本文選題：閉合性骨折　切入點：機械振動　出處：《陜西師范大學》2012年碩士論文

【摘要】：目的：探討機械振動協同骨髓干細胞(Bone marrow stem cells, BMSCs)動員對大鼠閉合性骨折愈合血管生成的影響及其可能機制。 方法：清潔級3月齡Sprague-Dawley雌性大鼠160只,體重200±20g。隨機分為4組,即骨折模型組(C組),骨折+動員劑組(M組),骨折+振動組(V組),骨折+動員劑+振動組(M+V組),每組40只。V組、M+V組骨折72h后接觸30Hz累計15min/d,5d/W的間歇振動。M組和M+V組骨折3h后背部皮下注射重組人粒細胞集落刺激因子(rhG-CSF,5μg/kg/d),共5d,各組分別在第1w/2w/4w/5w取材。免疫組化檢測組織中CD31、CD34、VEGF、ANG-1和FⅧ-Rag蛋白表達,免疫印跡法分析愈合期骨組織中ANG-1和VEGF蛋白表達。 實驗結果： 本實驗采用大鼠骨折后rhG-CSF動員的方法來動員干細胞,促進BMSCs向骨外周血遷移,增加外周血的BMSCs,加速血管再生,改善局部血液供應促進骨折愈合。采用機械振動,加速局部血液循環(huán),促進血管再生與重建,通過機械振動改善骨折周圍力學環(huán)境,為骨折愈合提供合適的微環(huán)境。其結果如下： 1用大鼠骨折造模支架造成大鼠股骨中段閉合性骨折,通過X光片判斷,成功建立大鼠骨折模型。 2機械振動和(或)rhG-CSF動員均可促進大鼠骨折愈合與功能恢復。但機械振動和rhG-CSF動員雙重作用效果更為顯著,優(yōu)于骨折后單純rhG-CSF動員作用或單純機械振動作用。 3機械振動和(或)rhG-CSF動員對大鼠骨折恢復過程中的血管再生及骨組織形成有一定促進作用。機械振動或rhG-CSF動員可使毛細血管數量增多,血管重建加速,較早地形成血管網；尤其以機械振動和機械振動協同rhG-CSF動員效果更明顯。提示：骨折后進行機械振動協同rhG-CSF動員可促進骨折愈合早期的血管形成,改善骨折部位的微環(huán)境,促進骨折愈合。 4機械振動和(或)rhG-CSF動員對大鼠骨折部位BMSCs動員顯著。CD34+在各組實驗大鼠骨折部位表達先升高后降低,第2周時是4次取材峰值,機械振動協同rhG-CSF動員組機械振動組rhG-CSF動員組模型組。提示：機械振動協同rhG-CSF動員雙重作用加強了BMSCs的動員,可趨化募集更多的BMSCs,加速骨折部位的損傷修復。 5機械振動和(或)rhG-CSF動員可促進VEGF的表達,機械振動協同rhG-CSF動員的雙重作用效果更加顯著。VEGF在各組實驗大鼠骨折部位表達先升高后降低,第2周時是4次取材峰值,機械振動協同rhG-CSF動員組機械振動組rhG-CSF動員組模型組。 6機械振動和(或)rhG-CSF動員均可促進Ang-1的表達,機械振動組、rhG-CSF動員組和模型組大鼠骨折部位表達規(guī)律為先升高后降低,第4周時是4次取材峰值,而機械振動和rhG-CSF動員雙重作用組第2周時是4次取材的峰值,之后逐次下降,表現為機械振動協同rhG-CSF動員組機械振動組rhG-CSF動員組模型組；機械振動和rhG-CSF動員雙重作用刺激Ang-1表達最顯著。 7機械振動和(或)rhG-CSF動員均可促進FⅧ-Rag、CD31的表達,可反映大鼠骨折愈合過程中的血管新生。骨折愈合的血管再生高峰期在第2-4周,且機械振動協同rhG-CSF動員組機械振動組rhG-CSF動員組模型組。提示,BMSCs動員和機械振動以及二者雙重作用均可促進缺血組織血管生成,改善缺血區(qū)的血液灌注,加速骨折愈合。 8血管生成素Ang-1的WB結果顯示,正常情況下Ang-1表達量較少,骨折1周達高峰；動員劑組表達量呈逐漸升高,表達量多于安靜組；振動組早期明顯升高后呈自然下降規(guī)律,但表達量較多；動員振動組則從第2周到4周明顯升高,5周時表達再次顯著下降,但各期Ang-1表達量明顯高于其他組。表明在骨折愈合期,機械振動和動員劑均可促進Ang-1表達,機械振動作用優(yōu)于動員劑,早期二者合用促進血管生成效果顯著。機械振動與動員劑合用效果更加顯著,作用優(yōu)于單一因素。 9血管內皮生長因子VEGF的WB結果顯示,正常情況下VEGF表達較少；動員劑組表達量逐漸升高,但表達量多于安靜組；振動組早期明顯升高后呈自然降低,但表達量較多；動員振動組表達量先降低后升高,但各期VEGF表達量明顯高于其他組。表明在骨折愈合期,機械振動和動員劑均可促進VEGF表達,機械振動作用優(yōu)于動員劑,在早期兩者合用促進血管生成效果顯著。機械振動與動員劑合用效果更加顯著,作用優(yōu)于單一因素。 結論： 機械振動和(或)rhG-CSF動員可顯著增加大鼠骨折部位BMSCs標志物、血管再生因子、血管新生因子的表達,但機械振動和rhG-CSF動員雙重作用效果更為顯著。提示,機械振動和(或)rhG-CSF動員可顯著促進BMSCs動員,促進血管再生,促進骨折修復。機械振動協同BMSCs動員是促進骨折愈合的較好方法。
[Abstract]:Objective : To investigate the effects of mechanical vibration and bone marrow stem cells ( BMSC ) mobilization on the angiogenesis of closed fracture healing in rats and its possible mechanism .

Methods : 160 male Sprague - Dawley female rats weighing 200 鹵 20g were randomly divided into four groups : fracture model group ( group C ) , fracture + mobilization agent group ( M group ) , fracture + vibration group ( group V ) , fracture + mobilization agent + vibration group ( M + V group ) .

Experimental results :

We use the method of rhG - CSF mobilization after fracture of rats to mobilize the stem cells , promote the migration of bone marrow into the peripheral blood of the bone , increase the peripheral blood cells , accelerate the blood vessel regeneration , improve the local blood supply and promote the fracture healing . The mechanical vibration is used to accelerate the local blood circulation , promote the vascular regeneration and reconstruction , improve the surrounding mechanical environment of the fracture by mechanical vibration , and provide a suitable microenvironment for the fracture healing .

1 Rat fracture model was successfully established by X - ray film .

2 Mechanical vibration and ( or ) rhG - CSF mobilization can promote fracture healing and functional recovery in rats , but the effects of mechanical vibration and rhG - CSF mobilization are more remarkable than that of simple rhG - CSF mobilization or simple mechanical vibration after fracture .

3 . Mechanical vibration and / or rhG - CSF mobilization can promote the regeneration of blood vessels and the formation of bone tissue during the recovery of fracture of rats . Mechanical vibration or rhG - CSF mobilization can increase the number of capillaries and accelerate the reconstruction of blood vessels , thus forming the vascular network early .
It is suggested that mechanical vibration and rhG - CSF mobilization can accelerate the formation of blood vessels in the early stage of fracture healing , improve the microenvironment of fracture site and promote fracture healing .

4 mechanical vibrations and ( or ) rhG - CSF mobilization significantly increased the mobilization of bone marrow cells in the fracture site of rats . The expression of CD34 + in the fracture sites of the rats was increased first and then decreased at the second week . The mechanical vibration was combined with rhG - CSF to mobilize the rhG - CSF mobilization group model group .

5 Mechanical vibration and ( or ) rhG - CSF mobilization could promote the expression of VEGF . The effects of mechanical vibration and rhG - CSF mobilization were more significant .

Both mechanical and rhG - CSF mobilization could promote the expression of Ang - 1 , the mechanical vibration group , the rhG - CSF mobilization group and the model group rats .
Mechanical vibration and rhG - CSF mobilized double action to stimulate the expression of Ang - 1 .

7 mechanical vibration and ( or ) rhG - CSF mobilization can promote the expression of F鈪，

本文編號：1729247