脊髓完全性损伤大鼠胫骨骨折愈合的实验研究及神经生长因子对失神经骨折愈合的影响
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摘要
背景：
在临床实践中观察到一个现象，截瘫和颅脑损伤患者伴随的四肢骨折往往可以见到大量骨痂过度生长，甚至在肌肉中可以出现异位骨化，但是这种骨痂强度差，编织骨向板层骨转化慢，因此认为神经系统参与了骨折愈合过程，推测神经系统最终与骨生成和骨转化有关。采用脱钙的免疫组织化学方法对骨组织进行研究中发现，骨不连的断端骨痂、异位成骨及失神经状态下的骨痂在组织结构上有一个共同点，就是少有或者没有神经支配，那么能否通过研究神经对骨再生支配与调控机理，最终找到解决骨不连及异位骨化的治疗方法。
神经生长因子(NGF)是于1952年发现的第一个神经营养因子，在神经系统中调节中枢和外周神经元生长、发育、分化以及维持正常存活的作用已得到公认。NGF广泛分布于全身器官，除营养神经外，还参与一些其他的生理活动，如刺激单核细胞、肥大细胞的增殖和分化，促进皮肤溃疡、角膜溃疡和皮肤切口的愈合，刺激毛细血管生长以增加局部血供等。近来，研究发现在正常骨组织和骨折骨痂中有NGF及其受体的表达，同时局部给予外源性NGF对骨折愈合具有促进作用。这说明NGF可能也是骨折愈合复杂因子网络中的一员。在失神经状态下能否应用神经生长因子改善骨折过程中骨痂钙化骨力差从而为神经生长因子促骨折愈合找到理论依据还是一个新的课题。
目的：
1、对T_(10)脊髓完全性横断损伤状态下大鼠胫骨骨折模型的后肢感觉运动情况，动物的存活率，骨痂生成情况进行探讨研究，从而确立较为理想的失神经状态下的骨折愈合实验动物模型。
2、比较失神经状态下与正常骨折愈合过程中骨折断端刚度与强度，骨形态计量学中动力性参数与静力性参数，及骨密度的变化规律验证失神经状态对骨折愈合有重要影响作用。
3、探讨失神经状态下骨痂组织形态结构、骨钙素的表达、成骨细胞超微结
Background:
There were mass calluses appeared at bone fractured ends of the patient who were suffered with injury of brain or SCI,even meanwhile ectopic ossification often appeared in muscle.The significant decease were showed in calluses strength ,speed of transformation from waven bone to lamellar bone.Therefore ,it is presumed that the nerve system not noly take part in fracture healing ,but also play important role in bone generation and bone transformation.With decalcifying immunohistochemistry method , it is the interesting note that the nonunited diaphyseal fracture、ectopic ossification bone have been observed to have an almost total lack of innervation , as denervated calluses .Then ,perhaps the above-mentioned three diseases would be treated eventually finally when the mechanism of nerve system what may dominate and regulate osteanagenesis is confirmed .
Nerve growth factor was found in 1952 ,NGF as the first neurotrophic factor is a neutotrophic substance produced by most tissues of the body ,.Now it is known that NGF can regulate nerve center and promote the growth development differentiation of peripheral neurone and maintain normal survive of neurone. NGF may be found in all tissues of the body ,it not only nourish nerve ,but also take part in some physiological functions such as, the production and differentiation of monocyte and mastocyte ;healing of dermal ulcer、corneal ulcer、skin incision; growth of blood capillary ,at last, increase of topical blood supply.
At present ,the research have testified the expression of NGF and NGF acceptor in normal bone tissue and calluses .the effect of NGF on fracture healing is obvious after topical and extrinsic source application of NGF.the result demonstrate that NGF is also a member of complicated factor net of fractured bone healing .it is new research
在临床实践中观察到一个现象，截瘫和颅脑损伤患者伴随的四肢骨折往往可以见到大量骨痂过度生长，甚至在肌肉中可以出现异位骨化，但是这种骨痂强度差，编织骨向板层骨转化慢，因此认为神经系统参与了骨折愈合过程，推测神经系统最终与骨生成和骨转化有关。采用脱钙的免疫组织化学方法对骨组织进行研究中发现，骨不连的断端骨痂、异位成骨及失神经状态下的骨痂在组织结构上有一个共同点，就是少有或者没有神经支配，那么能否通过研究神经对骨再生支配与调控机理，最终找到解决骨不连及异位骨化的治疗方法。
神经生长因子(NGF)是于1952年发现的第一个神经营养因子，在神经系统中调节中枢和外周神经元生长、发育、分化以及维持正常存活的作用已得到公认。NGF广泛分布于全身器官，除营养神经外，还参与一些其他的生理活动，如刺激单核细胞、肥大细胞的增殖和分化，促进皮肤溃疡、角膜溃疡和皮肤切口的愈合，刺激毛细血管生长以增加局部血供等。近来，研究发现在正常骨组织和骨折骨痂中有NGF及其受体的表达，同时局部给予外源性NGF对骨折愈合具有促进作用。这说明NGF可能也是骨折愈合复杂因子网络中的一员。在失神经状态下能否应用神经生长因子改善骨折过程中骨痂钙化骨力差从而为神经生长因子促骨折愈合找到理论依据还是一个新的课题。
目的：
1、对T_(10)脊髓完全性横断损伤状态下大鼠胫骨骨折模型的后肢感觉运动情况，动物的存活率，骨痂生成情况进行探讨研究，从而确立较为理想的失神经状态下的骨折愈合实验动物模型。
2、比较失神经状态下与正常骨折愈合过程中骨折断端刚度与强度，骨形态计量学中动力性参数与静力性参数，及骨密度的变化规律验证失神经状态对骨折愈合有重要影响作用。
3、探讨失神经状态下骨痂组织形态结构、骨钙素的表达、成骨细胞超微结
Background:
There were mass calluses appeared at bone fractured ends of the patient who were suffered with injury of brain or SCI,even meanwhile ectopic ossification often appeared in muscle.The significant decease were showed in calluses strength ,speed of transformation from waven bone to lamellar bone.Therefore ,it is presumed that the nerve system not noly take part in fracture healing ,but also play important role in bone generation and bone transformation.With decalcifying immunohistochemistry method , it is the interesting note that the nonunited diaphyseal fracture、ectopic ossification bone have been observed to have an almost total lack of innervation , as denervated calluses .Then ,perhaps the above-mentioned three diseases would be treated eventually finally when the mechanism of nerve system what may dominate and regulate osteanagenesis is confirmed .
Nerve growth factor was found in 1952 ,NGF as the first neurotrophic factor is a neutotrophic substance produced by most tissues of the body ,.Now it is known that NGF can regulate nerve center and promote the growth development differentiation of peripheral neurone and maintain normal survive of neurone. NGF may be found in all tissues of the body ,it not only nourish nerve ,but also take part in some physiological functions such as, the production and differentiation of monocyte and mastocyte ;healing of dermal ulcer、corneal ulcer、skin incision; growth of blood capillary ,at last, increase of topical blood supply.
At present ,the research have testified the expression of NGF and NGF acceptor in normal bone tissue and calluses .the effect of NGF on fracture healing is obvious after topical and extrinsic source application of NGF.the result demonstrate that NGF is also a member of complicated factor net of fractured bone healing .it is new research
引文
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16. Grills BL, Schuijer S JA, Ward J Orthop Res, 1997; 15(2): 235-242.
17. Lwata E, Nakanishi T, Ogawa N, et al Biochem Biophys Acta, 1996; 1 311(2): 85-92.
18. Yada M. YamaguchiK. Tsuji T.Biochem Biophys Res Commun, 1994; 2 05(2): 1187—1193.
19. Frenkel SR. Guerra LA, Mitchell OG, et al Cell Tissue Re-s, 1990; 2 60(3): 507-511.
20. Yao M, Dooley PC, Schuijers JA, et aL J Musculoskelet euronal Interact, 2002; 2(4): 327-334
21. Grills, Bl Schuijers JA Acta Orthop Scand, 1998; 69(4): 415-419.
22. Asaumi K, Nakanishi T, Asahara H, et al. Bone, 2000; 26(6): 625— 633.
23. Nakanishi T, Takahashi K, Aoki C, et al. Biochem Biophys Res Commun, 1994; 198(3): 891—897.
24. Iannone F, de Bari C, Dell'Accio F et al Rheumatology ,2002;41(12): 14 13-1418.
25. Eppley BL, Snyders RV, Winkkelmann T, et al J Oral Maxi-llofac Surg, 1991; 49(1)61-68.
26. Grills BL, Schuijers JA, Ward AR J Orthop Res, 1997; 15(2): 235— 242.
27. Abe T,Morgan DA, Gutterman DD.Protective role of nerve growth fractor against postischemic dysfunction of sympathetic coronary in nervation circulation 1997,95:213-220.
28. Lee TH,Kato H,Kogure k,etal.Temporal profile of nerve growth factor-like immunoreactivity after transient focal cerebral ischemia in rats, Brain Res , 1996,713:199-210.
29. Woolf CJ,Alchorne A,Safieh-Garabedian B ,et al ,Cytokines ,nerve growth factor and inflammatory hyperalgesia: the contribution of tumour necrosis factor alpha.Br J Pharmacol,1997,121:417-424
30. Imai S, Rauvala H, Konttinen YT, et al. J Bone Miner Res, 1997; 12(7): 1018-1027.
31. Hukkanen M ,Kontinen YT,Santavirta S, et al. Neuroscience, 1993; 54(4): 969—979.
31. Mukohyama H, Ransjo M , Taniquchj H, et al. Biochem Biophys Res Commun, 2000;271(1);158-163.
32. Togari A , Arai M , M izutani S, et al. Neurosi Lett, 1997; (2-3) ; 125—128.
33. Ransjo M,Lie A,Mukohyama H, et al. Biochem Biophys Re-s Commun,2000;274(2): 400-404.
34. Vignery A, McCarthy T L. Bone, 1996; 18(4):331-335
35. Eppley BL, Snyders RV,W,inkelmann TM, et al. J Oral Maxill-ofac Surg, 1991;49(1):61~68.
36. Ransjo M ,Lie A, Mukohyama H, et al. Biochem Biophys Res Commun, 2000;274(2): 400—404.
37. Madsen JE, Hukkanen M, Aune AK, et al. Fracture healing and callus innervation after peripheral nerve resection in rats [J]. Cl in Orthop, 1998, 351: 230-240.