Littérature scientifique sur le sujet « Tubular regeneration »

The proximal tubule epithelium regenerates following nephrotoxic damage. To determine the role of fibroblast growth factors (FGFs) in the regeneration of rat proximal tubule epithelial (RPTE) cells, we investigated proliferation, differentiation, and FGF-1 expression in vivo in rat kidney before and after nephrotoxic damage to the proximal tubule epithelium caused by S-(1,1,2,2-tetrafluoroethyl)-L-cysteine administration. In undamaged kidneys, FGF-1 was expressed in distal tubule elements, including cortical and medullary collecting ducts, as well as in blood vessels and glomeruli, but was absent in RPTE. One day after damage, there was an increase in proliferation of surviving proximal tubule epithelial cells and a coincident increase in FGF-1 expression in invading mononuclear cells. After this initial burst of proliferation, FGF-1 expression increased in poorly differentiated vimentin-positive regenerative epithelial cells, indicating that autocrine FGF-1 expression in the regenerative epithelium is a later event in the regeneration process. FGF-1 staining persisted in foci of macrophages, interstitial cells, and nephropathic tubules within areas of interstitial expansion 2 wk after damage. We concluded that transient paracrine and autocrine expression of FGF-1 could play mitogenic and/or morphogenic roles during tubular regeneration. Persistent expression in macrophages, fibroblasts, and nephropathic tubules may be associated with tubular degeneration. FGF-1 expression may be an important contributor to both tubular regeneration and degenerative disease following toxicant exposure.

Renal tubular epithelium has the capacity to regenerate, repair, and reepithelialize in response to a variety of insults. Previous studies with several kidney injury models demonstrated that various growth factors, transcription factors, and extracellular matrices are involved in this process. Surviving tubular cells actively proliferate, migrate, and differentiate in the kidney regeneration process after injury, and some cells express putative stem cell markers or possess stem cell properties. Using fate mapping techniques, bone marrow-derived cells and endothelial progenitor cells have been shown to transdifferentiate into tubular components in vivo or ex vivo. Similarly, it has been demonstrated that, during tubular cell regeneration, several inflammatory cell populations migrate, assemble around tubular cells, and interact with tubular cells during the repair of tubular epithelium. In this review, we describe recent advances in understanding the regeneration mechanisms of renal tubules, particularly the characteristics of various cell populations contributing to tubular regeneration, and highlight the targets for the development of regenerative medicine for treating kidney diseases in humans.

Acute kidney injury has a high global morbidity associated with an increased risk of death and chronic kidney disease. Renal tubular epithelial cell regeneration following injury may be a decisive factor in renal repair or the progression of acute kidney injury to chronic kidney disease, but the underlying mechanism of abnormal renal tubular repair remains unclear. In the present study, we investigated the role of heterotrimeric G stimulatory protein α-subunit (Gsa) in renal tubular epithelial cell regeneration. We generated renal tubule epithelium-specific Gsa knockout (GsaKspKO) mice to show the essential role of Gsa in renal tubular epithelial cell regeneration in two AKI models: acute aristolochic acid nephropathy (AAN) and unilateral ischemia-reperfusion injury (UIRI). GsaKspKO mice developed more severe renal impairment after AAN and UIRI, higher serum creatinine levels, and more substantial tubular necrosis than wild-type mice. More importantly, Gsa inactivation impaired renal tubular epithelial cell proliferation by reducing bromodeoxyuridine+ cell numbers in the AAN model and inhibiting cyclin-dependent kinase 2/cyclin E1 expression in the UIRI model. This reduced proliferation was further supported in vitro with Gsa-targeting siRNA. Downregulation of Gsa inhibited tubular epithelial cell proliferation in HK-2 and mIMCD-3 cells. Furthermore, Gsa downregulation inhibited cyclin-dependent kinase 2/cyclin E1 expression, which was dependent on the Raf-MEK-ERK signaling pathway. In conclusion, Gsa is required for tubular epithelial cell regeneration during kidney repair after AKI. Loss of Gsa impairs renal tubular epithelial cell regeneration by blocking the Raf-MEK-ERK pathway.

Differential display-polymerase chain reaction (DD-PCR) was used to identify genes that are expressed in kidney following induction of acute ischemic renal injury. The receptor for activated C kinase (RACK1) mRNA expression in kidneys obtained from rats 12 h following ischemia is enhanced twofold compared with sham-operated rats. The maximal enhancement of expression (3.3-fold) is at 7 days following reperfusion. Expression remains elevated at 14 days. RACK1 transcripts and protein are localized to the damaged and regenerating segments of proximal tubules. At 1 day following injury, RACK1 protein is present in the epithelial cells of the damaged S3 segment and in cells sloughed into the tubular lumen. By 5 days following injury, RACK1 protein expression is enhanced in the regenerating cells relining the injured tubules of the S3 segment and in papillary proliferations within regenerating tubules. Increased expression of RACK1 could enhance the activity of PKC and, in so doing, regulate the process of regeneration of the proximal tubule following ischemic renal injury.

Background: Acute kidney injury (AKI) is a common cause of morbidity and mortality. It mainly targets the renal tubular epithelium with pathological changes, referred to as acute tubular injury. The latter is followed by a regenerative response that is difficult to visualize on routine hematoxylin and eosin (H&E) stains. In this study, we examined the regenerative capacity of renal tubules by correlating vimentin (VIM) immunohistochemical (IHC) expression and pathological findings of AKI and renal tubular regeneration (RTR) on H&E.Methods: We reviewed 23 autopsies performed in the clinical setting of AKI and RTR. VIM expression was scored in the renal cortical tubular epithelium using a statistical cutoff ≥ 3% for high expression and < 3% for low expression.Results: Of the 23 kidney tissues examined, seven (30.4%) had low VIM expression, and 16 (69.6%) had high VIM expression. Kidney tissues with evidence of AKI and RTR had significantly higher VIM expression. Renal peritubular microenvironment features showing regenerative changes on H&E were associated with high VIM expression. In the univariate model, kidney tissues with RTR were 18-fold more likely to have high VIM expression.Conclusions: In conclusion, our findings suggest that VIM could serve as an IHC marker for RTR following AKI. However, correlation with H&E findings remains critical to excluding chronic tubular damage. Collectively, our preliminary results pave the way for future studies including a larger sample size to validate the use of VIM as a reliable biomarker for RTR.

Renal proximal tubular epithelium can regenerate after various insults. To examine whether the tubular repair process is regulated by surrounding peritubular capillaries, we established an in vitro human tubulogenesis model that mimics in vivo tubular regeneration after injury. In this model, HGF, a potent renotropic factor, dose dependently induced tubular structures in human renal proximal tubular epithelial cells cultured in gels. Consistent with regenerating tubular cells after injury, HGF-induced tubular structures expressed a developmental gene, Pax-2, and a mesenchymal marker, vimentin, and formed a lumen with aquaporin-1 expression. Electron microscopic analysis showed the presence of microvilli on the apical site of the lumen, suggesting that these structures are morphologically equivalent to renal tubules in vivo. When cocultured with human umbilical vein endothelial cells (HUVEC), HGF-induced tubular formation was significantly enhanced. This could not be reproduced by the addition of VEGF, basic FGF, or PDGF. Protein array revealed that HUVEC produced various matrix metalloproteinases (MMPs). The stimulatory effects of coculture with HUVEC or HUVEC-derived conditional medium were almost completely abolished by addition of the tissue inhibitor of metalloproteinase (TIMP)-1 or TIMP-2. These data suggest that endothelial cell-derived factors including MMPs play a critical role in tubulogenesis and imply a potential role of peritubular capillary endothelium as a source of factor(s) required for tubular recovery after injury.

BackgroundThe kidney is considered to be a structurally stable organ with limited baseline cellular turnover. Nevertheless, single cells must be constantly replaced to conserve the functional integrity of the organ. PDGF chain B (PDGF-BB) signaling through fibroblast PDGF receptor-β (PDGFRβ) contributes to interstitial-epithelial cell communication and facilitates regenerative functions in several organs. However, the potential role of interstitial cells in renal tubular regeneration has not been examined.MethodsIn mice with fluorescent protein expression in renal tubular cells and PDGFRβ-positive interstitial cells, we ablated single tubular cells by high laser exposure. We then used serial intravital multiphoton microscopy with subsequent three-dimensional reconstruction and ex vivo histology to evaluate the cellular and molecular processes involved in tubular regeneration.ResultsSingle-tubular cell ablation caused the migration and division of dedifferentiated tubular epithelial cells that preceded tubular regeneration. Moreover, tubular cell ablation caused immediate calcium responses in adjacent PDGFRβ-positive interstitial cells and the rapid migration thereof toward the injury. These PDGFRβ-positive cells enclosed the injured epithelium before the onset of tubular cell dedifferentiation, and the later withdrawal of these PDGFRβ-positive cells correlated with signs of tubular cell redifferentiation. Intraperitoneal administration of trapidil to block PDGFRβ impeded PDGFRβ-positive cell migration to the tubular injury site and compromised the recovery of tubular function.Conclusions Ablated tubular cells are exclusively replaced by resident tubular cell proliferation in a process dependent on PDGFRβ-mediated communication between the renal interstitium and the tubular system.

Objective: The aim of the study was to evaluate whether Vernonia cinerea (VC) regenerates the proximal renal tubular cells in cisplatin-induced necrosis in male Swiss albino mice.Methods: The crude aqueous extract (CAE) of VC was fractionated from non-polar to polar using different solvents. Mice were injected a single dose of cisplatin (15 mg/kg) on day 1, which took 5 days to cause maximal renal damage. From day 6, CAE and all fractions were orally administered (200, 300, and 400 mg/kg) for 5 continuous days. On day 11, blood was collected to estimate urea and creatinine. Kidney was collected for histology and grading was done.Results: Cisplatin induced proximal renal tubular damage (grade 5) in corticomedullary junction, characterized by necrosis, proximal tubular dilatation, inflammation and vasodilation. Aqueous fraction (AF) did not show any regeneration; whereas, 400 mg/kg dose of CAE and butanol fraction (BF) showed a significant reduction (p<0.001) in proximal tubular damage (Grade 3) and 50–75% regeneration of proximal tubular epithelial cells.Conclusion: This is the first study to demonstrate the regenerative potential of Neichitti kashayam (CAE of VC) and its BF in cisplatin-induced proximal tubular damage in kidney. Further study is warranted to find out the dose regimen for complete regeneration, lead compounds, and molecular mechanism.

Objective: The aim of the study was to evaluate whether Vernonia cinerea (VC) regenerates the proximal renal tubular cells in cisplatin-induced necrosis in male Swiss albino mice.Methods: The crude aqueous extract (CAE) of VC was fractionated from non-polar to polar using different solvents. Mice were injected a single dose of cisplatin (15 mg/kg) on day 1, which took 5 days to cause maximal renal damage. From day 6, CAE and all fractions were orally administered (200, 300, and 400 mg/kg) for 5 continuous days. On day 11, blood was collected to estimate urea and creatinine. Kidney was collected for histology and grading was done.Results: Cisplatin induced proximal renal tubular damage (grade 5) in corticomedullary junction, characterized by necrosis, proximal tubular dilatation, inflammation and vasodilation. Aqueous fraction (AF) did not show any regeneration; whereas, 400 mg/kg dose of CAE and butanol fraction (BF) showed a significant reduction (p<0.001) in proximal tubular damage (Grade 3) and 50–75% regeneration of proximal tubular epithelial cells.Conclusion: This is the first study to demonstrate the regenerative potential of Neichitti kashayam (CAE of VC) and its BF in cisplatin-induced proximal tubular damage in kidney. Further study is warranted to find out the dose regimen for complete regeneration, lead compounds, and molecular mechanism.

ABSTRACT Peripheral nerve injury is a common clinical problem significantly affecting the patients’ quality of life. In case of severe transections, the bridging of the gap between the proximal and distal nerve stumps is required and autologous nerve grafts using sensory nerves (i.e. the sural nerve or antebrachial cutaneous nerve) are the current criterion standard. Nevertheless, donor-site morbidities, permanent loss of function, size mismatch between the donor nerve and the injured nerve and poor functional recovery rates have prompted the interest towards the identification of an alternative to this technique. To date, surgeons and researchers are turning their attention towards different grafts made of biological or artificial polymers. In fact, the development of hollow nerve guide conduits a) creating an adequate microenvironment for nutritional support/axons regeneration; b) acting as a barrier against the surrounding tissue infiltration; c) matching the effectiveness of the autologous nerve graft, would be beneficial to the field of peripheral nerve surgery. Over the years, many biomaterials of natural or synthetic origin and with different characteristics in terms of biodegradability have been studied. However, it has not been identified yet a prosthesis able to guarantee a better regenerated tissue than the others.
The aim of the present study was to manufacture and investigate in vitro and in vivo the characteristics and the regenerative potential of three different nerve conduits made up of polyvinyl alcohol (PVA); 1% Oxidized PVA (1% Ox PVA) and Silk-Fibroin (SF). While the use of PVA and SF for the realization of neuro-guides has already been studied in the past, oxidized PVA (recently patented by our research group) is a new material for this purpose. In parallel, this study also allowed to assess the quality of axonal regeneration guaranteed by neuro-guides with different origin (synthetic vs. natural) and biodegradation properties (vs non-biodegradable biodegradable). After preparing the three different polymer solutions, disk-shaped and tubular supports were manufactured. These were employed for in vitro and in vivo studies respectively. Considering in vitro analysis, a morphological characterization of supports was performed by Scanning Electron Microscopy (SEM). Thereafter, the biocompatibility and the biological activity of the three different scaffolds was assessed using a Schwann-cell line (SH-SY5Y). Cells were seeded on supports and their adhesion and proliferation was evaluated by SEM and MTT assay at two different end-points (3 and 7 days from seeding). Regarding in vivo tests, nerve conduits were implanted in animal models (Sprague-Dawley rats) of peripheral nerve injury with loss of substance (nerve gap: 5 mm). At 12-weeks from surgery, the functional recovery of the sciatic nerve was assessed; thereafter, the animals were euthanized and the dissection occurred. Prior to explant, the gross appearance of grafts was carefully observed in situ. Specimens were than processed for histological (hematoxylin and eosin staining) and immunohistochemical analysis (anti-CD3; anti-S100) as well as for further Transmission Electron Microscopy (TEM) analysis. The objective was to assess the quality of the regenerated nerve-tissue highlighting any differences in efficacy between the three types of nerve-conduits; to this end, the histomorphological analysis has been fundamental allowing to quantify the axons (myelinated vs unmyelinated nerve fibers) at different levels of the explant (proximal vs central vs distal portion); the controlateral sciatic nerve was used as control. Considering the in vitro results, SEM micrographs showed that PVA and SF supports have a smooth and regular surface; conversely, a certain roughness was noticed observing the ultrastructure of 1% Ox PVA disk-shaped scaffolds. Despite the superficial appearance of supports, it does not seem to affect the interaction with the cells. In fact, PVA-based scaffolds do not support cell adhesion and proliferation; SEM analysis and the MTT assay do not identified the presence of SH-SY5Y cells after 3 and 7 days from seeding. This result can be attributed to the high hydrophilic nature of the hydrogels. Conversely, SF scaffolds are adequate to promote SH-SY5Y cells growth. Regarding the in vivo study, all nerve conduits showed good characteristics in terms of handiness, being easy-suturing and demonstrating also an adequate tear-resistance feature; PVA-based scaffolds appear more flexible than SF guides. After 12 weeks from surgery, all animals showed a sciatic nerve functional recovery; in particular, all of them supported their body weight on the hind leg even though animals implanted with PVA and SF nerve conduits sometimes showed spasms during the walk while not limping. On the contrary, animals implanted with 1% Ox PVA nerve conduits exhibited a normal movement. At the time of dissection, the three scaffolds were still clearly identifiable. Any dislocation of the grafts or neuroma formation at the stumps was observed; moreover, the transparency of the three scaffolds allowed to identify the presence of a regenerated tissue inside. Thereafter, histological and immunohistochemical analysis were performed to evaluate the quality of axonal regeneration. Preliminarily, the haematoxylin and eosin staining of the specimens (cross-section of the central portion) highlighted the morphological integrity of the structure. In fact, three layers are recognizable proceeding from the periphery to the inside of the sections: an external fibrous capsule; a layer corresponding to the nerve conduit; a homogeneous and dense regenerated tissue in the middle. The biocompatibility of the grafts was verified by immunohistochemical analysis; anti-CD3 immunohistochemistry demonstrated the absence of severe inflammatory reactions. At the same time, several S100+ cells were identified suggesting the extensive presence of Schwann-cells. In parallel, the typical peripheral nerve morphology was highlighted also by Toluidine Blue staining by means of was considered also the appearance of the proximal and distal stumps. Although all samples support the recovery of the lesion, some differences can be found between the three experimental groups; these results were confirmed also by TEM micrographs. The histomorphometric analysis of samples evaluated the total axons number per nerve and axon density (axons/μm2); for each graft were considered the proximal, the central and the distal section. The collected data showed that 1% Ox PVA conduits assure a better outcome in nerve regeneration than the non-biodegradable PVA grafts which among the three groups proved to be the ones with the lower outcomes.
The results of this study showed that all nerve conduits considered (PVA; 1% Ox PVA and SF) promote peripheral nerve regeneration in case of neurotmesis with loss of substance. Considering the quality of regenerates, better outcomes were observed analyzing the 1% Ox PVA explants compared to PVA and Silk-Fibroin ones.
RIASSUNTO Le lesioni nervose periferiche costituiscono un problema clinico piuttosto comune, il quale inficia in modo significativo la qualità della vita dei pazienti. In caso di lesioni gravi con perdita di sostanza, al fine di colmare il gap tra il moncone prossimale ed il distale, il gold- standard prevede l’impianto di innesti nervosi autologhi utilizzando nervi sensoriali (ad es., nervo surale o nervo cutaneo antibrachiale). Tuttavia, criticità quali la morbidità del sito donatore, la perdita in funzionalità, la mancata corrispondenza dimensionale tra il nervo donatore ed il nervo lesionato oltre ad uno scarso recupero funzionale hanno spinto l'interesse verso l'identificazione di un approccio alternativo. Allo stato dell’arte, chirurghi e ricercatori stanno volgendo la loro attenzione verso innesti polimerici diversi (grafts) di natura sia biologica che artificiale. Infatti, lo sviluppo di neuroguide capaci di: a) creare un microambiente ideale per la rigenerazione assonale; b) fornire una protezione dall'infiltrazione di tessuto circostante; c) possedere un’efficacia analoga a quella garantita dall’innesto nervoso autologo; costituirebbe un vantaggio significativo nell’ambito della chirurgia del nervo periferico. Nel corso degli anni, sono stati studiati molti biomateriali di origine sia naturale che sintetica aventi caratteristiche differenti in termini di biodegradabilità. Tuttavia, considerando la qualità del tessuto rigenerato, non è ancora stata individuata una protesi più performante rispetto alle altre. L’obiettivo di questo studio è stato quello di allestire e studiare, sia in vitro che in vivo, le caratteristiche ed il potenziale rigenerativo di tre diverse neuroguide rispettivamente costituite da: alcool polivinilico (PVA); PVA ossidato 1% (PVA Ox 1%) e Fibroina della Seta (FS). Mentre l’impiego di PVA e FS per la realizzazione di grafts è già stato investigato in passato, il PVA Ox 1% (recentemente brevettato dal nostro gruppo di ricerca) costituisce un nuovo materiale per questo scopo. In parallelo, questo studio ha anche consentito di confrontare la qualità della rigenerazione assonale sostenuta da neuroguide diverse sia per origine (sintetica vs naturale) che per proprietà biodegradative (biodegradabili vs nonbiodegradabili).
Dopo aver allestito le tre diverse soluzioni polimeriche, sono stati quindi preparati scaffolds sia discoidali che in forma di graft tubulare, utilizzati rispettivamente per i successivi studi in vitro e in vivo. Nell’ambito degli studi in vitro, è stata effettuata una caratterizzazione morfologica dei supporti mediante microscopia elettronica a scansione (SEM). Successivamente, la biocompatibilità e l'attività biologica dei tre differenti scaffolds è stata valutata utilizzando una linea di cellule di Schwann (SH-SY5Y). Le cellule sono state seminate sui supporti e la loro adesione e la proliferazione è stata valutata mediante saggio MTT oltre che SEM a due differenti end-point (3 e 7 giorni dalla semina). Per quanto riguarda lo studio in vivo, i graft tubulari sono stati impiantati in modelli animali (ratti Sprague- Dawley) di lesione nervosa periferica con perdita di sostanza (gap tra moncone prossimale e distale: 5 mm). A 12 settimane dalla chirurgia, è stato valutato il recupero funzionale del nervo sciatico; successivamente, gli animali sono stati sacrificati. Dopo dissezione, prima di procedere all’espianto, l'aspetto macroscopico degli innesti è stato attentamente osservato in situ. I campioni sono stati quindi prelevati e trattati per le successive analisi istologiche (ematossilina ed eosina) ed immunoistochimiche (anti-CD3; anti-S100) nonché per ulteriori analisi di microscopia elettronica a scansione (TEM). L'obiettivo è stato quello di valutare la qualità del tessuto rigenerato evidenziando eventuali differenze di efficacia tra i tre tipi di grafts; a tal fine, anche l'analisi istomorfologica si è rivelata fondamentale, permettendo essa di quantificare gli assoni (mielinici vs amielinici) in diverse porzioni del campione (porzione prossimale vs centrale vs distale). Il nervo sciatico controlaterale è stato usato come controllo. Considerando i risultati degli studi in vitro, le immagini al SEM hanno mostrato come i supporti in PVA e FS mostrino una superficie liscia e regolare; al contrario, una certa ruvidità è stata notata osservando l’ultrastruttura degli scaffold discoidali in PVA Ox 1%. Nonostante il diverso aspetto ultrastrutturale dei supporti, esso non sembra influenzare l'interazione con le cellule. Il PVA (sia nativo che ossidato) non sostiene l'adesione e la proliferazione cellulare; infatti, sia le analisi al SEM che il saggio MTT non hanno identificato la presenza di cellule SH-SY5Y dopo 3 e 7 giorni dalla semina. Questo risultato può essere attribuito alla elevata idrofilia degli idrogeli Al contrario, gli scaffold in FS sono adeguati per promuovere la crescita delle SH-SY5Y. Per quanto riguarda lo studio in vivo, tutti i graft mostrato buone caratteristiche in termini di manipolabilità, essendo facilmente suturabili e dimostrando anche una adeguata resistenza allo strappo; gli scaffold in PVA appaiono più flessibile rispetto alle guide in FS. Dopo 12 settimane dalla chirurgia, tutti gli animali hanno mostrato un certo recupero funzionale dell’arto operato; in particolare, tutti distribuivano il proprio peso corporeo anche sulla zampa posteriore. Pur non zoppicando, gli animali impiantati con PVA e SF mostravano talvolta degli spasmi durante la deambulazione, al contrario, gli animali impiantati con graft in PVA Ox 1% esibivano un movimento normale. Al momento della dissezione, i tre graft erano ancora chiaramente identificabili. Non è stata riscontrata alcuna dislocazione degli innesti o formazione di neuroma in corrispondenza dei monconi; inoltre, la trasparenza delle tre neuroguide ha permesso di identificare la presenza di un tessuto rigenerato al loro interno. Successivamente, sono state effettuate analisi istologiche ed immunoistochimiche per valutare la qualità della rigenerazione assonale. Preliminarmente, mediante colorazione con ematossilina ed eosina (sezione trasversale della porzione centrale) è stato possibile mettere in evidenza l'integrità morfologica della struttura. Procedendo dalla periferia della sezione verso l'interno sono riconoscibili: una capsula fibrosa esterna; il graft; ed il tessuto neo-rigenerato, omogeneo e denso, nel mezzo. La biocompatibilità degli innesti è stata verificata mediante analisi immunoistochimica; la scarsa presenza di cellule CD3+ ha dimostrato l'assenza di reazioni infiammatorie gravi riconducibili all’impianto. Contestualmente, l’elevata presenza di elementi S100+ riscontrata in tutti i campioni ha comprovato una evidente rigenerazione assonale. In parallelo, la morfologia tipica del tessuto nervoso periferico è stata altresì evidenziata mediante colorazione con Blu di Toluidina mediante la quale è stato considerato anche l'aspetto dei monconi prossimale e distale.
Sebbene tutti i campioni supportino il recupero della lesione, alcune differenze possono essere riscontrate tra i tre gruppi sperimentali; questi risultati sono stati confermati anche dalle micrografie al TEM. L'analisi morfometrica dei campioni ha valutato il numero totale di assoni/nervo e la loro densità (assoni / μm2); per ogni innesto sono state considerate le sezioni prossimale, centrale e distale. I dati raccolti hanno dimostrato che il PVA Ox 1% assicura un risultato migliore nella rigenerazione assonale rispetto agli innesti non biodegradabili in PVA, il quale tra i tre gruppi è risultato essere quello con l’outcome inferiore. I risultati di questo studio hanno mostrato che, in caso di neurotmesi con perdita di sostanza, tutti i graft allestiti (PVA; PVA Ox 1% e FS) promuovono la rigenerazione del nervo. Considerando la qualità del tessuto rigenerato, sono stati osservati dei risultati migliori con graft in PVA Ox 1% rispetto a quelli ottenuti da neuroguide in PVA e FS.

Thesis (Ph.D.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2000.
Includes bibliographical references (leaves 143-148).
Peripheral nerve injury affects nearly 200,000 patients annually in the United States and unless treated results in paralysis of skeletal muscle and loss of sensation. Previous studies in this laboratory have focused on comparing the effectiveness of various tubular devices in repairing experimental nerve injuries in an animal model. The devices were rank-ordered based on clinically relevant assays of regeneration such as number regenerated nerve fibers and electrophysiological conduction properties of the regenerated nerves. Such assays provide a useful measure of the clinical efficacy of devices but require long-term (up to 60-week) studies in order to obtain meaningful results. There exists a need for a short-term (less than 12-week) assay with which nerve repair devices can be compared. The overall goal of this thesis was to establish an experimental assay that can be used to detect statistically significant differences among nerve repair devices in short-term studies. In this thesis, four different assays of nerve regeneration were compared on the basis of their appropriateness to quantify the regeneration promoted by nerve repair devices in studies less than 12 weeks in duration. An acceptable assay must reach a plateau with time during short-term studies and must yield a quantitative metric with which nerve devices can be compared. The results of this thesis suggest that an assay based on ability of a nerve repair device to promote reinnervation across nerve gaps of various lengths meets the criteria for an acceptable assay. The data also indicate that the characteristic gap length (Lc), which is derived from curve-fitting the experimental data for reinnervation versus gap length, can be used as a quantitative metric of nerve regeneration. The experimental data indicate that for the silicone tube device, the value of Lc reached a plateau with time before 9 weeks, and the standard error in L was less than 5 percent of the value in two different nerve repair models (single-leg and crossanastomosis). The data also suggest that statistically significant differences between the silicone tube device and a collagen-based device (the CG device) are obtainable during short-term (12-week) studies.
by Mark H. Spilker.
Ph.D.

Orientador: Eliana Aparecida de Resende Duek
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Mecanica
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Mestrado
Materiais e Processos de Fabricação
Mestre em Engenharia Mecânica

During the central nervous system (CNS) morphogenesis, chemical gradients of morphogens such as retinoic acid (RA) or sonic hedgehog play a central role in regulating CNS patterning and differentiation of neural subtypes. Recapitulation of these gradients in a 3D hydrogel matrix may provide a model for CNS tissue formation in vitro. 3D Printing technology offers an opportunity to reproduce the complex architecture of cell microenvironment. We have developed a 3D-printable alginate hydrogel bioink suitable for extrusion-based bioprinting. The bioink was characterised by shear thinning, high printing resolution and minimal adverse effects on cell viability. The bioink was successfully used to print mouse embryonic stem cells (mESCs)-laden constructs and supported their differentiation into neural-like cells. Extrusion-based bioprinting was used to 3D-print hybrid polycaprolactone (PCL)-alginate tubular scaffolds functionalised with a fluorescein isothiocyanate-conjugated bovine serum albumin (FITC-BSA) concentration gradient pattern. Quantification of the FITC-BSA concentrations in the scaffold showed a linear reduction in concentration as a function of scaffold’s distance (length). Tubular scaffolds printed with fibroblast-laden alginate supported cell viability and proliferation up to 6 days after printing. Next, the developed model was used to replicate the in vivo RA-induced directed differentiation of mESCs into spinal cord neurons. RA-concentration-dependent acquisition of neural identity was investigated using immunocytochemistry and flow cytometry analysis. RA promoted the formation of neurons with hindbrain and spinal cord identity and supressed the forebrain identity in a concentration-dependent manner. Among the investigated hydrogels, gelatine methacrylate (GelMA) supported neural differentiation and neurite outgrowth of the mESCs-derived embryoid bodies (EBs). Subsequently EBs-laden GelMA (5%) was successfully used as a bioink to print the hybrid PCL-hydrogel scaffolds. 3D Printing of EBs and RA-loaded GelMA in PCL scaffold induced differentiation of EBs into neurons with spinal cord positional identity. In conclusion, the model can be used for effective morphogens gradients delivery to replicate some of the complex processes of CNS development in vitro.

Acute kidney injury (AKI), regardless of its aetiology, can elicit persistent or permanent kidney tissue changes that are associated with progression to end-stage renal disease and a greater risk of chronic kidney disease (CKD). In other cases, AKI may result in complete repair and restoration of normal kidney function. The pathophysiological mechanisms of renal injury and repair include vascular, tubular, and inflammatory factors. The initial injury phase is characterized by rarefaction of peritubular vessels and engagement of the immune response via Toll-like receptor binding, activation of macrophages, dendritic cells, natural killer cells, and T and B lymphocytes. During the recovery phase, cell adhesion molecules as well as cytokines and chemokines may be instrumental by directing the migration, differentiation, and proliferation of renal epithelial cells; recent data also suggest a critical role of M2 macrophage and regulatory T cell in the recovery period. Other processes contributing to renal regeneration include renal stem cells and the expression of growth hormones and trophic factors. Subtle deviations in the normal repair process can lead to maladaptive fibrotic kidney disease. Further elucidation of these mechanisms will help discover new therapeutic interventions aimed at limiting the extent of AKI and halting its progression to CKD or ESRD.

This chapter discusses renal disorders and includes discussion on prevention of acute kidney injury, including optimizing renal perfusion with the use of volume expansion, inotropic, vasopressor, and vasodilator medications; modulation of renal physiology, including renal metabolism, tubular obstruction, oxygen radical damage, and renal regeneration and repair. This chapter also discusses the diagnosis of acute kidney injury, including parameters of glomerular function, urine analyses, biomarkers, ultrasound, autoimmune profiling, and renal biopsy.

Abstract Heat pumps based on the vapor compression cycle account for a significant portion of energy use around the world. However, growing demands for energy efficient and environmentally friendly technologies have created a need for new space conditioning approaches. Novel systems which use elastocaloric material have shown potential to replace traditional vapor compression due to high energy efficiency and use of environmentally friendly, solid-state refrigerants. The solid-state refrigerants exhibit the elastocaloric effect, a phenomenon that occurs when metal alloys experience stress-induced reversible phase transformations resulting in latent heat release or absorption. Prototypes built in the Center for Environmental Energy Engineering have utilized the active elastocaloric regeneration (AER) operating method to develop high temperature gradients between the ends of a regenerative heat exchanger made of tubular elastocaloric material. Though this schema significantly increases the temperature span developed by elastocaloric cooling devices, the current heat pump design leads to temperature degradation as a result of conduction along the length of the tubes in the regenerator. The novel regenerator concept presented in this work mitigates that issue by using short, thermally insulated tubes layers which also enables fluid flow over external surface areas of the material.

Abstract A novel hybrid system for combustion air heating, including flue gas cooling, air heating and heat regeneration has been proposed. In the reformative scheme, the air gains energy from four tubular heat exchangers and the flue gas releases heat in four tubular heat exchangers as well, instead of the rotary regenerative air preheater (APH) that is used in the conventional scheme. Consequently, the temperature differences between the fluids during heat transmission can be diminished, and the mixing of the hot-cold primary air and the severe leakages are avoided, which remarkably reduces the exergy destruction and enhances the thermal performance of the power unit. The new design was evaluated based on a 670 MW coal-fired supercritical power unit. The results show that the additional net power output of the power unit can reach 8.57 MW with a net efficiency promotion of 0.57 percentage points due to the novel configuration. And the energy saving mechanism of the proposed concept was revealed on grounds of the first and second laws of thermodynamics.

Development of a small-diameter vascular graft (<6 mm) have been challenging due to thrombosis and intimal hyperplasia [1]. To overcome this problem, cardiovascular tissue engineers have attempted to construct a highly porous and biocompatible fibrous scaffold providing a sufficient mechanical strength for the regeneration of a functional tissue [2–5]. Herein, we present a 3D tubular-shaped micro/nanofibrous composite-layered scaffold for vascular tissue engineering. The surface of scaffold has high surface roughness by introducing nanofibrous layer and the biophysical properties have been fulfilled by using microfibrous layer. Moreover, the atomized spraying technique is applied to spray elastin proteins, which is well known as an antithrombogenic material, on the surface of micro/nanofibrous composite-layered scaffold to introduce an appropriate antithrombogenic surface.

A review of regenerative gas turbines operating at natural gas pipeline compressor stations across the Russia has been performed. Main performance characteristics, first of all, the power and efficiency of the recuperated gas turbines, many of those have been used up a design service life, can be recovered. Since a large negative effect on the performances is contributed by defective plate-type regenerators, their change seems to be essential when updating gas-pumping units. A tubular regenerator is developed and incorporated into the gas turbines for driving natural gas blowers. The regenerator being installed in place of the plate-type heat exchanger joints a rather simple fabrication process and high durability. Its design features are presented and discussed. Ways to enhance efficiency and decrease the weight of the regenerator are considered.

On the way to a new era of our society which will be based on hydrogen energy, it is needed to develop on-site hydrogen production systems to cover current insufficient infrastructures of hydrogen supply network systems. For this, a highly efficient compact reformer can be one of the most suitable solutions for on-site production of hydrogen which is supplied to distributed electric power-generation systems. But, the local and overall energy balance in the reformer should be precisely controlled since the reforming reaction processes of hydrocarbon fuels are very sensitive to reaction temperature in the reformer. For smaller reformers, in particular, the amount of heat loss through the outer surfaces is large enough to dominate the reactions. An appropriate way for thermal energy management, therefore, is necessary to accomplish highly efficient reformers. For these backgrounds, a compact tubular-typed fuel reformer was fabricated in this study, and was applied to produce hydrogen from methanol, focusing on the partial oxidation reaction (POR). The reformer was composed of a stainless steel pipe as the reactor exterior and ceramic honeycomb blocks inserted in two locations of the reactor. The honeycomb blocks are expected to assist the reforming reactions and transfer the thermal energy of the exhaust gas to the reaction region, acting as a heat regenerator. The upstream-side honeycomb block was aimed to perform an effective heat exchange from the reactor wall to the reactant gas. By inserting the block, the reforming reaction became stable at right after the block. The maximum hydrogen production was achieved in the condition of equivalence ratio, around 3.5. The other honeycomb block was inserted in the downstream of the reaction zone to convert the thermal energy of exhaust gas to radiation energy which can be transferred to the upstream reaction region. Comparing to the case without the downstream-side block, the temperature of the reaction region became higher. Gas temperatures in the downstream region, on the other hand, became lower. Methanol conversion ratio and hydrogen production ratio enhanced due to the higher temperature at the reaction region. These results indicate that the thermal energy possessed by the exhaust gas was regenerated in the reaction region by the downstream-side honeycomb block and contributes to enhance the efficiency of the fuel reformer.

A compact tubular-type fuel reformer was fabricated and operated under fuel-rich combustion conditions of methanol, focusing on the partial oxidation reaction (POR). Ceramic honeycomb strainer blocks were inserted in the reactor. In the authors’ previous study, Case-1 of only one honeycomb block insertion showed that the reaction region formed in the downstream of the block. This block worked as a reaction stabilizer. The other condition, Case-2, was operated with the secondary honeycomb block inserted in the downstream of the reaction region in addition to the first block. This geometrical structure sandwiched the reaction region between the two blocks, and the thermal energy possessed by the exhaust gas could be regenerated to the reaction region by radiation exchange between these two blocks, which resulted in enhancing the preheating of the premixed gas. By this effect, the methanol-conversion and hydrogen-production in Case-2 were enhanced by about 10% compared to Case-1. In the present study, the reaction characteristics of the fuel reformer were investigated in detail, by detecting the location of the reaction region. Detailed temperature profiles in the streamwise direction were measured with traversable thermocouples, and positive ion current distributions corresponding to the reaction region were measured with a Langmuir probe. It was confirmed by the both measurements that there exists a reaction region right after the first honeycomb block which accompanies with sharp temperature gradients. The estimated thickness of the reaction region, however, was as wide as several millimeters to a centimeter, which is believed to be a ‘mild reaction’ stabilized by the first honeycomb block. In Case-2, the high-temperature region became broader compared to Case-1, which indicates that the enhancement of preheating of premixed gas was achieved by the heat regenerated from the secondary honeycomb block.

Defects in combustion chambers of aircraft engines might have an impact on the reliability of the downstream turbine and the machine’s performance. Detecting failures in the combustion chamber of an aircraft engine during operation may improve the resource management and the availability of the system. Aim of the ongoing research project is to find an approach to evaluate the state of the jet engine by analyzing the temperature and emissions field in the exhaust jet. This investigation is part of the collaborative research center SFB 871. The SFB 871 deals with the improvement of the regeneration process of complex capital goods such as aircraft engines. Maintenance, repair, and overhaul processes would be more efficient if the internal status of the engine would be known while still on the wing before it is disassembled. The feasibility of this approach is investigated for a pilot scaled model combustor, which provides optical access and allows the selection of “defined errors” in the combustor. It consists of an atmospheric tubular combustor with an array of eight premixed swirl burners with a maximum output of 160 kW. The operating conditions of one of the eight burners concerning power and air-fuel ratio, can be controlled. A power distribution between the burners is typical fault in an aircraft combustor and will be investigated in this study. It is observed that it is possible to determine small deviations by measuring density profiles applying a tomographic background-oriented schlieren (BOS) technique behind the combustor. Additionally, particle image velocimetry is used to measure differences in the velocity field of the exhaust gases. This study shows that a minimum power deviation of one burner in an array of a total of eight burners is detectable in the exhaust plane with the above mentioned measurement techniques.

This paper presents the design, modeling, and Finite Element (FE) analysis of a novel Electromagnetic Damper (ED). This cost-effective, regenerative ED is based on the concept of the tubular, linear, brushless dc motor. The structure of the proposed passive ED is straightforward, and it does not require an external power supply. An analytical model of the system is obtained using the magnetic circuit method and used to optimize the non-dimensional geometry factors and to estimate the electromagnetic forces and flux induced in the system. The model can be used to design high-performance dampers for various applications. To confirm the design, dynamic FE simulations were conducted and compared with the analytical and experimental results.

Experimental investigations on heat transfer in tubular micro- or minichannel arrangements more often report on two-phase flow instabilities, pulsations or oscillations, which result in a remarkable influence on heat transfer efficiency. In order to explain the piston-like oscillations of the steam-plugs and water-slugs (-columns), the authors studied the somehow similar process which occurs in the worldwide known toy steam boat. Experiments have been performed which used a demonstration plant made of glass. By controlled electrical heating, high-speed video, pressure and local temperature measurements, the paths of energy have been disclosed. The results are as surprising as the effect of making gold from sand with respect to an equivalent axial heat-conductivity of the water-filled glass tube. Initiated by these results, an abstracting model is presented that analytically quantifies this new regenerating (oscillating and conducting) heat transfer mode e.g. concerning the combination of a heat recharging tube wall and an oscillating water column in a field of diminishing temperatures between the temperature of the boiler surface and the subcooled bulk water. By introducing these heat transfer details, the steam boat can give an answer, not only on frequency and amplitude of the oscillations, but on the steady state conditions for — or time-dependency of — the location of zero-crossing as well. Experimental results and model calculations are in good agreement and need no fitting factors. This is the base to discuss that process along with its physical parameters and compare it to the above mentioned observations in flow-boilers or pulsating heat pipes etc. which use microchannels or minichannels.