Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
Tehran University of Medical Sciences, Department of Tissue Engineering and Applied Cell Sciences, Tehran, Iran.
J Biomed Mater Res A. 2020 Apr;108(4):938-946. doi: 10.1002/jbm.a.36871. Epub 2020 Jan 8.
Cartilage tissue engineering is the interdisciplinary science that will help to improve cartilage afflictions, such as arthrosis, arthritis, or following joints traumatic injuries. In the present work, we developed an injectable hydrogel which derived from decellularized extracellular matrix of sheep cartilage. Successful decellularization was evaluated by measuring the DNA, glycosaminoglycans (GAG), collagen contents, and histological analyses. There was a minor difference in GAG and collagen contents among natural cartilage and decellularized tissue as well as ultimate hydrogel. Rheological analysis showed that the temperature and gelation time of prepared hydrogel were 37°C and between 5 and 7 min, respectively. Mechanical properties evaluation indicated a storage modulus of 20 kPa. The results show that prepared hydrogel possessed cell-friendly microenvironment as confirmed via calcein staining and MTT assay. Also, cells were able to proliferate which observed by H&E and alcian blue staining. Cell attachment and proliferation at the surface of the decellularized hydrogel was apparent by Scanning Electron Microscope (SEM) images and microphotographs. Furthermore, the cells embedded within the hydrogel were able to differentiate into chondrocyte with limited evidence of hypertrophy and osteogenesis in utilized cells which proved by SOX9, CoL2, ACAN, and also CoL1 and CoL10 gene expression levels. In summary, the results suggest that developed novel injectable hydrogel from decellularized cartilage could be utilized as a promising substrate for cartilage tissue engineering applications.
软骨组织工程是一门跨学科的科学，它将有助于改善软骨疾病，如骨关节炎、关节炎或关节创伤后。在本工作中，我们开发了一种可注射水凝胶，它来源于绵羊软骨的去细胞细胞外基质。成功的去细胞化通过测量 DNA、糖胺聚糖 (GAG)、胶原蛋白含量和组织学分析来评估。天然软骨和去细胞组织以及最终水凝胶之间的 GAG 和胶原蛋白含量存在微小差异。流变学分析表明，制备的水凝胶的温度和凝胶时间分别为 37°C 和 5-7 分钟。力学性能评估表明储存模量为 20 kPa。结果表明，所制备的水凝胶具有细胞友好的微环境，这通过钙黄绿素染色和 MTT 测定得到证实。此外，通过 H&E 和阿利新蓝染色观察到细胞能够增殖。通过扫描电子显微镜 (SEM) 图像和显微照片可以明显看出细胞在去细胞水凝胶表面的附着和增殖。此外，细胞在水凝胶内能够分化为软骨细胞，而在使用的细胞中几乎没有肥大和成骨的证据，这通过 Sox9、CoL2、ACAN 以及 CoL1 和 CoL10 基因表达水平得到证实。总之，结果表明，从去细胞软骨开发的新型可注射水凝胶可作为软骨组织工程应用的有前途的基质。
