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

Video：

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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.	纪庆明, 杨育晖, 陈昊, 等. 富血小板血浆辅助前交叉韧带重建治疗的临床疗效分析. 中国修复重建外科杂志, 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.	王鹏飞, 时长江, 王玉亮, 等. 兔富血小板血浆制备及其活性分析. 中国组织工程研究, 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, 唐康来. 肌腱病模型研究进展. 中国运动医学杂志, 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.	方家刘, 尹宗生, 王伟, 等. 富白细胞和血小板血浆对 BMSCs 在兔股骨头缺血性坏死中成骨作用的影响. 中国修复重建外科杂志, 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.	方建强, 朱文峰, 李维芝, 等. 声触诊组织成像量化技术在诊断急性跟腱炎中的应用价值. 临床超声医学杂志, 2017, 19(2): 99-101.
15.	李敏, 金力. 腱病动物模型研究进展. 现代预防医学, 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.	李辉, 唐康来, 邓银栓, 等. 外源性前列腺素 E2 对兔跟腱胶原含量的影响. 中国修复重建外科杂志, 2012, 26(3): 352-358.
18.	吴言, 杜国庆, 宗立秋, 等. 超声弹性弥散定量技术评价兔跟腱病的转归及生物力学特征. 中华超声影像学杂志, 2016, 25(5): 433-437.
19.	耿震, 王宸, 周海洋. 富血小板血浆对肌腱愈合影响的实验研究. 中国修复重建外科杂志, 2011, 25(3): 344-348.
20.	沈勇伟, 史晓伟. 低功率激光对末端病大鼠跟腱蛋白多糖和胶原的作用. 中国运动医学杂志, 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. 纪庆明, 杨育晖, 陈昊, 等. 富血小板血浆辅助前交叉韧带重建治疗的临床疗效分析. 中国修复重建外科杂志, 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. 王鹏飞, 时长江, 王玉亮, 等. 兔富血小板血浆制备及其活性分析. 中国组织工程研究, 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, 唐康来. 肌腱病模型研究进展. 中国运动医学杂志, 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. 方家刘, 尹宗生, 王伟, 等. 富白细胞和血小板血浆对 BMSCs 在兔股骨头缺血性坏死中成骨作用的影响. 中国修复重建外科杂志, 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. 方建强, 朱文峰, 李维芝, 等. 声触诊组织成像量化技术在诊断急性跟腱炎中的应用价值. 临床超声医学杂志, 2017, 19(2): 99-101.
15. 李敏, 金力. 腱病动物模型研究进展. 现代预防医学, 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. 李辉, 唐康来, 邓银栓, 等. 外源性前列腺素 E2 对兔跟腱胶原含量的影响. 中国修复重建外科杂志, 2012, 26(3): 352-358.
18. 吴言, 杜国庆, 宗立秋, 等. 超声弹性弥散定量技术评价兔跟腱病的转归及生物力学特征. 中华超声影像学杂志, 2016, 25(5): 433-437.
19. 耿震, 王宸, 周海洋. 富血小板血浆对肌腱愈合影响的实验研究. 中国修复重建外科杂志, 2011, 25(3): 344-348.
20. 沈勇伟, 史晓伟. 低功率激光对末端病大鼠跟腱蛋白多糖和胶原的作用. 中国运动医学杂志, 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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