Abstract
Objective To explore the effect of toll like receptor (TLR) on ossification of bone marrow mesenchymal stem cells of rat (BMSCs).Methods Mter isolation and purification,rat-BMSCs were identified by detecting surface makers,such as CD44,CD105,CD34,CD14,CD29,CD95,HLA-Ⅰand Vimentin,using immunofluorescence or flow cytometry (FCM).BMSCs of 3rd passage were randomly divided into 3 groups:control group,TLR3-Primed group (Poly ＜ I∶ C ＞,1 mg/L) and TLR4-primed group (LPS,10 μg/L).Alkaline phosphatase (ALP) and alizarin red staining were performed in 10th day,and 14th day.Expression of collagen-Ⅰ and osteocalcin were detected by Western blotting,then statistically analysed.Results CD44,CD105,Vimentin,CD29,CD95,human leukocyte antigens (HLA)-Ⅰwere positive,CD14,while CD34 were negative.Collagen-Ⅰ and Osteocalcin were all expressed in in three groups,strongest in TLR-4-primed group,weakest in Controlled group.ALP and alizarin red staining were all positive in all groups,stronger in TLR-3-primed group than other two groups,TLR-4-primed group and controlled group had no statistically difference.Conclusion The differentiation of BMSCs to osteoblasts (OB) can be promoted by TLR-3 and TLR-4.TLR-4 had stronger effect in this process.The ossification of OB to osteocytes was promoted by TLR-3,but inhibited by TLR-4. Key words: Toll like receptor; Bone marrow mesenchymal stem cells; Osteogenic differentiation
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