目的 富血小板血漿（platelet-rich plasma，PRP）具有刺激椎間盤細胞增殖、促進細胞外基質(zhì)合成代謝及抑制纖維環(huán)細胞凋亡等作用。通過觀察自體PRP干預(yù)兔早期椎間盤退變，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；隨機分為實驗組、對照組、假手術(shù)組（n=15）。取實驗組兔耳中央動脈血，采用Landesberg等方法制備PRP，同時對全血及PRP行血小板計數(shù)。實驗組及對照組采用纖維環(huán)針刺法建立L4、5及L5、6椎間盤退變模型，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，不作處理。觀察實驗動物造模后一般情況；造模2周及干預(yù)1、2周時各組取5只實驗動物行腰椎MRI、HE染色及Ⅱ型膠原免疫組織化學染色觀察，腰椎MRI退變程度分級及Ⅱ型膠原陽性積分吸光度（IA）值檢測。 結(jié)果兔PRP中血小板計數(shù)約為外周血的4.92倍。實驗動物均存活至實驗完成。造模2周時，與假手術(shù)組相比，實驗組和對照組椎間盤信號降低，髓核細胞減少，基質(zhì)退變，Ⅱ型膠原表達降低。腰椎MRI退變程度分級及Ⅱ型膠原陽性IA值結(jié)果顯示，各時間點實驗組、對照組與假手術(shù)組相比差異均有統(tǒng)計學意義（P lt; 0.05），干預(yù)1、2周時，實驗組MRI退變程度分級顯著低于對照組 （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，實驗組髓核細胞及軟骨樣基質(zhì)較對照組增多，基質(zhì)纖維化程度輕，Ⅱ型膠原表達明顯強于對照組（P lt; 0.05）。 結(jié)論椎間盤內(nèi)注射自體PRP可終止甚至一定程度逆轉(zhuǎn)兔早期椎間盤退變，可能與PRP含有多種生長因子調(diào)控細胞功能、改善組織微環(huán)境、促進組織再生修復有關(guān)。
引用本文： 胡新鋒,王宸,芮云峰. 自體富血小板血漿干預(yù)兔早期椎間盤退變的初步研究. 中國修復重建外科雜志, 2012, 26(8): 977-983. doi: 復制
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| 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. |
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| 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退變模型的比較研究. 中國修復重建外科雜志, 2010, 24(1): 41-45. |
| 13. | 張聞力, 劉浩, 李壇珠. 椎間盤退變動物模型的研究進展. 中國修復重建外科雜志, 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退變模型. 中國矯形外科雜志, 2010, 18(21): 1814-1816. |
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| 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.
- 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退變模型的比較研究. 中國修復重建外科雜志, 2010, 24(1): 41-45.
- 13. 張聞力, 劉浩, 李壇珠. 椎間盤退變動物模型的研究進展. 中國修復重建外科雜志, 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退變模型. 中國矯形外科雜志, 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.