| Background and Objective:For the purpose of increasing bone mass, autologous, allogeneic, heterogeneous andartificial bone materials are now widely used in dentistry and maxillofacial surgery.Angiogenesis in bone replacement graft is considered that play an important role insuccessful healing and bone formation. Interaction of osteogenesis and angiogenesis isregulated by a variety of factors produced by autocrine-paracrine networks, which areconsisted of osteoblasts, endothelial cells and their precursors etc. Effect of angiogenic factoris not only stimulating angiogenesis, but also promoting osteogenesis directly or indirectly.In this research two characteristic angiogenic factor，VEGF and PDGF，are chosen to be thestudy objects. We investigated effects of Choukroun’s PRF on osteoblast-like cell ofsecreting VEGF and PDGF and expressing VEGFR-2, to discuss the probability ofChoukroun’s PRF ’s application as a local source of angiogenic factors to vascularize bonegraft.Methods:In vitro cultured osteoblast, and strictly followed the protocol to prepare humanChoukroun’s PRF.There were three groups of the first experiment: MG-63cells were carried in standardconditions (control group), MG-63cells with a PRF in standard conditions (test group(PRF)), one PRF in standard conditions (control group (PRF)). The culture plates of controlgroup and test group (PRF) were removed for counting at experimental time, i e. on days2(D2),4(D4),6(D6),8(D8) and10(D10). And then,a curve of growth was drawn.Separately collected three groups’ culture mediums, using ELISA method to measurecontents of VEGF and PDGF.There were three groups of the second experiment: MG-63cells were carried instandard conditions (control group), MG-63cells with1/4PRF in standard conditions (testgroup (1/4PRF)), MG-63cells with one PRF in standard conditions (test group (1PRF)).Then the expression of VEGFR-2on the surface of MG-63cells was detected by Western Blotting at the sixth day.Results:1、Cell proliferationIn the presence of1PRF membrane, the number of MG-63in culture was significantlyhigher (p <0.05) compared to the control cultures at the5experimental times.2、Contents of VEGF and PDGFMeasured in the culture of test group were both significantly higher (P<0.05) comparedto control group (blank) and control group (PRF) at the5experimental times..3、Expression of VEGFR-2of MG-63In the presence of1and1/4PRF membranes, the expression of VEGFR-2of MG-63was significantly higher compared to the control cultures, while the test group (1PRF) ishigher than the test group (1/4PRF).Conclusion:In the experimental group,we compared the effect of Choukroun’s PRF on humanosteoblast-like cells(MG-63) proliferation, secretion of VEGF and PDGF and expression ofVEGFR-2by adding the Choukroun’s PRF in the medium and the blank control.Come to thefollowing conclusions:1、Choukroun ’ s PRF significantly promote the proliferation of human Osteoblast-likecells, and shorten the incubation period of growth.2、Choukroun ’ s PRF membrane release growth factors slowly and steadily during atleast10days.3、Preparation of Choukroun’s PRF effectively induced increasing VEGF concentrationin the environment, and stimulating the secretion of VEGF of human Osteoblast-like cells,providing evidence of probable mechanisms of action of Choukroun’s PRF invascularization.4、Preparation of Choukroun’s PRF effectively induced increasing PDGF concentrationin the environment and maintaining its high level, stimulating the secretion of VEGF ofhuman Osteoblast-like cells, prompting Choukroun’s PRF can improve angiogenesis andosteogenesis via strongly mitosis promoting.5、Choukroun ’ s PRF stimulate the expression of VEGFR-2on the suface of humanOsteoblast-like cell in a dose-dependent way, probably will promote the availability ofVEGF, make the reponse to VEGF more sensitive and enhance its biological effect. 6、In all clinical applications, Choukroun ’ s PRF has to be considered and used as afibrin-based living biomaterial, and not only as simple source of growth factors. |
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