Awang M A, Firdaus M A B, Busra M B, Chowdhury S R, Fadilah N R, Wan Hamirul W K, Reusmaazran M Y, Aminuddin M Y, Ruszymah B H I
Tissue Engineering Centre, UKM Medical Centre, Kuala Lumpur, Malaysia.
Department of Biomedical Science, Faculty of Allied Health Science, UKM, Kuala Lumpur, Malaysia.
Biomed Mater Eng. 2014;24(4):1715-24. doi: 10.3233/BME-140983.
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.
我们实验室早期的研究表明，从羊肌腱中提取的胶原蛋白对人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将这种胶原蛋白用作支架，需要一种机械强度更高的支架，以便在移植前能够承受操作。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。胶原海绵用1-乙基-3-(3-二甲基氨基丙基)碳二亚胺（EDC）和戊二醛（GA）进行交联。分别使用拉伸试验、傅里叶变换红外光谱（FTIR）研究和压汞法来评估机械性能、化学性质和孔隙率。进行MTT试验以评估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。用EDC和GA交联显著提高了胶原海绵的机械强度，其中GA更优。胶原海绵的交联显著降低了孔隙率，且这种影响在GA交联的胶原海绵中更为明显。与EDC交联的胶原（细胞活力80%）和未交联的胶原（细胞活力100%）相比，GA交联的胶原对人真皮成纤维细胞的细胞活力显著降低，仅为60%。尽管使用GA时机械强度更好，但对真皮成纤维细胞毒性更大，这使得EDC成为未来皮肤组织工程更合适的交联剂。
