Liao Sida, Meng Haoye, Zhao Jun, Lin Wancheng, Liu Xiuzhi, Tian Zhuang, Lan Lan, Yang Hanyu, Zou Yulan, Xu Yichi, Gao Xiao, Lu Shibi, Peng Jiang
Institute of Orthopedics/ Beijing Key Laboratory of Regenerative Medicine in Orthopedics/ Key Laboratory of Musculoskeletal Trauma & War Injuries PLA, Chinese PLA General Hospital, Beijing, 100853, China.
Department of Anesthesiology, The First Affiliated Hospital of Guangzhou Medical University, China (mainland), Guangzhou, 510120, China.
J Orthop Translat. 2022 Apr 11;33:174-185. doi: 10.1016/j.jot.2022.03.007. eCollection 2022 Mar.
To prepare adipose-derived stem cells (ADSCs)-embedded alginate-gelatinemicrospheres (Alg-Gel-ADSCs MSs) by electrospray and evaluate their feasibility for cartilage tissue engineering. To observe the efficacy of Alg-Gel-ADSCs MSs in repairing articular cartilage defects in SD rats.
ADSCs were isolated and characterized by performing induced differentiation and flow cytometry assays. Alginate-gelatine microspheres with different gelatine concentrations were manufactured by electrospraying, and the appropriate alginate-gelatine concentration and ratio were determined by evaluating microsphere formation. Alg-Gel-ADSCs MSs were compared with Alg-ADSCs MSs through the induction of chondrogenic differentiation and culture. Their feasibility for cartilage tissue engineering was analysed by performing Live/Dead staining, cell proliferation analysis, toluidine blue staining and a glycosaminoglycan (GAG) content analysis. Alg-Gel-ADSCs MSs were implanted in the cartilage defects of SD rats, and the cartilage repair effect was evaluated at different time points. The evaluation included gross observations and histological evaluations, fluorescence imaging tracking, immunohistochemical staining, microcomputed tomography (micro-CT) and a CatWalk evaluation.
The isolated ADSCs showed multidirectional differentiation and were used for cartilage tissue engineering. Using 1.5 w:v% alginate and 0.5 w:v% gelatine (Type B), we successfully prepared nearly spherical microspheres. Compared with alginate microspheres, alginate gel increased the viability of ADSCs and promoted the proliferation and chondrogenesis of ADSCs. In our experiments on knee cartilage defects in SD rats in vivo, the Alg-Gel-ADSCs MSs showed superior cartilage repair in cell resides, histology evaluation, micro-CT imaging and gait analysis.
Microspheres composed of 1.5 w:v% alginate-0.5 w:v% gelatine increase the viability of ADSCs and supported their proliferation and deposition of cartilage matrix components. ADSCs embedded in 1.5 w:v% alginate-0.5 w:v% gelatine microspheres show superior repair efficacy and prospective applications in cartilage tissue repair.
In this study, injectable adipose-derived stem cells-embedded alginate-gelatin microspheres (Alg-Gel-ADSCs MSs) were prepared by the electrospray . Compared with the traditional alginate microspheres, its support ability for ADSCs is better and shows a better repair effect. This study provides a promising strategy for cartilage tissue regeneration.
通过电喷雾法制备包埋脂肪来源干细胞（ADSCs）的海藻酸钠 - 明胶微球（Alg - Gel - ADSCs MSs），并评估其用于软骨组织工程的可行性。观察Alg - Gel - ADSCs MSs修复SD大鼠关节软骨缺损的效果。
分离ADSCs并通过诱导分化和流式细胞术分析进行鉴定。通过电喷雾法制备不同明胶浓度的海藻酸钠 - 明胶微球，并通过评估微球形成确定合适的海藻酸钠 - 明胶浓度和比例。通过软骨生成分化诱导和培养，将Alg - Gel - ADSCs MSs与Alg - ADSCs MSs进行比较。通过活/死染色、细胞增殖分析、甲苯胺蓝染色和糖胺聚糖（GAG）含量分析，分析其用于软骨组织工程的可行性。将Alg - Gel - ADSCs MSs植入SD大鼠的软骨缺损处，并在不同时间点评估软骨修复效果。评估包括大体观察和组织学评估、荧光成像跟踪、免疫组织化学染色、微计算机断层扫描（micro - CT）和CatWalk评估。
分离出的ADSCs表现出多向分化能力，并用于软骨组织工程。使用1.5 w:v%海藻酸钠和0.5 w:v%明胶（B型），成功制备出近乎球形的微球。与海藻酸钠微球相比，海藻酸钠凝胶提高了ADSCs的活力，促进了ADSCs的增殖和软骨生成。在我们对SD大鼠膝关节软骨缺损的体内实验中，Alg - Gel - ADSCs MSs在细胞驻留、组织学评估、micro - CT成像和步态分析方面显示出优异的软骨修复效果。
由1.5 w:v%海藻酸钠 - 0.5 w:v%明胶组成的微球提高了ADSCs的活力，并支持其增殖和软骨基质成分的沉积。包埋于1.5 w:v%海藻酸钠 - 0.5 w:v%明胶微球中的ADSCs在软骨组织修复中显示出优异的修复效果和潜在应用前景。
在本研究中，通过电喷雾法制备了可注射的包埋脂肪来源干细胞的海藻酸钠 - 明胶微球（Alg - Gel - ADSCs MSs）。与传统的海藻酸钠微球相比，其对ADSCs的支持能力更好，并显示出更好的修复效果。本研究为软骨组织再生提供了一种有前景的策略。
