Zoledronic acid promotes alveolar bone formation in ovariectomized rats

doi:10.12307/2023.518

Abstract

Abstract: BACKGROUND: Zoledronic acid mainly acts on the formation of osteoclasts and also inhibits osteoblast apoptosis, proliferation, and differentiation. However, the effect and mechanism of zoledronic acid on alveolar bone formation in ovariectomized rats are still controversial.
OBJECTIVE: To explore the osteogenic effect of zoledronic acid on alveolar bone of ovariectomized rats via alveolar nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/cysteinyl-aspartate protease-1 (Caspase-1)/interleukin-1β pathway.
METHODS: Totally 36 female Sprague-Dawley rats were randomly divided into control group, model group, and zoledronic acid group, with 12 rats in each group. Bilateral oophorectomy was used to establish an animal model of osteoporosis in the latter two groups. The zoledronic acid group was subcutaneously given 20 μg/kg zoledronic acid at 12 weeks after modeling, while the control and model groups were injected with the same dose of normal saline. All animals were anesthetized after 4-week drug intervention to collect serum samples from the abdominal aorta and alveolar bone samples. Serum alkaline phosphatase, osteocalcin and interleukin-1β levels were detected by ELISA. Hematoxylin-eosin staining was used to observe the pathological changes of rat alveolar bone. TUNEL staining was used to detect the apoptosis of osteoblasts in rat alveolar bone. The formation of alveolar bone was detected by calcein fluorescent labeling. Western blot assay was used to detect the levels of NLPR3, Caspase-1 and interleukin-1β in alveolar bone tissue as well the protein expression of Runt related transcription factor 2, alkaline phosphatase, and osteocalcin.
RESULTS AND CONCLUSION: Compared with the model group, the expression level of alkaline phosphatase in rat serum decreased in the zoledronic acid group (P < 0.01), while the expression level of osteocalcin increased significantly (P < 0.05). Compared with the model group, the pathological structure of alveolar bone in the zoledronic acid group was improved. The number of TUNEL positive cells in the zoledronic acid group was significantly lower than that in the model group (P < 0.001). The mineralization deposition rate and new bone formation rate of trabecular bone in the zoledronic acid group were higher than those in the model group (P < 0.05). (5) Western blot results showed that compared with the control group, NLRP3, Caspase-1, interleukin-1β proteins was highly expressed in the model group (P < 0.05), while the expressions of Runt related transcription factor 2, alkaline phosphatase, and osteocalcin proteins were decreased (P < 0.05). However, treatment with zoledronic acid decreased the protein expression of NLRP3, Caspase-1, and interleukin-1β (P < 0.05), but increased the protein expression of Runt related transcription factor 2, alkaline phosphatase, and osteocalcin (P < 0.05). The above results show that zoledronic acid can affect the expression of osteogenic related factors and inflammatory factors in serum to improve bone metabolism and inflammatory response, enhance the differentiation function of osteoblasts, and delay the formation of osteoporosis. Zoledronic acid can improve alveolar bone structure, reduce alveolar bone osteoblast apoptosis and the protein expression of NLRP3, Caspase-1, and interleukin-1β, increase the mineralization deposition rate and new bone formation rate of alveolar bone trabecula and the protein expression of Runt related transcription factor 2, alkaline phosphatase, and osteocalcin, which may be related to the inhibition of NLRP3/Caspase-1/interleukin-1β signaling pathway.

Key words: zoledronic acid, ovariectomized rat, alveolar bone, NLRP3 , inflammasome, osteoblast

CLC Number:

Song Na, Liu Guanjuan, Cheng Yuting, Xiong Yue, Luo Shanshan, Hong Wei, Liao Jian. Zoledronic acid promotes alveolar bone formation in ovariectomized rats[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(28): 4494-4501.

Figures/Tables 5

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