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Effects of recombinant human sclerostin on proliferation and migration in human cementoblast lineage cells
Naonobu Okimura1, Ryo Kunimatsu1, Isamu Kado1
1Department of Orthodontics and Craniofacial Development Biology, Graduate School of Biomedical & Health Sciences, Hiroshima University, Japan.
Objective:
To evaluate the effects of sclerostin on the proliferation and migration of human cementoblasts and periodontal ligament cells.
Design:
Sclerostin expression in human cementoblasts and periodontal ligament cells was assessed using immunochemical staining. Human cementoblasts and periodontal ligament cells were cultured and treated with 100 ng/mL of recombinant human sclerostin. Cell proliferation was evaluated using a 5-bromo-2-deoxyuridine enzyme-linked immunosorbent assay and quantified with a live-cell imaging and analysis platform (IncuCyte® S3 system). Furthermore, sclerostin's impact on apoptosis in human cementoblasts and periodontal ligament cells was evaluated using IncuCyte® Caspase-3/7 green dye. Additionally, cell migration was analyzed through quantitative wound healing assessment using the IncuCyte® S3 system. Polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blotting were then performed to confirm the effect of sclerostin on CEMP-1. The data obtained were statistically analyzed using the Mann-Whitney U test.
Results:
Intracellular sclerostin localization in human cementoblasts and periodontal ligament cells were confirmed form the immunochemical staining. The sclerostin-treated group showed suppressed proliferation and migration of human cementoblasts and periodontal ligament cells compared with the non-treated group. Furthermore, the sclerostin-treated group showed significantly elevated caspase-3/7 activity compared with the non-treated group. However, the addition of sclerostin did not result in any significant changes in CEMP-1.
Conclusion:
Sclerostin is crucial in regulating the proliferation and migration of cementoblasts and periodontal ligament cells. This highlights its importance in regenerating the cementum and periodontal ligament.
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