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使用间充质干细胞与聚二氧六环酮/聚乙烯醇支架构建物修复骨软骨缺损。

Repair of osteochondral defects with a construct of mesenchymal stem cells and a polydioxanone/poly(vinyl alcohol) scaffold.

作者信息

Jeong Woong-Kyo, Oh Se-Heang, Lee Jin-Ho, Im Gun-Il

机构信息

Department of Orthopedic Surgery, Korea University Anam Hospital, 126-1 Anam-Dong, Seoul 136-705, Korea.

出版信息

Biotechnol Appl Biochem. 2008 Feb;49(Pt 2):155-64. doi: 10.1042/BA20070149.

Abstract

The purpose of the present study was to test the potential of an MSC (mesenchymal stem cell)/PDO/PVA [polydioxanone/poly(vinyl alcohol)] hybrid scaffold construct to repair cartilage defects. For the in vitro study, MSCs were isolated from human bone marrow and cultured in PDO/PVA scaffolds for 4 weeks. Gross and microscopic examinations were performed, as well as RT-PCR (reverse-transcription PCR) analyses for cartilage-specific genes. For the in vivo study, MSCs isolated from rabbits were cultured in the PDO/PVA scaffolds and tissue-engineered into neocartilage, then implanted into the osteochondral defect on the distal femur of the same rabbits. Gross and histological evaluations were performed at 8 weeks after the implantation. The results of the in vitro study demonstrated that the physical stability of the cell-cultured hybrid scaffold was maintained until 4 weeks after initial placement. Scanning electronmicroscopy indicated the infiltration of the cells into, and appropriate interactions with, the scaffolds. RT-PCR showed an expression of cartilage-specific genes similar to that seen with pellet-cultured MSCs. From the in vivo study, the defect area of the experimental group showed smooth consistent glistening white tissue resembling articular cartilage, whereas the control group showed a red irregular tissue with surface depression. The regenerated cartilage of the experimental group showed metachromasia on both Safranin-O and dense staining for type II collagen, whereas the control group showed little metachromatic staining and less intense staining for type II collagen. A histological score of the quality of cartilage formation indicated that the MSC/PDO/PVA hybrid scaffold successfully produced neocartilage in vitro and enhanced the repair of the osteochondral defect in vivo. Further in vivo studies will be necessary to elucidate further the value of PDO/PVA as a scaffold material for cartilage regeneration.

摘要

本研究的目的是测试间充质干细胞(MSC)/聚对二氧环己酮/聚乙烯醇(PDO/PVA)混合支架构建体修复软骨缺损的潜力。在体外研究中,从人骨髓中分离出MSC,并在PDO/PVA支架中培养4周。进行大体和显微镜检查,以及针对软骨特异性基因的逆转录聚合酶链反应(RT-PCR)分析。在体内研究中,将从兔子分离出的MSC在PDO/PVA支架中培养并组织工程化为新软骨,然后植入同一只兔子股骨远端的骨软骨缺损处。在植入后8周进行大体和组织学评估。体外研究结果表明,细胞培养的混合支架的物理稳定性在初始放置后4周内得以维持。扫描电子显微镜显示细胞浸润到支架中并与支架进行了适当的相互作用。RT-PCR显示软骨特异性基因的表达与颗粒培养的MSC相似。在体内研究中,实验组的缺损区域显示出类似于关节软骨的光滑一致的闪亮白色组织,而对照组显示出红色不规则组织且表面凹陷。实验组再生软骨在番红O染色和II型胶原密集染色中均显示异染性,而对照组异染性染色较少且II型胶原染色强度较低。软骨形成质量的组织学评分表明,MSC/PDO/PVA混合支架在体外成功产生了新软骨,并在体内增强了骨软骨缺损的修复。需要进一步的体内研究来进一步阐明PDO/PVA作为软骨再生支架材料的价值。

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