Researches Of The Effect Of Zoledronate On Human Periodontal Ligament Stem Cells

ObjectiveBased on the previous animal model of Bisphosphonate-related osteonecrosis of the jaws(BRONJ)and the study of periodontal destroy in rats,we studied the periodontal changes in BRONJ patients.To explore the underlying mechanism,we analyzed the effects of Zoledronate(ZOL)on cell proliferation、adhesion、migration、cell apoptosis、cycle and osteogenic differentiation of human periodontal ligament stem cells(hPDLSCs),which provided a new strategy to reveal the pathogenesis of BRONJ.Material and methods 1.The study of periodontal lesions in BRONJ patients through imaging and histology;2.Isolation,purification and identification of hPDLSCs;3.To detect the effect of Zoledronate on the morphology,proliferation,migration,adhesion,cycle and apoptosis of hPDLSCs;4.The effects of Zoledronate on osteoblastic differentiation of hPDLSCs weredetected by alkaline phosphatase,Von Kossa,real-time PCR andimmunofluorescence;5.hPDLSCs treated with Zoledronate were combined with β-TCP.The complexeswere implanted subcutaneously in nude mice.The ectopic bone formations wereobserved with histological method.Results 1.We observed abnormal changes of periodontal tissue adjacent to the BRONJ lesionin patients,including malformed periodontal ligament(PDL)and significantperiodontal bone loss.2.hPDLSCs were found growing around the PDL tissues 4-7 days after initialincubation.The proliferation,osteoblastic phenotype and stem cell characteristicsof hPDLSCs were confirmed by Cell cloning,alkaline phosphatase,Von Kossa andflow cytometry.3.100μM Zoledronate significantly reduced the proliferation,adhesion,migration andosteogenic differentiation of PDLSCs;4.The percentage of newly formed bone area of subcutaneous implantation in nudemice were 38.19±1.90%,46.99±1.62%,46.77±1.19%,36.64±0.76%,5.74±0.70%for Group 0μM、0.1 μM、1 μM、10 μM、100 μM,respectively.Conclusions 1.Zoledronate induced periodontal destruction in BRONJ patients;2.High doses of Zoledronate remarkably inhibited PDLSCs proliferation,adhesion,migration,and induced cell apoptosis;3.High concentrations of Zoledronate significantly prevented osteogenesis ofPDLSCs in vivo and in vitro.