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富血小板血浆预处理的人软骨细胞种子 PLGA 网塞修复体内半月板。

Meniscal repair in vivo using human chondrocyte-seeded PLGA mesh scaffold pretreated with platelet-rich plasma.

机构信息

Department of Orthopaedic Surgery, Seoul National University College of Medicine, Korea.

Medical Research Centre, Seoul National University, Korea.

出版信息

J Tissue Eng Regen Med. 2017 Feb;11(2):471-480. doi: 10.1002/term.1938. Epub 2014 Jun 19.

Abstract

The objective of this study was to test the hypothesis that platelet-rich plasma (PRP) pretreatment on a poly-lactic-co-glycolic acid (PLGA) mesh scaffold enhances the healing capacity of the meniscus with human chondrocyte-seeded scaffolds in vivo, even when the seeded number of cells was reduced from 10 million to one million. A flexible PLGA mesh scaffold was pretreated with PRP using a centrifugal technique. One million human articular chondrocytes were seeded onto the scaffold by dynamic oscillation. After 7 days, scaffolds were placed between human meniscal discs and were implanted subcutaneously in nude mice for 6 weeks (n = 16/group). Fluorescence microscopy demonstrated uniform attachment of the chondrocytes throughout the scaffolds 24 h following seeding. Cell attachment analysis revealed a significantly increased number of chondrocytes on PRP-pretreated than non-treated scaffolds (p < 0.05). Field emission scanning electron microscopy revealed chondrocytes attached to the PRP-pretreated scaffolds interconnecting their cellular processes with the fibrin network at 24 h and day 7 of culture. Of the 16 constructs containing PRP-pretreated scaffolds implanted in mice, six menisci healed completely, nine healed incompletely and one did not heal. Histological results from the 16 control constructs containing non-treated scaffolds revealed that none had healed completely, four healed incompletely and 12 did not heal. The histological outcome between the groups was significantly different (p < 0.05). These findings suggest that human articular chondrocytes on PRP-pretreated PLGA mesh scaffolds demonstrate increased cell attachment and enhance the healing capacity of meniscus with a reduced number of seeding cells in a meniscal repair mouse model. Copyright © 2014 John Wiley & Sons, Ltd.

摘要

本研究旨在验证以下假设,即富血小板血浆(PRP)预处理聚丙交酯-乙交酯(PLGA)网片支架可增强经 PRP 预处理的 PLGA 网片支架中的人软骨细胞的黏附能力,从而增强半月板的愈合能力,即便细胞接种数从 1000 万减少至 100 万。采用离心技术对柔性 PLGA 网片支架进行 PRP 预处理。通过动态振荡将 100 万个关节软骨细胞接种到支架上。7 天后,将支架置于人半月板之间,并将其植入裸鼠皮下 6 周(每组 n=16)。荧光显微镜显示,接种后 24 小时,软骨细胞均匀地附着在支架上。细胞黏附分析显示,PRP 预处理组的软骨细胞数量明显多于未处理组(p<0.05)。场发射扫描电子显微镜显示,PRP 预处理组的软骨细胞在接种后 24 小时和第 7 天与纤维蛋白网络连接,其细胞突起相互连接。在植入小鼠的含有 PRP 预处理支架的 16 个构建体中,有 6 个半月板完全愈合,9 个不完全愈合,1 个未愈合。在含有未处理支架的 16 个对照构建体的组织学结果显示,没有一个完全愈合,4 个不完全愈合,12 个未愈合。组间的组织学结果差异有统计学意义(p<0.05)。这些发现表明,PRP 预处理 PLGA 网片支架上的人关节软骨细胞表现出增强的细胞黏附能力,并在半月板修复小鼠模型中,以减少细胞接种数增强半月板的愈合能力。版权所有©2014 约翰威立父子公司

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