Experimental study of platelet-rich plasma in treatment of Achilles tendinopathy in rabbits_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

GONG Dong , ZHAO Maosheng , SU Wencai , DONG Chenhui , DENG Yinshuan ,  ZHEN Ping

Department of Orthopedics, the 940th Hospital of Joint Logistics Support Force of Chinese People’s Liberation Army, Lanzhou Gansu, 730050, P.R.China;

Corresponding?author：

ZHEN Ping, Email: zhenpingok@163.com

Keywords：

Platelet-rich plasma; Achilles tendinopathy; rabbit

DOI：

10.7507/1002-1892.201809045

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Objective To explore the effect of platelet-rich plasma (PRP) in treatment of Achilles tendinopathy in rabbits, and provide experimental evidence for the clinical application of PRP in treatment of Achilles tendinopathy. Methods Forty-eight adult New Zealand white rabbits, weighing 2.5-3.0 kg, male or female, were randomly divided into model group (group A), model control group (group B), model+treatment control group (group C), model+treatment group (group D), with 12 in each group. The rabbits were injected with type Ⅰ collagenase to prepare Achilles tendinopathy models in groups A, C, and D, and with an equal dose of normal saline in group B. The blood from the central artery of rabbit ear was taken to preprare PRP by secondary centrifugation in group D. The results of platelet counts showed that PRP platelets reached 3 to 5 times the whole blood. After the model was prepared, the rabbits in groups C and D were injected with physiological saline and autologous PRP at the molding site respectively, once a week, 0.8 mL each time for 4 weeks. At 1 week after PRP injection, the relative hardness (expressed as HRD%) of Achilles tendon was evaluated by ultrasound elastic quantitative imaging detection technique; the maximum breaking load of Achilles tendon was measured by universal electronic tensile testing machine; the contents of collagen type Ⅰ and Ⅲ were determined by ELISA; and the morphology of Achilles tendon collagen fibers was observed by HE and Masson stainings. Results All animals survived during the experiment. The results of ultrasound elastic quantitative imaging and mechanical tests showed that the HRD% and the maximum breaking load were significantly lower in group A than in group B (P<0.05) and in group C than in group D (P<0.05). The results of ELISA showed that the content of collagen type Ⅰ was significantly lower in group A than in group B (P<0.05) and in group C than in group D (P<0.05); the content of collagen type Ⅲ was significantly higher in group A than in group B (P<0.05) and in group D than in group C (P<0.05). HE and Masson stainings showed that the Achilles tendon collagen fibers were irregularly curled and the structure was severely damaged in group A; the fibers were parallel and ordered, and the structure was complete in group B; the fibers were irregularly curled and structurally disordered in group C; the fibers were slightly curled and the structure was relatively complete in group D. Conclusion A rabbit model of Achilles tendinopathy can be reconstructed by type Ⅰ collagenase injection. PRP treatment can increase the Achilles tendon hardness and maximum breaking load, up-regulate the expression level of collagen type Ⅰ and Ⅲ, improve the structure of Achilles tendon collagen fiber, and promote the repair in rabbit Achilles tendinopathy model.

Citation： GONG Dong, ZHAO Maosheng, SU Wencai, DONG Chenhui, DENG Yinshuan, ZHEN Ping. Experimental study of platelet-rich plasma in treatment of Achilles tendinopathy in rabbits. Chinese Journal of Reparative and Reconstructive Surgery, 2019, 33(7): 871-876. doi: 10.7507/1002-1892.201809045 Copy

1.	Longo UG, Ronga M, Maffulli N. Achilles tendinopathy. Sports Med Arthrosc Rev, 2018, 26(1): 16-30.
2.	Smith WB, Melton W, Davies J. Midsubstance tendinopathy, percutaneous techniques (platelet-rich plasma, extracorporeal shock wave therapy, prolotherapy, radiofrequency ablation). Clin Podiatr Med Surg, 2017, 34(2): 161-174.
3.	紀慶明, 楊育暉, 陳昊, 等. 富血小板血漿輔助前交叉韌帶重建治療的臨床療效分析. 中國修復重建外科雜志, 2017, 31(4): 410-416.
4.	Le ADK, Enweze L, DeBaun MR, et al. Current clinical recommendations for use of patelet-rich plasma. Curr Rev Musculoskelet Med, 2018, 11(4): 624-634.
5.	Chen J, Yu Q, Wu B, et al. Autologous tenocyte therapy for experimental Achilles tendinopathy in a rabbit model. Tissue Eng Part A, 2011, 17(15-16): 2037-2048.
6.	Landesberg R, Roy M, Glickman RS. Quantification of growth factor levels using a simplified method of platelet-rich plasma gel preparation. J Oral Maxillofac Surg, 2000, 58(3): 297-301.
7.	王鵬飛, 時長江, 王玉亮, 等. 兔富血小板血漿制備及其活性分析. 中國組織工程研究, 2013, 17(8): 1411-1417.
8.	Jeong CD, Kim SE, Shim KS, et al. Exploring the in vivo anti-inflammatory actions of simvastatin-loaded porous microspheres on inflamed tenocytes in a collagenase-induced animal model of Achilles tendinitis. Int J Mol Sci, 2018, 19(3): 820-826.
9.	de Cesar Netto, Godoy-Santos AL, Augusto Pontin P, et al. Novel animal model for Achilles tendinopathy: Controlled experimental study of serial injections of collagenase in rabbits. PLoS One, 2018, 13(2): e0192769.
10.	周游, 唐康來. 肌腱病模型研究進展. 中國運動醫學雜志, 2013, 32(9): 829-833.
11.	Masoudi EA, Ribas J, Kaushik G, et al. Platelet-rich blood derivatives for stem cell-based tissue engineering and regeneration. Curr Stem Cell Reports, 2016, 2(1): 33-42.
12.	方家劉, 尹宗生, 王偉, 等. 富白細胞和血小板血漿對 BMSCs 在兔股骨頭缺血性壞死中成骨作用的影響. 中國修復重建外科雜志, 2015, 29(2): 227-233.
13.	Ooi CC, Schneider ME, Malliaras P, et al. Diagnostic performance of axial-strain sonoelastography in confirming clinically diagnosed Achilles tendinopathy: comparison with B-mode ultrasound and color Doppler imaging. Ultrasound Med Biol, 2015, 41(1): 15-25.
14.	方建強, 朱文峰, 李維芝, 等. 聲觸診組織成像量化技術在診斷急性跟腱炎中的應用價值. 臨床超聲醫學雜志, 2017, 19(2): 99-101.
15.	李敏, 金力. 腱病動物模型研究進展. 現代預防醫學, 2013, 40(3): 565-567.
16.	Fu SC, Chan KM, Rolf CG. Increased deposition of sulfated glycosaminoglycans in human patellar tendinopathy. Clin J Sport Med, 2007, 17(2): 129-134.
17.	李輝, 唐康來, 鄧銀栓, 等. 外源性前列腺素 E2 對兔跟腱膠原含量的影響. 中國修復重建外科雜志, 2012, 26(3): 352-358.
18.	吳言, 杜國慶, 宗立秋, 等. 超聲彈性彌散定量技術評價兔跟腱病的轉歸及生物力學特征. 中華超聲影像學雜志, 2016, 25(5): 433-437.
19.	耿震, 王宸, 周海洋. 富血小板血漿對肌腱愈合影響的實驗研究. 中國修復重建外科雜志, 2011, 25(3): 344-348.
20.	沈勇偉, 史曉偉. 低功率激光對末端病大鼠跟腱蛋白多糖和膠原的作用. 中國運動醫學雜志, 2014, 33(2): 125-129.

1. Longo UG, Ronga M, Maffulli N. Achilles tendinopathy. Sports Med Arthrosc Rev, 2018, 26(1): 16-30.
2. Smith WB, Melton W, Davies J. Midsubstance tendinopathy, percutaneous techniques (platelet-rich plasma, extracorporeal shock wave therapy, prolotherapy, radiofrequency ablation). Clin Podiatr Med Surg, 2017, 34(2): 161-174.
3. 紀慶明, 楊育暉, 陳昊, 等. 富血小板血漿輔助前交叉韌帶重建治療的臨床療效分析. 中國修復重建外科雜志, 2017, 31(4): 410-416.
4. Le ADK, Enweze L, DeBaun MR, et al. Current clinical recommendations for use of patelet-rich plasma. Curr Rev Musculoskelet Med, 2018, 11(4): 624-634.
5. Chen J, Yu Q, Wu B, et al. Autologous tenocyte therapy for experimental Achilles tendinopathy in a rabbit model. Tissue Eng Part A, 2011, 17(15-16): 2037-2048.
6. Landesberg R, Roy M, Glickman RS. Quantification of growth factor levels using a simplified method of platelet-rich plasma gel preparation. J Oral Maxillofac Surg, 2000, 58(3): 297-301.
7. 王鵬飛, 時長江, 王玉亮, 等. 兔富血小板血漿制備及其活性分析. 中國組織工程研究, 2013, 17(8): 1411-1417.
8. Jeong CD, Kim SE, Shim KS, et al. Exploring the in vivo anti-inflammatory actions of simvastatin-loaded porous microspheres on inflamed tenocytes in a collagenase-induced animal model of Achilles tendinitis. Int J Mol Sci, 2018, 19(3): 820-826.
9. de Cesar Netto, Godoy-Santos AL, Augusto Pontin P, et al. Novel animal model for Achilles tendinopathy: Controlled experimental study of serial injections of collagenase in rabbits. PLoS One, 2018, 13(2): e0192769.
10. 周游, 唐康來. 肌腱病模型研究進展. 中國運動醫學雜志, 2013, 32(9): 829-833.
11. Masoudi EA, Ribas J, Kaushik G, et al. Platelet-rich blood derivatives for stem cell-based tissue engineering and regeneration. Curr Stem Cell Reports, 2016, 2(1): 33-42.
12. 方家劉, 尹宗生, 王偉, 等. 富白細胞和血小板血漿對 BMSCs 在兔股骨頭缺血性壞死中成骨作用的影響. 中國修復重建外科雜志, 2015, 29(2): 227-233.
13. Ooi CC, Schneider ME, Malliaras P, et al. Diagnostic performance of axial-strain sonoelastography in confirming clinically diagnosed Achilles tendinopathy: comparison with B-mode ultrasound and color Doppler imaging. Ultrasound Med Biol, 2015, 41(1): 15-25.
14. 方建強, 朱文峰, 李維芝, 等. 聲觸診組織成像量化技術在診斷急性跟腱炎中的應用價值. 臨床超聲醫學雜志, 2017, 19(2): 99-101.
15. 李敏, 金力. 腱病動物模型研究進展. 現代預防醫學, 2013, 40(3): 565-567.
16. Fu SC, Chan KM, Rolf CG. Increased deposition of sulfated glycosaminoglycans in human patellar tendinopathy. Clin J Sport Med, 2007, 17(2): 129-134.
17. 李輝, 唐康來, 鄧銀栓, 等. 外源性前列腺素 E2 對兔跟腱膠原含量的影響. 中國修復重建外科雜志, 2012, 26(3): 352-358.
18. 吳言, 杜國慶, 宗立秋, 等. 超聲彈性彌散定量技術評價兔跟腱病的轉歸及生物力學特征. 中華超聲影像學雜志, 2016, 25(5): 433-437.
19. 耿震, 王宸, 周海洋. 富血小板血漿對肌腱愈合影響的實驗研究. 中國修復重建外科雜志, 2011, 25(3): 344-348.
20. 沈勇偉, 史曉偉. 低功率激光對末端病大鼠跟腱蛋白多糖和膠原的作用. 中國運動醫學雜志, 2014, 33(2): 125-129.

Chinese Journal of Reparative and Reconstructive Surgery

Experimental study of platelet-rich plasma in treatment of Achilles tendinopathy in rabbits

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