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体外和体内分化的间充质干细胞修复软骨缺损的比较。

Comparison of chondral defects repair with in vitro and in vivo differentiated mesenchymal stem cells.

作者信息

Fan Hongbin, Liu Haifeng, Zhu Rui, Li Xusheng, Cui Yuming, Hu Yunyu, Yan Yongnian

机构信息

Institute of Orthopaedics & Traumatology, Xijing Hospital, The Fourth Military Medical University, Xi'an, PR China.

出版信息

Cell Transplant. 2007;16(8):823-32. doi: 10.3727/000000007783465181.

Abstract

The purpose of this study was to compare chondral defects repair with in vitro and in vivo differentiated mesenchymal stem cells (MSCs). A novel PLGA-gelatin/chondroitin/hyaluronate (PLGA-GCH) hybrid scaffold with transforming growth factor-beta1 (TGF-beta1)-impregnated microspheres (MS-TGF) was fabricated to mimic the extracellular matrix. MS-TGF showed an initial burst release (22.5%) and a subsequent moderate one that achieved 85.1% on day 21. MSCs seeded on PLGA-GCH/MS-TGF or PLGA-GCH were incubated in vitro and showed that PLGA-GCH/MS-TGF significantly augmented proliferation of MSCs and glycosaminoglycan synthesis compared with PLGA-GCH. Then MSCs seeded on PLGA-GCH/MS-TGF were implanted and differentiated in vivo to repair chondral defect on the right knee of rabbit (in vivo differentiation repair group), while the contralateral defect was repaired with in vitro differentiated MSCs seeded on PLGA-GCH (in vitro differentiation repair group). The histology observation demonstrated that in vivo differentiation repair showed better chondrocyte morphology, integration, and subchondral bone formation compared with in vitro differentiation repair 12 and 24 weeks postoperatively, although there was no significant difference after 6 weeks. The histology grading score comparison also demonstrated the same results. The present study implies that in vivo differentiation induced by PLGA-GCH/MS-TGF and the host microenviroment could keep chondral phenotype and enhance repair. It might serve as another way to induce and expand seed cells in cartilage tissue engineering.

摘要

本研究的目的是比较体外和体内分化的间充质干细胞(MSCs)对软骨缺损的修复效果。制备了一种新型的聚乳酸-羟基乙酸共聚物-明胶/软骨素/透明质酸盐(PLGA-GCH)混合支架,并负载转化生长因子-β1(TGF-β1)微球(MS-TGF)以模拟细胞外基质。MS-TGF呈现出初始的突发释放(22.5%),随后在第21天达到85.1%的中度释放。接种在PLGA-GCH/MS-TGF或PLGA-GCH上的MSCs在体外培养,结果显示与PLGA-GCH相比,PLGA-GCH/MS-TGF显著增强了MSCs的增殖和糖胺聚糖的合成。然后将接种在PLGA-GCH/MS-TGF上的MSCs植入兔右膝以在体内分化修复软骨缺损(体内分化修复组),而对侧缺损则用接种在PLGA-GCH上的体外分化的MSCs修复(体外分化修复组)。组织学观察表明,术后12周和24周时,与体外分化修复相比,体内分化修复显示出更好的软骨细胞形态、整合以及软骨下骨形成,尽管术后6周时无显著差异。组织学分级评分比较也显示了相同的结果。本研究表明,PLGA-GCH/MS-TGF诱导的体内分化和宿主微环境可保持软骨表型并增强修复。这可能是软骨组织工程中诱导和扩增种子细胞的另一种方法。

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