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支架辅助的软骨组织工程,使用源自人髋关节软骨的婴儿软骨细胞。

Scaffold-assisted cartilage tissue engineering using infant chondrocytes from human hip cartilage.

机构信息

Department of Orthopaedic Surgery, University Medical Center Rostock, Doberanerstrasse 142, 18057 Rostock, Germany.

出版信息

Osteoarthritis Cartilage. 2013 Dec;21(12):1997-2005. doi: 10.1016/j.joca.2013.09.007. Epub 2013 Oct 2.

Abstract

OBJECTIVE

Studies about cartilage repair in the hip and infant chondrocytes are rare. The aim of our study was to evaluate the use of infant articular hip chondrocytes for tissue engineering of scaffold-assisted cartilage grafts.

METHOD

Hip cartilage was obtained from five human donors (age 1-10 years). Expanded chondrocytes were cultured in polyglycolic acid (PGA)-fibrin scaffolds. De- and re-differentiation of chondrocytes were assessed by histological staining and gene expression analysis of typical chondrocytic marker genes. In vivo, cartilage matrix formation was assessed by histology after subcutaneous transplantation of chondrocyte-seeded PGA-fibrin scaffolds in immunocompromised mice.

RESULTS

The donor tissue was heterogenous showing differentiated articular cartilage and non-differentiated tissue and considerable expression of type I and II collagens. Gene expression analysis showed repression of typical chondrocyte and/or mesenchymal marker genes during cell expansion, while markers were re-induced when expanded cells were cultured in PGA-fibrin scaffolds. Cartilage formation after subcutaneous transplantation of chondrocyte loaded PGA-fibrin scaffolds in nude mice was variable, with grafts showing resorption and host cell infiltration or formation of hyaline cartilage rich in type II collagen. Addition of human platelet rich plasma (PRP) to cartilage grafts resulted robustly in formation of hyaline-like cartilage that showed type II collagen and regions with type X collagen.

CONCLUSION

These results suggest that culture of expanded and/or de-differentiated infant hip cartilage cells in PGA-fibrin scaffolds initiates chondrocyte re-differentiation. The heterogenous donor tissue containing immature chondrocytes bears the risk of cartilage repair failure in vivo, which may be possibly overcome by the addition of PRP.

摘要

目的

关于髋关节和婴儿软骨细胞的软骨修复研究较为罕见。本研究旨在评估利用婴儿关节髋关节软骨细胞进行支架辅助软骨移植物的组织工程。

方法

从 5 名供体(年龄 1-10 岁)中获取髋关节软骨。扩增的软骨细胞在聚乙二醇酸(PGA)-纤维蛋白支架中培养。通过组织学染色和典型软骨细胞标志物基因的基因表达分析评估软骨细胞的去分化和再分化。在体内,通过免疫缺陷小鼠皮下移植软骨细胞接种 PGA-纤维蛋白支架评估软骨基质形成。

结果

供体组织不均一,表现出分化的关节软骨和未分化的组织,以及大量 I 型和 II 型胶原的表达。基因表达分析显示,在细胞扩增过程中典型软骨细胞和/或间充质标志物基因受到抑制,而当扩增细胞在 PGA-纤维蛋白支架中培养时,标志物被重新诱导。将负载软骨细胞的 PGA-纤维蛋白支架皮下移植到裸鼠后,软骨形成情况不一,移植物出现吸收和宿主细胞浸润,或形成富含 II 型胶原的透明软骨。向软骨移植物中添加富含血小板的血浆(PRP)可强有力地形成透明样软骨,表现出 II 型胶原和 X 型胶原区域。

结论

这些结果表明,在 PGA-纤维蛋白支架中培养扩增和/或去分化的婴儿髋关节软骨细胞可启动软骨细胞再分化。含有未成熟软骨细胞的异质供体组织存在体内软骨修复失败的风险,通过添加 PRP 可能克服这一风险。

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