Experimental Study Of Chondrogenic Differentiation Of Bone Marrow Stem Cells Of Rabbits In Different Concentrations Of Gentamicin In Vitro

Objective:Antibiotics is often used in clinical treatment to cartilage defect.Gentamicin is a broad spectrum of powerful antibiotics and commonly topical used in clinical work, which often gets a good efficacy.This experimental study is to explore the capability of bone marrow stem cells of rabbits differentiating into Chondrocytes in vitro. To study the effect of gentamicin in different concentrations (0μg/ml10μg/ml、50μg/ml、100μg/ml、200μg/ml) on proliferation and chondrogenic differentiation of bone marrow stem cells of rabbits in vitro.Methods:At first, Isolate femur,tibia and fibula from adopted eight-weeks old rabbit.Then digese with collagenase-I and placed in Dulbeco Modified Eagle Medium (DMEM) with fetal bovine serum in primary cultivation.Observe the growth and appearance of the cells by a inversion microscope.The third passage of BMSCs were divided into5groups:one control group(containing bone marrow stem cells complete medium include DMEM serum medium、10%fetal bovine serum、100U/ml Penicillin、100U/ml Streptomycin,without any gentamicin) and four experimental groups(containing bone marrow stem cells complete medium+four different concentrations of gentamicin:10μg/ml、50μg/ml、100μg/ml、200μg/ml).The cell proliferation was assessed by MTT methods, toluidine blue stain and type Ⅱ collagen immunohistochemistry were applied for chondrogenic identificatio.Results:1. BMSCs cultured in vitro were in flat fusiform、fibroblast-like cells shape and stably passaged, during which BMSCs kept the same shape.2.MTT experiments showed that gentamicin at100μg/ml,200μg/ml inhibited cell proliferation, the remaining concentrations had no side effect on cell proliferation.3.the toluidine blue staining with the cells after chondrogenic differentiation shows:200μg/ml and100μg/ml groups lots of cell out of stain, the other groups express positive, type Ⅱ collagen immunohistochemistry staining shows:200μg/ml and100μg/ml groups weak positive Expression, the other groups express positiveConclusions:1.BMSCs readily available, grown rapidly and stable after culture and passage;2.BMSCs have the potential to differentiate into chondrogenic lineage;3.gentamicin in lower concentrations(10μg/ml、50μg/ml)shows no side effect on BMSCs proliferation and differentiate into chondrogenic, gentamicin in higher concentrations(100μg/ml、200μg/ml)inhibited the BMSCs proliferation and differentiate into chondrogenic.