Objective

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.

Methods

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.

Results

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.

Conclusions

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.

The translational potential of this article

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.

中文翻译：

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电喷雾制备可注射脂肪干细胞包埋海藻酸盐-明胶微球用于软骨组织再生

客观的

采用电喷雾法制备脂肪源性干细胞（ADSCs）包埋海藻酸盐-明胶微球（Alg-Gel-ADSCs MSs）并评价其在软骨组织工程中的可行性。观察Alg-Gel-ADSCs MSs修复SD大鼠关节软骨缺损的疗效。

方法

通过进行诱导分化和流式细胞术测定分离和表征 ADSC。采用电喷雾法制备了不同明胶浓度的海藻酸盐-明胶微球，并通过评价微球的形成来确定合适的海藻酸盐-明胶浓度和比例。通过诱导软骨分化和培养，将 Alg-Gel-ADSCs MSs 与 Alg-ADSCs MSs 进行比较。通过进行活/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，分析了它们用于软骨组织工程的可行性。将Alg-Gel-ADSCs MSs植入SD大鼠软骨缺损处，评价不同时间点的软骨修复效果。评估包括总体观察和组织学评估，

结果

分离的ADSCs表现出多向分化，可用于软骨组织工程。使用 1.5 w:v% 的海藻酸盐和 0.5 w:v% 的明胶（B 型），我们成功地制备了近乎球形的微球。与海藻酸盐微球相比，海藻酸盐凝胶增加了ADSCs的活力，促进了ADSCs的增殖和软骨形成。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、显微 CT 成像和步态分析中显示出优异的软骨修复。

结论

由 1.5 w:v% 藻酸盐-0.5 w:v% 明胶组成的微球可提高 ADSC 的活力并支持其增殖和软骨基质成分的沉积。嵌入 1.5 w:v% alginate-0.5 w:v% 明胶微球中的 ADSC 在软骨组织修复中显示出卓越的修复功效和前瞻性应用。

本文的翻译潜力

在这项研究中，通过电喷雾制备了可注射的脂肪干细胞包埋的海藻酸盐-明胶微球（Alg-Gel-ADSCs MSs）。与传统的海藻酸盐微球相比，其对ADSCs的支持能力更好，修复效果更好。该研究为软骨组织再生提供了一种有前景的策略。