An oriented-collagen scaffold including Wnt5a promotes osteochondral regeneration and cartilage interface integration in a rabbit model
An oriented-collagen scaffold including Wnt5a promotes osteochondral regeneration and cartilage interface integration in a rabbit model
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包含 Wnt5a 的定向胶原支架促进兔模型中的骨软骨再生和软骨界面整合
DOI：
10.1096/fj.202000280r
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发表时间：
2020-07-06
期刊：
影响因子：
4.8
通讯作者：
Feng, Gang
中科院分区：
文献类型：
--
作者：
Qi, Yiying;Zhang, Wenkan;Feng, Gang
关节软骨再生仍然是骨科的主要挑战。在这种情况下，非经典Wnt5a是一种特别有吸引力的生长因子; Wnt5a抑制软骨细胞肥大，但维持软骨形成。我们设计了一种新型的垂直定向胶原支架。评价Wnt5a对MSC和软骨细胞的作用以及Wnt5a/定向胶原支架在骨软骨修复和软骨整合方面的治疗效果。在体外，评估用Wnt5a处理的MSC和软骨细胞的增殖、迁移和分化及其机制。进行mRNA微阵列分析以比较Wnt5a处理前后MSC的表达谱。在体内，在24只新西兰白色兔的髌骨沟中制造全层圆柱形骨软骨缺损（直径4 mm，深度3 mm），并植入定向胶原支架（n = 8）、Wnt 5a/定向胶原支架（n = 8）或不植入任何支架（n = 8）。6周和12周后，评价整合和组织反应。经Wnt5a处理后，MSC和软骨细胞的增殖、迁移、成软骨分化和细胞外基质形成得到了极大的改善。Western blotting结果显示PI3K/AKT/JNK信号通路被激活。微阵列分析显示，Wnt5a组表现出PI3K通路的显著上调。Reactome GSEA途径相互作用分析显示，这种上调与胶原蛋白和细胞外基质形成相关。在体内，与其他组相比，Wnt 5a/定向胶原支架组表现出最佳的界面整合、软骨再生和胶原纤维排列，伴随着再生和整合区域中糖胺聚糖和胶原积累的显著增加。基因表达分析显示，在Wnt 5a/导向的胶原支架组中，编码软骨形成相关基因的mRNA水平显著增加（均P <.05）。Wnt5a通过激活PI3K/AKT/JNK信号通路促进MSCs和软骨细胞的增殖、迁移和向软骨细胞分化。Wnt5a/定向胶原构建体增强了兔模型中透明软骨的结构特异性再生，并且可能是修复人类软骨缺损的有希望的治疗方法。
Articular cartilage regeneration remains a major challenge in orthopedics. Noncanonical Wnt5a is a particularly attractive growth factor in this context; Wnt5a inhibits chondrocyte hypertrophy but maintains chondrogenesis. We designed a novel, vertically oriented-collagen scaffold. The effect of Wnt5a on MSCs and chondrocytes and the therapeutic effects of the Wnt5a/oriented-collagen scaffold in terms of osteochondral repair and cartilage integration were evaluated. In vitro, the proliferation, migration, and differentiation of MSCs and chondrocytes treated with Wnt5a, and the mechanisms thereof, were assessed. mRNA microarray analysis was performed to compare the expression profiles of MSCs before and after Wnt5a treatment. In vivo, full-thickness cylindrical osteochondral defects (4 mm in diameter, 3 mm in depth) were created in the patellar grooves of 24 New Zealand white rabbits and implanted with oriented-collagen scaffolds (n = 8), Wnt5a/oriented-collagen scaffolds (n = 8), or nothing (n = 8). After 6 and 12 weeks, integration and tissue responses were evaluated. The proliferation, migration, chondrogenic differentiation, and extracellular matrix formation of/by MSCs and chondrocytes improved greatly after treatment with Wnt5a. Western blotting showed that the PI3K/AKT/JNK signaling pathway was activated. Microarray analysis revealed that the Wnt5a group exhibited a significant upregulation of the PI3K pathway. Reactome GSEA pathway interaction analysis revealed that such upregulation was associated with collagen and extracellular matrix formation. In vivo, the Wnt5a/oriented-collagen scaffold group exhibited optimal interface integration, cartilage regeneration, and collagenous fiber arrangement, accompanied by significantly increased glycosaminoglycan and collagen accumulations in the zones of regeneration and integration, compared to the other groups. Gene expression analysis showed that the levels of mRNAs encoding genes involved in cartilage formation were significantly increased in the Wnt5a/oriented, collagen scaffold group (allP < .05). Wnt5a promoted the proliferation, migration, and chondrogenic differentiation of MSCs and chondrocytes via the activation of the PI3K/AKT/JNK signaling pathway. The Wnt5a/oriented-collagen constructs enhanced the structure-specific regeneration of hyaline cartilage in a rabbit model and may be a promising treatment for the repair of human cartilage defects.