Repair of a Meniscal Defect in a Rabbit Model Through Use of a Thermosensitive, Injectable, In Situ Crosslinked Hydrogel With Encapsulated Bone Mesenchymal Stromal Cells and Transforming Growth Factor β1
Repair of a Meniscal Defect in a Rabbit Model Through Use of a Thermosensitive, Injectable, In Situ Crosslinked Hydrogel With Encapsulated Bone Mesenchymal Stromal Cells and Transforming Growth Factor β1
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DOI：
10.1177/0363546519898519
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发表时间：
2020-01-22
影响因子：
4.8
通讯作者：
Zhao, Jinzhong
中科院分区：
文献类型：
--
作者：
Chen, Chen;Song, Jialin;Zhao, Jinzhong
背景：组织工程技术修复半月板损伤具有广阔的应用前景。在众多的支架材料中，温敏性可注射水凝胶是近年来备受关注的一类。目的：（1）评价温敏性、可注射、原位交联水凝胶的生物相容性;（2）确定含有或不含有转化生长因子β 1（TGF-β 1）的水凝胶是否能够支持骨髓间充质干细胞（BMSCs）向纤维软骨分化并促进兔半月板临界尺寸缺损的修复。研究设计：对照实验室研究。方法：对该凝胶的流变学性质和缓释性能进行研究。分离培养骨髓间充质干细胞。体外检测细胞活力、定量聚合酶链反应（qPCR）和蛋白质印迹。在体内，一个临界尺寸的缺陷被引入到30只兔的半月板。将每个缺损随机分配植入磷酸盐缓冲盐水（PBS）;负载BMSC的水凝胶;或负载BMSC的TGF-β 1掺入的水凝胶。术后8周进行组织学和免疫组化分析。采用Ishida评分系统对愈合情况进行定量评价。结果：水凝胶的弹性模量约为1000 Pa。该水凝胶表现出持续释放的特性，并且在TGF-β 1掺入后可以促进BMSCs的增殖并诱导BMSCs向纤维软骨分化（P < .001）。术后8周，大量的纤维软骨组织，番红-O染色阳性，并表达强II型胶原蛋白与弱I型胶原蛋白混合，观察到在BMSC负载，TGF-β 1掺入水凝胶组的缺损区域。在BMSC负载水凝胶组中，缺损由纤维组织和少量纤维软骨填充。PBS、BMSC水凝胶和BMSC掺入TGF-β 1的水凝胶3组的平均+/- SD定量评分分别为1.00、3.20 +/- 0.84和5.00 +/- 0.71（P <0.001）。结论：该水凝胶具有生物相容性，并且在TGF-β 1掺入后可以刺激BMSCs强烈的纤维软骨分化。局部应用载BMSC的TGF-β 1水凝胶可促进兔桡骨损伤的愈合。
Background: Meniscal injury repair with tissue engineering technique is promising. Among various scaffolds, the thermosensitive injectable hydrogel has recently attracted much attention. Purpose: (1) Evaluate the biocompatibility of thermosensitive, injectable, in situ crosslinked hydrogel and (2) determine whether the hydrogel with or without transforming growth factor beta 1 (TGF-beta 1) could support the fibrochondrogenic differentiation of bone mesenchymal stromal cells (BMSCs) and promote the repair of a critical-sized defect in rabbit meniscus. Study Design: Controlled laboratory study. Methods: The rheological and sustained release properties of the hydrogel were demonstrated. BMSCs were isolated and cultured. Cell viability, quantitative polymerase chain reaction (qPCR), and Western blot were tested in vitro. In vivo, a critical-sized defect was introduced into the meniscus of 30 rabbits. Each defect was randomly assigned to be implanted with either phosphate-buffered saline (PBS); BMSC-laden hydrogel; or BMSC-laden, TGF-beta 1-incorporated hydrogel. Histological and immunohistochemical analyses were performed at 8 weeks after surgery. The Ishida scoring system was adopted to evaluate the healing quantitatively. Results: The elastic modulus of the hydrogel was about 1000 Pa. The hydrogel demonstrated a sustained-release property and could promote proliferation and induce fibrochondrogenic differentiation of BMSCs after the incorporation of TGF-beta 1 (P < .001). At 8 weeks after surgery, a large amount of fibrocartilaginous tissue, which was positive on safranin-O staining and expressed strong type II collagen intermingled with weak type I collagen, was observed in the defect region of the BMSC-laden, TGF-beta 1-incorporated hydrogel group. In the BMSC-laden hydrogel group, the defect was filled with fibrous tissue together with a small amount of fibrocartilage. The mean +/- SD quantitative scores obtained for the 3 groups-PBS; BMSC-laden hydrogel; and BMSC-laden, TGF-beta 1-incorporated hydrogel-were 1.00, 3.20 +/- 0.84, and 5.00 +/- 0.71, respectively (P < .001). Conclusion: The hydrogel was biocompatible and could stimulate strong fibrochondrogenic differentiation of BMSCs after the incorporation of TGF-beta 1. The local administration of the BMSC-laden, TGF-beta 1-incorporated hydrogel could promote the healing of rabbit meniscal injury.