AN EXPERIMENTAL STUDY ON EFFECT OF AUTOLOGOUS PLATELET-RICH PLASMA ON TREATMENT OF EARLY INTERVERTEBRAL DISC DEGENERATION_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

HU Xinfeng , WANG Chen , RUI Yunfeng.

Department of Orthopaedics, Affiliated Zhongda Hospital, Southeast University, Nanjing Jiangsu, 210009, P.R.China. Corresponding author: WANG Chen, E-mail: wangchen@medmail.com.cn;

Corresponding author：

Keywords：

Autologous platelet-rich plasma; Early intervertebral disc degeneration; Biological therapy; MRI; Collagen type II; Rabbit

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Objective Platelet-rich plasma (PRP) can stimulate intervertebral disc cell proliferation, promote extracellular matrix synthesis, and inhibit annulus fibrosus cell apoptosis. To investigate the effects of autologous PRP on the treatment of the early intervertebral disc degeneration (IDD) so as to provide the experimental basis for its clinical application. Methods Forty-five healthy New Zealand white rabbits (male or female, weighing 2.5-3.0 kg) were randomly divided into the experimental group (n=15), the control group (n=15), and the sham group (n=15). PRP was prepared from the arterial blood of rabbit’s ears of the experimental group with Landesberg’s method. The platelet concentrations in both whole blood and PRP were detected. The rabbit model of early IDD was established by annulus fibrosus puncture (L4, 5, L5, 6) in both the experimental group and the control group; 100 ?L autologous PRP and 100 ?L PBS were injected into the degenerative intervertebral discs respectively after 2 weeks of models creation. In sham group, intervertebral discs were separated and exposed without treatment. The general conditions of the rabbits were observed after building models; at 2 weeks after degeneration, 1 and 2 weeks after intervention, 5 rabbits were selected randomly from each group respectively for MRI observation, histological observation by using HE staining and collagen type II immunohistochemical staining. The signal of lumbar MRI was assessed and the contents of collagen type II were detected. Results The platelet concentration of PRP was about 4.92 times as much as that of the whole blood. All the animals survived to the end of the experiment. At 2 weeks after degeneration, a lower T2 signal was observed in both the experimental group and the control group; the nucleus pulposus cells decreased and extracellular matrix degenerated; and the expression of collagen type II decreased in both the experimental group and control group. The degenerative grade of lumbar MRI in the experimental group and control group were significantly higher than that in the sham group (P lt; 0.05), and the content of collagen type II were significantly lower than that in the sham group (P lt; 0.05). At 1, 2 weeks after intervention, disc degeneration in the experimental group was significantly lower than that in control group (P lt; 0.05), and significant difference was found between experimental group and sham group (P lt; 0.05). The nucleus pulposus cells and chondroid matrix in the experimental group were more than those in the control group, showing slight stromal fibrosis; but the expression of collage type II was significantly higher than that in the control group (P lt; 0.05). Conclusion The disc injection of autologous PRP may terminate or even reverse the progress of rabbit early IDD, which may be associated with the role of multiple growth factors of PRP in regulating cell function, improving the tissue microenvironment, and promoting tissue regeneration.

Citation： HU Xinfeng,WANG Chen,RUI Yunfeng.. AN EXPERIMENTAL STUDY ON EFFECT OF AUTOLOGOUS PLATELET-RICH PLASMA ON TREATMENT OF EARLY INTERVERTEBRAL DISC DEGENERATION. Chinese Journal of Reparative and Reconstructive Surgery, 2012, 26(8): 977-983. doi: Copy

1.	Adams MA, Roughley PJ. What is intervertebral disc degeneration, and what causes it. Spine (Phila Pa 1976), 2006, 31(18): 2151-2161.
2.	Karppinen J, Shen FH, Luk KD, et al. Management of degenerative disk disease and chronic low back pain. Orthop Clin North Am, 2011, 42(4): 513-528.
3.	Masuda K. Biological repair of the degenerated intervertebral disc by the injection of growth factors. Eur Spine, 2008, 17 Suppl 4: 441-451.
4.	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.
5.	Chen S, Emery SE, Pei M. Coculture of synovium-derived stem cells and nucleus pulposus cells in serum-free defined medium with supplementation of transforming growth factor-beta1: a potential application of tissue specific stem cells in disc regeneration. Spine (Phila Pa 1976), 2009, 34(12): 1272-1280.
6.	Gruber HE, Norton HJ, Hanley EN Jr. Anti-apoptotic effects of IGF-1 and PDGF on human interverterbral disc cells in vitro. Spine (Phila Pa 1976), 2000, 25(17): 2153-2157.
7.	Osada R, Ohshima H, Ishihara H, et al. Autocrine/paracrine mechanism of insulin-like growth factor-1 secretion, and the effect of insulin-like growth factor-1 on proteoglycan synthesis in bovine intervertebral discs. J Orthop Res, 1996, 14(5): 690-699.
8.	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): 297-300.
9.	Chen WH, Lo WC, Lee JJ, et al. Tissue-engineered intervertebral disc and chondrogenesis using human nucleus pulposus regulated through TGF-beta1 in platelet-rich plasma. J Cell Physiol, 2006, 209(3): 744-754.
10.	Masuda K, Aota Y, Muehleman C, et al. A novel rabbit model of mild, reproducible disc degeneration by an anulus needle puncture: correlation between the degree of disc injury and radiological and histological appearances of disc degeneration. Spine (Phila Pa 1976), 2005, 30(1): 5-41.
11.	Eppley BL, Woodell JE, Higgin J. Platelet quantification and growth factor analysis from platelet-rich plasma: implications for wound healing. Plast Reconstr Surg, 2004, 114(6): 1502-1508.
12.	张闻力, 李涛, 龚全, 等. 三种方法建立兔椎间盘退变模型的比较研究. 中国修复重建外科杂志, 2010, 24(1): 41-45.
13.	张闻力, 刘浩, 李坛珠. 椎间盘退变动物模型的研究进展. 中国修复重建外科杂志, 2007, 21(11): 1259-1262.
14.	Kim KS, Yoon ST, Li J, et al. Disc degeneration in the rabbit: a biochemical and radiological comparison between four disc injury models. Spine (Phila Pa 1976), 2005, 30(1): 33-37.
15.	桂柯科, 尹望平, 张飚, 等. 纤维环穿刺法建立兔椎间盘退变模型. 中国矫形外科杂志, 2010, 18(21): 1814-1816.
16.	Urban JP, Roberts S. Degeneration of the intervertebral disc. Arthritis Res Ther, 2003, 5(3): 120-130.
17.	Thompson JP, Pearce RH, Schechter MT, et al. Preliminary evaluation of a scheme for grading the gross morphology of the human intervertebral disc. Spine (Phila Pa 1976), 1990, 15(5): 411-415.
18.	Pfirrmann CW, Metzdorf A, Zanetti M, et al. Magnetic resonance classification of lumbar intervertebral disc degeneration. Spine (Phila Pa 1976), 2001, 26(17): 1873-1878.
19.	Zhang Y, Chee A, Thonar EJ, et al. Intervertebral disk repair by protein, gene, or cell injection: a framework for rehabilitation-focused biologics in the spine. PMR, 2011, 3(6 Suppl 1): S88-94.
20.	Nagae M, Ikeda T, Mikami Y, et al. Intervertebral disc regeneration using platelet-rich plasma and biodegradable gelatin hydrogel microspheres. Tissue Eng, 2007, 13(1): 147-158.
21.	Sawamura K, Ikeda T, Nagae M, et al. Characterization of in vivo effects of platelet-rich plasma and biodegradable gelatin hydrogel microspheres on degenerated intervertebral discs. Tissue Eng Part A, 2009, 15(12): 3719-3727.
22.	Chen WH, Liu HY, Lo WC, et al. Intervertebral disc regeneration in an ex vivo culture system using mesenchymal stem cells and platelet-rich plasma. Biomaterials, 2009, 30(29): 5523-5533.
23.	Akeda K, An HS, Pichika R, et al. Platelet-rich plasma (PRP) stimulates the extracellular matrix metabolism of porcine nucleus pulposus and anulus fibrosus cells cultured in alginate beads. Spine (Phila Pa 1976), 2006, 31(9): 959-966.
24.	Gruber HE, Hanley EN Jr. Analysis of aging and degeneration of the human intervertebral disc. Comparison of surgical specimens with normal controls. Spine (Phila Pa 1976), 1998, 23(7): 751-757.

1. Adams MA, Roughley PJ. What is intervertebral disc degeneration, and what causes it. Spine (Phila Pa 1976), 2006, 31(18): 2151-2161.
2. Karppinen J, Shen FH, Luk KD, et al. Management of degenerative disk disease and chronic low back pain. Orthop Clin North Am, 2011, 42(4): 513-528.
3. Masuda K. Biological repair of the degenerated intervertebral disc by the injection of growth factors. Eur Spine, 2008, 17 Suppl 4: 441-451.
4. 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.
5. Chen S, Emery SE, Pei M. Coculture of synovium-derived stem cells and nucleus pulposus cells in serum-free defined medium with supplementation of transforming growth factor-beta1: a potential application of tissue specific stem cells in disc regeneration. Spine (Phila Pa 1976), 2009, 34(12): 1272-1280.
6. Gruber HE, Norton HJ, Hanley EN Jr. Anti-apoptotic effects of IGF-1 and PDGF on human interverterbral disc cells in vitro. Spine (Phila Pa 1976), 2000, 25(17): 2153-2157.
7. Osada R, Ohshima H, Ishihara H, et al. Autocrine/paracrine mechanism of insulin-like growth factor-1 secretion, and the effect of insulin-like growth factor-1 on proteoglycan synthesis in bovine intervertebral discs. J Orthop Res, 1996, 14(5): 690-699.
8. 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): 297-300.
9. Chen WH, Lo WC, Lee JJ, et al. Tissue-engineered intervertebral disc and chondrogenesis using human nucleus pulposus regulated through TGF-beta1 in platelet-rich plasma. J Cell Physiol, 2006, 209(3): 744-754.
10. Masuda K, Aota Y, Muehleman C, et al. A novel rabbit model of mild, reproducible disc degeneration by an anulus needle puncture: correlation between the degree of disc injury and radiological and histological appearances of disc degeneration. Spine (Phila Pa 1976), 2005, 30(1): 5-41.
11. Eppley BL, Woodell JE, Higgin J. Platelet quantification and growth factor analysis from platelet-rich plasma: implications for wound healing. Plast Reconstr Surg, 2004, 114(6): 1502-1508.
12. 张闻力, 李涛, 龚全, 等. 三种方法建立兔椎间盘退变模型的比较研究. 中国修复重建外科杂志, 2010, 24(1): 41-45.
13. 张闻力, 刘浩, 李坛珠. 椎间盘退变动物模型的研究进展. 中国修复重建外科杂志, 2007, 21(11): 1259-1262.
14. Kim KS, Yoon ST, Li J, et al. Disc degeneration in the rabbit: a biochemical and radiological comparison between four disc injury models. Spine (Phila Pa 1976), 2005, 30(1): 33-37.
15. 桂柯科, 尹望平, 张飚, 等. 纤维环穿刺法建立兔椎间盘退变模型. 中国矫形外科杂志, 2010, 18(21): 1814-1816.
16. Urban JP, Roberts S. Degeneration of the intervertebral disc. Arthritis Res Ther, 2003, 5(3): 120-130.
17. Thompson JP, Pearce RH, Schechter MT, et al. Preliminary evaluation of a scheme for grading the gross morphology of the human intervertebral disc. Spine (Phila Pa 1976), 1990, 15(5): 411-415.
18. Pfirrmann CW, Metzdorf A, Zanetti M, et al. Magnetic resonance classification of lumbar intervertebral disc degeneration. Spine (Phila Pa 1976), 2001, 26(17): 1873-1878.
19. Zhang Y, Chee A, Thonar EJ, et al. Intervertebral disk repair by protein, gene, or cell injection: a framework for rehabilitation-focused biologics in the spine. PMR, 2011, 3(6 Suppl 1): S88-94.
20. Nagae M, Ikeda T, Mikami Y, et al. Intervertebral disc regeneration using platelet-rich plasma and biodegradable gelatin hydrogel microspheres. Tissue Eng, 2007, 13(1): 147-158.
21. Sawamura K, Ikeda T, Nagae M, et al. Characterization of in vivo effects of platelet-rich plasma and biodegradable gelatin hydrogel microspheres on degenerated intervertebral discs. Tissue Eng Part A, 2009, 15(12): 3719-3727.
22. Chen WH, Liu HY, Lo WC, et al. Intervertebral disc regeneration in an ex vivo culture system using mesenchymal stem cells and platelet-rich plasma. Biomaterials, 2009, 30(29): 5523-5533.
23. Akeda K, An HS, Pichika R, et al. Platelet-rich plasma (PRP) stimulates the extracellular matrix metabolism of porcine nucleus pulposus and anulus fibrosus cells cultured in alginate beads. Spine (Phila Pa 1976), 2006, 31(9): 959-966.
24. Gruber HE, Hanley EN Jr. Analysis of aging and degeneration of the human intervertebral disc. Comparison of surgical specimens with normal controls. Spine (Phila Pa 1976), 1998, 23(7): 751-757.

Chinese Journal of Reparative and Reconstructive Surgery

AN EXPERIMENTAL STUDY ON EFFECT OF AUTOLOGOUS PLATELET-RICH PLASMA ON TREATMENT OF EARLY INTERVERTEBRAL DISC DEGENERATION

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