Basiri Arefeh, Farokhi Mehdi, Azami Mahmoud, Ebrahimi-Barough Somayeh, Mohamadnia Abdolreza, Rashtbar Morteza, Hasanzadeh Elham, Mahmoodi Narges, Baghaban Eslaminejad Mohamadreza, Ai Jafar
Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
Prog Biomater. 2019 Mar;8(1):31-42. doi: 10.1007/s40204-019-0108-7. Epub 2019 Jan 31.
A hybrid hydrogel was obtained from decellularized extract from Wharton's jelly (DEWJ) and silk fibroin (SF) and characterized for cartilage tissue engineering. Wharton's jelly was used due to its similarity with articular cartilage in extracellular matrix composition. Also, silk fibroin has good mechanical properties which make this construct appropriate for cartilage repair. Decellularization of Wharton's jelly was verified by DAPI staining, DNA quantification, and PCR analysis. Then, the biochemical composition of DEWJ was determined by ELISA kits for total proteins, collagens, sulfated glycosaminoglycans (sGAG), and transforming growth factor β1 (TGF-β1). After fabricating pure SF and SF/DEWJ hybrid hydrogels, their physical and mechanical properties were characterized by FESEM, Fourier-transform infrared spectroscopy (FTIR) and rheological assays (amplitude and frequency sweeps). Furthermore, cell viability and proliferation were assessed by MTT assay. The results have shown that DEWJ in hybrid hydrogels enhances mechanical properties of the construct relative to pure SF hydrogels. Also, this extract at its 40% concentration in culture media and 20% or 40% concentrations in SF/DEWJ hybrid hydrogels significantly increases population of the cells compared to control and pure SF hydrogel after 7 days. In conclusion, this study proposes the potential of SF/DEWJ hybrid hydrogels for cartilage tissue engineering applications.
一种混合水凝胶由来自脐带华通氏胶的脱细胞提取物(DEWJ)和丝素蛋白(SF)制备而成,并对其进行了软骨组织工程特性表征。选用脐带华通氏胶是因为其细胞外基质组成与关节软骨相似。此外,丝素蛋白具有良好的机械性能,这使得该构建体适合用于软骨修复。通过DAPI染色、DNA定量和PCR分析验证了脐带华通氏胶的脱细胞效果。然后,使用ELISA试剂盒测定DEWJ的总蛋白、胶原蛋白、硫酸化糖胺聚糖(sGAG)和转化生长因子β1(TGF-β1)的生化组成。制备纯SF和SF/DEWJ混合水凝胶后,通过场发射扫描电子显微镜(FESEM)、傅里叶变换红外光谱(FTIR)和流变学测定(振幅和频率扫描)对其物理和机械性能进行了表征。此外,通过MTT法评估细胞活力和增殖情况。结果表明,相对于纯SF水凝胶,混合水凝胶中的DEWJ增强了构建体的机械性能。而且,与对照组和纯SF水凝胶相比,该提取物在培养基中浓度为40%以及在SF/DEWJ混合水凝胶中浓度为20%或40%时,7天后细胞数量显著增加。总之,本研究提出了SF/DEWJ混合水凝胶在软骨组织工程应用中的潜力。