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Mechanical Vibration Synergy Bone Marrow Stem Cells Mobilization Effect On Angiogenesis Of Closed Fracture Healing In Rats And Its Possible Mechanism

Posted on:2013-12-13Degree:MasterType:Thesis
Country:ChinaCandidate:L LiFull Text:PDF
GTID:2247330377457357Subject:Human Movement Science
Abstract/Summary:PDF Full Text Request
Objective:This paper is in order to study the impacts of mechanical vibration collaborative bone marrow stem cells mobilization to closed fracture healing angiogenesis and its possible mechanism.Methods:Clean level,3months old, weight nearby200g Sprague Dawley female rats, which are randomly divided into four groups:fracture group (group C), fracture with mobilization group (group M), fracture with vibration group (group V), fracture with mobilization and vibration group (group M+V), each group40rats. Group V and group M+V contact intermittent vibration of30Hz,15min/d,5d/W after fracture72h. Group M and group M+V inject at back subcutaneous recombinant human granulocyte colony-stimulating factor(5μg/kg/d) after fracture3h. Each group is killed respectively on lw,2w,4w,5w after fracture. Immunohistochemical method detects CD31, CD34, VEGF, Ang-1and FⅧ-Rag expression. Western Blot analysis method detects the expression of VEGF and Ang-1protein in bony callus tissue. Results:In this study rats rhG-CSF mobilization after fracture methods to mobilize stem cells to promote migration of peripheral bone marrow MSCs to increase the peripheral blood HSCs,accelerated angiogenesis,improve local blood supply to promote healing. mechanical vibration accelerate local blood circulation, promote blood vessel regeneration and reconstruction of fractures by mechanical vibration improve the mechanical environment in which the proliferation of bone cells,bone fracture healing arranged in order to achieve the purpose.The results are as follows:1After internal fixation with percutaneous needle, then make use of fracture-model-maker to build rats femur closed fracture model. We successfully design and make a vibration machine using the principle of inertia, whose frequency is adjustable among the range of0-50Hz, can choose continuous or intermittent vibration modes, and can also meet10rats vibration at the same time. These designs conform to the international standards. Stent fracture was made with rat femoral closed fracture caused by X-ray film to judge,successfully established a rat fracture model.2Mechanical vibration and (or) rhG-CSF mobilization may accelerate fracture and functional recovery in rats.However, mechanical vibration and the dual role of rhG-CSF mobilization is more effective,than a simple fracture or the role of rhG-CSF mobilization of muscle electrical stimulation alone.3Mechanical vibration and (or) rhG-CSF mobilization on rat blood vessels during the recovery process fracture regeneration and bone formation which stimulated.4Mechanical vibration, or rhG-CSF mobilization can increase the number of capillaries and blood vessels reconstruction speed,early vascular network formation;especially mechanical vibration and mechanical vibration of rhG-CSF mobilization coordination more effective.Tip:After the fracture mechanical vibration coordination rhG-CSF mobilization can promote angiogenesis in early fracture healing, to improve the fracture site within the environment,mobilizing repair fibroblasts and hematopoietic systems derived mononuclear cells,osteoclasts to promote bone healing.5Mechanical vibration and (or) rhG-CSF mobilization can promote cartilage fracture callus in rats increased number of bone tissue calcification occurs earlier,fracture repair is fairly good.Particularly in the rhG-CSF mobilization+mechanical vibration group and the mechanical vibration group is more effective.Suggest that mechanical vibration after fracture and (or) rhG-CSF increased the mobilization of early fracture healing and cartilage cells in blood vessel density,and promote endochondral bone formation and accelerate fracture healing.6Mechanical vibration and (or) rhG-CSF mobilization on rat HSCs mobilized significant fracture site.CD34+rats in each group expressed fracture site first and then decreased, the first4weeks,peak, mechanical vibration collaborative rhG-CSF mobilization group> mechanical vibration group>rhG-CSF mobilization group>model group.Tip:mechanical vibration synergies dual role of rhG-CSF mobilized to strengthen the mobilization of MSCs may be chemotactic to raise more HSCs/ESCs,to accelerate fracture repair parts.7Mechanical vibration and (or) rhG-CSF mobilization can promote VEGF expression, mechanical vibration collaborative rhG-CSF mobilization was more remarkable dual role. VEGF rats in each group expressed fracture site first and then decreased, the first4weeks,peak, mechanical vibration collaborative rhG-CSF mobilization group> mechanical vibration group>rhG-CSF mobilization group>model group.8Mechanical vibration and (or) rhG-CSF mobilization can increase the expression of Ang-1stimulation, mechanical vibration, rhG-CSF mobilization group and model group the expression of fracture site first and then decreased,the first4weeks peak value, while the mechanical vibration, and the dual role of rhG-CSF mobilization group2reached peak values, followed by successive decline in the performance of mechanical vibration synergies for the rhG-CSF mobilization group> mechanical vibration group>rhG-CSF mobilization Groups>model group; mechanical vibration and dual role of rhG-CSF mobilization of Ang-1expression stimulated the most significant.9Mechanical vibration and (or) rhG-CSF mobilization can promote F Ⅷ-Rag、 CD31expression,reflecting the process of fracture healing in rats angiogenesis.Angiogenesis in fracture healing peaked at2-6weeks, and mechanical vibration synergies rhG-CSF mobilization group> mechanical vibration group>rhG-CSF mobilization group>model group.Suggest that bone marrow stem cell mobilization and mechanical vibration,and the dual role of both ischemic tissue may promote angiogenesis and collateral circulation to establish, improve the blood perfusion ischemia to accelerate fracture healing.10Vascular endothelia growth factor, according to the results of the WB expression increases gradually,5w reach peak in normal; Group M expressions have not obvious differences among periods but are more than group C; Group V expression increases significantly in early then take on the natural buildup rules, but expression is more; Group M+V gradually declines from1w to4w, increases on5w, but the expression is much higher than other groups in each period(P<0.05).All the results show, in early stage of fracture healing, stress produced from mechanical vibration enhances the activity of osteoblasts, increases the expression of vascular endothelia growth factor, mechanical vibration can improve the activity of osteogenic stem and progenitor cells, increases osteogenesis, promotes bone formation and bone matrix mineralization processes, and the effects of mechanical vibration collaborative stem cell mobilization are more apparent and better than single factor.11Ang-1, according to the results of the WB expression increases gradually,5w reach peak in normal; Group M expressions have not obvious differences among periods but are more than group C; Group V expression increases significantly in early then take on the natural buildup rules, but expression is more; Group M+V gradually declines from1w to4w, increases on5w, but the expression is much higher than other groups in each period(P<0.05).All the results show, in early stage of fracture healing, stress produced from mechanical vibration enhances the activity of osteoblasts, increases the expression of Ang-1, mechanical vibration can promote angiogenesis, and the effects of mechanical vibration collaborative stem cell mobilization are more apparent and better than single factor.Conclusion:Rat fracture model was successfully established; mechanical vibration and (or) rhG-CSF mobilization can significantly increase the fracture site in rats stem cell markers, angiogenesis factor, angiogenic factor expression tag, but mechanical vibration and rhG-CSF dual effect of mobilizing more significant. Suggest that mechanical vibration and(or)rhG-CSF mobilization can significantly promote bone marrow stem cell mobilization, promoting angiogenesis and bone cell proliferation, promote fracture healing. mechanical vibration coordination of bone marrow stem cell mobilization is a good method to promote bone healing.
Keywords/Search Tags:Closed fractures, Mechanical vibration, Stem cell mobilization, angiogenesis
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