Osteogenic induction and 1,25-dihydroxyvitamin D3 oppositely regulate the proliferation and expression of RANKL and the vitamin D receptor of human periodontal ligament cells.
OBJECTIVE
Human periodontal ligament cells (hPDLCs) may play an important role in osteoclastogenesis in alveolar bone by expressing the receptor activator of NF-KappaB ligand (RANKL) and osteoprotegerin (OPG). The present study aimed to investigate the differences between the effects of osteogenic induction and 1,25-dihydroxyvitamin D(3) (VD(3)) on hPDLC proliferation and the expression of RANKL, osteoprotegerin, and the vitamin D receptor (VDR) in hPDLCs.


METHODS
Primary cultures of 11 hPDLC populations from 11 donors were obtained. Three samples of each hPDLC population from passage 3 were, respectively, treated with osteogenic induction medium, 10(-8)M VD(3), or vehicle as a control. Cell proliferation at days 0, 1, 3, 5, and 7 was estimated with the MTT method. At day 6, the mRNA levels of RANKL, OPG and VDR were determined with real-time RT-PCR.


RESULTS
Osteogenic induction significantly promoted hPDLC proliferation, while VD(3) inhibited proliferation. Osteogenic induction significantly down-regulated the mRNA level of RANKL by 1.61-fold (P = 0.033) and decreased the level of VDR by 2.13-fold (P = 0.003), while there was no change in the level of OPG and OPG/RANKL ratio with osteogenic induction. On the contrary, VD(3) significantly up-regulated the level of RANKL by 9.58-fold (P = 0.001) and increased the level of VDR by 3.15-fold (P = 0.004), while down-regulating the OPG/RANKL ratio by 7.14-fold (P=0.004).


CONCLUSIONS
Osteogenic induction and VD(3) exert opposite effects in regulating hPDLC proliferation and mRNA expression of RANKL and VDR. This may induce hPDLCs to play different roles in alveolar bone metabolism.
Tang
Xiaolin
X
Department of Periodontology, School of Stomatology, China Medical University, Shenyang, Liaoning Province 110002, China. xltang@mail.cmu.edu.cn
Meng
Huanxin
H
D000293:Adolescent; D002113:Calcification, Physiologic / Q000187:drug effects; D002117:Calcitriol / Q000494:pharmacology*; D002120:Calcium Channel Agonists / Q000494:pharmacology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D002648:Child; D004396:Coloring Agents; D015536:Down-Regulation; D006801:Humans; D007633:Keratins / Q000032:analysis; D010010:Osteoclasts / Q000187:drug effects / Q000502:physiology; D010012:Osteogenesis / Q000502:physiology*; D053244:Osteoprotegerin / Q000032:analysis / Q000187:drug effects; D010513:Periodontal Ligament / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D053245:RANK Ligand / Q000032:analysis / Q000187:drug effects*; D018167:Receptors, Calcitriol / Q000032:analysis / Q000187:drug effects*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors; D014158:Transcription, Genetic / Q000235:genetics; D015854:Up-Regulation; D014746:Vimentin / Q000032:analysis; D055815:Young Adult
D002120:Calcium Channel Agonists; D004396:Coloring Agents; D053244:Osteoprotegerin; D053245:RANK Ligand; D018167:Receptors, Calcitriol; D013778:Tetrazolium Salts; D013844:Thiazoles; D014746:Vimentin; D007633:Keratins; C022616:thiazolyl blue; D002117:Calcitriol