RESUMO
It has previously been demonstrated that the polybisphosphonate osteodex (ODX) inhibits bone resorption in organ-cultured mouse calvarial bone. In this study, we further investigate the effects by ODX on osteoclast differentiation, formation, and function in several different bone organ and cell cultures. Zoledronic acid (ZOL) was used for comparison. In retinoid-stimulated mouse calvarial organ cultures, ODX and ZOL significantly reduced the numbers of periosteal osteoclasts without affecting Tnfsf11 or Tnfrsf11b mRNA expression. ODX and ZOL also drastically reduced the numbers of osteoclasts in cell cultures isolated from the calvarial bone and in vitamin D3-stimulated mouse crude bone marrow cell cultures. These data suggest that ODX can inhibit osteoclast formation by inhibiting the differentiation of osteoclast progenitor cells or by directly targeting mature osteoclasts. We therefore assessed if osteoclast formation in purified bone marrow macrophage cultures stimulated by RANKL was inhibited by ODX and ZOL and found that the initial formation of mature osteoclasts was not affected, but that the bisphosphonates enhanced cell death of mature osteoclasts. In agreement with these findings, ODX and ZOL did not affect the mRNA expression of the osteoclastic genes Acp5 and Ctsk and the osteoclastogenic transcription factor Nfatc1. When bone marrow macrophages were incubated on bone slices, ODX and ZOL inhibited RANKL-stimulated bone resorption. In conclusion, ODX does not inhibit osteoclast formation but inhibits osteoclastic bone resorption by decreasing osteoclast numbers through enhanced cell death of mature osteoclasts.
Assuntos
Reabsorção Óssea , Osteoclastos , Animais , Camundongos , Osteoclastos/metabolismo , Osteogênese , Medula Óssea , Células Cultivadas , Reabsorção Óssea/tratamento farmacológico , Reabsorção Óssea/metabolismo , Macrófagos/metabolismo , Diferenciação Celular , Morte Celular , Ácido Zoledrônico/farmacologia , Ácido Zoledrônico/metabolismo , RNA Mensageiro/metabolismo , Ligante RANK/farmacologia , Ligante RANK/metabolismoRESUMO
We investigated the effects of zoledronate (ZA) and compressive force, separately and in combination, on the proliferation and differentiation of human gingival fibroblasts (HGFs) to verify the mechanism underlying medication-related osteonecrosis of the jaw (MRONJ). The addition of 100 µM ZA markedly inhibited cell proliferation. Expression of type I collagen, fibroblast growth factor 2, and connective tissue growth factor genes, was decreased by ZA and compressive force. Similar results were observed for collagen expression by using Sirius red staining. These results, together with clinical findings that MRONJ is more common in cases with excessive mechanical stress on the oral mucosa, suggest that bisphosphonates such as ZA and mechanical stress may act in conjunction as risk factors for the development of MRONJ by affecting homeostasis of the oral mucosal tissues, including HGFs.
Assuntos
Osteonecrose da Arcada Osseodentária Associada a Difosfonatos , Conservadores da Densidade Óssea , Humanos , Ácido Zoledrônico/farmacologia , Ácido Zoledrônico/metabolismo , Difosfonatos/efeitos adversos , Gengiva , Fibroblastos/metabolismo , Proliferação de Células , Conservadores da Densidade Óssea/efeitos adversosRESUMO
Periodontitis represents a severe inflammatory illness in tooth supporting tissue. It has been supported that cAMP response element binding protein 1 (CREB1), a common transcription factor, extensively participates in osteogenic differentiation. Here, the current study was to look into the impacts of CREB1 on the process of periodontitis and its possible action mechanism. After human periodontal ligament stem cells (PDLSCs) were challenged with zoledronic acid (ZA), CREB1 expression was examined with RT-qPCR and western blotting. CCK-8 assay appraised cell activity. Following CREB1 elevation or/and vascular endothelial growth factor (VEGF) silencing in ZA-treated PDLSCs, CCK-8 and TUNEL assays separately estimated cell viability and apoptosis. Western blotting tested the expression of apoptosis- and osteogenic differentiation-associated proteins. ALP staining measured PDLSCs osteogenic ability and ARS staining estimated mineralized nodule formation. JASPAR predicted the potential binding of CREB1 with VEGF promoter, which was then testified by ChIP and luciferase reporter assays. RT-qPCR and western blotting tested VEGF expression. CREB1 expression was declined in ZA-exposed PDLSCs and CREB1 elevation exacerbated the viability and osteogenic differentiation while obstructed the apoptosis of PDLSCs. Additionally, CREB1 bond to VEGF promoter and transcriptionally activated VEGF expression. Further, VEGF absence partially stimulated the apoptosis while suppressed the osteogenic differentiation of CREB1-overexpressing PDLSCs treated by ZA. To be concluded, CREB1 might activate VEGF transcription to obstruct the apoptosis while contribute to the osteogenic differentiation of ZA-treated PDLSCs.
Assuntos
Osteogênese , Periodontite , Humanos , Apoptose/genética , Diferenciação Celular/fisiologia , Proliferação de Células , Células Cultivadas , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Osteogênese/fisiologia , Ligamento Periodontal , Periodontite/metabolismo , Transdução de Sinais/fisiologia , Sincalida/metabolismo , Células-Tronco , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Ácido Zoledrônico/farmacologia , Ácido Zoledrônico/metabolismoRESUMO
Long-term usage of bisphosphonates, especially zoledronic acid (ZA), induces osteogenesis disorders and medication-related osteonecrosis of the jaw (MRONJ) in patients, thereby contributing to the destruction of bone remodeling and the continuous progression of osteonecrosis. Menaquinone-4 (MK-4), a specific vitamin K2 isoform converted by the mevalonate (MVA) pathway in vivo, exerts the promotion of bone formation, whereas ZA administration suppresses this pathway and results in endogenous MK-4 deficiency. However, no study has evaluated whether exogenous MK-4 supplementation can prevent ZA-induced MRONJ. Here we showed that MK-4 pretreatment partially ameliorated mucosal nonunion and bone sequestration among ZA-treated MRONJ mouse models. Moreover, MK-4 promoted bone regeneration and inhibited osteoblast apoptosis in vivo. Consistently, MK-4 downregulated ZA-induced osteoblast apoptosis in MC3T3-E1 cells and suppressed the levels of cellular metabolic stresses, including oxidative stress, endoplasmic reticulum stress, mitochondrial dysfunction, and DNA damage, which were accompanied by elevated sirtuin 1 (SIRT1) expression. Notably, EX527, an inhibitor of the SIRT1 signaling pathway, abolished the inhibitory effects of MK-4 on ZA-induced cell metabolic stresses and osteoblast damage. Combined with experimental evidences from MRONJ mouse models and MC3T3-E1 cells, our findings suggested that MK-4 prevents ZA-induced MRONJ by inhibiting osteoblast apoptosis through suppression of cellular metabolic stresses in a SIRT1-dependent manner. The results provide a novel translational direction for the clinical application of MK-4 for preventing MRONJ.
Assuntos
Conservadores da Densidade Óssea , Osteonecrose , Camundongos , Animais , Sirtuína 1/genética , Sirtuína 1/metabolismo , Ácido Zoledrônico/efeitos adversos , Ácido Zoledrônico/metabolismo , Difosfonatos/efeitos adversos , Osteonecrose/induzido quimicamente , Osteonecrose/tratamento farmacológico , Osteonecrose/genética , Osteoblastos , Apoptose , Transdução de Sinais , Estresse Fisiológico , Conservadores da Densidade Óssea/efeitos adversosRESUMO
In addition to reducing fracture risk, zoledronic acid has been found in some studies to decrease mortality in humans and extend lifespan and healthspan in animals. Because senescent cells accumulate with aging and contribute to multiple co-morbidities, the non-skeletal actions of zoledronic acid could be due to senolytic (killing of senescent cells) or senomorphic (inhibition of the secretion of the senescence-associated secretory phenotype [SASP]) actions. To test this, we first performed in vitro senescence assays using human lung fibroblasts and DNA repair-deficient mouse embryonic fibroblasts, which demonstrated that zoledronic acid killed senescent cells with minimal effects on non-senescent cells. Next, in aged mice treated with zoledronic acid or vehicle for 8 weeks, zoledronic acid significantly reduced circulating SASP factors, including CCL7, IL-1ß, TNFRSF1A, and TGFß1 and improved grip strength. Analysis of publicly available RNAseq data from CD115+ (CSF1R/c-fms+) pre-osteoclastic cells isolated from mice treated with zoledronic acid demonstrated a significant downregulation of senescence/SASP genes (SenMayo). To establish that these cells are potential senolytic/senomorphic targets of zoledronic acid, we used single cell proteomic analysis (cytometry by time of flight [CyTOF]) and demonstrated that zoledronic acid significantly reduced the number of pre-osteoclastic (CD115+/CD3e-/Ly6G-/CD45R-) cells and decreased protein levels of p16, p21, and SASP markers in these cells without affecting other immune cell populations. Collectively, our findings demonstrate that zoledronic acid has senolytic effects in vitro and modulates senescence/SASP biomarkers in vivo. These data point to the need for additional studies testing zoledronic acid and/or other bisphosphonate derivatives for senotherapeutic efficacy.
Assuntos
Senescência Celular , Fenótipo Secretor Associado à Senescência , Humanos , Animais , Camundongos , Senescência Celular/fisiologia , Ácido Zoledrônico/farmacologia , Ácido Zoledrônico/metabolismo , Senoterapia , Proteômica , Fibroblastos/metabolismoRESUMO
Zoledronic acid (ZA), one of the commonly used bisphosphonates, is mainly used for bone-metabolic diseases. Studies proved that ZA has adverse effects on oral soft tissues. As the first line of innate immunity, the gingival epithelium could be infected by periodontal pathogens, which is a key process of the initiation of periodontal diseases. Yet, how ZA affects the periodontal pathogens infecting the epithelial barrier remains unclear. This study aimed to investigate the influences of ZA on the process of Porphyromonas gingivalis (P. gingivalis) infecting the gingival epithelial barrier via in-vitro and in-vivo experiments. In the in-vitro experiments, under the condition of different concentrations of ZA (0, 1, 10, and 100 µM), P. gingivalis was used to infect human gingival epithelial cells (HGECs). The infections were detected by transmission electron microscope and confocal laser scanning microscope. Besides, the internalization assay was applied to quantify the P. gingivalis, which infected the HGECs, in the different groups. To evaluate the expression levels of pro-inflammatory cytokines, including interleukin (IL)-1ß, IL-6, and IL-8, by infected HGECs, real-time quantitative reverse transcription-polymerase chain reactions were applied. In the in-vivo experiments, rats were given ZA solution (ZA group) or saline (control group) by tail intravenous injection for 8 weeks. Subsequently, we put ligatures around the maxillary second molars of all the rats and inoculated P. gingivalis to the gingiva every other day from day 1 to day 13. The rats were sacrificed on days 3, 7, and 14 for micro-CT and histological analyses. The in-vitro results manifested that the quantity of P. gingivalis that had infected HGECs increased with the ZA concentrations. Pro-inflammatory cytokines expression by HGECs were significantly increased by 100 µM ZA. In the in-vivo study, compared to the control group, more P. gingivalis was detected in the superficial layer of gingival epithelium in the ZA group. Besides, ZA significantly increased the expression level of IL-1ß on day 14 and IL-6 on days 7 and 14 in gingival tissues. These findings suggest that the oral epithelial tissues of patients who receive high-dose ZA treatment may be more susceptible to periodontal infections, resulting in severe inflammatory conditions.
Assuntos
Interleucina-6 , Porphyromonas gingivalis , Humanos , Ratos , Animais , Interleucina-6/metabolismo , Porphyromonas gingivalis/metabolismo , Ácido Zoledrônico/farmacologia , Ácido Zoledrônico/metabolismo , Citocinas/metabolismo , Células Epiteliais , Gengiva/metabolismoRESUMO
Improving local bone mineral density (BMD) at fracture-prone sites of bone is a clinical concern for osteoporotic fracture prevention. In this study, a featured radial extracorporeal shock wave (rESW) responsive nano-drug delivery system (NDDS) is developed for local treatment. Based on a mechanic simulation, a sequence of hollow zoledronic acid (ZOL)-contained nanoparticles (HZNs) with controllable shell thickness that predicts various mechanical responsive properties is constructed by controlling the deposition time of ZOL and Ca2+ on liposome templates. Attributed to the controllable shell thickness, the fragmentation of HZNs and the release of ZOL and Ca2+ can be precisely controlled with the intervention of rESW. Furthermore, the distinct effect of HZNs with different shell thicknesses on bone metabolism after fragmentation is verified. In vitro co-culture experiments demonstrate that although HZN2 does not have the strongest osteoclasts inhibitory effect, the best pro-osteoblasts mineralization results are achieved via maintaining osteoblast-osteoclast (OB-OC) communication. In vivo, the HZN2 group also shows the strongest local BMD enhancement after rESW intervention and significantly improves bone-related parameters and mechanical properties in the ovariectomy (OVX)-induced osteoporosis (OP) rats. These findings suggest that an adjustable and precise rESW-responsive NDDS can effectively improve local BMD in OP therapy.
Assuntos
Osteoporose , Fraturas por Osteoporose , Feminino , Ratos , Animais , Osteoclastos , Fraturas por Osteoporose/metabolismo , Liberação Controlada de Fármacos , Osso e Ossos , Osteoporose/tratamento farmacológico , Osteoporose/prevenção & controle , Osteoporose/metabolismo , Osteoblastos , Ácido Zoledrônico/metabolismo , Ácido Zoledrônico/farmacologia , Ácido Zoledrônico/uso terapêuticoRESUMO
Although polyploid giant cancer cells (PGCCs) are known as a key source of failure of current therapies, sufficient drugs to target these cells are not yet introduced. Considering the similarities of polyploid cells in regeneration and cancer, we hypothesized that zoledronic acid (ZA), an osteoclast-targeting agent, might be used to eliminate PGCCs. The 5637-bladder cancer cell line was treated with various doses of cisplatin to enrich polyploid cells and the efficacy of different concentrations of ZA in reducing this population was assessed. The metabolic profile of PGCCs was investigated with gas chromatography-mass spectrometry. Lipid profiles, mitochondrial density, and ROS content were also measured to assess the response of the cells to ZA. Cancer cells surviving after three days of exposure with 6 µM cisplatin were mainly polyploid. These cells demonstrated special morphological features such as fusion with diploid or other polyploid cells and originated in daughter cells through budding. ZA could substantially eradicate PGCCs with the maximal effect observed with 50 µM which resulted in the drop of PGCC fraction from 60 ± 7.5 to 19 ± 1.7%. Enriched PGCCs after cisplatin-treatment demonstrated a drastic metabolic shift compared to untreated cancer cells with an augmentation of lipids. Further assays confirmed the high content of lipid droplets and cholesterol in these cells which were reduced after ZA administration. Additionally, the mitochondrial density and ROS increased in PGCCs both of which declined in response to ZA. Taken together, we propose that ZA is a potent inhibitor of PGCCs which alters the metabolism of PGCCs. Although this drug has been successfully exploited as adjuvant therapy for some malignancies, the current evidence on its effects on PGCCs justifies further trials to assess its potency for improving the success of current therapies for tackling tumor resistance and relapse.
Assuntos
Cisplatino , Neoplasias , Humanos , Cisplatino/farmacologia , Cisplatino/uso terapêutico , Cisplatino/metabolismo , Ácido Zoledrônico/farmacologia , Ácido Zoledrônico/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Linhagem Celular Tumoral , Células Gigantes/metabolismo , Poliploidia , Neoplasias/patologiaRESUMO
BACKGROUND: In a series of our preclinical studies, we have reported that conditioning of α/ß CD8+ T cells in vitro with interleukin-12 (IL-12) during their expansion improves their homing phenotype and anti-tumor cytolytic function upon their adoptive transfer in vivo. Vγ9+Vδ2+ T cells can also be expanded in vitro with amino bisphosphonates such as zoledronate (ZOL) for the purpose of adoptive therapy. AIM: We aimed in this study to use IL-12 to enhance the expansion and cytotoxic functions of ZOL-expanded Vγ9+Vδ2+T cells. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMCs) were separated from healthy donors and stage II breast cancer patients. PBMCs (1 × 106 cells/mL) were cultured and treated with ZOL/IL2, ZOL/IL2/IL12, or IL2/IL12. Cultured cells were harvested on days 7 and 14 of culture and their numbers, phenotype, and cytolytic activity were assessed. The levels of pro- and inflammatory cytokines/chemokines in the plasma and supernatants of the cultured cells were analyzed by Luminex. RESULTS: In healthy subjects, the addition of IL-12 to ZOL/IL2-stimulated PBMCs increased the expansion and the cytotoxic activity of Vγ9+Vδ2+ T cells on days 7 and 14 of culture. The latter was measured by the expression level of the cytolytic molecules granzyme B (GZB) and perforin (PER). Of note, αß CD8 + T cells were also activated under the same condition but with a lesser extent addition of IL-12 to ZOL/IL2-stimulated PBMCs from cancer patients also induced similar effects but were lower than in control subjects. Interestingly, ZOL/IL2/IL12-treated PBMCs showed higher levels of cytokines/chemokines, in particular, CCL, CCL4, GM-CSF, IL-1rα; IL-12, IL-13, TNF, and IFNγ measured on days 7 and 14. CONCLUSION: The addition of IL12 at the start of the expansion protocol can enhance the activity of γδ T cells which might be mediated in part by the activation of αß T cells.
Assuntos
Antineoplásicos , Neoplasias , Ácido Zoledrônico/farmacologia , Ácido Zoledrônico/metabolismo , Interleucina-12/metabolismo , Interleucina-2/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Leucócitos Mononucleares/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Imidazóis/farmacologia , Antineoplásicos/farmacologia , Citocinas/metabolismo , Células Cultivadas , Neoplasias/metabolismoRESUMO
Zoledronate (ZOL) were found to inhibit bone resorption in an animal model of diabetes, high glucose concentrations have been shown to decreased the osteogenesis-related gene expression. But the molecular mechanism by which high glucose levels affect osteoblasts and the effects of ZOL on osteoblast differentiation in a high-glucose environment remain unclear. Therefore, we aimed to investigate the effect of ZOL on osteoblast differentiation in a high-glucose environment and determine the responsible mechanism. Cell proliferation was detected by MTT assay, and cell differentiation was evaluated by immunofluorescence staining for alkaline phosphatase expression, alizarin red staining, cytoskeletal arrangement, and actin fiber formation. Real-time PCR and western blot analyses were performed to detect the mRNA and protein expression of p38MAPK, phosphorylated (p)-p38MAPK, CREB, p-CREB, collagen (COL) I, osteoprotegerin (OPG), and RANKL. The results showed that cell proliferation activity did not differ among the groups. But high glucose inhibited osteoblast differentiation; actin fiber formation; and p38MAPK, p-p38MAPK, CREB, p-CREB, COL I, and OPG expression, while promoting RANKL expression. However, we found that treatment with ZOL reversed these effects of high glucose. And further addition of a p38MAPK inhibitor led to inhibition of osteoblast differentiation and actin fiber formation, and lower p38MAPK, p-p38MAPK, CREB, p-CREB, COL I, and OPG expression than in the high glucose +ZOL group with higher RANKL expression than in the high glucose +ZOL group. Collectively, this study demonstrates that high glucose inhibits the differentiation of osteoblasts, and ZOL could partly overcome these effects by regulating p38MAPK pathway activity.
Assuntos
Osteogênese , Proteínas Quinases p38 Ativadas por Mitógeno , Animais , Ácido Zoledrônico/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Actinas/metabolismo , Diferenciação Celular , Osteoblastos/metabolismo , Osteoprotegerina/metabolismo , Glucose/metabolismo , Ligante RANK/metabolismoRESUMO
Osteoclasts are specialized cells that degrade and resorb bone. Bisphosphonates (BPs) are drugs with well-known capacity to inhibit the resorption of mineralized tissues. Nitrogen-containing BPs, like alendronate (ALN) and zoledronic acid (ZA), inactivate osteoclast activity mostly by alterations on the cytoskeleton architecture of the cell. In this study, we used an in vitro model to test the hypothesis that bisphosphonates may have inhibitory effects on the osteoclastogenesis and osteoclast activity after the therapy was discontinued. Primary osteoclasts were generated from mouse bone marrow in media supplemented with 1,25-dihydroxyvitamin D3 and cultivated over bones pre-treated with ALN and ZA. The pre-saturation of the bone slices with bisphosphonates did not affect cell viability. We found, however, that by disrupting the gene expression of RANKL and OPG the osteoclastogenesis and resorption activity of osteoclasts was significantly disturbed. These inhibitory effects were confirmed by scanning electron microscopy resorption assay, assessment of osteoclast ultrastructure, and by gene expression analysis of TRAP and Cathepsin K. In conclusion, ALN and ZA adhered to the bone matrix reduced the osteoclast activity in vitro.
Assuntos
Reabsorção Óssea , Osteogênese , Animais , Reabsorção Óssea/tratamento farmacológico , Reabsorção Óssea/metabolismo , Osso e Ossos/metabolismo , Difosfonatos/metabolismo , Difosfonatos/farmacologia , Camundongos , Osteoclastos/metabolismo , Ácido Zoledrônico/metabolismo , Ácido Zoledrônico/farmacologiaRESUMO
In this work, we investigated the in vitro and in vivo functions of bisphosphonate of zoledronate (Zd) in hypoxia/reoxygenation (H/R) injured human embryonic stem cell-derived cardiomyocytes (hES-CMs). In the in vitro setting, the effects of Zd on hES-CM survival and differentiation were examined. We found that low and medium concentrations (<2 µm) of Zd did not induce cell death of hES-CMs. 0.5 µm Zd protected H/R-induced hES-CM apoptosis but did not affect key differentiation proteins, including hcTnl, PECM-1 Cnx43 and Pan-Cadherin. In addition, Zd-induced TrkA/B phosphorylation and promoted VEGF to counter the apoptotic effect of H/R injury. In the in vivo animal model of myocardial infarction, Zd treatment promoted the survival of hES-CMs by inducing PECAM1 and hcTnl. Thus, we concluded that Zd protected H/R-induced hES-CM apoptosis in vitro and promoted hES-CM survival in vivo. These data may facilitate the development of human embryonic stem cells into clinical applications for patients with ischemic heart disease.
Assuntos
Células-Tronco Embrionárias Humanas , Traumatismo por Reperfusão Miocárdica , Animais , Apoptose , Difosfonatos/metabolismo , Difosfonatos/farmacologia , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Hipóxia/metabolismo , Traumatismo por Reperfusão Miocárdica/metabolismo , Miócitos Cardíacos , Transdução de Sinais , Ácido Zoledrônico/metabolismo , Ácido Zoledrônico/farmacologiaRESUMO
Targeting excessive osteoclast differentiation and activity is considered a valid therapeutic approach for osteoporosis. Zoledronic acid (ZOL) plays a pivotal role in regulating bone mineral density. However, the exact molecular mechanisms responsible for the inhibitory effects of ZOL on receptor activator of nuclear factor (NF)κB ligand (RANKL)induced osteoclast formation are not entirely clear. The present study aimed to investigate the role of ZOL in osteoclast differentiation and function, and to determine whether NFκB and mitogenactivated protein kinase, and their downstream signalling pathways, are involved in this process. RAW264.7 cells were cultured with RANKL for differentiation into osteoclasts, in either the presence or absence of ZOL. Osteoclast formation was observed by tartrateresistant acid phosphatase staining and bone resorption pit assays using dentine slices. The expression of osteoclastspecific molecules was analysed using reverse transcriptionquantitative polymerase chain reaction and western blotting assays to deduce the molecular mechanisms underlying the role of ZOL in osteoclastogenesis. The results showed that ZOL significantly attenuated osteoclastogenesis and bone resorptive capacity in vitro. ZOL also suppressed the activation of NFκB and the phosphorylation of cJun Nterminal kinase. Furthermore, it inhibited the expression of the downstream factors cJun, cFos and nuclear factor of activated T cells c1, thereby decreasing the expression of dendritic cellspecific transmembrane protein and other osteoclastspecific markers. In conclusion, ZOL may have therapeutic potential for osteoporosis.
Assuntos
Reabsorção Óssea/tratamento farmacológico , Reabsorção Óssea/metabolismo , MAP Quinase Quinase 4/metabolismo , NF-kappa B/metabolismo , Osteogênese/efeitos dos fármacos , Ácido Zoledrônico/metabolismo , Ácido Zoledrônico/farmacologia , Animais , Densidade Óssea/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Regulação da Expressão Gênica , Camundongos , Fatores de Transcrição NFATC/metabolismo , Fosforilação , Ligante RANK/metabolismo , Células RAW 264.7 , Transdução de SinaisRESUMO
BACKGROUND Concerns have been raised among clinicians and patients about the cardiovascular risks of bisphosphonates used in the treatment of osteoporosis. The goal of this study was to investigate the acute effect of zoledronic acid (ZA) infusion on arrhythmia development using an electrocardiograph (ECG). MATERIAL AND METHODS This prospective study was a self-controlled case series study that recruited 116 female patients with osteoporosis. The patients underwent standard 12-lead electrocardiography before and 1 day after zoledronic acid intravenous infusion to evaluate cardiac adverse effects and the change in ECG parameters after the infusion. Heart rhythm, atrial and ventricular premature contractions, atrial fibrillation, P wave, and QTc parameters were measured using an ECG. A blood biochemical examination was performed for all patients before the ZA infusion. Body temperature was measured twice per day. RESULTS Before ZA administration, ECG findings were normal in 47 patients and abnormal in 69 patients. After ZA administration, ECG findings were normal in 35 patients and abnormal in 81 patients. New onsets of premature atrial contractions and atrial fibrillation were observed in 1 patient each, and new onsets of premature ventricular contractions were observed in 2 patients. The heart rate was obviously higher, and the QT interval was obviously shorter after ZA administration, compared with before administration. No significant differences in P wave and QTc parameters were found between the 2 ECG measurements. CONCLUSIONS During the acute phase, 116 women with osteoporosis who were treated with zoledronic acid infusion did not develop significantly abnormal ECG changes.
Assuntos
Arritmias Cardíacas/etiologia , Ácido Zoledrônico/efeitos adversos , Ácido Zoledrônico/uso terapêutico , Doença Aguda , Idoso , Arritmias Cardíacas/fisiopatologia , Povo Asiático , Fibrilação Atrial/fisiopatologia , Difosfonatos/efeitos adversos , Eletrocardiografia/métodos , Feminino , Átrios do Coração/fisiopatologia , Frequência Cardíaca/efeitos dos fármacos , Humanos , Pessoa de Meia-Idade , Monitorização Fisiológica , Osteoporose/tratamento farmacológico , Estudos Prospectivos , Ácido Zoledrônico/metabolismoRESUMO
Bisphosphonates (BPs), especially zoledronic acid (ZOL), are clinically used to treat osteolytic bone lesions. However, serious side-effects may be also induced during the therapeutic process. To improve the BPs drugs, here, we investigated the effects of a series of ZOL derivatives with increasing number of methylene linker between the imidazole ring and the P-C-P backbone named IPrDP, IBDP, IPeDP, and IHDP on cell viability and receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation, function and apoptosis induction in mouse bone marrow-derived macrophages (BMMs). Our results suggested that IPeDP and IHDP, which contains 4 and 5 methylene linkers, respectively, exerted lower toxicity on BMMs compared with ZOL, IPrDP, and IBDP, which contains 1, 2, and 3 methylene linkers respectively. At concentrations below cytotoxicity threshold, IPeDP and IHDP possessed strong abilities of antiosteoclast formation, antibone absorption, and inducing osteoclast apoptosis, which were similar to ZOL and more powerful than IPrDP and IBDP. The mechanism behind these effects of IPeDP and IHDP might involve the interference of small GTPases prenylation through suppression of mevalonate pathway. The downregulation of JNK and Akt phosphorylation and subsequent inhibition of the expression of c-Fos and NFATc1 might also be involved. Our results supported the potential usage of IPeDP and IHDP to treat bone-related disorders involving increased osteoclastogenesis. Our attempt to extend the methylene linker between the imidazole ring and the P-C-P backbone of ZOL also reveals some regularities between the structure and properties of the BPs drugs.
Assuntos
Osteoclastos/metabolismo , Ácido Zoledrônico/farmacologia , Animais , Células da Medula Óssea/citologia , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Difosfonatos/metabolismo , Difosfonatos/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Fatores de Transcrição NFATC/metabolismo , Osteoclastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ligante RANK/metabolismo , Transdução de Sinais/efeitos dos fármacos , Ácido Zoledrônico/análogos & derivados , Ácido Zoledrônico/metabolismoRESUMO
Zoledronic acid (ZA) is an FDA-approved drug and a third-generation bisphosphonate (BPs). A systematic evaluation of the mechanisms of ZA has not previously been performed. In this study, validated targets of ZA were screened using PubChem, Herbal Ingredients' Targets Database (HIT), Binding Database (BindingDB), and ChemBank, and potential targets of ZA were identified based on structural characteristics of ligands and proteins. The candidate targets were then assessed using GeneMANIA, Gene Ontology (GO), and pathway analysis, and molecule-target-GO-pathway networks were visualized using Cytoscape. Nine validated targets and 26 potential targets were obtained. The networks generated via this analysis showed that the candidate targets were associated with cell proliferation and metabolism as well as other biological processes (BP) and pathways. In general, ZA appeared to play crucial roles in multiple functions, including metabolism, regulation of vascular smooth muscle cell proliferation, and chemical carcinogenesis; a great deal of additional research must be performed. Moreover, the current study showed that it is feasible to analyze the mechanisms of ZA via target prediction, which facilitates systematic pharmacological evaluation.
Assuntos
Algoritmos , Ácido Zoledrônico/química , Ontologia Genética , Ligantes , Ácido Zoledrônico/metabolismoRESUMO
A method for the extraction and quantification of zoledronic acid (ZA) from human bone was set up and validated. This method allowed the quantification of ZA from jawbone sequestrations of patients affected by bisphosphonate-related osteonecrosis of the jaw (BRONJ) associated with ZA treatment. The analyte was extracted from the bone tissues with phosphoric acid and derivatized using trimethylsilyl diazomethane (TMS-DAM). ZA tetramethyl phosphonate was then quantified by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), showing high accuracy, repeatability and selectivity. Lower limits of quantification and detection (LLOQ and LLQD) were 3.4 ng/mL and 1 ng/mg, respectively. This study fully described the analytical process for the determination of ZA in human bone sequestrations, representing a pivotal step for further biomedical research on ZA and BRONJ.
Assuntos
Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/metabolismo , Conservadores da Densidade Óssea/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Arcada Osseodentária/metabolismo , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem , Ácido Zoledrônico/metabolismo , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/diagnóstico , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/etiologia , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/cirurgia , Conservadores da Densidade Óssea/efeitos adversos , Feminino , Humanos , Arcada Osseodentária/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Ácido Zoledrônico/efeitos adversosRESUMO
Osteoporosis presents a challenge to the long-term success of osseointegration of endosseous implants. The bio-inspired 3,4-dihydroxy-L-phenylalanine (Dopa) coating is widely used as a basic layer to bind osteogenetic molecules that may improve osseointegration. To date, little attention has focused on application of Dopa alone or binding inhibitors of bone resorption in osteoporosis. Local use of a bisphosphonate such as zoledronic acid (ZA), an inhibitor of osteoclast-mediated bone resorption, has been proven to improve implant osseointegration. In this study, ovariectomized rats were divided into four groups and implanted with implants with different surface modifications: sandblasted and acid-etched (SLA), SLA modified with Dopa (SLA-Dopa), SLA modified with ZA (SLA-ZA), and SLA modified with Dopa and ZA (SLA-Dopa + ZA). Measurement of removal torque, micro-computed tomography and histology revealed a greater extent of bone formation around the three surface-modified implants than SLA-controls. No synergistic effect was observed for combined Dopa + ZA coating. Microarray analysis showed the Dopa coating inhibited expression of genes associated with osteoclast differentiation, similarly to the mechanism of action of ZA. Simple Dopa modification resulted in a similar improvement in osseointegration compared to ZA. Thus, our data suggest simple Dopa coating is promising strategy to promote osseointegration of implants in patients with osteoporosis.