Mechanical loading increases pro-inflammatory effects of nitrogen-containing bisphosphonate in human periodontal fibroblasts.

OBJECTIVES: There is increasing evidence that inflammation and biomechanical loading can influence the effects of bisphosphonates (BP). The aim of this study was to investigate the influence of tensile strain application combined with IL-1ß and clodronate or zoledronate on human periodontal ligament fibroblasts (HPdLF) in vitro. MATERIALS AND METHODS: HPdLF were cultured with 10 nM IL-1ß and 5 µM clodronate or zoledronate for 48 h. Cells were applied to cyclic tensile strain (CTS; 3% elongation) for 12 h in vitro. Cell number was analyzed directly after CTS by MTT assay. Gene expression of receptor activator of cyclooxygenase-2 (COX-2) was investigated using real-time PCR. MMP-8, TIMP-1, and PGE2 were measured by ELISA. Statistics were performed with SPSS (ANOVA, p < 0.05). RESULTS: Zoledronate reduced the cell number of HPdLF (60.3 vs. 100%), which was significant when combined with IL-1ß. Combined with 3% CTS, this effect was voided and cell number increased over the level of the control cells. IL-1ß led to a 10-fold increase of COX-2 gene expression. Combined with CTS and zoledronate, this increase was enhanced to a gene expression 70-fold that of control cells with related PGE2 synthesis. Clodronate neither reduce the cell number nor enhanced the COX-2 gene expression. CTS increased MMP-8 protein synthesis. Combined with BP, this increase was voided. TIMP-1 protein synthesis was increased at all conditions under CTS. CONCLUSIONS: Mechanical loading might activate cell metabolism and abolish BP- and inflammation-induced reduction of viability. Combination of mechanical loading, inflammation, and nitrogen-containing bisphosphonates can cause pro-inflammatory effects. CLINICAL RELEVANCE: Periodontal inflammation should be treated initially before BP intake to prevent decreased cell viability of the periodontium and increased inflammation, which might be enhanced by the addition of mastication forces.

Mechanical loading influences the effects of bisphosphonates on human periodontal ligament fibroblasts.

OBJECTIVES: There is increasing evidence that bisphosphonates affect orthodontic tooth movement. The object of the study was to investigate the changes produced by tensile strain on human periodontal ligament fibroblasts (HPdLFs) treated with clodronate or zoledronate. MATERIALS AND METHODS: HPdLF were cultured with 5 and 50 µM clodronate or zoledronate for 48 h and applied to tensile strain (TS) (5 and 10 %) for 12 h in vitro. Viability was verified by MTT assay and apoptosis rate via caspase 3/7 assay. Gene expression of receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG) was investigated using real-time PCR. OPG was also analyzed by ELISA and RANKL by immunocytochemical staining. RESULTS: Zoledronate (50 µM) reduced the viability of HPdLF (76 vs 100 %) and combined with 5 % TS to 53 %. TS of 10 % and clodronate reduced viability to 79 % with increased caspase 3/7 activity. Clodronate (5 µM) led to a slight increase of OPG gene expression, zoledronate (5 µM) to a slight decrease. Combined with 5 % TS, both increased OPG gene expression (2-3-fold) and OPG synthesis. Zoledronate increased gene expression of RANKL (4-fold). Combined with 5 % of TS, this increase was abolished. TS of 10 % in combination amplified increase of RANKL ending up with a 9-fold gene expression by clodronate and high RANKL protein synthesis. CONCLUSIONS: This study shows for the first time that mechanical loading alters the effects of bisphosphonates on viability, apoptosis rate, and OPG/RANKL system of HPdLF dependent on the applied strength. Low forces and bisphosphonates increase factors for bone apposition, whereas high forces combined with bisphosphonates stimulate osteoclastogenesis. CLINICAL RELEVANCE: Mechanical loading of periodontal ligament with high strengths should be avoided during bisphosphonate therapy.