It is well known that extensive osteoclast formation plays a key role in osteoporosis in post‑menopausal women and the elderly. The suppression of extensive osteoclastogenesis and bone resorption may be an effective preventive strategy for osteoporosis. Zoledronic acid (ZOL) has been indicated to play an essential role in regulating bone mineral density and has already been used in large clinical trials. However, the effects of ZOL on osteoclastogenesis remain to be fully elucidated. Therefore, the present study aimed to determine the effects of ZOL on osteoclastogenesis, and to explore the corresponding signalling pathways. By using a cell viability assay, as well as in vitro osteoclastogenesis, immunofluorescence and resorption pit assays, we demonstrated that ZOL (0.1‑5 µM) suppressed receptor activator of nuclear factor‑κB ligand (RANKL)‑induced osteoclast differentiation and bone resorptive activity. Furthermore, western blot analysis and reverse transcription‑quantitative PCR indicated that ZOL inhibited the RANKL‑induced activation of NF‑κB and the phosphorylation of JNK in RAW264.7 cells, and subsequently decreased the expression of osteoclastogenesis‑associated genes, including calcitonin receptor, tartrate‑resistant acid phosphatase and dendritic cell‑specific transmembrane protein. ZOL inhibited osteoclast formation and resorption in vitro by specifically suppressing NF‑κB and JNK signalling. On the whole, the findings of this study indicate that ZOL may serve as a potential agent for the treatment of osteoclast‑associated diseases, including osteoporosis.

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 NF‑κB and mitogen‑activated 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 tartrate‑resistant acid phosphatase staining and bone resorption pit assays using dentine slices. The expression of osteoclast‑specific molecules was analysed using reverse transcription‑quantitative 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 NF‑κB and the phosphorylation of c‑Jun N‑terminal kinase. Furthermore, it inhibited the expression of the downstream factors c‑Jun, c‑Fos and nuclear factor of activated T cells c1, thereby decreasing the expression of dendritic cell‑specific transmembrane protein and other osteoclast‑specific markers. In conclusion, ZOL may have therapeutic potential for osteoporosis.

With the aging population and the popularity of implant prostheses, an increasing number of postmenopausal osteoporosis (PMOP) patients require implant restorations; however, poor bone condition affects the long-term stability of implant prostheses. This study aimed to investigate the therapeutic effect of quercetin (QR) compared with alendronate (ALN), the primary treatment for PMOP, on mandibular osteoporosis (OP) induced by ovariectomy (OVX) in female rats. Adult female rats were treated with QR (50 mg/kg/day), ALN (6.25 mg/kg/week) by gavage for 8 weeks, chloroquine (CQ, 10 mg/kg/twice a week), and cytokine release inhibitory drug 3 (MCC950, 10 mg/kg/three times a week) by intraperitoneal injection for 8 weeks after bilateral OVX. Blood samples were collected prior to euthanasia; the mandibles were harvested and subjected to micro-computed tomography (micro-CT) and pathological analysis. QR administration controlled weight gain and significantly improved the bone microstructure in OVX rats, increasing bone mass, and bone mineral density (BMD), reducing bone trabecular spacing, and decreasing osteoclast numbers. Western blotting, real-time quantitative PCR (RT-qPCR), and serum markers confirmed that QR inhibited interleukin- 1β (IL-1β) and interleukin-18 (IL-18) on the nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) protein 3 (NLRP3) pathway thereby inhibiting osteoclast differentiation, immunofluorescence and western blotting also confirmed that QR inhibited autophagy in OVX rats and suppressed the number of tartrate-resistant acid phosphatase (TRAP)-stained positive osteoclasts. The findings suggest that QR may protect the bone structure and prevent bone loss in osteoporotic rats by inhibiting the NLRP3 pathway and autophagy in osteoclasts with comparable effects to ALN, thus QR may have the potential to be a promising alternative supplement for the preventive and therapeutic treatment of PMOP.

Ai Tian, DDS/Jun-jiang Zhai, DDS/Yan Peng, DDS/Li Zhang, BDS/Min-hua Teng, DDS/Jian Liao, DDS/Xu Sun, DDS/Xing Liang, PhD, DDS: Purpose: Improving the peri-implant bone quality has notable clinical value in cases of osteoporosis following implant insertion. Materials and Methods: In this study, strontium ranelate?loaded chitosan film was produced on a titanium surface in different concentrations of strontium ranelate (SR) (0, 2, 20, 40, and 80 mmol/L of the strontium ion [Sr2+]) with the expectation of utilizing the bone healing?enhancement effect of Sr2+. The physicochemical properties of SR-loaded chitosan films were characterized by x-ray diffraction (XRD), scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR). The release/dissolution mechanism was tested by inductively coupled plasma optical emission spectrometry (ICP-OES), and in vitro cell responses were evaluated using primary osteoblasts (POBs) in terms of cell proliferation, alkaline phosphatase (ALP) activity, and quantitative analysis of key osteoblastic gene expression. Results: XRD and FTIR observation showed that only a few SRs reacted chemically with chitosan through hydrogen bonds or conjugate action. The initial burst release (70% to 85%) of Sr2+ occurred in the first 3 days and was followed by a slower release stage. At a low concentration (2 mmol/L or 20 mmol/L), SR-loaded chitosan film promoted cell responses of POBs with enhanced proliferation, ALP activity, and expression levels of bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (Runx-2), ALP, and osteocalcin, but at a high concentration (40 mmol/L or 80 mmol/L), it inhibited POB growth. Conclusion: These results indicated that SR-loaded chitosan film on a titanium surface promotes osteoblast proliferation and differentiation in a dose-dependent manner, which may represent a new approach in the treatment of a titanium implant.