Observe thebone morphogenetic protein 4 (BMP4) isolated primary cultured rat brain stem cells (NSC) differentiation induced by cholinergic effect. From 2-month-old rat hippocampus, striatum region isolated cells were cultured in DMEM/F12 medium containing EGF and bFGF, to Nestin (of nestin in parallel morphological characteristics of the cells in the light microscope,) cytochemical staining. After 24h, a switch of BMP4-containing culture medium, the culture was continued for 7 or 8 days. Morphological changes of the cells in the light microscope, parallel choline acetyltransferase (choline acetyltransferase staining of ChAT) and nestin double-labeled immune cells. The results showed that, plus BMP4 cultured for 7 or 8 days, the light microscope to see 34% of the cultured cells with the morphological characteristics of neurons. The immunocytochemical staining visible ChAT-positive cells and nestin-positive cells coexist ChAT-positive cells accounted for 28% of nestin-positive cells accounted for 38%. In short, in the culture broth was added the BMP4 can induce NSC differentiation into cells having a cholinergic properties

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In this experiment, we fracture repair process a BMP4mRNA Expression and localization of detected using BMP4cDNA probe, in order to investigate their role in fracture healing. It was found that: (1) not detected in normal bone tissue and soft tissue and BMP4mRNA expression; (2) early fracture (fracture after 12 to 72 hours) that a BMP4mRNA expression; (3) BMP4 gene mainly in early fracture hematoma emerging within the cell and fracture the surrounding soft tissue, expressed in mesenchymal cells. Consistent with the the Nakase other reported results. Bone marrow stromal cells may be derived from these hematoma, osteoblast potential of stem cells, i.e. directed into the bone precursor cells, the emerging positive cells in muscle tissue compared can induce osteogenic precursor cells, i.e. mesenchymal cells.

Trauma to activate the BMP4 gene's expression and showed some regional, that is limited to the fracture callus formation around the area of soft tissue. Topical application of exogenous recombinant expression product BMP4 able to induce new bone formation in normal soft tissue ectopic, so they think, local BMP4 gene expression of the fracture hematoma the osteogenic ability is the very important factor in the process of fracture healing callus formation also shows that the fracture hematoma and soft tissue has a very important position in the process of fracture healing. BMP4 gene expression is also reported in the literature in embryonic tissue, organs and embryonic bone, endochondral bone repair fractures embryonic osteogenesis reproduction, BMP4 only bone injury repair related, are more likely to tissues and organs and embryos related.

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Histological changes in the process of fracture healing is a complex and involve a number of different cell activities. Fracture occurs, the presence of the different stages of osteoblast and osteoclast number and their synthetic cell interstitial far can not meet the demand of the fracture healing. Local cell regulation mechanisms (including precursor cells, mesenchymal cells, other synergy cells, capillaries, lymphatic, nerve growth factor autocrine and paracrine) to produce a sufficient number of osteoblasts and osteoclasts.

With the development of recombinant DNA technology, making it possible therapeutic areas. Such as growth factors and some other regulatory peptides for the regulation of cell metabolism, division and differentiation in the bone healing process. Part in the process of fracture healing growth factors and cytokines involved in the formation of bone cartilage has been reported in the literature.

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Post-traumatic bone tissue regeneration and repair is very complete, and its repair is often no scar residue. The reason why there is such a perfect bone tissue healing ability, because the memory of bone tissue factor in successful bone, bone growth factors. Bone morphogenetic proteins (bone morphogenetic pro-tein, BMP) because of its emphasis on direct induction of soft tissue into bone. Application BMP4cDNA probe to detect the fracture healing process foreign callus localization and distribution of BMP4 gene expression to explore the BMP4 gene expression outside in closed fracture healing callus formation. 64 healthy SD rats with closed tibial fractures animal models. 12 hours after fracture, 1,3,5,7,9,14 and 28 days after operation. Drawn underwent cryostat, the digoxigenin labeled BMP4cDNA probe situ hybridization. Rat fracture after 12 hours to three days, the detection of fractures around hematoma within cells and muscle emerging mesenchymal cells BMP4mRNA expression as a positive signal. Show that The trauma activates expression of BMP4mRNA, and was regional involved in the repair of fractures, the fracture hematoma and soft tissue has a very important role in the process of fracture healing.

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Follow-up period of 3 months to 6 years, with an average of 44 months follow-up of more than 3 years 28 patients (36 hips), 18 patients (22 hips) F ic cases at stage I, II, hip no significant pain and dysfunction, more than 3 years follow-up of 28 patients (36 hips) the hip Ha rris score, the mean preoperative (4 ± 9) minutes, mean postoperative (7 ± 7. 2) points (P <0. 0 1). The authors believe that the method of using bone morphogenetic protein can induce to enhance the osteogenic potential of bone marrow and allogeneic bone allograft fibula to provide direct mechanical support for block development and collapse of the femoral head lesions, bone allograft bone morphogenetic protein and autologous bone marrow can be used as the treatment of femoral F ic at Ⅰ, Ⅱ of avascular necrosis of a choice.

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Angiogenesis is an important part of the fracture healing process, a good blood supply to the fracture site of great significance in fracture repair. Vascular endothelial growth factor (vascular endothelial growth factor, VEGF) is a multifunctional cytokine, one of the most directly promotion of angiogenesis research has shown to play an important role of VEGF in bone formation and bone metabolism, the role is mainly by promoting angiogenesis involved in bone development in the form. Many recent studies have shown that VEGF is mainly derived from vascular endothelial cells is an important mediator of angiogenesis induced by multiple cytokines. On the other hand, increased expression levels of these cytokines will lead to increased expression of VEGF or synthetic. Various cytokines to promote bone efficacy and VEGF expression in varied ways, better application of what kind of cytokines, or combined with the feasibility of a variety of cytokines, there is no clear conclusion.

VEGF is a multifunctional cytokine secreted by platelets, megakaryocytes, endothelial cells and osteoblasts, and its role throughout human reproduction, development, tissue regeneration, repair, and tumor pathophysiological process. Many recent studies have shown that also play an important role of VEGF in bone formation and repair, the expression level directly affect the future into the bone's blood supply, affect the speed and quality of the osteoblast.

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BMP, bFGF, has long been studies have shown that bone formation in two very important cytokines. BMP has long been proved to have a strong osteoinductive capacity. From bone cells themselves synthesized bFGF in an autocrine or paracrine in the bone matrix, combining bFGF and osteoblast-like cell surface receptor can promote its conversion to osteogenic cells. In vitro experiments confirmed that of bFGF can directly stimulate rat calvaria bone cells increased DNA synthesis, thereby increasing the number of osteoblasts the synthesis of collagen. Recently, studies have found that BMP, bFGF, and VEGF secretion and expression of the existence of a very close relationship. This experiment on the basis of the results of previous studies, observation of VEGF, respectively, in the application of rhBMP-2 expression and distribution of bFGF stimulation of bone marrow stromal cells, and the corresponding performance for osteogenesis do a comparison and found that in vitro application of rhBMP-2 and bFGF can indeed promote the expression of important mediators of VEGF to promote vascular endothelial cell proliferation, while increasing osteoblast performance. The joint application of these two cytokines can not only accelerate the bone marrow stromal cell proliferation and promote cell conversion to osteoblasts direction, so as to achieve better osteogenesis. First of bFGF to promote the proliferation of bone marrow stromal cells and then apply to promote the expression of osteogenic phenotype rhBMP-2, then the effect on the proliferation of bFGF and rhBMP-2 to promote differentiation are unaffected, which can play two cytokines in maximum biological effectiveness.

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VEGF is a multifunctional cytokine secreted by platelets, megakaryocytes, endothelial cells and osteoblasts, and its role throughout human reproduction, development, tissue regeneration, repair, and tumor pathophysiological process. Many recent studies have shown that also play an important role of VEGF in bone formation and repair, the expression level directly affect the future into the bone's blood supply, affect the speed and quality of the osteoblast.

VEGF was the introduction of bone tissue engineering technology is bound to promote new bone formation, increase the speed and quality of new bone. Exogenous VEGF in vivo half-life is short, and expensive, its application bring great inconvenience. Bone marrow stromal cells is a commonly used seed cells for bone tissue engineering, it contains many cells such as endothelial cells into fiber cells, macrophages, etc. can be secreted expression of VEGF in. How to stimulate the bone marrow stromal cells to promote the secretion of VEGF is another important means of expression of VEGF

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In the early stage of fracture, soft tissue defects, systemic disease causes new blood vessels will lead to delayed union or nonunion. Bone after transplantation, the activity of vascular granulation tissue ingrowth of bone grafts, bone graft is absorbed, take the initiative to the formation of new bone. However, in the large segment of bone grafts, this process slow and incomplete. The key to these problems lies in the revascularization of bone problems that plagued mankind for a long time.

Angiogenesis is an important part of the fracture healing process, a good blood supply to the fracture site of great significance in fracture repair. Vascular endothelial growth factor (vascular endothelial growth factor, VEGF) is a multifunctional cytokine, one of the most directly promotion of angiogenesis research has shown to play an important role of VEGF in bone formation and bone metabolism, the role is mainly by promoting angiogenesis involved in bone development in the form. Many recent studies have shown that VEGF is mainly derived from vascular endothelial cells is an important mediator of angiogenesis induced by multiple cytokines. On the other hand, increased expression levels of these cytokines will lead to increased expression of VEGF or synthetic. Various cytokines to promote bone efficacy and VEGF expression in varied ways, better application of what kind of cytokines, or combined with the feasibility of a variety of cytokines, there is no clear conclusion.

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