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富含血小板的血浆与琼脂糖联合作为生物活性支架增强软骨修复:一项体外研究。

Platelet-rich plasma combined with agarose as a bioactive scaffold to enhance cartilage repair: an in vitro study.

机构信息

1Orthopaedic Laboratory of Nanjing First Hospital, Nanjing Medical University, Nanjing, China.

出版信息

J Biomater Appl. 2014 Mar;28(7):1039-50. doi: 10.1177/0885328213492573. Epub 2013 Jul 3.

Abstract

OBJECTIVE

The purpose of this study was to determine whether the platelet-rich plasma-agarose gel scaffold could be a bioactive scaffold capable of growth factors release for cartilage repair.

METHODS

Porcine chondrocytes were seeded in agarose gel and platelet-rich plasma-agarose gel. During the 28-days culture, microstructure of hydrogels and morphologies of chondrocytes seeded in the hydrogels were observed using scanning electron microscope; viability of chondrocytes in gels was examined by live/dead assay; qualitative and quantitative analysis of glycosaminoglycan, collagen and DNA were assessed by histological, immunohistochemical staining and biochemical assay; gene expression was measured by real-time polymerase chain reaction. In vitro cartilage ring models were used to evaluate the integration of the scaffolds, and the integration strength was analyzed by mechanical push-out tests.

RESULTS

Scanning electron microscope revealed both scaffolds had highly uniform porous structure. Live/dead scaffolds showed 100% cells alive in both groups. After 28-days culture, glycosaminoglycan, collagen, DNA content and chondrocyte-related genes expression in platelet-rich plasma-agarose gel were significantly higher than pure agarose gel. Integration strength in platelet-rich plasma-agarose gel was also higher compared to pure agarose gel.

CONCLUSION

Platelet-rich plasma showed a positive effect on chondrocytes proliferation, differentiation and integration between native cartilage and engineered tissue when combined with agarose gel. Our findings suggest that platelet-rich plasma-agarose gel scaffold is a promising bioactive scaffold for future cartilage tissue engineering and future clinical works.

摘要

目的

本研究旨在确定富含血小板的血浆-琼脂糖凝胶支架是否可以作为一种具有生物活性的支架,能够释放生长因子用于软骨修复。

方法

将猪软骨细胞接种在琼脂糖凝胶和富含血小板的琼脂糖凝胶中。在 28 天的培养过程中,使用扫描电子显微镜观察水凝胶的微观结构和水凝胶中接种的软骨细胞的形态;通过死活检测评估凝胶中软骨细胞的活力;通过组织学、免疫组织化学染色和生化分析评估糖胺聚糖、胶原和 DNA 的定性和定量分析;通过实时聚合酶链反应测量基因表达。体外软骨环模型用于评估支架的整合,通过机械推出试验分析整合强度。

结果

扫描电子显微镜显示两种支架均具有高度均匀的多孔结构。活/死支架显示两组细胞均 100%存活。培养 28 天后,富含血小板的琼脂糖凝胶中的糖胺聚糖、胶原、DNA 含量和软骨细胞相关基因表达均显著高于纯琼脂糖凝胶。与纯琼脂糖凝胶相比,富含血小板的琼脂糖凝胶的整合强度也更高。

结论

富含血小板的血浆与琼脂糖凝胶结合时,对软骨细胞的增殖、分化以及天然软骨与工程组织之间的整合具有积极作用。我们的研究结果表明,富含血小板的琼脂糖凝胶支架是一种有前途的生物活性支架,可用于未来的软骨组织工程和未来的临床工作。

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