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应用滑膜来源基质细胞和壳聚糖/Ⅰ型胶原支架进行软骨组织工程。

Use of synovium-derived stromal cells and chitosan/collagen type I scaffolds for cartilage tissue engineering.

机构信息

Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830054, People's Republic of China.

出版信息

Biomed Mater. 2010 Oct;5(5):055005. doi: 10.1088/1748-6041/5/5/055005. Epub 2010 Sep 9.

Abstract

The objective was to investigate synovium-derived stromal cells (SDSCs) coupled with chitosan/collagen type I (CS/COL-I) scaffolds for cartilage engineering. CS/COL-I scaffolds were fabricated through freeze-drying and cross-linked by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. SDSCs were isolated from synovium and cultured onto CS/COL-I scaffolds, constructs of which were incubated in serum-free chondrogenic medium with sequential application of TGF-β1 and bFGF for up to 21 days and then implanted into nude mice. The physical characteristics of the scaffolds were examined. The quality of the in vitro constructs was assessed in terms of DNA content by PicoGreen assay and cartilaginous matrix by histological examination. The implants of the constructs were evaluated by histological and immunohistochemical examinations and reverse transcription PCR. Results indicated that the CS/COL-I scaffold showed porous structures, and the DNA content of SDSCs in CS/COL-I scaffolds increased at 1 week culture time. Both of the constructs in vitro and the implants were examined with positive stained GAGs histologically and the implants with positive collagen type II immunohistochemically. RT-PCR of the implants indicated that aggrecan and collagen type II expressed. It suggested that SDSCs coupled with CS/COL-I scaffolds treated sequentially with TGF-β1 and bFGF in vitro were highly competent for engineered cartilage formation in vitro and in vivo.

摘要

目的是研究滑膜衍生基质细胞(SDSCs)与壳聚糖/胶原 I 型(CS/COL-I)支架相结合在软骨工程中的应用。CS/COL-I 支架通过冷冻干燥和 1-乙基-3-(3-二甲基氨基丙基)碳二亚胺交联制成。从滑膜中分离 SDSCs 并培养到 CS/COL-I 支架上,构建体在无血清软骨形成培养基中孵育,依次应用 TGF-β1 和 bFGF 长达 21 天,然后植入裸鼠体内。检查支架的物理特性。通过 PicoGreen 测定法评估体外构建体的质量,通过组织学检查评估软骨基质的质量。通过组织学和免疫组织化学检查以及逆转录 PCR 评估构建体的植入物。结果表明,CS/COL-I 支架具有多孔结构,CS/COL-I 支架中 SDSCs 的 DNA 含量在 1 周培养时间增加。体外的构建体和植入物均在组织学上用阳性染色的 GAGs 进行检查,植入物在免疫组织化学上用阳性的 II 型胶原进行检查。植入物的 RT-PCR 表明聚集蛋白聚糖和 II 型胶原表达。这表明,体外经 TGF-β1 和 bFGF 序贯处理后与 CS/COL-I 支架结合的 SDSCs 非常适合体外和体内工程软骨的形成。

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