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仿生一体化多孔PVA/凝胶/V-n-HA/pa6支架与骨髓间充质干细胞构建体的成骨与成软骨作用在关节骨软骨缺损修复中的应用

Osteogenesis and chondrogenesis of biomimetic integrated porous PVA/gel/V-n-HA/pa6 scaffolds and BMSCs construct in repair of articular osteochondral defect.

作者信息

Li Xiang, Li Yubao, Zuo Yi, Qu Dan, Liu Yiming, Chen Tao, Jiang Nan, Li Hui, Li Jihua

机构信息

State Key Laboratory of Oral Diseases and Department of Oral and Maxillofacial Surgery, West China College of Stomatology, Sichuan University, Chengdu, 610041, China.

Research Center for Nano-Biomaterials, Analytical and Testing Center, Sichuan University, Chengdu, 610064, China.

出版信息

J Biomed Mater Res A. 2015 Oct;103(10):3226-36. doi: 10.1002/jbm.a.35452. Epub 2015 Apr 1.

DOI:10.1002/jbm.a.35452
PMID:25772000
Abstract

A novel bi-layered osteochondral scaffold, including of PVA/Gel/V layer for the cartilage and n-HA/PA6 layer for the subchondral bone, has been proposed to evaluate the potential of the engineered of osteochondral grafts in repairing articular osteochondral defects in rabbits. The two different layers of the scaffolds were seeded with allogenic bone marrow-derived stem cells (BMSCs), which were chondrogenically and osteogenically induced respectively. The critical-size osteochondral defects were created in the knees of adult rabbits. The defects were treated with cell-bi-layered constructs (Group A), bi-layered constructs (Group B) and untreated group C as control group. The adhesion, proliferation and differentiation of BMSCs were demonstrated by immunohistochemical staining and scanning electron microscopy (SEM) in vitro. Cell survival was tracked via fluorescent labeling in vivo. Overall, the porous PVA/Gel/V-n-HA/PA6 scaffold was compatible and had no negative effects on the BMSCs in vitro culture. The cell-bi-layered scaffolds showed superior repair results as compared to the control group using gross examination and histological assessment. With BMSCs implantation, the two different layers of the composite biomimetic scaffolds provided a suitable environment for cells to form respective tissue. Simultaneously, the RT-PCR results confirmed the expression of specific extracellular matrix (ECM) markers for cartilaginous or osteoid tissue. This investigation showed that the porous PVA/Gel/V-n-HA/PA6 scaffold is a potential matrix for treatment of osteochondral defects, and the method of using chondrogenically and osteogenically differentiated BMSCs as seed cells on each layer might be a promising strategy in repair of articular osteochondral defect due to enhanced chondrogenesis and osteogenesis.

摘要

一种新型的双层骨软骨支架已被提出,该支架包括用于软骨的PVA/凝胶/ V层和用于软骨下骨的n - HA/PA6层,以评估工程化骨软骨移植物修复兔关节骨软骨缺损的潜力。将异体骨髓来源的干细胞(BMSCs)接种到支架的两个不同层上,分别进行软骨生成和骨生成诱导。在成年兔的膝关节中制造临界尺寸的骨软骨缺损。缺损分别用细胞双层构建体(A组)、双层构建体(B组)处理,未处理的C组作为对照组。通过免疫组织化学染色和扫描电子显微镜(SEM)在体外证明BMSCs的黏附、增殖和分化。通过体内荧光标记追踪细胞存活情况。总体而言,多孔PVA/凝胶/ V - n - HA/PA6支架具有相容性,对体外培养的BMSCs没有负面影响。使用大体检查和组织学评估,细胞双层支架显示出比对照组更好的修复效果。通过植入BMSCs,复合仿生支架的两个不同层为细胞形成各自的组织提供了合适的环境。同时,RT - PCR结果证实了软骨或类骨组织特异性细胞外基质(ECM)标志物的表达。这项研究表明,多孔PVA/凝胶/ V - n - HA/PA6支架是治疗骨软骨缺损的潜在基质,并且在每层上使用软骨生成和骨生成分化的BMSCs作为种子细胞的方法可能是修复关节骨软骨缺损的一种有前途的策略,因为它增强了软骨生成和骨生成。

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