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1
Yong, Hsin Nam Ernest, Kim Yeow Tshai, and Siew Shee Lim. "Aqueous Stability of Cross-Linked Thermal Responsive Tissue Engineering Scaffold Produced by Electrospinning Technique." Key Engineering Materials 897 (August 17, 2021): 39–44. http://dx.doi.org/10.4028/www.scientific.net/kem.897.39.
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Poly (N-isopropylacrylamide) (PNIPAm) has been one of the most widely studied thermal responsive polymer in tissue engineering owing to its reversible hydrophilic-hydrophobic phase transition across its lower critical solution temperature (~32°C) that is close to human physiological temperatures. Among tissue engineering constructs, nanofibrous scaffolds offer an added advantage in mimicking the morphology of the native extracellular matrix (ECM). Electrospinning has been reported as one of the most facile method to produce PNIPAm nanofibres and neat electrospun nanofibres scaffold is known to possess poor aqueous stability, limiting its use in tissue engineering applications. In contrast, numerous studies on PNIPAm hydrogels have shown relatively good aqueous stability owing to the hydrophilic 3D crosslinked structure of the hydrogel which resist instant dissolution but rather swell to a greater or lesser extent. However, the presence of crosslinkages in PNIPAm hydrogels causes it to be hardly electrospinnable into nanofibres. In the present work, crosslinker free PNIPAm was radical polymerized to a high molecular weight of 385 kDa. To produce nanofibers, electrospinning was carried out on a dedicated %wt of PNIPAm solution containing octaglycidyl polyhedral oligomeric silsesquioxane (OpePOSS) and 2-ethyl-4-methylimidazole (EMI). Resulting PNIPAm nanofibrous network was found to strongly resemble the ECM morphology with fiber diameter of 436.35 ± 187.04 nm, pore size 1.24 ± 1.27 μm and 63.6% total porosity. Aqueous stability was studied in cell culture media over the course of 28 days. The current result shows significant improvement with a gradual mass loss up to a maximum of 35% instead of the near immediate dissolution observed in the case of electrospun neat PNIPAm scaffold without crosslinks.
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Jeznach, Oliwia, Dorota Kołbuk, Tobias Reich, and Paweł Sajkiewicz. "Immobilization of Gelatin on Fibers for Tissue Engineering Applications: A Comparative Study of Three Aliphatic Polyesters." Polymers 14, no. 19 (October 4, 2022): 4154. http://dx.doi.org/10.3390/polym14194154.
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Immobilization of cell adhesive proteins on the scaffold surface has become a widely reported method that can improve the interaction between scaffold and cells. In this study, three nanofibrous scaffolds obtained by electrospinning of poly(caprolactone) (PCL), poly(L-lactide-co-caprolactone) (PLCL) 70:30, or poly(L-lactide) (PLLA) were subjected to chemical immobilization of gelatin based on aminolysis and glutaraldehyde cross-linking, as well as physisorption of gelatin. Two sets of aminolysis conditions were applied to evaluate the impact of amine group content. Based on the results of the colorimetric bicinchoninic acid (BCA) assay, it was shown that the concentration of gelatin on the surface is higher for the chemical modification and increases with the concentration of free NH2 groups. XPS (X-ray photoelectron spectroscopy) analysis confirmed this outcome. On the basis of XPS results, the thickness of the gelatin layer was estimated to be less than 10 nm. Initially, hydrophobic scaffolds are completely wettable after coating with gelatin, and the time of waterdrop absorption was correlated with the surface concentration of gelatin. In the case of all physically and mildly chemically modified samples, the decrease in stress and strain at break was relatively low, contrary to strongly aminolyzed PLCL and PLLA samples. Incubation testing performed on the PCL samples showed that a chemically immobilized gelatin layer is more stable than a physisorbed one; however, even after 90 days, more than 60% of the initial gelatin concentration was still present on the surface of physically modified samples. Mouse fibroblast L929 cell culture on modified samples indicates a positive effect of both physical and chemical modification on cell morphology. In the case of PCL and PLCL, the best morphology, characterized by stretched filopodia, was observed after stronger chemical modification, while for PLLA, there was no significant difference between modified samples. Results of metabolic activity indicate the better effect of chemical immobilization than of physisorption of gelatin.
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Phuegyod, Seubsakul, Sasivimon Pramual, Nungnit Wattanavichean, Supasuda Assawajaruwan, Taweechai Amornsakchai, Panithi Sukho, Jisnuson Svasti, Rudee Surarit, and Nuttawee Niamsiri. "Microbial Poly(hydroxybutyrate-co-hydroxyvalerate) Scaffold for Periodontal Tissue Engineering." Polymers 15, no. 4 (February 9, 2023): 855. http://dx.doi.org/10.3390/polym15040855.
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In this study, we fabricated three dimensional (3D) porous scaffolds of poly(hydroxybutyrate-co-hydroxyvalerate) with 50% HV content. P(HB-50HV) was biosynthesized from bacteria Cupriavidus necator H16 and the in vitro proliferation of dental cells for tissue engineering application was evaluated. Comparisons were made with scaffolds prepared by poly(hydroxybutyrate) (PHB), poly(hydroxybutyrate-co-12%hydroxyvalerate) (P(HB-12HV)), and polycaprolactone (PCL). The water contact angle results indicated a hydrophobic character for all polymeric films. All fabricated scaffolds exhibited a high porosity of 90% with a sponge-like appearance. The P(HB-50HV) scaffolds were distinctively different in compressive modulus and was the material with the lowest stiffness among all scaffolds tested between the dry and wet conditions. The human gingival fibroblasts (HGFs) and periodontal ligament stem cells (PDLSCs) cultured onto the P(HB-50HV) scaffold adhered to the scaffold and exhibited the highest proliferation with a healthy morphology, demonstrating excellent cell compatibility with P(HB-50HV) scaffolds. These results indicate that the P(HB-50HV) scaffold could be applied as a biomaterial for periodontal tissue engineering and stem cell applications.
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Lis-Bartos, Anna, Agnieszka Smieszek, Kinga Frańczyk, and Krzysztof Marycz. "Fabrication, Characterization, and Cytotoxicity of Thermoplastic Polyurethane/Poly(lactic acid) Material Using Human Adipose Derived Mesenchymal Stromal Stem Cells (hASCs)." Polymers 10, no. 10 (September 28, 2018): 1073. http://dx.doi.org/10.3390/polym10101073.
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Thermoplastic polyurethane (TPU) and poly(lactic acid) are types of biocompatible and degradable synthetic polymers required for biomedical applications. Physically blended (TPU+PLA) tissue engineering matrices were produced via solvent casting technique. The following types of polymer blend were prepared: (TPU+PLA) 7:3, (TPU+PLA) 6:4, (TPU+PLA) 4:6, and (TPU+PLA) 3:7. Various methods were employed to characterize the properties of these polymers: surface properties such as morphology (scanning electron microscopy), wettability (goniometry), and roughness (profilometric analysis). Analyses of hydrophilic and hydrophobic properties, thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC) of the obtained polymer blends were conducted. Tensile tests demonstrated that the blends exhibited a wide range of mechanical properties. Cytotoxicity of polymers was tested using human multipotent stromal cells derived from adipose tissue (hASC). In vitro assays revealed that (TPU+PLA) 3:7 matrices were the most cytocompatible biomaterials. Cells cultured on (TPU+PLA) 3:7 had proper morphology, growth pattern, and were distinguished by increased proliferative and metabolic activity. Additionally, it appeared that (TPU+PLA) 3:7 biomaterials showed antiapoptotic properties. hASC cultured on these matrices had reduced expression of Bax-α and increased expression of Bcl-2. This study demonstrated the feasibility of producing a biocompatible scaffold form based on (TPU+PLA) blends that have potential to be applied in tissue engineering.
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Chee, Tan Yong, Abdull Rahim Mohd Yusoff, and Nik Ahmad Nizam Nik Malek. "Characterisation of poly(vinyl alcohol)- polycaprolactone hybridized scaffold for potential skin tissue regeneration." Malaysian Journal of Fundamental and Applied Sciences 16, no. 1 (February 2, 2020): 6–9. http://dx.doi.org/10.11113/mjfas.v16n1.1469.
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The fabrication of a hybridized scaffold constituting hydrophobic and hydrophilic polymers for tissue engineering has received an increasing attention recently. Due to the high compatibility with water, a hydrophilic polymer, though is able to enhance cell affinity and proliferation, has a very high biodegradable rate and low stability in aqueous medium that eventually puncture its biomedical applications. Thereby, the addition of a hydrophobic polymer in the hydrophilic polymer scaffold is recommended to increase the hydrophobic property of the scaffold in order to reduce the limitation. Nonetheless, the fabrication of the hybridized scaffold is extremely challenging because the hydrophilic and the hydrophobic polymer tends to dissolve in different types of solvents, i.e. water and organic solvent, respectively, that subsequently restricts their blending process. In this work, a poly(vinyl alcohol) (PVA) scaffold, a polycaprolactone (PCL) scaffold, and their hybridized scaffold were produced through casting method for potential skin tissue regeneration. We found that the glacial acetic acid was an appropriate solvent used to prepare hydrophobic PCL solution with low molecular weight (16 kDa) for PCL-PVA blend, with mass ratio 1:1, without using any surfactant. The solvent was also used for the preparation of PCL scaffold with high molecular weight (80 kDa). The fabricated polymer scaffolds were then evaluated using FTIR-ATR, contact angle measurement, and tensile strength analysis. FESEM images of the PVA-PCL hybridized scaffold showed that the PCL was well dispersed in the PVA scaffold. FTIR-ATR spectra showed that the hybridized scaffold exhibited the crucial functional group of PVA and PCL at 3310.97, 1720.10, 1557.80, 1241.69, 1172.90, 1044.95, and 719.44 cm-1. The contact angle of the PVA, PCL, and PVA-PCL hybridized scaffold were 61.3o, 82.7o, and 75.9o, respectively, with tensile strength 16.5747, 2.4038, and 7.417 MPa, respectively.
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Cho, Kwang Joon, Dae Keun Song, Se Heang Oh, Young Joo Koh, Sahng Hoon Lee, Myung Chul Lee, and Jin Ho Lee. "Fabrication and Characterization of Hydrophilized Polydioxanone Scaffolds for Tissue Engineering Applications." Key Engineering Materials 342-343 (July 2007): 289–92. http://dx.doi.org/10.4028/www.scientific.net/kem.342-343.289.
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Porous polydioxanone (PDO)/polyvinyl alcohol (PVA) scaffolds were fabricated by blending PDO with a small amount of PVA to improve the hydrophilicity and cell/tissue compatibility of the scaffolds for tissue engineering applications. PDO/PVA scaffolds with different PVA compositions up to 10 wt% were fabricated by a melt-molding particulate-leaching method (non-solvent method). The prepared scaffolds exhibited highly porous, uniform open-cellular pore structures. The PDO/PVA scaffolds with PVA compositions more than 5 % were easily wetted in cell culture medium. The hydrophilized PDO/PVA (5 wt%) scaffold showed better cell adhesion and growth than the control hydrophobic PDO scaffold. The PDO/PVA (5 wt%) scaffold also showed faster tissue infiltration into the scaffold than the PDO scaffold. It seems that 5 wt% addition of PVA to PDO to fabricate PDO/PVA scaffolds is enough for improving the hydrophilicity and cell/tissue compatibility of the scaffolds.
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Lim, Mim Mim, Tao Sun, and Naznin Sultana. "In VitroBiological Evaluation of Electrospun Polycaprolactone/Gelatine Nanofibrous Scaffold for Tissue Engineering." Journal of Nanomaterials 2015 (2015): 1–10. http://dx.doi.org/10.1155/2015/303426.
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The fabrication of biocompatible and biodegradable scaffolds which mimic the native extracellular matrix of tissues to promote cell adhesion and growth is emphasized recently. Many polymers have been utilized in scaffold fabrication, but there is still a need to fabricate hydrophilic nanosized fibrous scaffolds with an appropriate degradation rate for skin tissue engineering applications. In this study, nanofibrous scaffolds of a biodegradable synthetic polymer, polycaprolactone (PCL), and blends of PCL with a natural polymer, gelatine (Ge), in three different compositions: 85 : 15, 70 : 30, and 50 : 50 were fabricated via an electrospinning technique. The nanofibrous scaffold prepared from 14% w/v PCL/Ge (70 : 30) exhibited more balanced properties of homogeneous nanofibres with an average fibre diameter of 155.60 ± 41.13 nm, 83% porosity, and surface roughness of 176.27 ± 2.53 nm.In vitrocell culture study using human skin fibroblasts (HSF) demonstrated improved cell attachment with a flattened morphology on the PCL/Ge (70 : 30) nanofibrous scaffold and accelerated proliferation on day 3 compared to the PCL nanofibrous scaffold. These results show that the PCL/Ge (70 : 30) nanofibrous scaffold was more favourable and has the potential to be a promising scaffold for skin tissue engineering applications.
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Ghaedamini, Sho'leh, Saeed Karbasi, Batool Hashemibeni, Ali Honarvar, and Abbasali Rabiei. "PCL/Agarose 3D-printed scaffold for tissue engineering applications: fabrication, characterization, and cellular activities." Research in Pharmaceutical Sciences 18, no. 5 (2023): 566–79. http://dx.doi.org/10.4103/1735-5362.383711.
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Background and purpose: Biomaterials, scaffold manufacturing, and design strategies with acceptable mechanical properties are the most critical challenges facing tissue engineering. Experimental approach: In this study, polycaprolactone (PCL) scaffolds were fabricated through a novel three-dimensional (3D) printing method. The PCL scaffolds were then coated with 2% agarose (Ag) hydrogel. The 3D-printed PCL and PCL/Ag scaffolds were characterized for their mechanical properties, porosity, hydrophilicity, and water absorption. The construction and morphology of the printed scaffolds were evaluated via Fourier-Transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The attachment and proliferation of L929 cells cultured on the scaffolds were investigated through MTT assay on the cell culture study upon the 1st, 3rd, and 7th days. Findings/Results: The incorporation of Ag hydrogel with PCL insignificantly decreased the mechanical strength of the scaffold. The presence of Ag enhanced the hydrophilicity and water absorption of the scaffolds, which could positively influence their cell behavior compared to the PCL scaffolds. Regarding cell morphology, the cells on the PCL scaffolds had a more rounded shape and less cell spreading, representing poor cell attachment and cell-scaffold interaction due to the hydrophobic nature of PCL. Conversely, the cells on the PCL/Ag scaffolds were elongated with a spindle-shaped morphology indicating a positive cell-scaffold interaction. Conclusion and implications: PCL/Ag scaffolds can be considered appropriate for tissue-engineering applications.
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Yang, Joseph, Masayuki Yamato, and Teruo Okano. "Cell-Sheet Engineering Using Intelligent Surfaces." MRS Bulletin 30, no. 3 (March 2005): 189–93. http://dx.doi.org/10.1557/mrs2005.51.
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AbstractThe possibility of recreating various tissues and organs for the purpose of regenerative medicine has received much interest. However, the field of tissue engineering has been restricted by the limitations of conventional approaches. A method to circumvent the need for traditional scaffold-based technologies is cell-sheet engineering, which uses temperature-responsive culture dishes. These surfaces, which are created by grafting the temperature-responsive polymer poly(N-isopropylacrylamide) onto ordinary culture dishes, enable the non-invasive harvesting of cells as intact sheets by simple temperature reduction. This article reviews current research on the applications of cell-sheet engineering for the reconstruction of various tissues, as well as the intelligent surfaces used by this novel technology.
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Pacilio, Serafina, Roberta Costa, Valentina Papa, Maria Teresa Rodia, Carlo Gotti, Giorgia Pagnotta, Giovanna Cenacchi, and Maria Letizia Focarete. "Electrospun Poly(L-lactide-co-ε-caprolactone) Scaffold Potentiates C2C12 Myoblast Bioactivity and Acts as a Stimulus for Cell Commitment in Skeletal Muscle Myogenesis." Bioengineering 10, no. 2 (February 11, 2023): 239. http://dx.doi.org/10.3390/bioengineering10020239.
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Tissue engineering combines a scaffold, cells and regulatory signals, reproducing a biomimetic extracellular matrix capable of supporting cell attachment and proliferation. We examined the role of an electrospun scaffold made of a biocompatible polymer during the myogenesis of skeletal muscle (SKM) as an alternative approach to tissue regeneration. The engineered nanostructure was obtained by electrospinning poly(L-lactide-co-ε-caprolactone) (PLCL) in the form of a 3D porous nanofibrous scaffold further coated with collagen. C2C12 were cultured on the PLCL scaffold, and cell morphology and differentiation pathways were thoroughly investigated. The functionalized PLCL scaffold recreated the SKM nanostructure and performed its biological functions, guiding myoblast morphogenesis and promoting cell differentiation until tissue formation. The scaffold enabled cell–cell interactions through the development of cellular adhesions that were fundamental during myoblast fusion and myotube formation. Expression of myogenic regulatory markers and muscle-specific proteins at different stages of myogenesis suggested that the PLCL scaffold enhanced myoblast differentiation within a shorter time frame. The functionalized PLCL scaffold impacts myoblast bioactivity and acts as a stimulus for cell commitment, surpassing traditional 2D cell culture techniques. We developed a screening model for tissue development and a device for tissue restoration.
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Dong, Yixiang, Thomas Yong, Susan Liao, Casey K. Chan, and S. Ramakrishna. "Long-term viability of coronary artery smooth muscle cells on poly( l -lactide- co -ϵ-caprolactone) nanofibrous scaffold indicates its potential for blood vessel tissue engineering." Journal of The Royal Society Interface 5, no. 26 (February 19, 2008): 1109–18. http://dx.doi.org/10.1098/rsif.2007.1354.
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Biodegradable polymer nanofibres have been extensively studied as cell culture scaffolds in tissue engineering. However, long-term in vitro studies of cell–nanofibre interactions were rarely reported and successful organ regeneration using tissue engineering techniques may take months (e.g. blood vessel tissue engineering). Understanding the long-term interaction between cells and nanofibrous scaffolds (NFS) is crucial in material selection, design and processing of the tissue engineering scaffolds. In this study, poly( l -lactide- co -ϵ-caprolactone) [P(LLA-CL)] (70 : 30) copolymer NFS were produced by electrospinning. Porcine coronary artery smooth muscle cells (PCASMCs) were seeded and cultured on the scaffold to evaluate cell–nanofibre interactions for up to 105 days. A favourable interaction between this scaffold and PCASMCs was demonstrated by cell viability assay, scanning electron microscopy, histological staining and extracellular matrix (ECM) secretion. Degradation behaviours of the scaffolds with or without PCASMC culture were determined by mechanical testing and gel permeation chromatography (GPC). The results showed that the PCASMCs attached and proliferated well on the P(LLA-CL) NFS. Large amount of ECM protein secretion was observed after 50 days of culture. Multilayers of aligned oriented PCASMCs were formed on the scaffold after two months of in vitro culture. In the degradation study, the PCASMCs were not shown to significantly increase the degradation rate of the scaffolds for up to 105 days of culture. The in vitro degradation time of the scaffold could be as long as eight months by extrapolating the results from GPC. These observations further supported the potential use of the P(LLA-CL) nanofibre in blood vessel tissue engineering.
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Zhao, Min Li, Gang Sui, Xu Liang Deng, Ji Gui Lu, Seung Kon Ryu, and Xiao Ping Yang. "PLLA/HA Electrospin Hybrid Nanofiber Scaffolds: Morphology, In Vitro Degradation and Cell Culture Potential." Advanced Materials Research 11-12 (February 2006): 243–46. http://dx.doi.org/10.4028/www.scientific.net/amr.11-12.243.
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Electrospinning has recently emerged as a potential technique for fabricating biomimetic tissue engineering scaffolds. In this study, Poly (l-lactic acid) (PLLA) /Nano-hydroxyapatite (HA) hybrid nanofibers scaffolds were prepared by electrospinning. The relationship between process parameters and fiber diameter has been investigated. The fiber diameter decreased with decreasing polymer concentration and with increasing electrospinning voltage; After 6 weeks of in vitro degradation, the mass, viscosity-average molecular weight of the nanofibers scaffolds and the pH value of the degradation solution were changed, the fibers lost their surface smoothness and a regular rough topology was generated after 32d of degradation, the degradation rates of PLLA/HA hybrid nanofibers were slower than those of pure PLLA fibers; The biocompatibility of the nanofibers scaffold has also been investigated by culturing cells on the nanofibers scaffold, elementary results showed that the cells adhered and proliferated well on the PLLA/HA hybrid nanofibers scaffolds.
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Meng, Di, Xiongxin Lei, Yang Li, Yingjun Kong, Dawei Huang, and Guifeng Zhang. "Three dimensional polyvinyl alcohol scaffolds modified with collagen for HepG2 cell culture." Journal of Biomaterials Applications 35, no. 4-5 (June 24, 2020): 459–70. http://dx.doi.org/10.1177/0885328220933505.
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The creation of in vitro functional hepatic tissue simulating micro environmental niche of the native liver is a keen area of research due to its demand in bioartificial liver. However, it is still unclear how to maintain benign cell function while achieving the sufficient cell quantity. In this work, we aim to prepare a novel scaffold for the culture of HepG2 cells, a liver cell line, by modifying polyvinyl alcohol (PVA) scaffold with collagen (COL). PVA is a kind of synthetic biostable polymer with high hydrophilicity in the human body, has been widely used in the biomedical field. However, the use of PVA is limited in cell cultures due to lack of biologically active functional groups. In this study, amino silane (KH-550), glutaraldehyde and native type I collagen were used to modify three-dimensional PVA scaffold to establish a suitable composite scaffold for hepatocyte culture. Three types of composite scaffolds were prepared for different collagen content, named as PVA/COL (0.2%), PVA/COL (0.5%) and PVA/COL (0.8%), respectively. The composite scaffolds were characterized by SEM, XPS, FTIR, MS, porosity estimation and water contact angle measurement. The PVA/COL (0.8%) scaffolds had the highest collagen content of 12.13%. The composite scaffold showed high porosity with interconnected pores. Furthermore, the biocompatibility between HepG2 cells and scaffolds was evaluated by the ability of cell proliferation, albumin secretion, as well as urea synthesis. The coating of collagen on PVA scaffolds promoted hydrophilicity and HepG2 cell adhesion. Additionally, enhanced cell proliferation, increased albumin secretion and urea synthesis were observed in HepG2 cells growing on collagen-coated three-dimensional PVA scaffolds.
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Fan, Daniel, Urs Staufer, and Angelo Accardo. "Engineered 3D Polymer and Hydrogel Microenvironments for Cell Culture Applications." Bioengineering 6, no. 4 (December 13, 2019): 113. http://dx.doi.org/10.3390/bioengineering6040113.
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The realization of biomimetic microenvironments for cell biology applications such as organ-on-chip, in vitro drug screening, and tissue engineering is one of the most fascinating research areas in the field of bioengineering. The continuous evolution of additive manufacturing techniques provides the tools to engineer these architectures at different scales. Moreover, it is now possible to tailor their biomechanical and topological properties while taking inspiration from the characteristics of the extracellular matrix, the three-dimensional scaffold in which cells proliferate, migrate, and differentiate. In such context, there is therefore a continuous quest for synthetic and nature-derived composite materials that must hold biocompatible, biodegradable, bioactive features and also be compatible with the envisioned fabrication strategy. The structure of the current review is intended to provide to both micro-engineers and cell biologists a comparative overview of the characteristics, advantages, and drawbacks of the major 3D printing techniques, the most promising biomaterials candidates, and the trade-offs that must be considered in order to replicate the properties of natural microenvironments.
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Hahn, Judith, Gundula Schulze-Tanzil, Michaela Schröpfer, Michael Meyer, Clemens Gögele, Mariann Hoyer, Axel Spickenheuer, Gert Heinrich, and Annette Breier. "Viscoelastic Behavior of Embroidered Scaffolds for ACL Tissue Engineering Made of PLA and P(LA-CL) After In Vitro Degradation." International Journal of Molecular Sciences 20, no. 18 (September 19, 2019): 4655. http://dx.doi.org/10.3390/ijms20184655.
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A rupture of the anterior cruciate ligament (ACL) is the most common knee ligament injury. Current applied reconstruction methods have limitations in terms of graft availability and mechanical properties. A new approach could be the use of a tissue engineering construct that temporarily reflects the mechanical properties of native ligament tissues and acts as a carrier structure for cell seeding. In this study, embroidered scaffolds composed of polylactic acid (PLA) and poly(lactic-co-ε-caprolactone) (P(LA-CL)) threads were tested mechanically for their viscoelastic behavior under in vitro degradation. The relaxation behavior of both scaffold types (moco: mono-component scaffold made of PLA threads, bico: bi-component scaffold made of PLA and P(LA-CL) threads) was comparable to native lapine ACL. Most of the lapine ACL cells survived 32 days of cell culture and grew along the fibers. Cell vitality was comparable for moco and bico scaffolds. Lapine ACL cells were able to adhere to the polymer surfaces and spread along the threads throughout the scaffold. The mechanical behavior of degrading matrices with and without cells showed no significant differences. These results demonstrate the potential of embroidered scaffolds as an ACL tissue engineering approach.
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Liu, Zheng, and Jun Wang. "Biological Influence of Nonswelling Microgels on Cartilage Induction of Mouse Adipose-Derived Stem Cells." BioMed Research International 2019 (October 13, 2019): 1–10. http://dx.doi.org/10.1155/2019/6508094.
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In cartilage tissue engineering, the target cells’ functional performance depends on the biomaterials. However, it is difficult to develop an appropriate scaffold to differentiate mouse adipose-derived stem cells (mADSCs) into chondrocyte despite an increasing number of studies on biological scaffold materials. The purpose of this study was to create a novel scaffold for mADSC culture and chondrogenic differentiation with a new series of microgels based on polyethyleneimine (PEI), polyethylene glycol (PEG), and poly (L-lactic acid) (PLLA) and able to resist swelling with changes in temperature, pH, and polymer concentration. The biocompatibility and ability of the nonswelling microgels were then examined and served as scaffolds for cell culture and for cartilage differentiation. The results show that the new microgels are a novel biomaterial that both retains its nonswelling properties under various conditions and facilitates important scaffold functions such as cell adhesion, proliferation, and cartilage induction.
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Niemczyk-Soczynska, Beata, Arkadiusz Gradys, and Pawel Sajkiewicz. "Hydrophilic Surface Functionalization of Electrospun Nanofibrous Scaffolds in Tissue Engineering." Polymers 12, no. 11 (November 10, 2020): 2636. http://dx.doi.org/10.3390/polym12112636.
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Electrospun polymer nanofibers have received much attention in tissue engineering due to their valuable properties such as biocompatibility, biodegradation ability, appropriate mechanical properties, and, most importantly, fibrous structure, which resembles the morphology of extracellular matrix (ECM) proteins. However, they are usually hydrophobic and suffer from a lack of bioactive molecules, which provide good cell adhesion to the scaffold surface. Post-electrospinning surface functionalization allows overcoming these limitations through polar groups covalent incorporation to the fibers surface, with subsequent functionalization with biologically active molecules or direct deposition of the biomolecule solution. Hydrophilic surface functionalization methods are classified into chemical approaches, including wet chemical functionalization and covalent grafting, a physiochemical approach with the use of a plasma treatment, and a physical approach that might be divided into physical adsorption and layer-by-layer assembly. This review discusses the state-of-the-art of hydrophilic surface functionalization strategies of electrospun nanofibers for tissue engineering applications. We highlighted the major advantages and drawbacks of each method, at the same time, pointing out future perspectives and solutions in the hydrophilic functionalization strategies.
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Tsai, Wei-Bor, and Ibrahim Nasser Ahmed. "The Impact of Polyethylene Glycol-Modified Chitosan Scaffolds on the Proliferation and Differentiation of Osteoblasts." International Journal of Biomaterials 2023 (January 3, 2023): 1–8. http://dx.doi.org/10.1155/2023/4864492.
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The objective of this study was to investigate the influence of polyethylene glycol (PEG) incorporated chitosan scaffolds on osteoblasts proliferation and differentiation. The chitosan polymer was initially modified by the predetermined concentration of the photoreactive azido group for UV-crosslinking and with RGD peptides (N-acetyl-GRGDSPGYG-amide). The PEG was mixed at different ratios (0, 10, and 20 wt%) with modified chitosan in 96-well tissue culture polystyrene plates to prepare CHI-100, CHI-90, and CHI-80 scaffolds. PEG-containing scaffolds exhibited bigger pore size and higher water content compared to unmodified chitosan scaffolds. After 10 days of incubation, the cell number of CHI-90 (1.1 × 106 cells/scaffold) surpasses that of CHI-100 (9.2 × 105 cells/scaffold) and the cell number of CHI-80 (7.6 × 105 cells/scaffold) were significantly lower. The ALP activity of CHI-90 was the highest on the fifth day indicating the favored osteoblasts' early-stage differentiation. Moreover, after 14 days of osteogenic culture, calcium deposition in the CHI-90 scaffolds (2.7 μmol Ca/scaffold) was significantly higher than the control (2.2 μmol Ca/scaffold) whereas on CHI-80 was 1.9 μmol/scaffold. The results demonstrate that PEG-incorporated chitosan scaffolds favored osteoblasts proliferation and differentiation; however, mixing relatively excess PEG (≥20% wt.) had a negative impact on osteoblasts proliferation and differentiation.
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Pazhanimala, Shaleena K., Driton Vllasaliu, and Bahijja T. Raimi-Abraham. "Engineering Biomimetic Gelatin Based Nanostructures as Synthetic Substrates for Cell Culture." Applied Sciences 9, no. 8 (April 17, 2019): 1583. http://dx.doi.org/10.3390/app9081583.
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There is a need for synthetic substrates that replicate the natural environment for in vitro intestinal models. Electrospinning is one of the most versatile and cost-effective techniques to produce nanofibrous scaffolds mimicking the basement membrane topography. In this study, three different novel electrospun nanofibrous scaffolds made of a polycaprolactone (PCL), gelatin, and poloxamer 188 (P188) blend were produced and compared with PCL and PCL/gelatin fibers produced using the same solvent system and electrospinning parameters. Each polymer solution used in this experiment was electrospun at four different voltages to study its influence on fiber diameter. The morphology and physical characteristics of the fibers were studied using scanning electron microscopy and atomic force microscopy. The average fiber diameter of all scaffolds was within 200–600 nm and no significant decrease in diameter with an increase in voltage was observed. Attenuated total reflection Fourier transform infrared spectroscopy was used to determine the chemical characteristics of the nanofibrous scaffold. The conductivity of the polymer solutions was also analyzed. Biocompatibility of the scaffolds was determined by a cell proliferation study performed using colorectal carcinoma (Caco-2) cells. PCL/gelatin/P188 scaffolds exhibited higher cell proliferation compared to PCL, PCL/gelatin scaffolds, and the control (tissue culture multi-well plate) with PCL/gelatin/P188 80:10:10 sample showing the highest cell proliferation.
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Pangesty, Azizah Intan, and Mitsugu Todo. "Improvement of Mechanical Strength of Tissue Engineering Scaffold Due to the Temperature Control of Polymer Blend Solution." Journal of Functional Biomaterials 12, no. 3 (August 14, 2021): 47. http://dx.doi.org/10.3390/jfb12030047.
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Polymeric scaffolds made of PCL/PLCL (ratio 1:3, respectively) blends have been developed by using the Thermally Induced Phase Separation (TIPS) process. A new additional technique has been introduced in this study by applying pre-heat treatment to the blend solution before the TIPS process. The main objective of this study is to evaluate the influence of the pre-heat treatment on mechanical properties. The mechanical evaluation showed that the mechanical strength of the scaffolds (including tensile strength, elastic modulus, and strain) improved as the temperature of the polymer blend solution increased. The effects on the microstructure features were also observed, such as increasing strut size and differences in phase separation morphology. Those microstructure changes due to temperature control contributed to the increasing of mechanical strength. The in vitro cell study showed that the PCL/PLCL blend scaffold exhibited better cytocompatibility than the neat PCL scaffold, indicated by a higher proliferation at 4 and 7 days in culture. This study highlighted that the improvement of the mechanical strength of polymer blends scaffolds can be achieved using a very versatile way by controlling the temperature of the polymer blend solution before the TIPS process.
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Khoramgah, Maryam Sadat, Javad Ranjbari, Hojjat-Allah Abbaszadeh, Fatemeh Sadat Tabatabaei Mirakabad, Shadie Hatami, Simzar Hosseinzadeh, and Hossein Ghanbarian. "Freeze-dried multiscale porous nanofibrous three dimensional scaffolds for bone regenerations." BioImpacts 10, no. 2 (February 8, 2020): 73–85. http://dx.doi.org/10.34172/bi.2020.10.
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Introduction: Simulating hydrophobic-hydrophilic composite face with hierarchical porous and fibrous architectures of bone extracellular matrix (ECM) is a key aspect in bone tissue engineering. This study focused on the fabrication of new three-dimensional (3D) scaffolds containing polytetrafluoroethylene (PTFE), and polyvinyl alcohol (PVA), with and without graphene oxide (GO) nanoparticles using the chemical cross-linking and freeze-drying methods for bone tissue application. The effects of GO on physicochemical features and osteoinduction properties of the scaffolds were evaluated through an in vitro study. Methods: After synthesizing the GO nanoparticles, two types of 3D scaffolds, PTFE/PVA (PP) and PTFE/PVA/GO (PPG), were developed by cross-linking and freeze-drying methods. The physicochemical features of scaffolds were assessed and the interaction of the 3D scaffold types with human adipose mesenchymal stem cells (hADSCs) including attachment, proliferation, and differentiation to osteogenic like cells were investigated. Results: GO nanoparticles were successfully synthesized with no agglomeration. The blending of PTFE as a hydrophobic polymer with PVA polymer and GO nanoparticles (hydrophilic compartments) were successful. Two types of 3D scaffolds had nano topographical structures, good porosities, hydrophilic surfaces, thermal stabilities, good stiffness, as well as supporting the cell attachments, proliferation, and osteogenic differentiation. Notably, GO incorporating scaffolds provided a better milieu for cell behaviors. Conclusion: Novel multiscale porous nanofibrous 3D scaffolds made from PTFE/ PVA polymers with and without GO nanoparticles could be an ideal candidate for bone tissue engineering as a 3D template.
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WANG, LU, YANNI CHEN, JUN QIAN, YANYAN TAN, SHAOHUA HUANGFU, YIJIANG DING, SHUQING DING, and BIN JIANG. "A BOTTOM-UP METHOD TO BUILD 3D SCAFFOLDS WITH PREDEFINED VASCULAR NETWORK." Journal of Mechanics in Medicine and Biology 13, no. 05 (October 2013): 1340008. http://dx.doi.org/10.1142/s0219519413400083.
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Tissue engineering is a promising approach to regenerate transplantable tissue or organ substitutes in vitro. However, the existing methods are based on seeding cells on macroscale polymer scaffolds, which are associated with several challenges including limited control over cell microenvironment, limited nutrient diffusion, directed cell alignment. The emerging bottom-up tissue engineering methods hold great potential to address these challenges by assembling building blocks into complex 3D tissue constructs. In this study, we developed a layer-by-layer assembly approach to recreate 3D cell-laden constructs. Our experiment showed the predefined channels form a vascular system and help the transplant cells to transport the requirement of culture cells in early case of cells attaching and growing up. It is an original concept to demonstrate the feasibility of forming a network with a vascular geometry in a biocompatible polymer and fabricated different scaffold with different cells. The concept was developed to create a complete branching vascular circulation in 3D on surface of mixture of chitosan and gelatin structures and pre-define the structure of channel for culturing smooth muscle for controlling the SMC growing up as smooth muscle fiber.
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García-Cerna, Sandra, Uriel Sánchez-Pacheco, Angélica Meneses-Acosta, José Rojas-García, Bernardo Campillo-Illanes, Daniel Segura-González, and Carlos Peña-Malacara. "Evaluation of Poly-3-Hydroxybutyrate (P3HB) Scaffolds Used for Epidermal Cells Growth as Potential Biomatrix." Polymers 14, no. 19 (September 26, 2022): 4021. http://dx.doi.org/10.3390/polym14194021.
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Advances in tissue engineering have made possible the construction of organs and tissues with the use of biomaterials and cells. Three important elements are considered: a specific cell culture, an adequate environment, and a scaffold. The present study aimed to develop P3HB scaffolds by 3D printing and evaluate their biocompatibility with HaCaT epidermal cells, as a potential model that allows the formation of functional tissue. By using a method of extraction and purification with ethanol and acetone, a biopolymer having suitable properties for use as a tissue support was obtained. This polymer exhibited a higher molecular weight (1500 kDa) and lower contact angle (less than 90°) compared to the material obtained using the conventional method. The biocompatibility analysis reveals that the scaffold obtained using the ethanol–acetone method and produced by 3D printing without pores was not cytotoxic, did not self-degrade, and allowed high homogenous cell proliferation of HaCaT cells. In summary, it is possible to conclude that the P3HB scaffold obtained by 3D printing and a simplified extraction method is a suitable support for the homogeneous development of HaCaT keratinocyte cell lineage, which would allow the evaluation of this material to be used as a biomatrix for tissue engineering.
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Iwanaga, Shintaroh, Yuta Hamada, Yoshinari Tsukamoto, Kenichi Arai, Taketoshi Kurooka, Shinji Sakai, and Makoto Nakamura. "Design and Fabrication of Mature Engineered Pre-Cardiac Tissue Utilizing 3D Bioprinting Technology and Enzymatically Crosslinking Hydrogel." Materials 15, no. 22 (November 9, 2022): 7928. http://dx.doi.org/10.3390/ma15227928.
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The fabrication of mature engineered cardiac tissue is one of the major challenges in cardiac tissue engineering. For this purpose, we attempted to apply the 3D bioprinting approach. Aiming to construct an oriented tissue, a fine fiber-shaped scaffold with a support structure was first designed using CAD software. Then, a 3D bioprinter and cell-adhesive bio-inks were utilized to fabricate this structure. The cell-adhesive bio-inks were synthesized by combining sodium alginate and gelatin with tyramine, respectively, to form pre-gel materials that allow enzymatic crosslinking by horseradish peroxidase. By absorbance measurements, we confirmed that the tyramine modification rate of each polymer was 0.535 mmol/g-alginate and 0.219 mmol/g-gelatin. The width of the fiber-shaped scaffold was 216.8 ± 24.3 μm for the fabricated scaffold, while the design value was 200 μm. After 3D printing and adhesion-adding treatment of the scaffold with these bio-ink materials, cardiomyocytes were seeded and cultured. As a result, the cells spread onto the scaffold, and the entire pre-tissue contracted synchronously by day 6 of culture, showing a greater pulsatility than in the early days. Video analysis showed that the beating rate of pre-myocardial tissue on day 6 was 31 beats/min. In addition, we confirmed that the cardiomyocytes partially elongated along the long axis of the fiber-shaped scaffold in the pre-tissue cultured for 15 days by staining actin, suggesting the possibility of cell orientation. Furthermore, treatment with adrenaline resulted in a 7.7-fold increase in peak beating rate compared to that before treatment (from 6 beats/min to 46 beats/min), confirming the responsiveness of the pre-tissues to the drug. These results indicate that 3D bioprinting effectively produces mature cultured myocardial tissue that is oriented, contracts synchronously, and is responsive to drugs.
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Maibohm, Christian, Alberto Saldana-Lopez, Oscar F. Silvestre, and Jana B. Nieder. "3D Polymer Architectures for the Identification of Optimal Dimensions for Cellular Growth of 3D Cellular Models." Polymers 14, no. 19 (October 4, 2022): 4168. http://dx.doi.org/10.3390/polym14194168.
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Organ-on-chips and scaffolds for tissue engineering are vital assay tools for pre-clinical testing and prediction of human response to drugs and toxins, while providing an ethical sound replacement for animal testing. A success criterion for these models is the ability to have structural parameters for optimized performance. Here we show that two-photon polymerization fabrication can create 3D test platforms, where scaffold parameters can be directly analyzed by their effects on cell growth and movement. We design and fabricate a 3D grid structure, consisting of wall structures with niches of various dimensions for probing cell attachment and movement, while providing easy access for fluorescence imaging. The 3D structures are fabricated from bio-compatible polymer SZ2080 and subsequently seeded with A549 lung epithelia cells. The seeded structures are imaged with confocal microscopy, where spectral imaging with linear unmixing is used to separate auto-fluorescence scaffold contribution from the cell fluorescence. The volume of cellular material present in different sections of the structures is analyzed, to study the influence of structural parameters on cell distribution. Furthermore, time-lapse studies are performed to map the relation between scaffold parameters and cell movement. In the future, this kind of differentiated 3D growth platform, could be applied for optimized culture growth, cell differentiation, and advanced cell therapies.
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Ghasemi, Sanaz, and Hamed Ghomi. "Investigation of applying chitosan coating on antibacterial and biocompatibility properties of bredigite/titanium dioxide composite scaffolds." Journal of Biomaterials Applications 36, no. 3 (February 16, 2021): 406–18. http://dx.doi.org/10.1177/0885328221994290.
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In this study by considering the advantages of bredigite (Br) and titanium dioxide (TiO2) bioceramics, composite scaffolds of bredigite/titanium dioxide were prepared by the gelcasting method, then, to improve the mechanical, biological and antibacterial properties, scaffolds were coated with chitosan (Ch) polymer phase. The phase structure, fundamental groups, chemical composition, and elemental distribution analysis, morphology and the form of porosity were respectively characterized by XRD, FTIR, EDS, and SEM. Mechanical properties and porosity percentage of scaffolds were also measured by the compressive strength test and Archimedean method, respectively. In order to verify the cell compatibility, MG63 bone marrow cells were cultured on the surface of the specimens. The results showed that the addition of titanium dioxide to the scaffold of bredigite resulted in decrease of porosity and increase of compressive strength of scaffolds from 0.299 to 0.687 MPa. Furthermore, the coated scaffold with chitosan polymer reduced porosity from 83 to 63 percent and a remarkable improvement in compressive strength from 0.585 to 2.339 MPa. The results of the antibacterial test showed that in composite scaffolds, The diameter of the inhibition zone is 22 and 29 mm, in the culture media of Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive), respectively. On the other hand, the results of cell compatibility and cell adhesion tests showed that the scaffolds had no toxicity and the growth, proliferation, and adhesion of MG63 bone cells adjacent to the scaffolds was desirable. Therefore, the scaffold in this study can be used as an ideal scaffold for use in bone tissue engineering.
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Rodrigues, Leonardo Ribeiro, Cecília Amélia de Carvalho Zavaglia, and Christiane Bertachini Lombello. "HA/TCP Scaffolds Coated by PLA and Gelatin: Preliminary In Vitro Evaluation." Key Engineering Materials 631 (November 2014): 289–94. http://dx.doi.org/10.4028/www.scientific.net/kem.631.289.
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The importance of this work is development of two methods to try to improve the bone tissue regeneration. The surface of scaffold was modified in order to favor cell interaction, through adhesion and proliferation. PLA and gelatin were used. PLA has shown in literature good results in bone tissue engineering. The gelatin is used as coating in cell culture plates to improve cell attachment for a variety of cell types, including osteoblast. Hydroxyapatite (HA) and tricalcium phosphate (TCP) were used due to its known properties in tissue engineering. It was used the polymer foam replication technique to produce the scaffolds. For characterization were used: scanning electron microscopy (SEM), optical microscopy (OP), stereoscopy, transmission electron microscopy (TEM), X-ray microtomography (Micro-CT), X-ray diffraction (XRD) and X-ray fluorescence (XRF). The scaffolds showed morphology with adequate porosity for tissue engineering and thein vitrotest showed evidence of not being cytotoxic.
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Choi, Dong Jin, Kyoung Choi, Sang Jun Park, Young-Jin Kim, Seok Chung, and Chun-Ho Kim. "Suture Fiber Reinforcement of a 3D Printed Gelatin Scaffold for Its Potential Application in Soft Tissue Engineering." International Journal of Molecular Sciences 22, no. 21 (October 27, 2021): 11600. http://dx.doi.org/10.3390/ijms222111600.
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Gelatin has excellent biological properties, but its poor physical properties are a major obstacle to its use as a biomaterial ink. These disadvantages not only worsen the printability of gelatin biomaterial ink, but also reduce the dimensional stability of its 3D scaffolds and limit its application in the tissue engineering field. Herein, biodegradable suture fibers were added into a gelatin biomaterial ink to improve the printability, mechanical strength, and dimensional stability of the 3D printed scaffolds. The suture fiber reinforced gelatin 3D scaffolds were fabricated using the thermo-responsive properties of gelatin under optimized 3D printing conditions (−10 °C cryogenic plate, 40–80 kPa pneumatic pressure, and 9 mm/s printing speed), and were crosslinked using EDC/NHS to maintain their 3D structures. Scanning electron microscopy images revealed that the morphologies of the 3D printed scaffolds maintained their 3D structure after crosslinking. The addition of 0.5% (w/v) of suture fibers increased the printing accuracy of the 3D printed scaffolds to 97%. The suture fibers also increased the mechanical strength of the 3D printed scaffolds by up to 6-fold, and the degradation rate could be controlled by the suture fiber content. In in vitro cell studies, DNA assay results showed that human dermal fibroblasts’ proliferation rate of a 3D printed scaffold containing 0.5% suture fiber was 10% higher than that of a 3D printed scaffold without suture fibers after 14 days of culture. Interestingly, the supplement of suture fibers into gelatin biomaterial ink was able to minimize the cell-mediated contraction of the cell cultured 3D scaffolds over the cell culture period. These results show that advanced biomaterial inks can be developed by supplementing biodegradable fibers to improve the poor physical properties of natural polymer-based biomaterial inks.
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Nakashima, Yoshiki, Hiroki Iguchi, Kenta Takakura, Yuta Nakamura, Kenji Izumi, Naoya Koba, Satoshi Haneda, and Masayoshi Tsukahara. "Adhesion Characteristics of Human Pancreatic Islets, Duct Epithelial Cells, and Acinar Cells to a Polymer Scaffold." Cell Transplantation 31 (January 2022): 096368972211205. http://dx.doi.org/10.1177/09636897221120500.
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We reported in 2018 that among several extracellular matrices, fibronectin, type I collagen, type IV collagen, laminin I, fibrinogen, and bovine serum albumin, fibronectin is particularly useful for adhesion of porcine pancreatic tissue. Subsequently, we developed a technology that enables the chemical coating of the constituent motifs of fibronectin onto cell culture dishes. In this experiment, we used islets (purity ≥ 90%), duct epithelial cells (purity ≥ 60%), and acinar cells (purity ≥ 99%) isolated from human pancreas according to the Edmonton protocol published in 2000 and achieved adhesion to the constituent motifs of fibronectin. A solution including cGMP Prodo Islet Media was used as the assay solution. In islets, adhesion was enhanced with the constitutive motifs of fibronectin compared with uncoated islets. In the functional evaluation of islets, insulin mRNA expression and insulin secretion were enhanced by the constitutive motif of fibronectin compared with non-coated islets. The stimulation index was comparable between non-coated islets and fibronectin motifs. In duct epithelial cells, adhesion was mildly promoted by the fibronectin component compared with non-coated component, while in acinar cells, adhesion was inhibited by the fibronectin component compared with the non-coated component. These data suggest that the constitutive motifs of fibronectin are useful for the adhesion of islets and duct epithelial cells.
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Rode, Michele Patricia, Addeli Bez Batti Angulski, Felipe Azevedo Gomes, Maiara Marques da Silva, Talita da Silva Jeremias, Rafael Guzella de Carvalho, Daniel Gonçalves Iucif Vieira, et al. "Carrageenan hydrogel as a scaffold for skin-derived multipotent stromal cells delivery." Journal of Biomaterials Applications 33, no. 3 (September 2018): 422–34. http://dx.doi.org/10.1177/0885328218795569.
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Carrageenan is a thermoreversible polymer of natural origin widely used in food and pharmaceutical industry that presents a glycosaminoglycan-like structure. Herein, we show that kappa-type carrageenan extracted by a semi-refined process from the red seaweed Kappaphycus alvarezii displayed both chemical and structural properties similar to a commercial carrageenan. Moreover, both extracted carrageenan hydrogel and commercial carrageenan hydrogel can serve as a scaffold for in vitro culture of human skin-derived multipotent stromal cells, demonstrating considerable potential as cell-carrier materials for cell delivery in tissue engineering. Skin-derived multipotent stromal cells cultured inside the carrageenan hydrogels showed a round shape morphology and maintained their growth and viability for at least one week in culture. Next, the effect of the extracted carrageenan hydrogel loaded with human skin-derived multipotent stromal cells was evaluated in a mouse model of full-thickness skin wound. Macroscopic and histological analyses revealed some pointed ameliorated features, such as reduced inflammatory process, faster initial recovery of wounded area, and improved extracellular matrix deposition. These results indicate that extracted carrageenan hydrogel can serve as a scaffold for in vitro growth and maintenance of human SD-MSCs, being also able to act as a delivery system of cells to wounded skin. Thus, evaluation of the properties discussed in this study contribute to a further understanding and specificities of the potential use of carrageenan hydrogel as a delivery system for several applications, further to skin wound healing.
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Mousavi, Seyyed Mojtaba, Seyyed Alireza Hashemi, Masoomeh Yari Kalashgrani, Navid Omidifar, Sonia Bahrani, Neralla Vijayakameswara Rao, Aziz Babapoor, Ahmad Gholami, and Wei-Hung Chiang. "Bioactive Graphene Quantum Dots Based Polymer Composite for Biomedical Applications." Polymers 14, no. 3 (February 5, 2022): 617. http://dx.doi.org/10.3390/polym14030617.
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Today, nanomedicine seeks to develop new polymer composites to overcome current problems in diagnosing and treating common diseases, especially cancer. To achieve this goal, research on polymer composites has expanded so that, in recent years, interdisciplinary collaborations between scientists have been expanding day by day. The synthesis and applications of bioactive GQD-based polymer composites have been investigated in medicine and biomedicine. Bioactive GQD-based polymer composites have a special role as drug delivery carriers. Bioactive GQDs are one of the newcomers to the list of carbon-based nanomaterials. In addition, the antibacterial and anti-diabetic potentials of bioactive GQDs are already known. Due to their highly specific surface properties, π-π aggregation, and hydrophobic interactions, bioactive GQD-based polymer composites have a high drug loading capacity, and, in case of proper correction, can be used as an excellent option for the release of anticancer drugs, gene carriers, biosensors, bioimaging, antibacterial applications, cell culture, and tissue engineering. In this paper, we summarize recent advances in using bioactive GQD-based polymer composites in drug delivery, gene delivery, thermal therapy, thermodynamic therapy, bioimaging, tissue engineering, bioactive GQD synthesis, and GQD green resuscitation, in addition to examining GQD-based polymer composites.
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Chung, Johnson H. Y., Sepidar Sayyar, and Gordon G. Wallace. "Effect of Graphene Addition on Polycaprolactone Scaffolds Fabricated Using Melt-Electrowriting." Polymers 14, no. 2 (January 13, 2022): 319. http://dx.doi.org/10.3390/polym14020319.
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Melt-electrowriting (MEW) is an emerging method that combines electrospinning and extrusion printing, allowing the fabrication of micron-scale structures suitable for tissue engineering. Compared to other additive fabrication methods, melt-electro written structures can offer more appropriate substrates for cell culture due to filament size and mechanical characteristics of the fabricated scaffolds. In this study, polycaprolactone (PCL)/graphene composites were investigated for fabrication of micron-size scaffolds through MEW. It was demonstrated that the addition of graphene can considerably improve the processability of PCL to fabricate micron-scale scaffolds with enhanced resolution. The tensile strength of the scaffold prepared from PCL/graphene composite (with only 0.5 wt.% graphene) was proved significantly (by more than 270%), better than that of the pristine PCL scaffold. Furthermore, graphene was demonstrated to be a suitable material for tailoring the degradation process to avoid undesirable bulk degradation, rapid mass loss and damage to the internal matrix of the polymer. The findings of this study offer a promising route for the fabrication of high-resolution scaffolds with micron-scale resolution for tissue engineering.
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Mouchati, Abdullah, and Najet Yagoubi. "Mechanical Performance and Cytotoxicity of an Alginate/Polyacrylamide Bipolymer Network Developed for Medical Applications." Materials 16, no. 5 (February 22, 2023): 1789. http://dx.doi.org/10.3390/ma16051789.
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Several hydrogels could be used as scaffolds for tissue engineering and a model of extracellular matrices for biological studies. However, the scope of alginate in medical applications is often severely limited by its mechanical behavior. In the present study, the modification of the mechanical properties of the alginate scaffold is obtained by its combination with polyacrylamide in order to obtain a multifunctional biomaterial. The advantage of this double polymer network is due to an improvement in the mechanical strength with regard to the alginate alone, and in particular, its Young’s modulus values. The morphological study of this network was carried out by scanning electron microscopy (SEM). The swelling properties were also studied over several time intervals. In addition to mechanical property requirements, these polymers must meet several biosafety parameters as part of an overall risk management strategy. Our preliminary study illustrates that the mechanical property of this synthetic scaffold depends on the ratio of the two polymers (alginate, polyacrylamide) which allows us to choose the appropriate ratio to mimic replaceable body tissue and be used in various biological and medical uses, including 3D cell culture, tissue engineering, and protection against local shocks.
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Hashemi, Seyedeh-Sara, Seyedeh-Somayeh Rajabi, Reza Mahmoudi, Amir Ghanbari, Kazem Zibara, and Mehrzad Jafari Barmak. "Polyurethane/chitosan/hyaluronic acid scaffolds: providing an optimum environment for fibroblast growth." Journal of Wound Care 29, no. 10 (October 2, 2020): 586–96. http://dx.doi.org/10.12968/jowc.2020.29.10.586.
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Objective: Biodegradable polymers can replace damaged tissue components using tissue engineering techniques. The objective of this study is to determine an optimum environment for polymer scaffolds to improve the proliferation of fibroblast cells capable of wound repair. Method: In this study, the addition of polysaccharides, such as chitosan (CH) or hyaluronic acid (HA), to a polyurethane (PU) polymer was evaluated using different methods to determine if they affect scaffold morphology and cell activity of fibroblasts prepared from human foreskin tissues. Mechanical properties, such as tensile strength, contact angle and swelling test, were used to check the physical and mechanical properties of the scaffold. Fibroblast growth was also measured at 24, 48 and 72 hours. Results: Scanning electron microscopy (SEM) determined that a 3:1 ratio of PU/CH scaffold, developed by electrospinning, allowed the formation of a uniform structure in scaffold fibres. Physical mechanical tests showed that PU electrospun scaffolds were not modified by the addition of CH. The mean stretch and mean water absorption increased significantly using the PU/CH scaffold, compared with the PU scaffold. However, the mean tensile strength and the mean contact angle, used to study space and porosity, did not differ between scaffolds. Fourier transform infrared spectroscopy confirmed the functional groups (–OH, –NH and –C=O) in the PU/CH scaffold, compared with PU or CH chemical structures alone. HA was then added to CH and PU/CH scaffolds to evaluate the growth of fibroblast cells. Results showed that cell viability and the number of cells, using MTT and trypan blue exclusion assay, respectively, increased significantly at 24, 48 and 72 hours of culture in PU/CH/HA scaffold compared to HA, CH/HA, and PU/HA. Moreover, PU/HA at 48 and 72 hours also increased cell viability and cell numbers compared to HA and CH/HA scaffolds. However, scaffolds at 72 hours had limited space for cell growth. Moreover, SEM data demonstrated that fibroblasts were able to proliferate, penetrate, migrate and survive on PU/HA and PU/CH/HA three-dimensional scaffolds, especially during the first 48 hours. Furthermore, 4′,6-diamidino-2-phenylindole (DAPI) staining confirmed that fibroblasts could penetrate PU scaffolds and showed higher cell viability and lower cellular damage in PU/CH/HA, compared to CH/HA and PU/HA scaffolds. Finally, flow cytometry using CD90 and CD105 surface markers revealed that >90% of cells isolated from the human dermis were fibroblasts. Conclusion: In summary, PU/HA and PU/CH/HA scaffolds were found to be biocompatible and provided a suitable environment for the growth and proliferation of fibroblasts, which filled and covered all pores between the fibres. The new scaffold used in this study, made of synthetic and natural polymers, is a good candidate for applications in tissue engineering. It is therefore recommended to use PU in grafts or in wound dressing.
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Carvalho, Estela O., Clarisse Ribeiro, Daniela M. Correia, Gabriela Botelho, and Senentxu Lanceros-Mendez. "Biodegradable Hydrogels Loaded with Magnetically Responsive Microspheres as 2D and 3D Scaffolds." Nanomaterials 10, no. 12 (December 3, 2020): 2421. http://dx.doi.org/10.3390/nano10122421.
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Scaffolds play an essential role in the success of tissue engineering approaches. Their intrinsic properties are known to influence cellular processes such as adhesion, proliferation and differentiation. Hydrogel-based matrices are attractive scaffolds due to their high-water content resembling the native extracellular matrix. In addition, polymer-based magnetoelectric materials have demonstrated suitable bioactivity, allowing to provide magnetically and mechanically activated biophysical electrical stimuli capable of improving cellular processes. The present work reports on a responsive scaffold based on poly (L-lactic acid) (PLLA) microspheres and magnetic microsphere nanocomposites composed of PLLA and magnetostrictive cobalt ferrites (CoFe2O4), combined with a hydrogel matrix, which mimics the tissue’s hydrated environment and acts as a support matrix. For cell proliferation evaluation, two different cell culture conditions (2D and 3D matrices) and two different strategies, static and dynamic culture, were applied in order to evaluate the influence of extracellular matrix-like confinement and the magnetoelectric/magneto-mechanical effect on cellular behavior. MC3T3-E1 proliferation rate is increased under dynamic conditions, indicating the potential use of hydrogel matrices with remotely stimulated magnetostrictive biomaterials for bone tissue engineering.
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Guo, Yongjian, Rouba Ghobeira, Sheida Aliakbarshirazi, Rino Morent, and Nathalie De Geyter. "Polylactic Acid/Polyaniline Nanofibers Subjected to Pre- and Post-Electrospinning Plasma Treatments for Refined Scaffold-Based Nerve Tissue Engineering Applications." Polymers 15, no. 1 (December 24, 2022): 72. http://dx.doi.org/10.3390/polym15010072.
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Composite biopolymer/conducting polymer scaffolds, such as polylactic acid (PLA)/ polyaniline (PAni) nanofibers, have emerged as popular alternative scaffolds in the electrical-sensitive nerve tissue engineering (TE). Although mimicking the extracellular matrix geometry, such scaffolds are highly hydrophobic and usually present an inhomogeneous morphology with massive beads that impede nerve cell-material interactions. Therefore, the present study launches an exclusive combinatorial strategy merging successive pre- and post-electrospinning plasma treatments to cope with these issues. Firstly, an atmospheric pressure plasma jet (APPJ) treatment was applied on PLA and PLA/PAni solutions prior to electrospinning, enhancing their viscosity and conductivity. These liquid property changes largely eliminated the beaded structures on the nanofibers, leading to uniform and nicely elongated fibers having average diameters between 170 and 230 nm. After electrospinning, the conceived scaffolds were subjected to a N2 dielectric barrier discharge (DBD) treatment, which significantly increased their surface wettability as illustrated by large decreases in water contact angles for values above 125° to values below 25°. X-ray photoelectron spectroscopy (XPS) analyses revealed that 3.3% of nitrogen was implanted on the nanofibers surface in the form of C–N and N–C=O functionalities upon DBD treatment. Finally, after seeding pheochromocytoma (PC-12) cells on the scaffolds, a greatly enhanced cell adhesion and a more dispersive cell distribution were detected on the DBD-treated samples. Interestingly, when the APPJ treatment was additionally performed, the extension of a high number of long neurites was spotted leading to the formation of a neuronal network between PC-12 cell clusters. In addition, the presence of conducting PAni in the scaffolds further promoted the behavior of PC-12 cells as illustrated by more than a 40% increase in the neurite density without any external electrical stimulation. As such, this work presents a new strategy combining different plasma-assisted biofabrication techniques of conducting nanofibers to create promising scaffolds for electrical-sensitive TE applications.
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Gögele, Clemens, Silvana Müller, Svetlana Belov, Andreas Pradel, Sven Wiltzsch, Armin Lenhart, Markus Hornfeck, et al. "Biodegradable Poly(D-L-lactide-co-glycolide) (PLGA)-Infiltrated Bioactive Glass (CAR12N) Scaffolds Maintain Mesenchymal Stem Cell Chondrogenesis for Cartilage Tissue Engineering." Cells 11, no. 9 (May 7, 2022): 1577. http://dx.doi.org/10.3390/cells11091577.
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Regeneration of articular cartilage remains challenging. The aim of this study was to increase the stability of pure bioactive glass (BG) scaffolds by means of solvent phase polymer infiltration and to maintain cell adherence on the glass struts. Therefore, BG scaffolds either pure or enhanced with three different amounts of poly(D-L-lactide-co-glycolide) (PLGA) were characterized in detail. Scaffolds were seeded with primary porcine articular chondrocytes (pACs) and human mesenchymal stem cells (hMSCs) in a dynamic long-term culture (35 days). Light microscopy evaluations showed that PLGA was detectable in every region of the scaffold. Porosity was greater than 70%. The biomechanical stability was increased by polymer infiltration. PLGA infiltration did not result in a decrease in viability of both cell types, but increased DNA and sulfated glycosaminoglycan (sGAG) contents of hMSCs-colonized scaffolds. Successful chondrogenesis of hMSC-colonized scaffolds was demonstrated by immunocytochemical staining of collagen type II, cartilage proteoglycans and the transcription factor SOX9. PLGA-infiltrated scaffolds showed a higher relative expression of cartilage related genes not only of pAC-, but also of hMSC-colonized scaffolds in comparison to the pure BG. Based on the novel data, our recommendation is BG scaffolds with single infiltrated PLGA for cartilage tissue engineering.
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Maibohm, Christian, Alberto Saldana-Lopez, Oscar F. Silvestre, and Jana B. Nieder. "3D Polymer Structures for the Identification of Optimal Dimensions for Cellular Growth for 3D Lung Alveolar Models." Engineering Proceedings 4, no. 1 (April 16, 2021): 33. http://dx.doi.org/10.3390/micromachines2021-09596.
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Organ-on-chips and scaffolds for tissue engineering are vital assay tools for pre-clinical testing and prediction of human response to drugs and toxins, while providing an ethically sound alternative to animal testing and a low-cost alternative to expensive clinical studies. An important success criterion for these models is the ability to have structural parameters for optimized performance. In this study we show how the two-photon polymerization fabrication method can be used to create 3D test platforms made for analyzing optimal scaffold parameters for cell growth. We design and fabricate a 3D grid structure, designed as a set of wall structures with niches of various dimensions for probing the optimal niche for cell attachment. The 3D grid structures are fabricated from bio-compatible polymer SZ2080 and subsequently seeded with A549 lung epithelia cells. The seeded structures are incubated and imaged with multi-color spectral confocal microscopy at several time points, to determine the volume of cell material present in the different niches of the grid structure. Spectral imaging with linear unmixing is used to separate the auto-fluorescence contribution from the scaffold from the fluorescence of the cells and use it to determine the volume of cell material present in the different sections of the grid structure. The variation in structural parameters influences the incubated A549 cells’ distribution and morphology. In future, this kind of differentiated 3D growth platform could be applied for optimized culture growth, cell differentiation and advanced cell therapies.
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Garcia-Sanchez, Mayra Elizabeth, Ines Jimenez Palomar, Yolanda Gonzalez-Garcia, and Jorge R. Robledo-Ortiz. "Bacterial Cellulose Produced by Gluconacetobacter xylinus Culture Using Complex Carbon Sources for Biomedical Applications." MRS Advances 1, no. 36 (2016): 2563–67. http://dx.doi.org/10.1557/adv.2016.462.
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ABSTRACTTissue engineering scaffolding is the external media or structure in which cell growth, migration and reproduction is enabled in order to stimulate tissue regeneration. In order to promote tissue regeneration, scaffolding materials are required to have certain properties such as biocompatibility, adequate mechanical properties and surface topographical features in order to provide specific biological signals to promote cell attachment and proliferation [1].Cellulose is the most abundant, inexpensive and readily available carbohydrate polymer in the world and it is traditionally extracted from plants or their wastes [2]. Although the plant itself is the major contributor of cellulose, various types of bacteria are able to produce cellulose and it is termed bacterial cellulose [3]. Bacterial cellulose is a well suited scaffold for tissue regeneration due to its biocompatibility, mechanical properties and its ability to be combined with other structures such calcium phosphates [4], which can create composites with intrinsic properties that meet the requirements of the different tissues of the human body [5].Through additive manufacturing, highly complex structures can be created which are similar to those found in nature. This work will explore the different ways to produce biomimetic structures for tissue engineering applications through the combination of bacterial cellulose and additive manufacturing producing complex structures of a highly a biocompatible material for a range of different biomedical applications [6]. In addition to the manufacturing and processing techniques, the use of mango (juice/peel) as a complex carbon source for the production of bacterial cellulose was investigated.
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Lee, Dongjin, and Chaenyung Cha. "The Combined Effects of Co-Culture and Substrate Mechanics on 3D Tumor Spheroid Formation within Microgels Prepared via Flow-Focusing Microfluidic Fabrication." Pharmaceutics 10, no. 4 (November 13, 2018): 229. http://dx.doi.org/10.3390/pharmaceutics10040229.
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Tumor spheroids are considered a valuable three dimensional (3D) tissue model to study various aspects of tumor physiology for biomedical applications such as tissue engineering and drug screening as well as basic scientific endeavors, as several cell types can efficiently form spheroids by themselves in both suspension and adherent cell cultures. However, it is more desirable to utilize a 3D scaffold with tunable properties to create more physiologically relevant tumor spheroids as well as optimize their formation. In this study, bioactive spherical microgels supporting 3D cell culture are fabricated by a flow-focusing microfluidic device. Uniform-sized aqueous droplets of gel precursor solution dispersed with cells generated by the microfluidic device are photocrosslinked to fabricate cell-laden microgels. Their mechanical properties are controlled by the concentration of gel-forming polymer. Using breast adenocarcinoma cells, MCF-7, the effect of mechanical properties of microgels on their proliferation and the eventual spheroid formation was explored. Furthermore, the tumor cells are co-cultured with macrophages of fibroblasts, which are known to play a prominent role in tumor physiology, within the microgels to explore their role in spheroid formation. Taken together, the results from this study provide the design strategy for creating tumor spheroids utilizing mechanically-tunable microgels as 3D cell culture platform.
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Ching, Kuan Yong, Orestis Andriotis, Bram Sengers, and Martin Stolz. "Genipin crosslinked chitosan/PEO nanofibrous scaffolds exhibiting an improved microenvironment for the regeneration of articular cartilage." Journal of Biomaterials Applications 36, no. 3 (March 17, 2021): 503–16. http://dx.doi.org/10.1177/08853282211002015.
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Towards optimizing the growth of extracellular matrix to produce repair cartilage for healing articular cartilage (AC) defects in joints, scaffold-based tissue engineering approaches have recently become a focus of clinical research. Scaffold-based approaches by electrospinning aim to support the differentiation of chondrocytes by providing an ultrastructure similar to the fibrillar meshwork in native cartilage. In a first step, we demonstrate how the blending of chitosan with poly(ethylene oxide) (PEO) allows concentrated chitosan solution to become electrospinnable. The chitosan-based scaffolds share the chemical structure and characteristics of glycosaminoglycans, which are important structural components of the cartilage extracellular matrix. Electrospinning produced nanofibrils of ∼100 nm thickness that are closely mimicking the size of collagen fibrils in human AC. The polymer scaffolds were stabilized in physiological conditions and their stiffness was tuned by introducing the biocompatible natural crosslinker genipin. We produced scaffolds that were crosslinked with 1.0% genipin to obtain values of stiffness that were in between the stiffness of the superficial zone human AC of 600 ± 150 kPa and deep zone AC of 1854 ± 483 kPa, whereas the stiffness of 1.5% genipin crosslinked scaffold was similar to the stiffness of deep zone AC. The scaffolds were degradable, which was indicated by changes in the fibril structure and a decrease in the scaffold stiffness after seven months. Histological and immunohistochemical analysis after three weeks of culture with human articular chondrocytes (HACs) showed a cell viability of over 90% on the scaffolds and new extracellular matrix deposited on the scaffolds.
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Dimida, Simona, Amilcare Barca, Nadia Cancelli, Vincenzo De Benedictis, Maria Grazia Raucci, and Christian Demitri. "Effects of Genipin Concentration on Cross-Linked Chitosan Scaffolds for Bone Tissue Engineering: Structural Characterization and Evidence of Biocompatibility Features." International Journal of Polymer Science 2017 (2017): 1–8. http://dx.doi.org/10.1155/2017/8410750.
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Genipin (GN) is a natural molecule extracted from the fruit of Gardenia jasminoides Ellis according to modern microbiological processes. Genipin is considered as a favorable cross-linking agent due to its low cytotoxicity compared to widely used cross-linkers; it cross-links compounds with primary amine groups such as proteins, collagen, and chitosan. Chitosan is a biocompatible polymer that is currently studied in bone tissue engineering for its capacity to promote growth and mineral-rich matrix deposition by osteoblasts in culture. In this work, two genipin cross-linked chitosan scaffolds for bone repair and regeneration were prepared with different GN concentrations, and their chemical, physical, and biological properties were explored. Scanning electron microscopy and mechanical tests revealed that nonremarkable changes in morphology, porosity, and mechanical strength of scaffolds are induced by increasing the cross-linking degree. Also, the degradation rate was shown to decrease while increasing the cross-linking degree, with the high cross-linking density of the scaffold disabling the hydrolysis activity. Finally, basic biocompatibility was investigated in vitro, by evaluating proliferation of two human-derived cell lines, namely, the MG63 (human immortalized osteosarcoma) and the hMSCs (human mesenchymal stem cells), as suitable cell models for bone tissue engineering applications of biomaterials.
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Peng, Yi-Yang, Qiuli Cheng, Meng Wu, Wenda Wang, Jianyang Zhao, Diana Diaz-Dussan, Michelle McKay, Hongbo Zeng, Sarute Ummartyotin, and Ravin Narain. "Highly Stretchable, Self-Healing, Injectable and pH Responsive Hydrogel from Multiple Hydrogen Bonding and Boron-Carbohydrate Interactions." Gels 9, no. 9 (September 1, 2023): 709. http://dx.doi.org/10.3390/gels9090709.
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A simple and cost-effective method for the fabrication of a safe, dual-responsive, highly stretchable, self-healing and injectable hydrogel is reported based on a combination of dynamic boronate ester bonds and hydrogen bonding interactions. The mechanical properties of the hydrogel are tunable by adjusting the molar ratios between sugar moieties on the polymer and borax. It was remarkable to note that the 2:1 ratio of sugar and borate ion significantly improves the mechanical strength of the hydrogel. The injectability, self-healing and stretchability properties of the hydrogel were also examined. In addition, the impact of the variation of the pH and the addition of free sugar responsiveness of the hydrogel was studied. High MRC-5 cell viability was noticed by the 3D live/dead assay after 24 h cell culture within the hydrogel scaffold. Hence, the developed hydrogels have desirable features that warrant their applications for drug delivery, scaffolds for cell and tissue engineering.
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Higa, Camila Fernandes, Thatyanne Gradowski, Selene Elifio-Esposito, Marcelo Fernandes de Oliveira, Paulo Inforçatti, Jorge Vicente Lopes da Silva, Fred Lacerda Amorim, and Michelle Sostag Meruvia. "Influence of selective laser sintering process parameters on microstructure and physicochemical properties of poly(vinyl alcohol) for the production of scaffolds." Rapid Prototyping Journal 26, no. 6 (June 10, 2020): 1155–64. http://dx.doi.org/10.1108/rpj-01-2019-0021.
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Purpose This study aims to investigate the production of scaffolds by selective laser sintering (SLS) using poly(vinyl alcohol) (PVA) polymer, for in vitro studies, a relatively new and growing area in which scaffolds could be used in the design of three-dimensional models for in vitro disease model or tissue equivalent for safety and effectiveness tests. Design/methodology/approach The influence of the SLS process parameters laser power, 26 W and 32 W, and number of laser scans, 1, 2, 4 and 6, on the surface microstructure of the samples and on the degree of crystallinity and chemical stability of PVA material, was investigated using powder with particle size of 20-320 µm. Laser sintered PVA samples were subjected to cell culture tests using osteoblastic cells derived from human osteosarcoma (SaOs-2). Findings The laser power has no significant influence on the microstructure of the laser-sintered samples, however the number of scans has a considerable influence on the sintering degree; the SLS process causes a decrease in the degree of crystallinity and changes the chemical structure of the as-received PVA, especially when using higher laser power and more number of scans. Preliminary in vitro cell culture tests show that the laser-sintered PVA material is biocompatible with SaOs-2 cells. Originality/value SLS offers good potential for the fabrication of scaffolds and thus, may be applied as an alternative to conventional scaffold fabrication processes to overcome their limitations.
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Careta, Oriol, Asier Salicio-Paz, Eva Pellicer, Elena Ibáñez, Jordina Fornell, Eva García-Lecina, Jordi Sort, and Carme Nogués. "Electroless Palladium-Coated Polymer Scaffolds for Electrical Stimulation of Osteoblast-Like Saos-2 Cells." International Journal of Molecular Sciences 22, no. 2 (January 7, 2021): 528. http://dx.doi.org/10.3390/ijms22020528.
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Three-dimensional porous scaffolds offer some advantages over conventional treatments for bone tissue engineering. Amongst all non-bioresorbable scaffolds, biocompatible metallic scaffolds are preferred over ceramic and polymeric scaffolds, as they can be used as electrodes with different electric field intensities (or voltages) for electric stimulation (ES). In the present work we have used a palladium-coated polymeric scaffold, generated by electroless deposition, as a bipolar electrode to electrically stimulate human osteoblast-like Saos-2 cells. Cells grown on palladium-coated polyurethane foams under ES presented higher proliferation than cells grown on foams without ES for up to 14 days. In addition, cells grown in both conditions were well adhered, with a flat appearance and a typical actin cytoskeleton distribution. However, after 28 days in culture, cells without ES were filling the entire structure, while cells under ES appeared rounded and not well adhered, a sign of cell death onset. Regarding osteoblast differentiation, ES seems to enhance the expression of early expressed genes. The results suggest that palladium-coated polyurethane foams may be good candidates for osteoblast scaffolds and demonstrate that ES enhances osteoblast proliferation up to 14 days and upregulate expression genes related to extracellular matrix formation.
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Careta, Oriol, Asier Salicio-Paz, Eva Pellicer, Elena Ibáñez, Jordina Fornell, Eva García-Lecina, Jordi Sort, and Carme Nogués. "Electroless Palladium-Coated Polymer Scaffolds for Electrical Stimulation of Osteoblast-Like Saos-2 Cells." International Journal of Molecular Sciences 22, no. 2 (January 7, 2021): 528. http://dx.doi.org/10.3390/ijms22020528.
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Three-dimensional porous scaffolds offer some advantages over conventional treatments for bone tissue engineering. Amongst all non-bioresorbable scaffolds, biocompatible metallic scaffolds are preferred over ceramic and polymeric scaffolds, as they can be used as electrodes with different electric field intensities (or voltages) for electric stimulation (ES). In the present work we have used a palladium-coated polymeric scaffold, generated by electroless deposition, as a bipolar electrode to electrically stimulate human osteoblast-like Saos-2 cells. Cells grown on palladium-coated polyurethane foams under ES presented higher proliferation than cells grown on foams without ES for up to 14 days. In addition, cells grown in both conditions were well adhered, with a flat appearance and a typical actin cytoskeleton distribution. However, after 28 days in culture, cells without ES were filling the entire structure, while cells under ES appeared rounded and not well adhered, a sign of cell death onset. Regarding osteoblast differentiation, ES seems to enhance the expression of early expressed genes. The results suggest that palladium-coated polyurethane foams may be good candidates for osteoblast scaffolds and demonstrate that ES enhances osteoblast proliferation up to 14 days and upregulate expression genes related to extracellular matrix formation.
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Guarino, Vincenzo, Francesco Urciuolo, Marco A. Alvarez-Perez, Benedetto Mele, Paolo A. Netti, and Luigi Ambrosio. "Osteogenic differentiation and mineralization in fibre-reinforced tubular scaffolds: theoretical study and experimental evidences." Journal of The Royal Society Interface 9, no. 74 (March 7, 2012): 2201–12. http://dx.doi.org/10.1098/rsif.2011.0913.
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The development of composite scaffolds with well-organized architecture and multi-scale properties (i.e. porosity, degradation) represents a valid approach for achieving a tissue-engineered construct capable of reproducing the medium- and long-term in vitro behaviour of hierarchically complex tissues such as spongy bone. To date, the implementation of scaffold design strategies able to summarize optimal scaffold architecture as well as intrinsic mechanical, chemical and fluid transport properties still remains a challenging issue. In this study, poly ɛ -caprolactone/polylactid acid (PCL/PLA) tubular devices (fibres of PLA in a PCL matrix) obtained by phase inversion/salt leaching and filament winding techniques were proposed as cell instructive scaffold for bone osteogenesis. Continuous fibres embedded in the polymeric matrix drastically improved the mechanical response as confirmed by compression elastic moduli, which vary from 0.214 ± 0.065 to 1.174 ± 0.143 MPa depending on the relative fibre/matrix and polymer/solvent ratios. Moreover, computational fluid dynamic simulations demonstrated the ability of composite structure to transfer hydrodynamic forces during in vitro culture, thus indicating the optimal flow rate conditions that, case by case, enables specific cellular events—i.e. osteoblast differentiation from human mesenchymal stem cells (hMSCs), mineralization, etc. Hence, we demonstrate that the hMSC differentiation preferentially occurs in the case of higher perfusion rates—over 0.05 ml min –1 —as confirmed by the expression of alkaline phosphate and osteocalcin markers. In particular, the highest osteopontin values and a massive mineral phase precipitation of bone-like phases detected in the case of intermediate flow rates (i.e. 0.05 ml min –1 ) allows us to identify the best condition to stimulate the bone extracellular matrix in-growth, in agreement with the hydrodynamic model prediction. All these results concur to prove the succesful use of tubular composite as temporary device for long bone treatment.
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Kunz, Regina Inês, Rose Meire Costa Brancalhão, Lucinéia de Fátima Chasko Ribeiro, and Maria Raquel Marçal Natali. "Silkworm Sericin: Properties and Biomedical Applications." BioMed Research International 2016 (2016): 1–19. http://dx.doi.org/10.1155/2016/8175701.
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Silk sericin is a natural polymer produced by silkworm,Bombyx mori, which surrounds and keeps together two fibroin filaments in silk thread used in the cocoon. The recovery and reuse of sericin usually discarded by the textile industry not only minimizes environmental issues but also has a high scientific and commercial value. The physicochemical properties of the molecule are responsible for numerous applications in biomedicine and are influenced by the extraction method and silkworm lineage, which can lead to variations in molecular weight and amino acid concentration of sericin. The presence of highly hydrophobic amino acids and its antioxidant potential make it possible for sericin to be applied in the food and cosmetic industry. The moisturizing power allows indications as a therapeutic agent for wound healing, stimulating cell proliferation, protection against ultraviolet radiation, and formulating creams and shampoos. The antioxidant activity associated with low digestibility of sericin that expands the application in the medical field, such as antitumour, antimicrobial and anti-inflammatory agent, anticoagulant, acts in colon health, improving constipation and protects the body from obesity through improved plasma lipid profile. In addition, the properties of sericin allow its application as a culture medium and cryopreservation, in tissue engineering and for drug delivery, demonstrating its effective use, as an important biomaterial.
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Grigoriev, A. M., Yu B. Basok, A. D. Kirillova, V. A. Surguchenko, N. P. Shmerko, V. K. Kulakova, R. V. Ivanov, V. I. Lozinsky, A. M. Subbot, and V. I. Sevastianov. "Cryogenically structured gelatin-based hydrogel as a resorbable macroporous matrix for biomedical technologies." Russian Journal of Transplantology and Artificial Organs 24, no. 2 (May 13, 2022): 83–93. http://dx.doi.org/10.15825/1995-1191-2022-2-83-93.
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Objective: to investigate the biological properties of a matrix made of cryogenically structured hydrogel in the form of a macroporous gelatin sponge, as well as the possibility of creating cell-engineered constructs (CECs) on its basis. Materials and methods. The main components of the cryogenically structured hydrogel were gelatin (type A) obtained from porcine skin collagen, N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide, (EDC) and urea (all from Sigma-Aldrich, USA). Surface morphology was examined using scanning electron microscopy (SEM). The degree of swelling in water of the samples was determined by gravimetric method. Cytotoxicity was studied on NIH3T3, a fibroblast cell line isolated from a mouse, and on human adipose-derived mesenchymal stem/stromal cells (hAMSCs) using IncuCyte ZOOM (EssenBioscience, USA). The metabolic activity of hAMSCs was assessed using PrestoBlue™ reagents (Invitrogen™, USA). To create CECs, we used hAMSCs, human hepatocellular carcinoma cell line HepG2 or human umbilical vein endothelial cell lines EA.hy926. Albumin content in the culture medium was determined by enzyme immunoassay. Ammonia metabolism rate was assessed after 90 minutes of incubation with 1 mM ammonium chloride (Sigma-Aldrich, USA) diluted in a culture medium on day 15 of the experiment. Results. Obtaining a cryogenically structured hydrogel scaffold in the form of macroporous gelatin sponge included freezing an aqueous solution of a gelatin+urea mixture, removal of polycrystals of frozen solvent by lyophilization, extraction of urea with ethanol and treatment of the cryostructurate with an ethanol solution of EDC. Scanning electron microscopy identified three types of pores on the carrier surface: large (109 ± 17 μm), medium (39 ± 10 μm), and small (16 ± 6 μm). The degree of swelling in water of the matrix samples was 3.8 ± 0.2 g H2O per 1 g of dry polymer. The macroporous gelatin sponge as a part of CEC was found to have the ability to support adhesion and proliferation of hAMSCs, EA.hy926 and HepG2 for 28, 15 and 9 days, respectively. Albumin secretion and ammonia metabolism when HepG2 cells were cultured on the gelatin sponge were detected. Conclusion. The use of a matrix made from macroporous cryogenically structured gelatin-based hydrogel for tissue engineering products is shown to be promising using a cell-engineered liver construct as a case.
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Zaman, Zara. "Exploring Bone Cell Research Using Bone-on-a-Chip Models and Microfluidics: A Literature Review." Undergraduate Research in Natural and Clinical Science and Technology (URNCST) Journal 7, no. 6 (June 12, 2023): 1–7. http://dx.doi.org/10.26685/urncst.477.
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Introduction: Organ-on-a-chip models are becoming popular due to its success in modeling human tissues and organs, to mimic human physiology and understand how diseases or drugs affect organs. Traditional 2-dimensional in vitro models are limited in recreating complicated bone structure and examining cell-cell interactions. Alternatively, bone-on-a-chip models establish biomimetic conditions to accurately recapitulate the complexity of the bone. However, bone-on-a-chip models as 3D culture systems do not accurately replicate the bone microenvironment. Rather, microfluidic devices allow for fluid control on a microscale or nanoscale level and the incorporation of fluid shear stress normally experienced by bone cells. The goal of this review paper is to summarize advancements to bone-on-a-chip models. Methods: Relevant articles were selected through a computerized search using GEOBASE and PubMED. Search terms included ‘microfluidic devices AND bones’, ‘organ-on-a-chip models’, ‘bone-on-a-chip models’, ‘PDMS AND bone regeneration’, ‘PolyHIPE AND bone regeneration’ and ‘bone scaffolds’. Results: Microfluidic chips are fabricated using soft lithography and poly-di-methyl siloxane (PDMS) which is a biocompatible, synthetic polymer that is used as a cell culture substrate but is too stiff to facilitate bone regeneration. Hydroxyapatite (HA), lined with PDMS, is commonly used, but the substrate degrades at a much slower rate. Moreover, β-tricalcium-phosphate (β-TCP) as a bone scaffold is both porous and degrades faster hence existing studies have used it to generate a dense extracellular matrix. Discussion: The studies examined in this paper highlight contributions made to scaffolds and microfluidics using bone-on-a-chip models. Notably, scaffolds must be osteoconductive to allow bone cells to adhere, proliferate and form an extracellular matrix on its surface and pore. While PDMS is both osteoconductive and biocompatible, its rigidity poses a concern. Both β-TCP and HA have capabilities for cell-mediated resorption and are more favourable substrates. Additionally, by incorporating microfluidics with bone-on-a-chip models, cells experience greater fluid shear stress similar to that of loading within the bone. Conclusion: In sum, advancements to bone-on-a-chip platforms are ongoing and the many published studies discussed in this paper aim to optimize both the design and materials used to create long lasting impacts on the rapidly growing field of cell and tissue engineering.