The regenerative effects of platelet-rich plasma on meniscal cells in vitro and its in vivo application with biodegradable gelatin hydrogel
The regenerative effects of platelet-rich plasma on meniscal cells in vitro and its in vivo application with biodegradable gelatin hydrogel
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DOI：
10.1089/ten.2006.0193
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发表时间：
2007-05-01
影响因子：
--
通讯作者：
Kurosaka, Masahiro
中科院分区：
文献类型：
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作者：
Ishida, Kazunari;Kuroda, Ryosuke;Kurosaka, Masahiro
本研究的目的是检验富血小板血浆（PRP）在体外和体内增强血管组织再生的假设。在体外研究中，制备单层胰岛细胞培养物，并进行3-（4，5-二甲基噻唑-2基）-5-（3-羧基甲氧基苯基）-2-（4-磺基苯基）-2H-四唑鎓内盐测定和5-溴-2 '-脱氧尿苷测定以评估在PRP存在下的增殖行为。进行阿新蓝测定以评估细胞外基质（ECM）合成。实时荧光定量聚合酶链反应检测纤维软骨相关信使核糖核酸（mRNA）的表达。在体内研究中，在兔半月板的无血管区域中创建1.5 mm直径的全层缺损。明胶水凝胶（GH）被用作PRP生长因子的药物递送系统。A组：GH + PRP; B组：GH+贫血小板血浆; C组：单纯GH。每组在术后4、8和12周进行组织学评价。PRP刺激脱氧核糖核酸合成和ECM合成（p < 0.05）。用富血小板血浆培养的半月板细胞显示双糖蛋白聚糖和核心蛋白聚糖的mRNA表达更高（p < 0.05）。组织学发现显示明胶水凝胶的残余物在4周时存在，表明水凝胶可以控制释放约4周。12周时缺损部位的组织学评分显示，接受PRP + GH的动物的尿道修复显著优于其他两组。这些发现表明PRP促进了骨缺损的愈合。
The objective of the study was to test the hypothesis that platelet-rich plasma (PRP) enhances meniscal tissue regeneration in vitro and in vivo. In the in vitro study, monolayer meniscal cell cultures were prepared, and 3-( 4,5- dimethylthiazol-2yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H- tetrazolium inner salt assay and 5-bromo-2 '- deoxyuridine assay were performed to assess proliferative behavior in the presence of PRP. Alcian blue assay was performed to assess extracellular matrix (ECM) synthesis. To detect the fibrocartilage-related messenger ribonucleic acid (mRNA) expressions, real-time polymerase chain reaction was performed. In the in vivo study, 1.5-mm-diameter full-thickness defects were created in the avascular region of rabbit meniscus. Gelatin hydrogel (GH) was used as the drug delivery system for PRP growth factors. The defects were filled as follows: Group A, GH with PRP; Group B, GH with platelet-poor plasma; Group C, GH only. Each group was evaluated histologically at 4, 8, and 12 weeks after surgery. PRP stimulated deoxyribonucleic acid synthesis and ECM synthesis ( p < 0.05). Meniscal cells cultured with PRP showed greater mRNA expression of biglycan and decorin (p < 0.05). Histological findings showed that remnants of gelatin hydrogels existed at 4 weeks, indicating that the hydrogels could control release for approximately 4 weeks. Histological scoring of the defect sites at 12 weeks revealed significantly better meniscal repair in animals that received PRP with GH than in the other two groups. These findings suggest that PRP enhances the healing of meniscal defects.