both SSc subpopulations compared to HC fibroblast populations, suggesting that these reflect a more generic SSc phenotype.ConclusionMigratory and resident SSc fibroblast populations exhibit distinct functional and transcriptional differences that are much less apparent for HC biopsies. Further work is required to understand the precise contribution these distinct SSc fibroblast populations in pathogenesis, and how they might be targeted therapeutically. Our findings also highlight that conventional explant culture techniques may ignore important fibroblasts populations and highlight the importance of more detailed analysis such as single cell analysis to better understand pathobiology of SSc.Table 1.Table of the most significantly differentially expressed genes with highest fold change and adjusted p-value (FDR) between the migratory and resident SSc fibroblast populationSignificantly upregulated genes in SSc migratory fibroblastsSignificantly upregulated genes in SSc resident fibroblastsFold changeAdjusted p valueFold changeAdjusted p valueGPR15.990.01CCL238.66<0.0001PPME14.860.011CXCL824.360.0001KIF20A4.340.015ICAM115.38<0.0001CENPF4.310.018EGR115.190.0002STC24.180.01EGR212.770.0002ASPM3.880.02IFI44L11.50.002TRIB33.600.011CXCL611.43<0.0001HAPLN13.360.039PDGF-D10.140.0003CCNB23.290.025OAS38.940.0009KIF143.240.027IFIT18.580.004Disclosure of InterestsKristina Clark: None declared, Alice Cole: None declared, Xu Shiwen: None declared, Voon Ong: None declared, Christopher D Buckley Employee of: founder of Mestag Therapeutics https://mestagtherapeutics.com/, Christopher P Denton: None declared.
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Zhang, Zhentao, Gabriella Shayani, Yanping Xu, Ashley Kim, Yurim Hong, Haiyue Feng, and Hua Zhu. "Induction of Senescence by Loss of Gata4 in Cardiac Fibroblasts." Cells 12, no. 12 (June 17, 2023): 1652. http://dx.doi.org/10.3390/cells12121652.
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Cardiac fibroblasts are a major source of cardiac fibrosis during heart repair processes in various heart diseases. Although it has been shown that cardiac fibroblasts become senescent in response to heart injury, it is unknown how the senescence of cardiac fibroblasts is regulated in vivo. Gata4, a cardiogenic transcription factor essential for heart development, is also expressed in cardiac fibroblasts. However, it remains elusive about the role of Gata4 in cardiac fibroblasts. To define the role of Gata4 in cardiac fibroblasts, we generated cardiac fibroblast-specific Gata4 knockout mice by cross-breeding Tcf21-MerCreMer mice with Gata4fl/fl mice. Using this mouse model, we could genetically ablate Gata4 in Tcf21 positive cardiac fibroblasts in an inducible manner upon tamoxifen administration. We found that cardiac fibroblast-specific deletion of Gata4 spontaneously induces senescence in cardiac fibroblasts in vivo and in vitro. We also found that Gata4 expression in both cardiomyocytes and non-myocytes significantly decreases in the aged heart. Interestingly, when αMHC-MerCreMer mice were bred with Gata4fl/fl mice to generate cardiomyocyte-specific Gata4 knockout mice, no senescent cells were detected in the hearts. Taken together, our results demonstrate that Gata4 deficiency in cardiac fibroblasts activates a program of cellular senescence, suggesting a novel molecular mechanism of cardiac fibroblast senescence.
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Cheng, Maye F., Faizah S. Abdullah, and Matthew B. Buechler. "Essential growth factor receptors for fibroblast homeostasis and activation: Fibroblast Growth Factor Receptor (FGFR), Platelet Derived Growth Factor Receptor (PDGFR), and Transforming Growth Factor β Receptor (TGFβR)." F1000Research 13 (May 21, 2024): 120. http://dx.doi.org/10.12688/f1000research.143514.2.
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Fibroblasts are cells of mesenchymal origin that are found throughout the body. While these cells have several functions, their integral roles include maintaining tissue architecture through the production of key extracellular matrix components, and participation in wound healing after injury. Fibroblasts are also key mediators in disease progression during fibrosis, cancer, and other inflammatory diseases. Under these perturbed states, fibroblasts can activate into inflammatory fibroblasts or contractile myofibroblasts. Fibroblasts require various growth factors and mitogenic molecules for survival, proliferation, and differentiation. While the activity of mitogenic growth factors on fibroblasts in vitro was characterized as early as the 1970s, the proliferation and differentiation effects of growth factors on these cells in vivo are unclear. Recent work exploring the heterogeneity of fibroblasts raises questions as to whether all fibroblast cell states exhibit the same growth factor requirements. Here, we will examine and review existing studies on the influence of fibroblast growth factor receptors (FGFRs), platelet-derived growth factor receptors (PDGFRs), and transforming growth factor β receptor (TGFβR) on fibroblast cell states.
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Nie, Wenyang, Zhijie Zhao, Yuhang Liu, Youcao Wang, Jingwen Zhang, Ying Hu, Yang Liu, Yong Wang, and Zhen Wang. "Integrative Single-Cell Analysis of Cardiomyopathy Identifies Differences in Cell Stemness and Transcriptional Regulatory Networks among Fibroblast Subpopulations." Cardiology Research and Practice 2024 (May 18, 2024): 1–22. http://dx.doi.org/10.1155/2024/3131633.
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Background. Cardiomyopathy encompasses a broad spectrum of diseases affecting myocardial tissue, characterized clinically by abnormalities in cardiac structure, heart failure, and/or arrhythmias. Clinically heterogeneous, major types include dilated cardiomyopathy (DCM), hypertrophic cardiomyopathy (HCM), restrictive cardiomyopathy (RM), ischemic cardiomyopathy (ICM), among which DCM is more prevalent, while ICM exhibits higher incidence and mortality rates. Myocardial injury during cardiomyopathy progression may lead to myocardial fibrosis. Failure to intervene early and inhibit the process of myocardial fibrosis may culminate in heart failure. Cardiac fibroblasts constitute crucial cellular components determining the extent and quality of myocardial fibrosis, with various subpopulations exerting diverse roles in cardiomyopathy progression. Despite this, understanding of the cellular plasticity and transcriptional regulatory networks of cardiac fibroblasts in cardiomyopathy remains limited. Therefore, in this study, we conducted comprehensive single-cell analysis of cardiac fibroblasts in cardiomyopathy to explore differences in cellular plasticity and transcriptional regulatory networks among fibroblast subpopulations, with the aim of providing as many useful references as possible for the diagnosis, prognosis, and treatment of cardiomyopathy. Materials and Methods. Cells with mitochondrial gene expression comprising >20% of total expressed genes were excluded. Differential expression genes (DEGs) and stemness genes within cardiac fibroblast subpopulations were subjected to Gene Ontology (GO) analysis of biological processes (BP) and AUCell analysis. Monocle software was employed to analyze the pseudo-temporal trajectory of cardiac fibroblasts in cardiomyopathy. Additionally, the Python package SCENIC was utilized to assess enrichment of transcription factors and activity of regulators within cardiac fibroblast subpopulations in cardiomyopathy. Results. Following batch effect correction, 179,927 cells were clustered into 32 clusters, designated as T_NK cells, endothelial cells, myeloid cells, fibroblasts, pericytes, SMCs, CMs, proliferating cells, EndoCs, and EPCs. Among them, 8148 fibroblasts were further subdivided into 4 subpopulations, namely C0 THBS4+ Fibroblasts, C1 LINC01133+ Fibroblasts, C2 FGF7+ Fibroblasts, and C3 AGT + Fibroblasts. Results from GO_BP and AUCell analyses suggest that C3 AGT + Fibroblasts may be associated with immune response activation, protein transport, and myocardial contractile function, correlating with disease progression in cardiomyopathy. Transcription factor enrichment analysis indicates that FOS is the most significant TF in C3 AGT + Fibroblasts, also associated with the M1 module, possibly implicated in protein hydrolysis, intracellular DNA replication, and cell proliferation. Moreover, correlation analysis of transcriptional regulatory activity between fibroblast subpopulations reveals a more pronounced heterogeneity within C3 AGT + Fibroblasts in cardiomyopathy. Conclusion. C3 AGT + Fibroblasts exhibit increased sensitivity towards adverse outcomes in cardiomyopathy, such as myocardial fibrosis and impaired cardiac contractile function, compared to other cardiac fibroblast subpopulations. The differential cellular plasticity and transcriptional regulatory activity between C3 AGT + Fibroblasts and other subgroups offer new perspectives for targeting fibroblast subpopulation activity to treat cardiomyopathy. Additionally, stemness genes EPAS1 and MYC, along with the regulator FOS, may play roles in modulating the biological processes of cardiac fibroblasts in cardiomyopathy.
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Hou, Jiwei, Yanru Yang, and Xin Han. "Machine Learning and Single-Cell Analysis Identify Molecular Features of IPF-Associated Fibroblast Subtypes and Their Implications on IPF Prognosis." International Journal of Molecular Sciences 25, no. 1 (December 20, 2023): 94. http://dx.doi.org/10.3390/ijms25010094.
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Idiopathic pulmonary fibrosis (IPF) is a devastating lung disease of unknown cause, and the involvement of fibroblasts in its pathogenesis is well recognized. However, a comprehensive understanding of fibroblasts’ heterogeneity, their molecular characteristics, and their clinical relevance in IPF is lacking. In this study, we aimed to systematically classify fibroblast populations, uncover the molecular and biological features of fibroblast subtypes in fibrotic lung tissue, and establish an IPF-associated, fibroblast-related predictive model for IPF. Herein, a meticulous analysis of scRNA-seq data obtained from lung tissues of both normal and IPF patients was conducted to identify fibroblast subpopulations in fibrotic lung tissues. In addition, hdWGCNA was utilized to identify co-expressed gene modules associated with IPF-related fibroblasts. Furthermore, we explored the prognostic utility of signature genes for these IPF-related fibroblast subtypes using a machine learning-based approach. Two predominant fibroblast subpopulations, termed IPF-related fibroblasts, were identified in fibrotic lung tissues. Additionally, we identified co-expressed gene modules that are closely associated with IPF-fibroblasts by utilizing hdWGCNA. We identified gene signatures that hold promise as prognostic markers in IPF. Moreover, we constructed a predictive model specifically focused on IPF-fibroblasts which can be utilized to assess disease prognosis in IPF patients. These findings have the potential to improve disease prediction and facilitate targeted interventions for patients with IPF.
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Kurose, Hitoshi. "Cardiac Fibrosis and Fibroblasts." Cells 10, no. 7 (July 6, 2021): 1716. http://dx.doi.org/10.3390/cells10071716.
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Cardiac fibrosis is the excess deposition of extracellular matrix (ECM), such as collagen. Myofibroblasts are major players in the production of collagen, and are differentiated primarily from resident fibroblasts. Collagen can compensate for the dead cells produced by injury. The appropriate production of collagen is beneficial for preserving the structural integrity of the heart, and protects the heart from cardiac rupture. However, excessive deposition of collagen causes cardiac dysfunction. Recent studies have demonstrated that myofibroblasts can change their phenotypes. In addition, myofibroblasts are found to have functions other than ECM production. Myofibroblasts have macrophage-like functions, in which they engulf dead cells and secrete anti-inflammatory cytokines. Research into fibroblasts has been delayed due to the lack of selective markers for the identification of fibroblasts. In recent years, it has become possible to genetically label fibroblasts and perform sequencing at single-cell levels. Based on new technologies, the origins of fibroblasts and myofibroblasts, time-dependent changes in fibroblast states after injury, and fibroblast heterogeneity have been demonstrated. In this paper, recent advances in fibroblast and myofibroblast research are reviewed.
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Rook, M. B., A. C. van Ginneken, B. de Jonge, A. el Aoumari, D. Gros, and H. J. Jongsma. "Differences in gap junction channels between cardiac myocytes, fibroblasts, and heterologous pairs." American Journal of Physiology-Cell Physiology 263, no. 5 (November 1, 1992): C959—C977. http://dx.doi.org/10.1152/ajpcell.1992.263.5.c959.
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Cultures of neonatal rat heart cells contain predominantly myocytes and fibroblastic cells. Most abundant are groups of synchronously contracting myocytes, which are electrically well coupled through large gap junctions. Cardiac fibroblasts may be electrically coupled to each other and to adjacent myocytes, be it with low intercellular conductances. Nevertheless, synchronously beating myocytes interconnected via a fibroblast were present, demonstrating that nonexcitable cardiac cells are capable of passive impulse conduction. In fibroblast pairs as well as in myocyte-fibroblast cell pairs, no sensitivity to junctional voltage could be detected when transjunctional conductance was > 1-2 nS. However, in pairs coupled by a conductance of < 1 nS, complex voltage-dependent gating was evident; gap junction channel open probability decreased with increasing junctional voltage but a nongated residual conductance remained at all voltages tested. Single gap junction channel conductance between fibroblasts was approximately 21 pS, very similar to an approximately 18-pS channel conductance that was found between myocytes next to the major conductance of 43 pS. Single-channel conductance in heterologous myocyte-fibroblast gap junctions was approximately 32 pS, which matches the theoretical value of 29 pS for gap junction channels composed of a fibroblast connexon and the major myocyte connexon. A site-directed antibody against rat heart gap junction protein connexin43 recognized gap junctions between neonatal cardiomyocytes, as demonstrated by immunocytochemical labeling. In contrast, junctions between fibroblasts showed no labeling, while in myocyte-fibroblast junctions labeling occasionally was present. Our results suggest the existence of two gap junction proteins between neonatal rat cardiocytes, connexin43 and another yet unidentified connexin. An alternative explanation (cell-specific regulation of the conductance of connexin43 channels) is discussed.
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Ortiz, A., C. Lorz, S. González-Cuadrado, R. Garcia del Moral, F. O'Valle, and J. Egido. "Cytokines and Fas regulate apoptosis in murine renal interstitial fibroblasts." Journal of the American Society of Nephrology 8, no. 12 (December 1997): 1845–54. http://dx.doi.org/10.1681/asn.v8121845.
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Renal fibrosis is characterized by an increased number of fibroblasts and excessive deposition of extracellular matrix. Apoptotic cell death is a physiological mechanism to limit cell numbers, and an insufficient rate of death may contribute to fibroblast accumulation. However, little is known about the regulation of renal fibroblast survival. The authors have studied the interaction of cytokines and the Fas receptor in the regulation of apoptosis of renal fibroblasts and have observed that murine renal fibroblasts express Fas and the Fas ligand. Tumor necrosis factor alpha (TNFalpha) and agonistic anti-Fas antibodies induce apoptosis of renal fibroblasts in a time- and dose-dependent manner. Serum contains survival factors for renal fibroblasts. Both serum deprivation and TNFalpha increase the sensitivity to Fas-induced death and the expression of fas mRNA and Fas receptor. By contrast, insulin-like growth factor-1 decreases apoptosis induced by both serum deprivation and Fas activation and partially prevents the increase in Fas receptor expression induced by serum deprivation. Murine renal fibroblasts express constitutively both fas ligand mRNA and cell-surface Fas ligand, but the authors could not demonstrate a role for Fas ligand in the autocrine regulation of fibroblast survival. These data suggest that Fas and other cytokines cooperate to regulate renal fibroblast apoptosis. Modulation of the Fas death-signaling pathway in renal fibroblasts could represent a new therapeutic target for renal fibrosis.
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Han, Kyu-Yeon, Jin-Hong Chang, and Dimitri T. Azar. "Proteomics-Based Characterization of the Effects of MMP14 on the Protein Content of Exosomes from Corneal Fibroblasts." Protein & Peptide Letters 27, no. 10 (November 2, 2020): 979–88. http://dx.doi.org/10.2174/0929866527666200408142827.
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Background: Exosomes secreted by corneal fibroblasts contain matrix metalloproteinase (MMP) 14, which is known to influence pro-MMP2 accumulation on exosomes. Accordingly, we hypothesized that the enzymatic activity of MMP14 may alter the protein content of corneal fibroblast- secreted exosomes. Objective: The aim of this study was to investigate the effects of MMP14 on the composition and biological activity of corneal fibroblast-derived exosomes. Methods: Knock out of the catalytic domain (ΔExon4) of MMP14 in corneal fibroblasts was used to determine the effect of MMP14 expression on the characteristics of fibroblast-secreted exosomes. The amount of secreted proteins and their size distribution were measured using Nano Tracking Analysis. Proteins within exosomes from wild-type (WT) and ΔExon4-deficient fibroblasts were identified by liquid chromatography-tandem mass spectrometry (MS/MS) proteomics analysis. The proteolytic effects of MMP14 were evaluated in vitro via MS identification of eliminated proteins. The biological functions of MMP14-carrying exosomes were investigated via fusion to endothelial cells and flow cytometric assays. Results: Exosomes isolated from WT and ΔExon4-deficient fibroblasts exhibited similar size distributions and morphologies, although WT fibroblasts secreted a greater amount of exosomes. The protein content, however, was higher in ΔExon4-deficient fibroblast-derived exosomes than in WT fibroblast-derived exosomes. Proteomics analysis revealed that WT-derived exosomes included proteins that regulated cell migration, and ΔExon4 fibroblast-derived exosomes contained additional proteins that were cleaved by MMP14. Conclusion: Our findings suggest that MMP14 expression influences the protein composition of exosomes secreted by corneal fibroblasts, and through those biological components, MMP14 in corneal fibroblasts derived-exosomes may regulate corneal angiogenesis.
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Parry, Dylan, and Keith Allison. "Is the future scarless? – Fibroblasts as targets for scarless wound healing: a narrative review." Scars, Burns & Healing 8 (January 2022): 205951312210953. http://dx.doi.org/10.1177/20595131221095348.
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Introduction: Scarless healing is the ideal outcome of wound healing and is exhibited in some species. This narrative review assembles the current understanding of fibroblast heterogenicity along with the latest fibroblast-related targets for scar reduction therapies. Human regenerative wound healing is deemed possible due to the wound regeneration already seen in the early gestation foetus. Methods: This literature narrative review was undertaken by searching PubMed and Web of Science databases and Google Scholar to find articles concerning the fibroblast involvement in wound healing. We evaluated and collated these articles to form a consensus of the current understanding of the field. Discussion: This article describes current understanding of fibroblast heterogenicity and involvement in wound healing, focusing on the role of fibroblasts during physiological scarring. We also present the current most promising targets involving fibroblasts in the reduction of scarring and how we can manipulate the behaviour of fibroblasts to mimic the wound regeneration models in the human foetus. These targets include the pro-fibrotic EN1 positive fibroblast lineage, TGFβ1 inhibition, and genetic therapies utilising miRNAs and siRNAs. Conclusion: No therapies are currently available to eradicate scarring; however, treatment options are available to reduce the appearance of scarring. Further research into the heterogenicity and interactions of fibroblasts in both the foetus and adult is needed, and this may lead to the development of novel treatments against scarring. Lay Summary Scarless healing refers to the repair of a wound with minimal residual scarring. The main cell responsible for the repair process is the fibroblast. It is now understood that there are different types of fibroblasts. Simply, some of these fibroblasts lead to scarring and some lead to regeneration. The early human foetus has mainly regenerative fibroblasts, but during aging the number of scarring fibroblasts increase to become the majority in the adult . Understanding how we can modify this process may ultimately result in the reduction in scarring. Currently, scar reduction therapies are aimed at optimal wound healing, surgical removal of abnormal scars, and using steroids and other drugs to encourage better wound repair by limiting the effect of scarring fibroblasts. Future therapies aim to target specific groups of fibroblasts to encourage regenerative wound healing. This narrative review aims to cover the current understanding of the different groups of fibroblasts and their effect on wound healing. We also cover the current and potential therapies that can be used to reduce scarring and suggest further areas for research in this field.
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Rettig, W. J., H. P. Erickson, A. P. Albino, and P. Garin-Chesa. "Induction of human tenascin (neuronectin) by growth factors and cytokines: cell type-specific signals and signalling pathways." Journal of Cell Science 107, no. 2 (February 1, 1994): 487–97. http://dx.doi.org/10.1242/jcs.107.2.487.
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The extracellular matrix protein tenascin (TN) is expressed with precise temporo-spatial patterns during embryonic and fetal development and is induced in healing wounds, inflammatory lesions and solid tumors. These tissue patterns suggest that TN synthesis may be modulated by soluble factors present in developing tissues or released from injured, inflammatory or neoplastic cells. To characterize the extrinsic control of human TN we examined the effects of several signalling molecules on cultured neural, melanocytic and fibroblastic cells. Results obtained with alpha TN antibodies in enzyme-linked immunosorbent and immunoprecipitation assays indicate that TN expression is tightly regulated in a cell type-specific manner: (1) Primitive neuroectodermal tumor (PNET) cells grown in chemically defined, serum-free media show up to > 100-fold TN induction in response to fibroblast growth factors (aFGF, bFGF, K-FGF) and phorbol ester, independent of changes in cell proliferation or total protein synthesis; no induction is seen in PNET cultures stimulated with serum or other growth and differentiation factors. (2) Normal melanocytes, which require FGF and phorbol ester for survival in vitro, fail to express TN; however, they produce TN following oncogenic transformation. (3) Fibroblasts derived from disparate tissues differ up to 100-fold in basal TN production; for example, fetal lung fibroblasts are TNhigh, but conjunctival fibroblasts derived from the same donors and fetal leptomeningeal cells are TNlow. (4) TNlow fibroblasts treated with interleukin-1, tumor necrosis factor-alpha, and interleukin-4 show up to > 100-fold increased TN secretion and TN incorporation into their extracellular matrix. Transforming growth factor-beta, which acts as an inducer of fibronectin, collagen, and integrin-type matrix receptors, has variable effects on fibroblast TN, ranging from increased deposition in the extracellular matrix of fetal conjunctival fibroblasts to reduced secretion in newborn foreskin fibroblasts. In contrast, FGFs (which are potent fibroblast mitogens), phorbol ester, bone morphogenetic proteins, and several other factors tested produced no discernible effects on fibroblast TN expression. These findings suggest that discrete sets of extrinsic signals modify TN expression in specific cell types, with the effects of a given ligand/receptor system determined by cell type-specific signalling pathways that may be linked to unique cis-regulatory elements of the TN gene. As a result, a limited set of regulatory peptides may produce highly diversified TN distribution patterns in developing and lesional tissues.
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Sanford-Crane, Hannah, Jaime Abrego, and Mara H. Sherman. "Fibroblasts as Modulators of Local and Systemic Cancer Metabolism." Cancers 11, no. 5 (May 3, 2019): 619. http://dx.doi.org/10.3390/cancers11050619.
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Fibroblast activation is an accompanying feature of solid tumor progression, resembling a conserved host response to tissue damage. Cancer-associated fibroblasts (CAFs) comprise a heterogeneous and plastic population with increasingly appreciated roles in tumor growth, metastatic capacity, and response to therapy. Classical features of fibroblasts in a wound-healing response, including profound extracellular matrix production and cytokine release, are recapitulated in cancer. Emerging evidence suggests that fibroblastic cells in the microenvironments of solid tumors also critically modulate cellular metabolism in the neoplastic compartment through mechanisms including paracrine transfer of metabolites or non-cell-autonomous regulation of metabolic signaling pathways. These metabolic functions may represent common mechanisms by which fibroblasts stimulate growth of the regenerating epithelium during a wound-healing reaction, or may reflect unique co-evolution of cancer cells and surrounding stroma within the tumor microenvironment. Here we review the recent literature supporting an important role for CAFs in regulation of cancer cell metabolism, and relevant pathways that may serve as targets for therapeutic intervention.
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Portnoy, Joshua, Tianli Pan, Charles A. Dinarello, John M. Shannon, Jay Y. Westcott, Lening Zhang, and Robert J. Mason. "Alveolar type II cells inhibit fibroblast proliferation: role of IL-1α." American Journal of Physiology-Lung Cellular and Molecular Physiology 290, no. 2 (February 2006): L307—L316. http://dx.doi.org/10.1152/ajplung.00102.2005.
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Alveolar type II (ATII) cells inhibit fibroblast proliferation in coculture by releasing or secreting a factor(s) that stimulates fibroblast production of prostaglandin E2(PGE2). In the present study, we sought to determine the factors released from ATII cells that stimulate PGE2production in fibroblasts. Exogenous addition of rat IL-1α to cultured lung fibroblasts induced PGE2secretion in a dose-response manner. When fibroblasts were cocultured with rat ATII cells, IL-1α protein was detectable in ATII cells and in the coculture medium between days 8 and 12 of culture, correlating with the highest levels of PGE2. Furthermore, under coculture conditions, IL-1α gene expression increased in ATII cells (but not fibroblasts) compared with either cell cultured alone. In both mixed species (human fibroblasts-rat ATII cells) and same species cocultures (rat fibroblasts and ATII cells), PGE2secretion was inhibited by the presence of IL-1 receptor antagonist (IL-1Ra) or selective neutralizing antibody directed against rat IL-1α (but not IL-1β). Conditioned media from cocultures inhibited fibroblast proliferation, and this effect was abrogated by the addition of IL-1Ra. Addition of keratinocyte growth factor (KGF) resulted in an earlier increase in PGE2secretion and fibroblast inhibition ( day 8 of coculture). This effect was inhibited by indomethacin but was not altered by IL-1Ra. We conclude that in this coculture system, IL-1α secretion by ATII cells is one factor that stimulates PGE2production by lung fibroblasts, thereby inhibiting fibroblast proliferation. In addition, these studies demonstrate that KGF enhances ATII cell PGE2production through an IL-1α-independent pathway.
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Wei, Juan, Junhui Zhan, Hui Ji, Yitong Xu, Qingfeng Xu, Xiaoyan Zhu, and Yujian Liu. "Fibroblast Upregulation of Vitamin D Receptor Represents a Self-Protective Response to Limit Fibroblast Proliferation and Activation during Pulmonary Fibrosis." Antioxidants 12, no. 8 (August 18, 2023): 1634. http://dx.doi.org/10.3390/antiox12081634.
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Dysregulation of vitamin D receptor (VDR) is implicated in chronic obstructive pulmonary disease. However, whether VDR dysregulation contributes to the development of pulmonary fibrosis remains largely unknown. Analysis of bulk and single-cell RNA profiling datasets revealed VDR upregulation in lung fibroblasts from patients with pulmonary fibrosis or fibrotic mice, which was validated in lung fibroblasts from bleomycin-exposed mice and bleomycin-treated fibroblasts. Stable VDR knockdown promoted, whereas the VDR agonist paricalcitol suppressed lung fibroblast proliferation and activation. Gene set enrichment analysis (GSEA) showed that the JAK/STAT pathway and unfolded protein response (UPR), a process related to endoplasmic reticulum (ER) stress, were enriched in lung fibroblasts of fibrotic lungs. Stable VDR knockdown stimulated, but paricalcitol suppressed ER stress and JAK1/STAT3 activation in lung fibroblasts. The STAT3 inhibitor blocked bleomycin- or stable VDR knockdown-induced ER stress. Paricalcitol inhibited the bleomycin-induced enrichment of STAT3 to the ATF6 promoter, thereby suppressing ATF6 expression in fibroblasts. Paricalcitol or intrapulmonary VDR overexpression inactivated JAK1/STAT3 and suppressed ER stress in bleomycin-treated mice, thus resulting in the inhibition of fibroblast proliferation and activation. Collectively, this study suggests that fibroblast VDR upregulation may be a self-protective response to limit fibroblast proliferation and activation during pulmonary fibrosis by suppressing the JAK1/STAT3/ER stress pathway.
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Volin, Michael, Brian Zanotti, Karolina Klosowska, Kelley Smith, Jennifer Birly, Manuel Rodriguez, and James Woods. "Rheumatoid arthritis synovial tissue fibroblast Mucin 3 expression and function (54.27)." Journal of Immunology 188, no. 1_Supplement (May 1, 2012): 54.27. http://dx.doi.org/10.4049/jimmunol.188.supp.54.27.
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Abstract Mucin 3 (MUC3) is elevated in rheumatoid arthritis (RA) synovial tissues (STs). We hypothesized that MUC3 is expressed by RA synovial fibroblasts, that it is cytokine inducible and chemotactic for RA synovial fibroblasts. To test this, RA synovial fibroblast MUC3 expression was determined by immunohistochemistry, flow cytometry and Western blot analysis. Levels of MUC3 from arthritic synovial fluids (SFs) were measured by a novel MUC3 ELISA. RA synovial fibroblast chemotaxis to RA SFs was assessed using a modified Boyden chamber assay. RA ST cells stained for both MUC3 and vimentin indicating that RA synovial fibroblasts express MUC3. Flow cytometry experiments showed that most of the RA synovial fibroblast MUC3 is cytoplasmic. Cytokine stimulation of these cells significantly increased MUC3 production (P<0.05). RA SFs contain on average 8.7μg/ml (n=8) of MUC3. Immunodepletion of MUC3 from RA SFs resulted in significantly reduced RA synovial fibroblast chemotaxis (P<0.05, n=5 RA SFs and n=8 RA synovial fibroblasts). These results show that MUC3 is produced by synovial fibroblasts in RA STs, that MUC3 is inducible by cytokine stimulation and that RA SF induced synovial fibroblast chemotaxis can be inhibited by depleting MUC3. Thus, MUC3 synovial fibroblast production may be induced by the inflammatory milieu of the RA joint and may drive synovial fibroblast chemotaxis within the RA ST potentially aiding pannus formation and cartilage destruction.
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Anggraeni, Rezky, and Moehamad Orliando Roeslan. "Effects of Clinacanthus nutans and Aloe vera Extracts on bFGF Synthesis in Fibroblasts." Journal of Indonesian Dental Association 5, no. 1 (May 17, 2022): 21. http://dx.doi.org/10.32793/jida.v5i1.726.
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Introduction: Wound healing is an important but complicated process, containing a multifaceted process. The growth factor involved in the wound healing process is basic fibroblast growth factor (bFGF). Clinacanthus nutans (C. nutans) and A. vera are traditional plants that have the ability to induce fibroblast migration. However, no study has yet compared the effects of C. nutans and A. vera on bFGF protein synthesis associated with wound healing.
Objective: The aim of this study was to evaluate the effect of extracts of C. nutans and A. vera on the fibroblast synthesis of bFGF in fibroblasts.
Method: Clinacanthus nutans leaf powder was extracted using a solution of hexane and chloroform sequentially. While the A. vera gel is taken. Fibroblasts were treated with several concentrations (10, 50, and 100 µg/mL) within 24 hours. Synthesis of bFGF protein was tested using enzyme linked immunoabsorbent assay (ELISA).
Result: The chloroform extract of C. nutans and A. vera up regulated the synthesis of bFGF. Concentration of 10 µg/mL C. nutans showed the highest bFGF protein synthesis compared to other treatment groups.
Conclusion: Chloroform extract of C. nutans and A. vera can up regulated the synthesis of bFGF in fibroblasts.
ABSTRAK
Pendahuluan: Penyembuhan luka adalah proses penting namun rumit, yang terdiri dari beberapa fase penyembuhan. Faktor pertumbuhan yang berhubungan dengan proses penyembuhan luka adalah, basic fibroblast growth factor (bFGF). Clinacanthus nutans (C. nutans) dan Aloe vera (A. vera) adalah tumbuhan tradisional yang memiliki kemampuan untuk menginduksi migrasi fibroblas. Namun, belum ada penelitian yang telah membandingkan efek C. nutans dan A. vera terhadap sintesis protein bFGF yang berhubungan dengan luka penyembuhan.
Tujuan: Tujuan dari penelitian ini adalah untuk mengetahui efek ekstrak C. nutans dan A. vera terhadap sintesis bFGF pada fibroblas.
Metode: Bubuk daun C. nutans di ektraksi menggunakan larutan heksana dan kloroform secara berurutan. Sedangkan A. vera diambil gel nya. Fibroblas diberi perlakuan dengan beberapa konsentrasi (10, 50, dan 100 µg/mL) dalam waktu 24 jam. Sintesis protein bFGF diuji menggunakan enzym linked immunoabsorbent assay (ELISA).
Hasil: Ekstrak kloroform C. nutans dan A. vera dapat meningkatkan sintesis protein bFGF lebih tinggi dibandingkan dengan kontrol negatif pada uji ELISA. C. nutans konsentrasi 10 µg/mL menunjukkan sintesis protein bFGF tertinggi dibandingkan dengan kelompok perlakuan lain.
Kesimpulan: Ekstrak kloroform C. nutans dan A. vera dapat meningkatkan sintesis protein bFGF pada fibroblas.
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Heinzelmann, Katharina, Mareike Lehmann, Michael Gerckens, Nina Noskovičová, Marion Frankenberger, Michael Lindner, Rudolf Hatz, et al. "Cell-surface phenotyping identifies CD36 and CD97 as novel markers of fibroblast quiescence in lung fibrosis." American Journal of Physiology-Lung Cellular and Molecular Physiology 315, no. 5 (November 1, 2018): L682—L696. http://dx.doi.org/10.1152/ajplung.00439.2017.
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Fibroblasts play an important role in lung homeostasis and disease. In lung fibrosis, fibroblasts adopt a proliferative and migratory phenotype, with increased expression of α-smooth muscle actin (αSMA) and enhanced secretion of extracellular matrix components. Comprehensive profiling of fibroblast heterogeneity is limited because of a lack of specific cell-surface markers. We have previously profiled the surface proteome of primary human lung fibroblasts. Here, we sought to define and quantify a panel of cluster of differentiation (CD) markers in primary human lung fibroblasts and idiopathic pulmonary fibrosis (IPF) lung tissue, using immunofluorescence and FACS analysis. Fibroblast function was assessed by analysis of replicative senescence. We observed the presence of distinct fibroblast phenotypes in vivo, characterized by various combinations of Desmin, αSMA, CD36, or CD97 expression. Most markers demonstrated stable expression over passages in vitro, but significant changes were observed for CD36, CD54, CD82, CD106, and CD140a. Replicative senescence of fibroblasts was observed from passage 10 onward. CD36- and CD97-positive but αSMA-negative cells were present in remodeled areas of IPF lungs. Transforming growth factor (TGF)-β treatment induced αSMA and collagen I expression but repressed CD36 and CD97 expression. We identified a panel of stable surface markers in human lung fibroblasts, applicable for positive-cell isolation directly from lung tissue. TGF-β exposure represses CD36 and CD97 expression, despite increasing αSMA expression; we therefore identified complex surface protein changes during fibroblast-myofibroblast activation. Coexistence of quiescence and activated fibroblast subtypes in the IPF lung suggests dynamic remodeling of fibroblast activation upon subtle changes to growth factor exposure in local microenvironmental niches.
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Hartati Wahyuningsih, Mae Sri, Dwi Aris Agung Nugrahaningsih, and Arief Budiyanto. "ETHANOLIC EXTRACT OF TITHONIA DIVERSIFOLIA (HEMSLEY) A. GRAY INHIBITS MIGRATION ACTIVITY AND DECREASE THE TRANSFORMING GROWTH FACTOR-BETA1, VEGF EXPRESSION ON KELOID FIBROBLASTS." Asian Journal of Pharmaceutical and Clinical Research 12, no. 1 (January 7, 2019): 342. http://dx.doi.org/10.22159/ajpcr.2018.v12i1.29850.
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Objectives: Keloid occurred due to abnormal wound healing, characterized by massive fibroblast proliferation and excessive collagen accumulation. Tithonia diversifolia Hemsley A. Gray has been known to show antiproliferative effect against some cancer cells in vitro. However, its potential as anti-keloid has not been explored. This study aims to assess the T. diversifolia ethanolic extract effect on fibroblast migration activity, transforming growth factor-beta1 (TGF-β1), and vascular endothelial growth factor (VEGF) expression of keloid fibroblasts in vitro.Methods: Fibroblasts were isolated from keloid collected from patient keloid tissue. The migration activity of keloid fibroblasts was assessed using scratch assay. The TGF-β1 and VEGF expression examination was done using ELISA.Results: Ethanolic extract of T. diversifolia treatment at a concentration of 20 μg/ml, 10 μg/ml, and 5 μg/mL for 24 h on keloid fibroblasts culture showed slower migration activity compare to those on keloid fibroblast culture without treatment (p<0.05). The TGF-β1 and VEGF expression was significantly lower in ethanolic extract of T. diversifolia treatment group compared to those on keloid fibroblast without treatment (p<0.05).Conclusion: Ethanolic extract of T. diversifolia inhibits fibroblast migration activity and decrease the expression of VEGF and TGF-β1 on keloid fibroblasts in vitro.
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Hartati Wahyuningsih, Mae Sri, Dwi Aris Agung Nugrahaningsih, and Arief Budiyanto. "ETHANOLIC EXTRACT OF TITHONIA DIVERSIFOLIA (HEMSLEY) A. GRAY INHIBITS MIGRATION ACTIVITY AND DECREASE THE TRANSFORMING GROWTH FACTOR-BETA1, VEGF EXPRESSION ON KELOID FIBROBLASTS." Asian Journal of Pharmaceutical and Clinical Research 12, no. 1 (January 7, 2019): 342. http://dx.doi.org/10.22159/ajpcr.2019.v12i1.29850.
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Objectives: Keloid occurred due to abnormal wound healing, characterized by massive fibroblast proliferation and excessive collagen accumulation. Tithonia diversifolia Hemsley A. Gray has been known to show antiproliferative effect against some cancer cells in vitro. However, its potential as anti-keloid has not been explored. This study aims to assess the T. diversifolia ethanolic extract effect on fibroblast migration activity, transforming growth factor-beta1 (TGF-β1), and vascular endothelial growth factor (VEGF) expression of keloid fibroblasts in vitro.Methods: Fibroblasts were isolated from keloid collected from patient keloid tissue. The migration activity of keloid fibroblasts was assessed using scratch assay. The TGF-β1 and VEGF expression examination was done using ELISA.Results: Ethanolic extract of T. diversifolia treatment at a concentration of 20 μg/ml, 10 μg/ml, and 5 μg/mL for 24 h on keloid fibroblasts culture showed slower migration activity compare to those on keloid fibroblast culture without treatment (p<0.05). The TGF-β1 and VEGF expression was significantly lower in ethanolic extract of T. diversifolia treatment group compared to those on keloid fibroblast without treatment (p<0.05).Conclusion: Ethanolic extract of T. diversifolia inhibits fibroblast migration activity and decrease the expression of VEGF and TGF-β1 on keloid fibroblasts in vitro.
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Pablos, José L., Patricia E. Carreira, Lourdes Serrano, Pedro Del Castillo, and Juan J. Gomez-Reino. "Apoptosis and Proliferation of Fibroblasts During Postnatal Skin Development and Scleroderma in the Tight-skin Mouse." Journal of Histochemistry & Cytochemistry 45, no. 5 (May 1997): 711–19. http://dx.doi.org/10.1177/002215549704500509.
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Tight-skin (Tsk) is a dominant gene mutation that causes a fibrotic skin disease in mice, similar to human scleroderma. Both conditions are characterized by increased numbers of dermal fibroblasts containing high levels of procollagen mRNA. Whether this fibroblast population arises from fibroblast growth or fibroblast transcriptional activation is debated. Proliferation and apoptosis of fibroblasts of normal and Tsk mice were studied in skin sections before, at onset, and in established fibrosis. Tissue sections were immunostained with proliferating cell nuclear antigen (PCNA) as proliferation marker. Apoptosis was investigated by in situ end-labeling of fragmented DNA and nuclear staining with propidium iodide. The expression of the apoptosis inhibitor Bcl-2 was investigated by immuno-histochemistry. We demonstrate differences in fibroblast proliferation and apoptosis related to postnatal skin growth and development. Neonatal skin exhibits the highest levels of proliferation and apoptosis in fibroblasts. In contrast, low proliferation and absence of apoptosis characterizes adult fibroblasts. Skin fibroblasts express Bcl-2 only in newborns, and at other ages Bcl-2 was restricted to epithelial cells. Our results also suggest that neither increased fibroblast proliferation nor defective apoptosis accounts for the fibrotic phenotype of Tsk. Therefore, transcriptional activation of extracellular matrix genes appears more relevant in the pathogenesis of Tsk fibrosis.
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Caniggia, I., I. Tseu, R. N. Han, B. T. Smith, K. Tanswell, and M. Post. "Spatial and temporal differences in fibroblast behavior in fetal rat lung." American Journal of Physiology-Lung Cellular and Molecular Physiology 261, no. 6 (December 1, 1991): L424—L433. http://dx.doi.org/10.1152/ajplung.1991.261.6.l424.
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Fibroblast-epithelial interactions were investigated in cells from late-gestation fetal rat lung. Fibroblasts from the pseudoglandular stage of lung development stimulated epithelial cell proliferation, whereas fibroblasts from the saccular stage promoted epithelial cell differentiation. The developmental switch from proliferation to differentiation seemed to be controlled by both cell types. Fibroblast-derived epithelial cell growth-promoting activity, evident in cells from the pseudoglandular period, decreased during development and almost disappeared in cells from the saccular stage. Interestingly, the response of epithelial cells to this growth-promoting activity declined with advancing gestational age as epithelial cells became more responsive to fibroblast-derived differentiation factor(s). Production of differentiation factor(s) by fibroblasts increased during the canalicular stage of lung development. Platelet-derived growth factor (PDGF) and low concentrations of transforming growth factor-beta (TGF-beta) stimulated epithelial cell proliferation. PDGF did not affect differentiation, whereas TGF-beta was inhibitory. Dependent on their proximity to the epithelium, two subpopulations of fibroblasts that differed in their ability to promote epithelial cell proliferation or differentiation were isolated. Fibroblasts in close proximity to the epithelium mainly produced differentiation factors, whereas more distant fibroblasts primarily stimulated proliferation.
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Herawati, Hillda, Rhabiah El Fithriyah, and Ahda A. Q. Ruswandi. "Efektivitas Ekstrak Etanol Biji Anggur Merah (Vitis vinifera) terhadap Jumlah Fibroblas pada Perawatan Ortodonti." e-GiGi 13, no. 1 (June 23, 2024): 138–44. http://dx.doi.org/10.35790/eg.v13i1.55057.
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Abstract: Malocclusion can be solved with orthodontic treatment. In orthodontic treatment, there will be movements motivated by inflammatory responses and tissue responses in the form of resorption and apposition or bone remodeling. Some of the cells responsible for the process are fibroblasts, osteoblasts, and osteoclasts. Grape seeds contain many active compounds that have high levels of antioxidants and phytoestrogens that can increase the division and proliferation of fibroblasts. This study aimed to find out whether grape seed ethanol extract was effective in increasing fibroblast cell count in orthodontic treatment. This was an experimental laboratory study conducted by attaching an elastomer separator and giving 4% ethanol extract of red grape seeds 0.2 ml in seven groups of guinea pigs that met the inclusion criteria and then were evaluated on days 1, 7, and 14. The Anova test showed a p value of < 0.001 (p<0.05) which meant that there was a statistically significant difference between the treatment groups and the test groups on days 7 and 14. The number of fibroblasts increased from day 1, 7 to day 14. The lowest number of fibroblasts was obtained in the control group (11.7±0.70) while the highest number of fibroblasts was found in the guinea pig group applied rubber separator and given 4% red grape seed extract as much as 0.2ml for 14 days (21.75±1.29). In conclusion, grape seed extract is effective in increasing the number of fibroblasts in orthodontic treatment and is most effective on day 14. Keywords: grape seed; fibroblasts; malocclusion; orthodontic treatment Abstrak: Masalah maloklusi dapat diselesaikan dengan melakukan perawatan ortodonti. Pada perawatan ortodonti akan terjadi pergerakan yang dilatarbelakangi oleh respon inflamasi dan respon jaringan berupa resorpsi dan aposisi atau remodeling tulang. Beberapa jenis sel yang bertanggung jawab atas proses tersebut ialah fibroblas, osteoblas, dan osteoklas. Biji anggur mengandung banyak senyawa aktif yang memiliki tingkat antioksidan tinggi dan fitoestrogen yang mampu membantu meningkatkan pembelahan serta proliferasi dari fibroblas. Penelitian ini bertujuan untuk mengetahui apakah ekstrak etanol biji anggur efektif dalam meningkatkan jumlah sel fibroblas pada perawatan ortodonti. Jenis penelitian ialah eksperimental laboratoris dengan memasangkan separator elastomer dan memberikan ekstrak etanol biji anggur merah 4% sebanyak 0,2 ml pada tujuh kelompok marmut sesuai kriteria inklusi dan dievaluasi pada hari ke-1, hari ke-7, dan hari ke-14. Uji Anova menunjukkan nilai p <0,001 (p<0,05) yang berarti terdapat perbedaan bermakna antara kelompok perlakuan dan kelompok uji dihari ke-7 dan ke-14. Jumlah fibroblas meningkat dari hari ke-1, ke-7 sampai hari ke-14. Jumlah fibroblas paling rendah didapatkan pada kelompok kontrol (11,7±0,70) sedangkan jumlah fibroblas yang paling meningkat terdapat pada kelompok marmut yang diaplikasikan karet separator dan diberi 4% ekstrak biji anggur merah sebanyak 0,2 ml selama 14 hari (21,75±1,29). Simpulan penelitian ini ialah ekstrak biji anggur efektif dalam meningkatkan jumlah fibroblas dalam perawatan ortodonti, paling efektif pada hari ke-14. Kata kunci: biji anggur; fibroblas; malokusi; perawatan ortodonti
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Lim, Ivor Jiun, Toan-Thang Phan, Boon-Huat Bay, Robert Qi, Hung Huynh, Walter Tiang-Lee Tan, Seng-Teik Lee, and Michael Thornton Longaker. "Fibroblasts cocultured with keloid keratinocytes: normal fibroblasts secrete collagen in a keloidlike manner." American Journal of Physiology-Cell Physiology 283, no. 1 (July 1, 2002): C212—C222. http://dx.doi.org/10.1152/ajpcell.00555.2001.
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Keloid scars represent a pathological response to cutaneous injury, reflecting a new set point between synthesis and degradation biased toward extracellular matrix (ECM) collagen accumulation. Using a serum-free two-chamber coculture model, we recently demonstrated a significant increase in normal fibroblast proliferation when cocultured with keloid-derived keratinocytes. We hypothesized that similar keratinocyte-fibroblast interactions might influence fibroblast collagen production and examined conditioned media and cell lysate from coculture for collagen I and III production by Western blot, allied with Northern analysis for procollagen I and III mRNA. Normal fibroblasts cocultured with keloid keratinocytes produced increased soluble collagen I and III with a corresponding increase in procollagen I and III mRNA transcript levels. This was associated with decreased insoluble collagen from cell lysate. When keloid fibroblasts were cocultured with keloid keratinocytes, both soluble and insoluble collagen were increased with associated procollagen III mRNA upregulation. Transmission electron microscopy of normal fibroblasts cocultured with keloid keratinocytes showed an ECM appearance similar to in vivo keloid tissue, an appearance not seen when normal fibroblasts were cocultured with normal keratinocytes.
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Göke, Michael, Michiyuki Kanai, and Daniel K. Podolsky. "Intestinal fibroblasts regulate intestinal epithelial cell proliferation via hepatocyte growth factor." American Journal of Physiology-Gastrointestinal and Liver Physiology 274, no. 5 (May 1, 1998): G809—G818. http://dx.doi.org/10.1152/ajpgi.1998.274.5.g809.
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Although the presence of subepithelial intestinal fibroblasts has been well recognized, the effects of fibroblasts on intestinal epithelial cell (IEC) growth are incompletely understood. In vitro studies were undertaken to evaluate the effects of fibroblasts on the proliferation of model IEC lines. IECs (Caco-2, T84, and IEC-6) were grown alone or in the presence of human intestinal (CCD-18), lung (CCD-37), or skin explant-derived fibroblasts. Cocultures were carried out directly on irradiated fibroblasts or by Transwell coculture technique with fibroblasts and epithelial cells separated by a porous filter. Cell proliferation was assessed by [3H]thymidine incorporation and cell counts. Hepatocyte growth factor (HGF) and c- met transcript expression in IECs and fibroblasts was examined by RT-PCR and Northern blotting; protein expression was evaluated by immunoblotting. Intestinal as well as lung and skin fibroblasts substantially stimulated proliferation of Caco-2, T84, and IEC-6 cells in both direct and Transwell cocultures. In addition, fibroblast-conditioned medium stimulated IEC proliferation, suggesting a paracrine mechanism. Anti-human HGF-neutralizing antibodies blocked the growth-promoting effects in both fibroblasts and fibroblast-conditioned medium. Recombinant human HGF dose dependently promoted IEC proliferation. HGF mRNA and protein expression was restricted to fibroblasts. High levels of c- met expression were found in Caco-2 and T84 cells; in contrast, expression in fibroblasts was weak. In summary, fibroblasts stimulate IEC proliferation through a paracrine mechanism mediated predominantly by HGF.
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Zinman, H. M., M. G. Joneja, P. Davies, and B. T. Smith. "Cell Culture of Embryonic Chick Duodenal Cells." Journal of Pediatric Gastroenterology and Nutrition 4, no. 1 (February 1985): 107–17. http://dx.doi.org/10.1002/j.1536-4801.1985.tb08798.x.
Full textAbstract:
A method for the primary cell culture of trypsin‐dissociated embryonic chick duodenum is described. Both heterotypic (epithelial cells and fibroblasts together) and homotypic (highly enriched cultures of epithelial cells or fibroblasts alone) cell cultures were established. Dispersed duodenal epithelial cells and fibroblasts grown in 10% fetal bovine serum (FBS) spontaneously aggregated and proliferated as a bilayer of cells with the epithelial cells growing on top of the fibroblasts. Changing the serum supplement to 6% chicken serum (CS) and 4% FBS when the fibroblast monolayer reached confluence resulted in epithelial cell proliferation. Homotypic cultures of epithelial cells and fibroblasts were prepared and analyzed by scanning electron microscopy and transmission electron microscopy. Fibroblasts, isolated by differential adhesion and grown in 10% FBS, did not demonstrate measurable alkaline phosphatase activity. Homotypic epithelial cell cultures, isolated by floating them off the fibroblasts with collagenase, and maintained on collagen in 6% CS/4% FBS, demonstrated higher alkaline phosphatase‐specific activity (16.1 β 2.3 U/mg protein) compared with epithelial‐fibroblast bilayer cell cultures (12.1 β 1.3 U/mg protein).
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Kuzmicheva, Valeriya I., Larisa T. Volova, Frida N. Gilmiyarova, Ilya M. Bykov, Elena V. Avdeeva, and Natalya A. Kolotieva. "Fibroblasts as the subject of proliferative activity research in vitro." Science and Innovations in Medicine 5, no. 3 (October 20, 2020): 210–15. http://dx.doi.org/10.35693/2500-1388-2020-5-3-210-215.
Full textAbstract:
This review presents the data devoted to anatomical and functional diversity of fibroblasts, peculiarities of metabolic processes and energy exchange in these cells.
In particular, the changes in fibroblast proliferative activity depending on various factors are discussed. The review shows the influence of the malate dehydrogenase shuttle system on the activity of metabolic processes and the life span of fibroblastsin vitro. The increase of cell cultivation timein vitrois associated with the cytosolic isoform of this enzyme.
The stability of fibroblast cell culture to the activation of free-radical processes and peroxidation with addition of biologically active compounds is described and followed by a discussion of the role of separate metabolites in providing free-radical protection and maintenance of the proliferative potential of cells.