Influence of different concentrations of enamel matrix proteins on bioactivity of human periodontal ligament cells

Qu Zhe; Zhang Jing-ying; Guo Ying; Ma Wei-dong; Ma Lan

doi:10.3969/j.issn.2095-4344

Abstract：BACKGROUND:Numerous studies have confirmed that enamel matrix proteins can promote the regeneration of osteoblasts and cementoblast, and then it can achieve approaching physiological periodontal regeneration in the treatment of periodontal defects. <br> OBJECTIVE: To observe the effects of different concentrations of enamel matrix proteins on proliferation, viability, differentiation and migration of human periodontal ligament cels. <br> METHODS: The human periodontal ligament cels at the third generation were gained, and then cultured in serum-free DMEM containing different concentrations of enamel matrix proteins (0, 12.5, 25, 50, 100, 250 mg/L). After 24 hours of culture, proliferation and viability of periodontal ligament cels were measured using [(3)H]-thymidine uptake and MTT assay. After 48 hours of culture, alkaline phosphatase activity and osteocalcin production were detected with commercial available test kits. When the cels grew as a monolayer, the cel culture fluid was removed, and then with a pipette head, a cel incision, 1 mm wideness, was prepared in a monolayer of <br> cels to further observe the cel fusion continuously within 24 hours. <br> RESULTS AND CONCLUSION:The proliferation, viability and differentiation of periodontal ligament cels were gradualy increased with the concentration increase of enamel matrix proteins (0-100 mg/L). When the concentration of enamel matrix proteins was 250 mg/L, these parameters began to decrease, but the levels were stil higher than those in the 0 mg/L enamel matrix protein group. In the 100 mg/L group, the cels in the wound edge began to grow towards the center in the initial 6 hours, then became confluent at 12 hours, and until the 20th hour of culture, the cels in the two sides of the wound edge were completely fused to fuly close the wound edge, indicating a better wound healing in the 100 mg/L group than the other groups. These findings suggest that enamel matrix proteins can stimulate proliferation, viability, differentiation and migration of periodontal ligament cels.