Effect Of Recombination Human Bone Morphogenetic Protein7on Proliferation In Cultured Primary Open-angle Glaucoma Trabecular Meshwork Cells

Purpose To investigate the effect of recombination human bone morphogeneticprotein7(rhBMP7)on proliferation on the basis of culturing the trabecular meshworkcells from primary open-angle glaucoma eye，to study the relationship betweenrhBMP7and beginning and progression of POAG.Methods The trabecular tissues from trabeculectomy without mitomycin werecultured in vitro and passaged for3times. The cellular morphology, morphologicfeature and growing characteristics of culture cells were observed by invertedmicroscope and immunocy to chemical method. The third passage cells wereincubated with different final concentrations of rhBMP7(eg:0ng/ml(control group)，20ng/ml,50ng/ml,80ng/ml,100ng/ml,200ng/ml) for24h,48h respectively, theproliferation effect of rhBMP7on trabecular meshwork cells from POAG wasdetected by Cell Counting Kit-8assay（CCK-8），fluorescence microscopy and flowcytometry.Results1.The primarily trabecular meshwork cells from POAG were obtained fromthe edge of tissue after about10days, the shapes of cells showedcyclotrilete,fusiform and polygon, etal. Through continuous cell culture, invertedmicroscope and immunocy to chemical method, the cells in vitro were identified aspurified trabecular meshwork cells;2. The OD value of trabecular meshwork cells from POAG which weretreated with different final concentrations of rhBMP7(eg:0ng/ml(control group),20ng/ml,50ng/ml,80ng/ml,100ng/ml,200ng/ml)for24h were0.5612±0.0269、0.7242±0.0393、1.4160±0.0162、1.7404±0.0392、1.8538±0.0145、1.9364±0.0546by CCK-8assay. The difference between experimental groups and control group wasstatistically significant(P <0.05).The difference among experimental groups wasstatistically significant(P <0.05);3.To observe by fluorescence microscopy:as the rhBMP7final concentrationincreased,the counts of the signed POAG trabecular meshwork cells by CFDA SEwere enhanced and the height of fluorescence was decreased;4. After the treatment of20ng/ml,50ng/ml,80ng/ml，100ng/ml,200ng/mlrhBMP7for48h,the proliferation rate in the experimental groups of POAG trabecular meshwork cells was17.85%、18.63%、20.10%、27.45%、72.41%compared with the control group. And with the increasing final concentration of therhBMP7, the height of fluorescence was decreased.Conclusion The proliferation ability of POAG trabecular meshwork cells wereaccelerated by rhBMP7in a certain range. And the degree of the proliferation showsa dose-time dependent manner.