| Objective:Dental follicle is loose and connective tissue surrounding growing dental germ in the early development of teeth. Dental follicle is closely associated with the formation of periodontal tissue. Dental follicle stem cells are cells inside dental follicle and have been proved that human dental follicle stem cells(HDFSCs) have the potential of differentiating into cementoblasts or osteoblasts under appropriate conditions. However, the mechanism(s) for osteogenic differentiation of DFCs are still unclear till now. The main cells in the dental pulp is odontoblasts,it really exists interactive signal transduction between Odontoblasts and DFCs in the early stages of tooth development which may be involved in the interaction. Hence, the aim of this study was to explore the influence of human dental pulp stem cells(HDPSCs) on proliferation and osteogenic differentiation of HDFSCs in vitro.Methods:1.HDFSCs and HDPSCs were isolated,cultured and identified by a modified tissue block digestion,cell immunohistochemistry,flow cytometry and multilineage inducement.2.The establishment of co-culture system in vitro:Cell culture chamber(Transwell) with pore size of 0.4 um in diameter were applied to establish the co-culture system of HDFSCs and HDPSCs in vitro.3.The changes of proliferative activity of HDFSCs were detected: The co-culture system was fabricated with 24 well-plate Transwell. Cell Counting Kit-8 assay was applied to detect the proliferation of HDFSCs.4. The genes expression related osteogenesis of HDFSCs were tested:The co-culture system was established with 6 well-plate Transwell.In one week after in vitro co-culture system of HDFSCs and HDPSCs,the expression of several genes related osteogenesis including the alkaline phosphatase(ALP),runt-related transcription factor 2(Runx2),osteopontin(OPN),bone sialoprotein(BSP) and collagen type 1(Col-1) were measured with qRT-PCR and 2-??Ct method was adopted to analyze the results.Results:1.HDFSCs and HDPSCs were isolated,cultured and identified successfully.2. The test results of HDFSCs' proliferation activity showed in the first 3 days co-cultured with HDPSCs,there was no obvious change compared with the control group. Starting from the fourth day,HDFSCs in the co-culture group showed a higher proliferative activity than that in the control group( P < 0.05).3.The results of genes expression related osteogenesis of HDFSCs indicated that compared with the control group,there were significant increases in the mRNA levels of ALP,Runx2,OPN,BSP and Col-1(P < 0.05) in the co-culture group.Conclusion:Co-culturing with HDPSCs could stimulate the proliferous activity and the expression of osteogenesis genes including ALP,Runx2,OPN,BSP and Col-1 in HDFSCs.The co-culture of HDFSCs and HDPSCs has potential application value in the field of bone regeneration. |
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