BACKGROUND:Cartilage tissue lacks the ability to heal. Cartilage tissue engineering using cell-free scaffolds has been increasingly used in recent years. OBJECTIVE:This study describes the use of a type I collagen scaffold combined with WNT5A plasmid to promote chondrocyte proliferation and differentiation in a rabbit osteochondral defect model. METHODS:Type I collagen was extracted and fabricated into a collagen scaffold. To improve gene transfection efficiency, a cationic chitosan derivative N,N,N-trimethyl chitosan chloride (TMC) vector was used. A solution of TMC/WNT5A complexes was adsorbed to the collagen scaffold to prepare a WNT5A scaffold. Osteochondral defects were created in the femoral condyles of rabbits. The rabbits were divided into defect, scaffold, and scaffold with WNT5A groups. At 6 and 12 weeks after creation of the osteochondral defects, samples were collected from all groups for macroscopic observation and gene expression analysis. RESULTS:Samples from the defect group exhibited incomplete cartilage repair, while those from the scaffold and scaffold with WNT5A groups exhibited “preliminary cartilage” covering the defect. Cartilage regeneration was superior in the scaffold with WNT5A group compared to the scaffold group. Safranin O staining revealed more proteoglycans in the scaffold and scaffold with WNT5A groups compared to the defect group. The expression levels of aggrecan, collagen type II, and SOX9 genes were significantly higher in the scaffold with WNT5A group compared to the other two groups. CONCLUSIONS:Type I collagen scaffold showed effective adsorption and guided the three-dimensional arrangement of stem cells. WNT5A plasmid promoted cartilage repair by stimulating the expression of aggrecan, type II collagen, and SOX9 genes and proteins, as well as inhibiting cartilage hypertrophy.

中文翻译：

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用于原位软骨组织工程的具有 WNT5A 质粒的 I 型胶原支架

背景：软骨组织缺乏自愈能力。近年来，使用无细胞支架的软骨组织工程得到了越来越多的应用。目的：本研究描述了使用I型胶原支架联合WNT5A质粒促进兔骨软骨缺损模型中软骨细胞增殖和分化。方法：提取I型胶原蛋白并制成胶原蛋白支架。为了提高基因转染效率，使用了阳离子壳聚糖衍生物 N,N,N-三甲基壳聚糖氯化物 (TMC) 载体。将 TMC/WNT5A 复合物溶液吸附到胶原支架上以制备 WNT5A 支架。在兔子的股骨髁中产生骨软骨缺损。将兔分为缺损组、支架组和带有WNT5A的支架组。在骨软骨缺损产生后 6 周和 12 周，从所有组中收集样本进行宏观观察和基因表达分析。结果：缺损组的样本软骨修复不完全，而支架和WNT5A组的样本表现出覆盖缺损的“初步软骨”。与支架组相比，具有 WNT5A 组的支架中的软骨再生更好。与缺陷组相比，番红 O 染色显示支架和具有 WNT5A 组的支架中的蛋白聚糖更多。与其他两组相比，WNT5A 组支架中蛋白聚糖、II 型胶原蛋白和 SOX9 基因的表达水平显着升高。结论：I型胶原支架表现出有效吸附并引导干细胞的三维排列。WNT5A 质粒通过刺激聚集蛋白聚糖、II 型胶原蛋白和 SOX9 基因和蛋白质的表达以及抑制软骨肥大来促进软骨修复。