Effect of recombinant human bone morphogenetic protein-2and osteoprotegerin-Fc in MC3T3-E1 cells
- Author:
Sang-Hyon KIM
1
;
Hye-Jung CHOI
;
Sang-Min LEE
;
Dae Sung YOON
;
Chang-Nam SON
Author Information
- Publication Type:Original Article
- From:Journal of Rheumatic Diseases 2024;31(2):79-85
- CountryRepublic of Korea
- Language:EN
-
Abstract:
Objective:We compared the osteoblastogenesis by serially administrating recombinant human bone morphogenetic protein-2 (rhBMP-2) and osteoprotegerin-immunoglobulin Fc segment complex (OPG-Fc).
Methods:The MC3T3-E1 preosteoblast cell line was differentiated for 1, 3, and 7 days with a treatment of OPG-Fc in 10~200 ng/mL concentration and the cell viability was evaluated by Cell Counting Kit-8 analysis. The level of differentiation from MC3T3-E1 cells to osteoblasts was determined by alkaline phosphatase activity. The level of runt domain-containing transcription factor 2 (Runx2) and osteopontin (OPN) manifestation, involved in osteoblast differentiation, was examined by real-time polymerase chain reaction and western blotting.
Results:During MC3T3-E1 cell differentiation, the differentiation level was high with 1-day treatment using 100 ng/mL OPGFc. The treatment with 50 ng/mL rhBMP-2 for 7 days, followed by 1-day treatment with 100 ng/mL OPG-Fc produced the highest differentiation level, which was approximately 5.3 times that of the control group (p<0.05). The expression of Runx2 mRNA significantly increased, reaching 2.5 times the level of the control group under the condition of 7-day treatment with rhBMP-2 and 1-day treatment with OPG-Fc (p<0.001). The expression of Runx2 protein significantly increased to approximately 5.7 times that of the control group under the condition of 7-day treatment with rhBMP-2, followed by 1-day treatment with OPG-Fc (p<0.01).The expression of OPN protein showed no change from that of the control group under various conditions of rhBMP-2 and OPGFc combinations.
Conclusion:These results imply that the treating preosteoblasts with rhBMP-2 first and then with OPG-Fc increased osteoblast differentiation efficacy.
