Department of Orthopedics, the First Affiliated Hospital of Nanjing Medical University, Nanjing, People's Republic of China.
Department of Orthopedics, Affiliated Hospital of Jiangsu University, Zhenjiang, People's Republic of China.
Int J Nanomedicine. 2019 Jul 2;14:4755-4765. doi: 10.2147/IJN.S199306. eCollection 2019.
Many techniques and methods have been used clinically to relieve pain from cartilage repair, but the long-term effect is still unsatisfactory. The objective of this study was to form an artificial chondroid tissue gene enhanced tissue engineering system to repair cartilage defects via nanosized liposomes. Cationic nanosized liposomes were prepared and characterized using transmission electron microscope (TEM) and dynamic laser light scattering (DLS). The rat mesenchymal stem cells (rMSCs) were isolated, cultivated, and induced by SRY (Sex-Determining Region Y)-Box 9 (Sox9) via cationic nanosized liposomes. The induced rMSCs were mixed with a thermo-sensitive chitosan hydrogel and subcutaneously injected into the nude mice. Finally, the newly-formed chondroid tissue obtained in the injection parts, and the transparent parts were detected by HE, collagen II, and safranin O. It was found that the presently prepared cationic nanosized liposomes had the diameter of 85.76±3.48 nm and the zeta potential of 15.76±2.1 mV. The isolated rMSCs proliferation was fibroblast-like, with a cultivated confluence of 90% confluence in 5-8 days, and stained positive for CD29 and CD44 while negative for CD34 and CD45. After transfection with cationic nanosized liposomes, we observed changes of cellular morphology and a higher expression of SOX9 compared with control groups, which indicated that rMSCs could differentiate into chondrocyte in vitro. By mixing transfected rMSCs with the thermo-sensitive hydrogel of chitosan in nude mice, chondroid tissue was successfully obtained, demonstrating that rMSCs can differentiate into chondrogenic cells in vivo. This study explored new ways to improve the quality of tissue engineered cartilage, thus accelerating clinical transformation and reducing patient pain.
许多技术和方法已被用于临床缓解软骨修复后的疼痛，但长期效果仍不理想。本研究的目的是构建人工软骨样组织基因增强组织工程系统，通过纳米脂质体修复软骨缺损。采用透射电子显微镜（TEM）和动态激光光散射（DLS）制备和表征阳离子纳米脂质体。分离、培养大鼠间充质干细胞（rMSCs），并通过阳离子纳米脂质体诱导 SRY（性别决定区 Y）-盒 9（Sox9）。将诱导的 rMSCs 与温敏壳聚糖水凝胶混合，皮下注射入裸鼠。最后，在注射部位和透明部位通过 HE、胶原 II 和番红 O 检测新形成的软骨样组织。结果发现，目前制备的阳离子纳米脂质体的粒径为 85.76±3.48nm，Zeta 电位为 15.76±2.1mV。分离的 rMSCs 增殖呈成纤维细胞样，培养 5-8 天达到 90%融合，CD29 和 CD44 染色阳性，CD34 和 CD45 染色阴性。阳离子纳米脂质体转染后，与对照组相比，细胞形态发生变化，SOX9 表达水平升高，表明 rMSCs 可在体外向软骨细胞分化。将转染的 rMSCs 与壳聚糖温敏水凝胶混合后，在裸鼠体内成功获得软骨样组织，表明 rMSCs 可在体内向软骨细胞分化。本研究探索了提高组织工程软骨质量的新途径，从而加速临床转化，减轻患者痛苦。
