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使用人间充质干细胞和生物反应器的基于模具形状的纳米纤维支架的软骨工程。

Mold-shaped, nanofiber scaffold-based cartilage engineering using human mesenchymal stem cells and bioreactor.

作者信息

Janjanin Sasa, Li Wan-Ju, Morgan Meredith T, Shanti Rabie M, Tuan Rocky S

机构信息

Cartilage Biology and Orthopaedics Branch, National Institute of Arthritis, and Musculoskeletal and Skin Diseases, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland 20892-8022, USA.

出版信息

J Surg Res. 2008 Sep;149(1):47-56. doi: 10.1016/j.jss.2007.12.788. Epub 2008 Jan 28.

Abstract

BACKGROUND

Mesenchymal stem cell (MSC)-based tissue engineering is a promising future alternative to autologous cartilage grafting. This study evaluates the potential of using MSCs, seeded into electrospun, biodegradable polymeric nanofibrous scaffolds, to engineer cartilage with defined dimensions and shape, similar to grafts used for subcutaneous implantation in plastic and reconstructive surgery.

MATERIALS AND METHODS

Human bone marrow derived MSCs seeded onto nanofibrous scaffolds and placed in custom-designed molds were cultured for up to 42 days in bioreactors. Chondrogenesis was induced with either transforming growth factor-beta1 (TGF-beta1) alone or in combination with insulin-like growth factor-I (IGF-I).

RESULTS

Constructs exhibited hyaline cartilage histology with desired thickness and shape as well as favorable tissue integrity and shape retention, suggesting the presence of elastic tissue. Time-dependent increase in cartilage matrix gene expression was seen in both types of culture: at Day 42, TGF-beta1/IGF-I treated cultures showed higher collagen Type 2 and aggrecan expression. Both culture conditions showed significant time-dependent increase in sulfated glycosaminoglycan and hydroxyproline contents. TGF-beta1/IGF-I-treated samples were significantly stiffer; with equilibrium compressive Young's modulus values reaching 17 kPa by Day 42.

CONCLUSIONS

The successful ex vivo development of geometrically defined cartilaginous construct using customized molding suggests the potential of cell-based cartilage tissue for reconstructive surgery.

摘要

背景

基于间充质干细胞(MSC)的组织工程是自体软骨移植未来有前景的替代方法。本研究评估了将MSC接种到电纺可生物降解聚合物纳米纤维支架中,以构建具有特定尺寸和形状的软骨的潜力,该软骨类似于整形和重建手术中用于皮下植入的移植物。

材料与方法

将人骨髓来源的MSC接种到纳米纤维支架上,并置于定制设计的模具中,在生物反应器中培养长达42天。单独使用转化生长因子-β1(TGF-β1)或与胰岛素样生长因子-I(IGF-I)联合诱导软骨形成。

结果

构建物呈现透明软骨组织学特征,具有所需的厚度和形状,以及良好的组织完整性和形状保持性,提示存在弹性组织。在两种培养类型中均观察到软骨基质基因表达随时间增加:在第42天,TGF-β1/IGF-I处理的培养物显示出更高的Ⅱ型胶原蛋白和聚集蛋白聚糖表达。两种培养条件下硫酸化糖胺聚糖和羟脯氨酸含量均随时间显著增加。TGF-β1/IGF-I处理的样品硬度显著更高;到第42天,平衡压缩杨氏模量值达到17 kPa。

结论

使用定制成型成功地在体外构建了具有几何定义的软骨构建物,表明基于细胞的软骨组织在重建手术中的潜力。

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