机械振动协同骨髓干细胞动员对大鼠闭合性骨折愈合血管生成的影响及其可能机制

发布时间：2018-04-10 02:23

  本文选题：闭合性骨折　切入点：机械振动　出处：《陕西师范大学》2012年硕士论文

【摘要】：目的：探讨机械振动协同骨髓干细胞(Bone marrow stem cells, BMSCs)动员对大鼠闭合性骨折愈合血管生成的影响及其可能机制。 方法：清洁级3月龄Sprague-Dawley雌性大鼠160只,体重200±20g。随机分为4组,即骨折模型组(C组),骨折+动员剂组(M组),骨折+振动组(V组),骨折+动员剂+振动组(M+V组),每组40只。V组、M+V组骨折72h后接触30Hz累计15min/d,5d/W的间歇振动。M组和M+V组骨折3h后背部皮下注射重组人粒细胞集落刺激因子(rhG-CSF,5μg/kg/d),共5d,各组分别在第1w/2w/4w/5w取材。免疫组化检测组织中CD31、CD34、VEGF、ANG-1和FⅧ-Rag蛋白表达,免疫印迹法分析愈合期骨组织中ANG-1和VEGF蛋白表达。 实验结果： 本实验采用大鼠骨折后rhG-CSF动员的方法来动员干细胞,促进BMSCs向骨外周血迁移,增加外周血的BMSCs,加速血管再生,改善局部血液供应促进骨折愈合。采用机械振动,加速局部血液循环,促进血管再生与重建,通过机械振动改善骨折周围力学环境,为骨折愈合提供合适的微环境。其结果如下： 1用大鼠骨折造模支架造成大鼠股骨中段闭合性骨折,通过X光片判断,成功建立大鼠骨折模型。 2机械振动和(或)rhG-CSF动员均可促进大鼠骨折愈合与功能恢复。但机械振动和rhG-CSF动员双重作用效果更为显著,优于骨折后单纯rhG-CSF动员作用或单纯机械振动作用。 3机械振动和(或)rhG-CSF动员对大鼠骨折恢复过程中的血管再生及骨组织形成有一定促进作用。机械振动或rhG-CSF动员可使毛细血管数量增多,血管重建加速,较早地形成血管网；尤其以机械振动和机械振动协同rhG-CSF动员效果更明显。提示：骨折后进行机械振动协同rhG-CSF动员可促进骨折愈合早期的血管形成,改善骨折部位的微环境,促进骨折愈合。 4机械振动和(或)rhG-CSF动员对大鼠骨折部位BMSCs动员显著。CD34+在各组实验大鼠骨折部位表达先升高后降低,第2周时是4次取材峰值,机械振动协同rhG-CSF动员组机械振动组rhG-CSF动员组模型组。提示：机械振动协同rhG-CSF动员双重作用加强了BMSCs的动员,可趋化募集更多的BMSCs,加速骨折部位的损伤修复。 5机械振动和(或)rhG-CSF动员可促进VEGF的表达,机械振动协同rhG-CSF动员的双重作用效果更加显著。VEGF在各组实验大鼠骨折部位表达先升高后降低,第2周时是4次取材峰值,机械振动协同rhG-CSF动员组机械振动组rhG-CSF动员组模型组。 6机械振动和(或)rhG-CSF动员均可促进Ang-1的表达,机械振动组、rhG-CSF动员组和模型组大鼠骨折部位表达规律为先升高后降低,第4周时是4次取材峰值,而机械振动和rhG-CSF动员双重作用组第2周时是4次取材的峰值,之后逐次下降,表现为机械振动协同rhG-CSF动员组机械振动组rhG-CSF动员组模型组；机械振动和rhG-CSF动员双重作用刺激Ang-1表达最显著。 7机械振动和(或)rhG-CSF动员均可促进FⅧ-Rag、CD31的表达,可反映大鼠骨折愈合过程中的血管新生。骨折愈合的血管再生高峰期在第2-4周,且机械振动协同rhG-CSF动员组机械振动组rhG-CSF动员组模型组。提示,BMSCs动员和机械振动以及二者双重作用均可促进缺血组织血管生成,改善缺血区的血液灌注,加速骨折愈合。 8血管生成素Ang-1的WB结果显示,正常情况下Ang-1表达量较少,骨折1周达高峰；动员剂组表达量呈逐渐升高,表达量多于安静组；振动组早期明显升高后呈自然下降规律,但表达量较多；动员振动组则从第2周到4周明显升高,5周时表达再次显著下降,但各期Ang-1表达量明显高于其他组。表明在骨折愈合期,机械振动和动员剂均可促进Ang-1表达,机械振动作用优于动员剂,早期二者合用促进血管生成效果显著。机械振动与动员剂合用效果更加显著,作用优于单一因素。 9血管内皮生长因子VEGF的WB结果显示,正常情况下VEGF表达较少；动员剂组表达量逐渐升高,但表达量多于安静组；振动组早期明显升高后呈自然降低,但表达量较多；动员振动组表达量先降低后升高,但各期VEGF表达量明显高于其他组。表明在骨折愈合期,机械振动和动员剂均可促进VEGF表达,机械振动作用优于动员剂,在早期两者合用促进血管生成效果显著。机械振动与动员剂合用效果更加显著,作用优于单一因素。 结论： 机械振动和(或)rhG-CSF动员可显著增加大鼠骨折部位BMSCs标志物、血管再生因子、血管新生因子的表达,但机械振动和rhG-CSF动员双重作用效果更为显著。提示,机械振动和(或)rhG-CSF动员可显著促进BMSCs动员,促进血管再生,促进骨折修复。机械振动协同BMSCs动员是促进骨折愈合的较好方法。
[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 卤 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，

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