Regenerative Medicine and Stem Cell Laboratory, Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, NH-65, Kandi, Sangareddy 502285, Telangana, India.
Biofab TE Laboratory, Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, NH-65, Kandi, Sangareddy 502285, Telangana, India.
Int J Biol Macromol. 2022 Jul 31;213:768-779. doi: 10.1016/j.ijbiomac.2022.06.010. Epub 2022 Jun 7.
Researchers have always tried expensive in vitro tests to show the 3D usability of dECM. The use of tissue-specific hydrogels in a microfluidic device is rarely studied. In this study, we have used ECM obtained from goat digital flexor tendons by decellularization technique. The tdECM was characterized for its structural properties using Scanning Electron Microscopy (SEM). Collagen, dsDNA, GAGs, and protein contents were quantified using spectrophotometric assays. The cell viability and proliferation of human umbilical cord-derived mesenchymal stem cells (hUMSCs) encapsulated in the tdECM hydrogel inside the microfluidic device were checked using Calcein-AM/PI. The FTIR data showed prominent peaks of the amide group, indicating the presence of collagen. The SEM data showed intact fiber morphology after the decellularization process. There was a 95 % reduction in double-stranded DNA (dsDNA) content, proving the effectiveness of the decellularization technique. There was no significant difference in the collagen content of tdECM and the GAGs were also in the acceptable range compared to the native tissue. Over 90 % cell viability in hUMSCs was observed qualitatively and quantitatively in vitro and inside a microfluidic device. In conclusion, we characterized the tdECM hydrogel and demonstrated its compatibility with the microfluidic device.
研究人员一直在尝试昂贵的体外测试，以展示 dECM 的 3D 可用性。很少研究在微流控装置中使用组织特异性水凝胶。在本研究中，我们使用脱细胞技术从山羊数字屈肌腱获得 ECM。使用扫描电子显微镜 (SEM) 对 tdECM 的结构特性进行了表征。使用分光光度法测定了胶原蛋白、dsDNA、GAG 和蛋白质含量。使用 Calcein-AM/PI 检查了包封在微流控装置内的 tdECM 水凝胶中的人脐带间充质干细胞 (hUMSC) 的细胞活力和增殖。FTIR 数据显示酰胺基团的特征峰，表明存在胶原蛋白。SEM 数据显示脱细胞过程后纤维形态完整。双链 DNA (dsDNA) 含量减少了 95%，证明了脱细胞技术的有效性。tdECM 中的胶原蛋白含量没有差异，与天然组织相比，GAG 也在可接受的范围内。体外和微流控装置内 hUMSC 的细胞活力超过 90%。总之，我们对 tdECM 水凝胶进行了表征，并证明了其与微流控装置的兼容性。
