Ishida Kazunari, Kuroda Ryosuke, Miwa Masahiko, Tabata Yasuhiko, Hokugo Akishige, Kawamoto Teruya, Sasaki Ken, Doita Minoru, Kurosaka Masahiro
Department of Orthopedic Surgery, Kobe University Graduate School of Medicine, Kobe, Japan.
Tissue Eng. 2007 May;13(5):1103-12. doi: 10.1089/ten.2006.0193.
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.
本研究的目的是检验富血小板血浆(PRP)在体外和体内增强半月板组织再生的假设。在体外研究中,制备了半月板细胞单层培养物,并进行了3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-磺基苯基)-2H-四唑内盐测定和5-溴-2'-脱氧尿苷测定,以评估在PRP存在下的增殖行为。进行阿尔辛蓝测定以评估细胞外基质(ECM)合成。为了检测纤维软骨相关信使核糖核酸(mRNA)表达,进行了实时聚合酶链反应。在体内研究中,在兔半月板的无血管区域制造了直径1.5毫米的全层缺损。明胶水凝胶(GH)用作PRP生长因子的药物递送系统。缺损填充如下:A组,含PRP的GH;B组,含贫血小板血浆的GH;C组,仅GH。每组在手术后4、8和12周进行组织学评估。PRP刺激了脱氧核糖核酸合成和ECM合成(p<0.05)。用PRP培养的半月板细胞显示双糖链蛋白聚糖和核心蛋白聚糖的mRNA表达更高(p<0.05)。组织学结果显示,4周时存在明胶水凝胶残余物,表明水凝胶可控制释放约4周。12周时缺损部位的组织学评分显示,接受含PRP的GH的动物的半月板修复明显优于其他两组。这些发现表明PRP可增强半月板缺损的愈合。