| Objective: In the orthopedic field, how to accelerate fracture healing, treatment of nonunion and delayed union fractures is a hot issue. This paper aims to investigate the effect of percutaneous electrical stimulation of skeletal muscle combine stem cell mobilization on bone remodeling and cell proliferation in rat fracture healing process and its possible mechanismMethod: we used 80 male SD rats, 3-month-age(provided by Xi'an JiaoTong University Medical School animal administrative center), and the weight was about 280～320g. then randomly divided them into 4 groups, each 20: Fracture model group (A group); model + mobilizing agent group (B group); model + electrical stimulation group (C group); model + mobilizing agent + electrical stimulation group (D group). All of them used the national standards murine dry feed, free drink and eat. Find support from the manufacturer with the establishment of rat model of closed fracture of tibia and stimulate lv xiao yu reference model and the Peng Liang stimulation mode and slightly changes the combination of a rat model of electrical stimulation. After fracture at 2,4,6,8 weeks to produce non-decalcified hard tissue and decalcified bone slices, the use of tetracycline and alizarin red double-fluorescent labeling, special staining, immunohistochemical SABC assay of bone tissue in the fracture healing process of structural change in calcium deposition and osteoblast cell proliferation and cytokine-related protein expression.Results:1 Rats closed fracture model caused by stent fracture of the middle tibia in rats by X-ray to determine: successful establish a rat closed fracture model of the middle tibia2 rhG-CSF and (or) electrical stimulation of skeletal muscle can promote functional recovery after fracture healing in rats. After fracture,the role of rhG-CSF alone and a simple electrical stimulation of skeletal muscle, as well as rhG-CSF combined of skeletal muscle stimulation all promote functional recovery after fracture healing, and the effect of rhG-CSF combined electrical stimulation of skeletal muscle is the most significant3 rhG-CSF and (or) electrical stimulation of skeletal muscle to cause the increase in the number of cartilage after fracture healing in rat. Bone tissue calcification occurs earlier, better fracture healing, especially to electrical stimulation and electrical stimulation + mobilizing agent group more obvious effects. Suggest that: after fracture electrical stimulation and bone marrow stem cell mobilization synergy, to increase the early fracture healing of the cartilage cells in the promotion of cartilage into bone, to accelerate fracture healing.4 pure rhG-CSF mobilization and simple electrical stimulation make the affected limb 2 labeling line spacing with the healing time and gradually increased, and compared to the model group significant increase,the unit area relative fluorescence intensity showed that: a simple intervention group can affect the fracture healing process of the deposition of calcium to promote bone matrix calcification. 5 rhG-CSF mobilization combined electrical stimulation of skeletal muscle can make 2 labeling line spacing is gradually increasing, and the peak to appeared in the six weeks after fracture. It suggests that calcium deposition in the latter half of the largest; compared with the pure intervention group 2 tag line spacing increased significantly, indicating that associate intervention is more than simply contribute to fracture healing.6 rhG-CSF mobilization and skeletal muscle electrical stimulation on fractured rats the expression of cell growth factor have an impact. Early fracture healing, in cartilage and bone cells, bFGF and c-kit significantly increased; simple electrical stimulation and electrical stimulation of joint mobilization intervention can promote cartilage and bone cells of c-kit expression in the fourth weeks to reach the peak, and the simple mobilization agent group reach peak in 6 weeks; each group TGF-β1 and bFGF to reach peak expression in the 6 weeks; fracture healing in the latter part of the expression of cytokines has declined, but still higher than control group. This showed that the simple way to stimulate and co-stimulation may be different levels to promote the expression of bone-inducing factor to promote fracture healing.7 rhG-CSF mobilization and skeletal muscle electrical stimulation have an impact on bone fracture in rats the expression of proliferation-related proteins.After electrical stimulation and combine with mobilization agents, PCNA, BrdU, and ki-67 in fracture healing process of a gradual increase in expression, indicating that cell proliferation-related proteins in fracture healing are the expression of the period, but at different times in different types of cells, the early period of the main division of mesenchymal stem cells and cartilage cells, the latter is the osteoblast. RhG-CSF in the role of early fracture healing in rats the expression of BrdU-positive is not significant, the latter has increased, PCNA and ki-67 expression continued high; simple electrical stimulation of group cell proliferation-associated protein expression with the progressive fracture healing increased, particularly, increased expression of BrdU with a very significant difference. Showed that electrical stimulation alone and compared to pure stem cell mobilization of bone marrow stem cells is more conducive to the proliferation and differentiation. After the joint role of both, PCNA, BrdU, and ki-67 expression peak appeared in the first six weeks, suggesting a dual role in promoting cell proliferation increase in the number of mitosis and in view of the cells are more conducive to the bone remodeling.Conclusions:Electrical stimulation of skeletal muscle to improve the fracture mechanics of the environment around the fracture site.The formation of micro-effect role for osteoblast-mechanical conditions of fracture healing in rats can callus after the increase in the number of cartilage, bone tissue calcification occurred in early, limb 2nd bone markers with the line spacing to extend the healing time and a gradual increase in the degree of fracture healing is better, the promotion of fracture healing after functional recovery. In this process, TGF-81, PCNA and BrdU expression increased significantly, and promote local cytokine secretion and release, to accelerate fracture healing. Therefore, the use of early forms of micro-fracture healing better than the traditional way of plaster splint is absolutely fixed.rhG-CSF mobilized autologous bone marrow stem cells to promote bone marrow-derived mesenchymal stem cells and the release of hematopoietic stem cells migrate to the fracture site, increasing stem cells into osteoblasts, the direction of differentiation into cartilage, promoting cartilage endochondral bone, bone matrix calcification and fracture of parts of new the number of blood vessels, accelerate fracture healing, fracture healing in rats after the promotion of functional recovery. During this process, the early fracture healing of cartilage and bone cells, TGF-β1, bFGF, PCNA, BrdU, and ki-67 in the first six weeks to reach the peak; fracture healing in the latter part of the expression of cytokines has declined, but still higher than the control group. Therefore, early use of autologous bone marrow stem cell mobilization to promote fracture healing is better than the traditional plaster splint means absolutely fixed.rhG-CSF mobilization and the joint role of skeletal muscle after electrical stimulation, can promote bone marrow-derived mesenchymal stem cells and the release of hematopoietic stem cells migrate to the fracture site, increasing stem cells into osteoblasts, the direction of differentiation into cartilage and bone growth by ensuring the needs for the mechanical environment, improve local blood circulation, speed up the process of bone remodeling; both of which will enhance bone-inducing factor and cell proliferation-related protein expression, thereby shortening the time to fracture healing. Therefore, the skeletal muscle caused by electrical stimulation of the micro-effects and the mobilization of stem cells to promote fracture healing better than the effect of electrical stimulation of skeletal muscle, rhG-CSF mobilization and traditional plaster splint, such as absolutely fixed. |
PDF Full Text Request