• 東南大學(xué)附屬中大醫(yī)院骨科（南京，210009）;

目的 富血小板血漿（platelet-rich plasma，PRP）具有刺激椎間盤(pán)細(xì)胞增殖、促進(jìn)細(xì)胞外基質(zhì)合成代謝及抑制纖維環(huán)細(xì)胞凋亡等作用。通過(guò)觀察自體PRP干預(yù)兔早期椎間盤(pán)退變，明確其治療效果，為臨床應(yīng)用提供理論依據(jù)。 方法取健康成年新西蘭大白兔45只，體重2.5～3.0 kg，雌雄不限；隨機(jī)分為實(shí)驗(yàn)組、對(duì)照組、假手術(shù)組（n=15）。取實(shí)驗(yàn)組兔耳中央動(dòng)脈血，采用Landesberg等方法制備PRP，同時(shí)對(duì)全血及PRP行血小板計(jì)數(shù)。實(shí)驗(yàn)組及對(duì)照組采用纖維環(huán)針刺法建立L4、5及L5、6椎間盤(pán)退變模型，造模2周后于L4、5及L5、6椎間隙分別注入100 ?L自體PRP及100 ?L PBS液；假手術(shù)組僅分離暴露椎間盤(pán)，不作處理。觀察實(shí)驗(yàn)動(dòng)物造模后一般情況；造模2周及干預(yù)1、2周時(shí)各組取5只實(shí)驗(yàn)動(dòng)物行腰椎MRI、HE染色及Ⅱ型膠原免疫組織化學(xué)染色觀察，腰椎MRI退變程度分級(jí)及Ⅱ型膠原陽(yáng)性積分吸光度（IA）值檢測(cè)。 結(jié)果兔PRP中血小板計(jì)數(shù)約為外周血的4.92倍。實(shí)驗(yàn)動(dòng)物均存活至實(shí)驗(yàn)完成。造模2周時(shí)，與假手術(shù)組相比，實(shí)驗(yàn)組和對(duì)照組椎間盤(pán)信號(hào)降低，髓核細(xì)胞減少，基質(zhì)退變，Ⅱ型膠原表達(dá)降低。腰椎MRI退變程度分級(jí)及Ⅱ型膠原陽(yáng)性IA值結(jié)果顯示，各時(shí)間點(diǎn)實(shí)驗(yàn)組、對(duì)照組與假手術(shù)組相比差異均有統(tǒng)計(jì)學(xué)意義（P  lt; 0.05），干預(yù)1、2周時(shí)，實(shí)驗(yàn)組MRI退變程度分級(jí)顯著低于對(duì)照組 （P  lt; 0.05），但仍與假手術(shù)組有差異（P  lt; 0.05）；干預(yù)1、2周時(shí)，實(shí)驗(yàn)組髓核細(xì)胞及軟骨樣基質(zhì)較對(duì)照組增多，基質(zhì)纖維化程度輕，Ⅱ型膠原表達(dá)明顯強(qiáng)于對(duì)照組（P  lt; 0.05）。 結(jié)論椎間盤(pán)內(nèi)注射自體PRP可終止甚至一定程度逆轉(zhuǎn)兔早期椎間盤(pán)退變，可能與PRP含有多種生長(zhǎng)因子調(diào)控細(xì)胞功能、改善組織微環(huán)境、促進(jìn)組織再生修復(fù)有關(guān)。

引用本文： 胡新鋒,王宸,芮云峰. 自體富血小板血漿干預(yù)兔早期椎間盤(pán)退變的初步研究. 中國(guó)修復(fù)重建外科雜志, 2012, 26(8): 977-983. doi: 復(fù)制

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. 富血小板血漿對(duì)肌腱愈合影響的實(shí)驗(yàn)研究. 中國(guó)修復(fù)重建外科雜志, 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. 張聞力, 李濤, 龔全, 等. 三種方法建立兔椎間盤(pán)退變模型的比較研究. 中國(guó)修復(fù)重建外科雜志, 2010, 24(1): 41-45.
13. 張聞力, 劉浩, 李壇珠. 椎間盤(pán)退變動(dòng)物模型的研究進(jìn)展. 中國(guó)修復(fù)重建外科雜志, 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, 張飚, 等. 纖維環(huán)穿刺法建立兔椎間盤(pán)退變模型. 中國(guó)矯形外科雜志, 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. 1. Adams MA, Roughley PJ. What is intervertebral disc degeneration, and what causes it. Spine (Phila Pa 1976), 2006, 31(18): 2151-2161.
  2. 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. 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. 4. 耿震, 王宸, 周海洋. 富血小板血漿對(duì)肌腱愈合影響的實(shí)驗(yàn)研究. 中國(guó)修復(fù)重建外科雜志, 2011, 25(3): 344-348.
  5. 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. 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. 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. 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. 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. 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. 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. 12. 張聞力, 李濤, 龔全, 等. 三種方法建立兔椎間盤(pán)退變模型的比較研究. 中國(guó)修復(fù)重建外科雜志, 2010, 24(1): 41-45.
  13. 13. 張聞力, 劉浩, 李壇珠. 椎間盤(pán)退變動(dòng)物模型的研究進(jìn)展. 中國(guó)修復(fù)重建外科雜志, 2007, 21(11): 1259-1262.
  14. 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. 15. 桂柯科, 尹望平, 張飚, 等. 纖維環(huán)穿刺法建立兔椎間盤(pán)退變模型. 中國(guó)矯形外科雜志, 2010, 18(21): 1814-1816.
  16. 16. Urban JP, Roberts S. Degeneration of the intervertebral disc. Arthritis Res Ther, 2003, 5(3): 120-130.
  17. 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. 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. 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. 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. 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. 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. 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. 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.