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Cytotoxic evaluation of biomechanically improved crosslinked ovine collagen on human dermal fibroblasts

机译:生物力学改良的交联绵羊胶原对人真皮成纤维细胞的细胞毒性评估

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摘要

Earlier studies in our laboratory demonstrated that collagen extracted from ovine tendon is biocompatible towards human dermal fibroblast. To be able to use this collagen as a scaffold in skin tissue engineering, a mechanically stronger scaffold is required that can withstand manipulation before transplantation. This study was conducted to improve the mechanical strength of this collagen sponge using chemical crosslinkers, and evaluate their effect on physical, chemical and biocompatible properties. Collagen sponge was crosslinked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and glutaraldehyde (GA). Tensile test, FTIR study and mercury porosimetry were used to evaluate mechanical properties, chemical property and porosity, respectively. MTT assay was performed to evaluate the cytotoxic effect of crosslinked collagen sponge on human dermal fibroblasts. The FTIR study confirmed the successful crosslinking of collagen sponge. Crosslinking with EDC and GA significantly increased the mechanical strength of collagen sponge, with GA being more superior. Crosslinking of collagen sponge significantly reduced the porosity and the effect was predominant in GA-crosslinked collagen sponge. The GA-crosslinked collagen showed significantly lower, 60 cell viability towards human dermal fibroblasts compared to that of EDC-crosslinked collagen, 80 and non-crosslinked collagen, 100. Although the mechanical strength was better when using GA but the more toxic effect on dermal fibroblast makes EDC a more suitable crosslinker for future skin tissue engineering.
机译:我们实验室的早期研究表明,从绵羊肌腱中提取的胶原蛋白对人体真皮成纤维细胞具有生物相容性。为了能够在皮肤组织工程中使用这种胶原蛋白作为支架,需要一种机械强度更强的支架,该支架可以承受移植前的操作。本研究旨在使用化学交联剂提高这种胶原海绵的机械强度,并评估它们对物理、化学和生物相容性的影响。海绵胶原蛋白与1-乙基-3-(3-二甲氨基丙基)碳二亚胺(EDC)和戊二醛(GA)交联。拉伸试验、傅里叶变换红外光谱(FTIR)和汞孔隙率测定法分别用于评估力学性能、化学性能和孔隙率。MTT法评价交联胶原海绵对人真皮成纤维细胞的细胞毒性作用。FTIR研究证实了胶原海绵的成功交联。EDC和GA交联显著提高了海绵胶原蛋白的机械强度,其中GA更胜一筹。胶原海绵的交联显著降低了孔隙率,其效果在GA交联的海绵胶原中占主导地位。与EDC交联胶原蛋白(80%)和非交联胶原蛋白(100%)相比,GA交联胶原蛋白对人真皮成纤维细胞的细胞活力显著降低(60%)。虽然使用GA时机械强度更好,但对真皮成纤维细胞的毒性作用更大,使EDC成为未来皮肤组织工程中更合适的交联剂。

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