Zou Shengtao, Xu Guochao, Zheng Zhenyu, Chen Tianming, Huang Yixing
Department of Orthopedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China.
Key Laboratory of Orthopedics of Zhejiang Province, Wenzhou, China.
Tissue Eng Part A. 2025 Jul;31(13-14):1015-1025. doi: 10.1089/ten.tea.2024.0231. Epub 2024 Dec 5.
In the present study, acellular cartilage matrix (ACM) was modified with poly-l-lysine/hyaluronic acid (PLL/HA) multilayers via detergent-enzyme chemical digestion and layer-by-layer self-assembly technology. This modified ACM was then loaded with Transforming Growth Factor Beta 3 (TGF-β3) and incorporated into a thermosensitive hydrogel (TH) to create a HA/PLL-ACM/TH composite scaffold with sustained-release function. This study aimed to evaluate the efficacy of this novel composite scaffold in promoting chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and facilitating osteochondral defect repair. , isolated, and cultured rat BMSCs were inoculated in equal amounts into TH, ACM/TH, and HA/PLL-ACM/TH groups, with or without TGF-β3 supplementation, for 21 days. Western blot (WB) analysis and immunofluorescence staining were employed to assess the expression levels of collagen II, aggrecan, and SOX-9. , osteochondral defect was created in the Sprague-Dawley rat trochlea using microdrilling. TH, ACM/TH, and HA/PLL-ACM/TH scaffolds, with or without TGF-β3, were implanted into the defect. After 6 weeks, the repairs were evaluated macroscopically, using Micro computed tomography (micro-CT), histological analysis, and immunohistochemistry. The results demonstrated that the HA/PLL-ACM/TH scaffold loaded with TGF-β3 significantly upregulated the expression of collagen II, aggrecan, and SOX-9 compared with the control and other experimental groups. Furthermore, at 6 weeks postsurgery, the HA/PLL-ACM/TH group loaded with TGF-β3 exhibited superior tissue formation on the joint surface, as confirmed by micro-CT and histological evidence, indicating improved osteochondral repair. These findings suggest that the HA/PLL-ACM/TH scaffold loaded with TGF-β3 holds promise as a therapeutic strategy for osteochondral defect and offers a novel approach for utilizing acellular cartilage microfilaments.
在本研究中,通过去污剂 - 酶化学消化和层层自组装技术,用聚 - L - 赖氨酸/透明质酸(PLL/HA)多层修饰无细胞软骨基质(ACM)。然后将这种修饰的ACM负载转化生长因子β3(TGF-β3),并掺入热敏水凝胶(TH)中,以创建具有缓释功能的HA/PLL - ACM/TH复合支架。本研究旨在评估这种新型复合支架在促进骨髓间充质干细胞(BMSCs)软骨分化和促进骨软骨缺损修复方面的功效。将分离并培养的大鼠BMSCs等量接种到TH、ACM/TH和HA/PLL - ACM/TH组中,添加或不添加TGF-β3,培养21天。采用蛋白质印迹(WB)分析和免疫荧光染色评估Ⅱ型胶原蛋白、聚集蛋白聚糖和SOX - 9的表达水平。使用微钻在Sprague-Dawley大鼠滑车中制造骨软骨缺损。将含或不含TGF-β3的TH、ACM/TH和HA/PLL - ACM/TH支架植入缺损处。6周后,通过微型计算机断层扫描(micro-CT)、组织学分析和免疫组织化学对修复情况进行宏观评估。结果表明,与对照组和其他实验组相比,负载TGF-β3的HA/PLL - ACM/TH支架显著上调了Ⅱ型胶原蛋白、聚集蛋白聚糖和SOX - 9的表达。此外,术后6周,micro-CT和组织学证据证实,负载TGF-β3的HA/PLL - ACM/TH组在关节表面表现出更好的组织形成,表明骨软骨修复得到改善。这些发现表明,负载TGF-β3的HA/PLL - ACM/TH支架有望成为骨软骨缺损的治疗策略,并为利用无细胞软骨微丝提供了一种新方法。