Chen Lei, Liu Jun-Peng, Tang Kang-Lai, Wang Qing, Wang Guo-Dong, Cai Xian-Hua, Liu Xi-Ming
Department of Orthopaedics, Wuhan General Hospital of Guangzhou Military Region, Wuhan, China.
Cell Physiol Biochem. 2014;34(6):2153-68. doi: 10.1159/000369659. Epub 2014 Dec 2.
BACKGROUND/AIMS: Tendon injuries are common, difficult to cure and usually healed with fibrosis and scar tissue. The aim of this study was to evaluate tendon derived stem cells (TDSCs) and platelet rich plasma (PRP) in the treatment of collagenase induced Achilles tendinopathy in rat.
Four and 8 weeks (n=18) after TDSCs, PRP, PRP with TDSC or PBS (control) injection into collagenase or saline (sham) injected rat Achilles tendon, tendon tissue was harvested and tendon quality was evaluated by histology and biomechanical testing. TDSCs were cultured and treated by 10% PRP, and the FAK/ERK1/2 signaling pathway and tenocyte-related genes were detected by western blot analysis.
Compared to the control, PRP treatment resulted in better healing of injured tendons with improved histological outcomes and biomechanical functions. The addition of TDSCs to PRP treatment significantly enhanced the effects of PRP treatment alone. TDSC injection alone had little effect on tendon healing. PRP and PRP with TDSC treatments of collagenase induced tendon injuries also increased the mRNA and protein expression of tenocyte-related genes (type I collagen, SCX, Tenascin C) and activated the focal adhesion kinase (FAK) and extracellular-regulated kinase (ERK) 1/2 signaling pathways. Treatment of TDSCs in vitro with 10% PRP significantly increased the phosphorylation levels of FAK and ERK1/2 and the protein levels of tenocyte-related genes (Col I, SCX and Tenascin C). Inhibition of the FAK and ERK1/2 signaling pathways abolished the effect of PRP.
This study concludes that PRP combined with TDSCs is potentially effective for the treatment of tendinopathy. The PRP induced, FAK and ERK1/2 dependent activation of tenocyte related genes in TDSCs in vitro suggests that the beneficial healing effect of the PRP with TDSC combination might occur by means of an improved TDSC differentiation toward the tenocyte lineage. Thus, a PRP with TDSC combination therapy may be clinically useful.
背景/目的：肌腱损伤很常见，难以治愈，通常会形成纤维化和瘢痕组织而愈合。本研究的目的是评估肌腱源性干细胞（TDSCs）和富血小板血浆（PRP）在治疗大鼠胶原酶诱导的跟腱病中的作用。
将TDSCs、PRP、TDSCs与PRP联合或PBS（对照）注射到胶原酶或生理盐水（假手术）注射的大鼠跟腱中，4周和8周后（n = 18），采集肌腱组织，通过组织学和生物力学测试评估肌腱质量。培养TDSCs并用10% PRP处理，通过蛋白质印迹分析检测粘着斑激酶（FAK）/细胞外信号调节激酶1/2（ERK1/2）信号通路和肌腱细胞相关基因。
与对照组相比，PRP治疗导致损伤肌腱愈合更好，组织学结果和生物力学功能得到改善。在PRP治疗中加入TDSCs显著增强了单独PRP治疗的效果。单独注射TDSCs对肌腱愈合影响不大。PRP以及PRP与TDSCs联合治疗胶原酶诱导的肌腱损伤也增加了肌腱细胞相关基因（I型胶原、SCX、肌腱蛋白C）的mRNA和蛋白表达，并激活了粘着斑激酶（FAK）和细胞外调节激酶（ERK）1/2信号通路。用10% PRP体外处理TDSCs显著增加了FAK和ERK1/2的磷酸化水平以及肌腱细胞相关基因（Col I、SCX和肌腱蛋白C）的蛋白水平。抑制FAK和ERK1/2信号通路消除了PRP的作用。
本研究得出结论，PRP联合TDSCs对肌腱病的治疗可能有效。PRP在体外诱导TDSCs中FAK和ERK1/2依赖性激活肌腱细胞相关基因，表明PRP与TDSCs联合的有益愈合作用可能是通过改善TDSCs向肌腱细胞谱系的分化而实现的。因此，PRP与TDSCs联合治疗可能具有临床应用价值。
