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三维聚合物-透明质酸支架中扩增的人半月板衍生细胞用于半月板修复。

Expanded human meniscus-derived cells in 3-D polymer-hyaluronan scaffolds for meniscus repair.

机构信息

TransTissue Technologies GmbH, Charitéplatz 1/Virchowweg 11, 10117 Berlin, Germany.

出版信息

Acta Biomater. 2012 Feb;8(2):677-85. doi: 10.1016/j.actbio.2011.10.007. Epub 2011 Oct 11.

DOI:10.1016/j.actbio.2011.10.007
PMID:22023746
Abstract

Treatment options for lesions of the avascular region of the meniscus using regenerative medicine approaches based on resorbable scaffolds are rare. Recent approaches using scaffold-based techniques for tissue regeneration known from cartilage repair may be a promising treatment option for meniscal tears. The aim of the study was the investigation of meniscus matrix formation of in vitro expanded human meniscus-derived cells in a three-dimensional (3-D) bioresorbable polymer graft for meniscal repair approaches. Cultivation of the human meniscus cells was performed in a resorbable scaffold material made of polyglycolic acid (PGA) and hyaluronic acid, stabilized with fibrin glue. Cell viability and distribution of human meniscus cells in PGA-hyaluronan scaffolds were evaluated by fluorescein diacetate and propidium iodide staining. Verification of typical meniscal extracellular matrix molecules like type I and type III collagen was performed histologically, immunohistochemically and by gene expression analysis. In results, 3-D scaffold-based meniscus cultures showed high cell viability over an observational period of 21 days in PGA-hyaluronan scaffolds. On the protein level, type I collagen and proteoglycans were evident. Gene expression analysis confirmed the re-expression of meniscus-specific markers in PGA-hyaluronan scaffolds. This study demonstrated that in vitro expanded human meniscus cells allow for formation of meniscal matrix components when cultured in 3-D PGA-hyaluronan scaffolds stabilized with fibrin. These results encourage scaffold-based approaches for the treatment of meniscal lesions.

摘要

基于可吸收支架的再生医学方法治疗半月板无血管区病变的选择很少。最近,基于已知用于软骨修复的支架技术的组织再生方法可能是半月板撕裂的一种有前途的治疗选择。本研究的目的是研究在用于半月板修复方法的三维(3-D)可生物吸收聚合物移植物中体外扩增的人半月板来源细胞的半月板基质形成。用人半月板细胞在聚乙二醇酸(PGA)和透明质酸可吸收支架材料中培养,用纤维蛋白胶稳定。通过荧光二乙酸酯和碘化丙啶染色评估人半月板细胞在 PGA-透明质酸支架中的活力和分布。通过组织学、免疫组织化学和基因表达分析验证典型的半月板细胞外基质分子,如 I 型和 III 型胶原。结果显示,在 PGA-透明质酸支架中,3-D 支架培养的半月板细胞在观察期 21 天内具有较高的活力。在蛋白质水平上,I 型胶原和蛋白聚糖明显。基因表达分析证实了在 PGA-透明质酸支架中半月板特异性标志物的重新表达。本研究表明,在纤维蛋白稳定的 3-D PGA-透明质酸支架中培养时,体外扩增的人半月板细胞允许形成半月板基质成分。这些结果鼓励基于支架的方法治疗半月板病变。

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