• Journal of Veterinary Science
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• v.23 no.4
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• pp.47.1-47.11
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• 2022

Background: In lipopolysaccharide-induced RAW264.7 cells, Aster tataricus (AT) inhibits the nuclear factor kappa-light-chain-enhancer of activated B cells and MAPKs pathways and critical pathways of osteoclast development and bone resorption. Objectives: This study examined how aster saponin A2 (AS-A2) isolated from AT affects the processes and function of osteoclastogenesis induced by receptor activator of nuclear factor kappa-B ligand (RANKL) in RAW264.7 cells and bone marrow macrophages (BMMs). Methods: The cell viability, tartrate-resistant acid phosphatase staining, pit formation assay, polymerase chain reaction, and western blot were carried out to determine the effects of AS-A2 on osteoclastogenesis. Results: In RAW264.7 and BMMs, AS-A2 decreased RANKL-initiated osteoclast differentiation in a concentration-dependent manner. In AS-A2-treated cells, the phosphorylation of ERK1/2, JNK, and p38 protein expression were reduced considerably compared to the control cells. In RAW264.7 cells, AS-A2 suppressed the RANKL-induced activation of osteoclast-related genes. During osteoclast differentiation, AS-A2 suppressed the transcriptional and translational expression of NFATc1 and c-Fos. AS-A2 inhibited osteoclast development, reducing the size of the bone resorption pit area. Conclusion: AS-A2 isolated from AT appears to be a viable therapeutic therapy for osteolytic illnesses, such as osteoporosis, Paget's disease, and osteogenesis imperfecta.

• Journal of Periodontal and Implant Science
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• v.52 no.5
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• pp.383-393
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• 2022

Purpose: Aloe-emodin (AE), a natural anthraquinone abundant in aloe plants and rhubarb (Rheum rhabarbarum), has long been used to treat chronic inflammatory diseases. However, AE's underlying mechanisms in periodontal inflammation have not been fully elucidated. Acidic mammalian chitinase (AMCase) is a potential biomarker involved in bone remodeling. This study aimed to evaluate AE's effect on periodontitis in rats and investigate AMCase expression. Methods: Eighteen Sprague-Dawley rats were separated into the following groups: healthy (group 1), disease (group 2), vehicle (group 3), AE high-dose (group 4), and AE low-dose (group 5). Porphyromonas gingivalis ligatures were placed in rats (groups 2-5) for 7 days. Groups 4 and 5 were then treated with AE for an additional 14 days. Saliva was collected from all groups, and probing pocket depth was measured in succession. Periodontal pocket tissues were subjected to histomorphometric analysis after the rats were sacrificed. Bone marrow-derived macrophages and murine macrophages were stimulated with receptor activator of nuclear factor-κB ligand (RANKL) and treated with different concentrations of AE. AMCase expression was detected from the analysis of saliva, periodontal pocket tissues, and differentiated osteoclasts. Results: Among rats with P. gingivalis-induced periodontitis, the alveolar bone resorption levels and periodontal pocket depth were significantly reduced after treatment with AE. AMCase protein expression was significantly higher in the disease group than in the healthy control (P<0.05). However, AE inhibited periodontal inflammation by downregulating AMCase expression in saliva and periodontal pocket tissue. AE significantly reduced RANKL-stimulated osteoclastogenesis by modulating AMCase (P<0.05). Conclusions: AE decreases alveolar bone loss and periodontal inflammation, suggesting that this natural anthraquinone has potential value as a novel therapeutic agent against periodontal disease.

• Journal of Physiology & Pathology in Korean Medicine
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• v.35 no.3
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• pp.87-96
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• 2021

The purpose of this study was to investigate the effect of single herbal medicines and Korean herbal medicine complex prescriptions for osteoporosis in Korea. We targeted studies published in domestic journals from January 1, 2013 to April 1, 2021. For domestic databases, RISS, NDSL, OASIS, DBpia, KISS, KISTI, and Korean Traditional Knowledge Portal were used. Words such as "osteoporosis", "herbal medicine", "herbal combination prescription" and "herbal" were combined and searched. A total of 32 studies were selected. Among them, 8 studies were conducted exclusively for animal studies, 18 studies were conducted on cell experiments, and 6 studies were conducted on both animal and cell experiments. There were 18 studies using a single herb and 14 studies using a combination of Herbal medicine. As a result of the investigation of a single herb, it was found that 補陽藥 was most widely used, and it was found that it was mainly used in the herbal combination prescription, mainly for 補陽藥, 補氣藥, and 補陰藥. Studies in animal experiments revealed significant results mainly in bone mineral density, serological indicators, and morphological indicators of bone tissue. In the study of cell experiments, the effects were found in factors related to the differentiation of osteoclasts and inhibitors of inflammation. Through the results of this study, we are able to confirm the effective single herbal medicines and herbal medicine complex prescriptions for the treatment of osteoporosis, and it is expected that they will contribute to clinical treatment.

• Journal of the Korean Chemical Society
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• v.67 no.2
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• pp.120-128
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• 2023

Osteoporosis is a disease that causes the weakening of bone by increasing porosity, which often results in fractures. Osteoporosis treatment measures include the use of Bisphosphonates and estrogen. However, these treatments cannot be used in the long term as these treatments have adverse side effects. Therefore, there is a need to identify better and safer treatment options. For this, 63 plant extracts were screened and among them, six extracts showed high anti-osteoclastic activity with low cytotoxicity. Of these six extracts, three extracts, Cudrania tricuspidata (P371), Ulmus davidiana var. japonica (P401), and Torilis japonica (P411), showed more than 50 percent osteoclast inhibition. While the remaining, Stewartia pseudocamellia extracts I and II (P370, P397) and Cuscuta chinensis (P418), showed moderate or between 40-50 percent osteoclast inhibition. Among all the extracts, Torilis japonica (P411) showed the highest inhibitory action against osteoclast development. Torilis japonica (P411) primary components include Kaempferol, Quercetin, and Luteolin, all proven to inhibit osteoclastogenesis. Stewartia pseudocamellia extracts I and II (P370 and P397) showed moderate or 44% osteoclast inhibition. Stewartia pseudocamellia extract II (P397) enhanced the growth of RAW 264.7 cells by 19%. Torilis japonica (P411) and Stewartia pseudocamellia extract II (P397) suppressed the expression of osteoclast-specific genes in RANKL-induced osteoclastogenesis in RAW 246.7 cells. Torilis japonica (P411) extracts even increased osteoblast-specific RUNX2 gene expression. This results provide that six extracts could be used as a potential treatment option for osteoporosis disease with the extracts of Torilis japonica (P411) and Stewartia pseudocamellia (P397) as an ideal candidates. However, the combination of the extract with higher osteoclastic inhibition and less toxic effects with further analysis should be recommended.

• Restorative Dentistry and Endodontics
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• v.48 no.2
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• pp.21.1-21.15
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• 2023

Objectives: This study evaluated the effects of Biodentine (BD), Bio-C Repair (BCR), and mineral trioxide aggregate (MTA) plug on the fracture resistance of simulated immature teeth with replacement root resorption (RRR) and in vitro-induced osteoclastogenesis. Materials and Methods: Sixty bovine incisors simulating immature teeth and RRR were divided into 5 groups: BD and BCR groups, with samples completely filled with the respective materials; MTA group, which utilized a 3-mm apical MTA plug; RRR group, which received no root canal filling; and normal periodontal ligament (PL) group, which had no RRR and no root canal filling. All the teeth underwent cycling loading, and compression strength testing was performed using a universal testing machine. RAW 264.7 macrophages were treated with 1:16 extracts of BD, BCR, and MTA containing receptor activator of nuclear factor-kappa B ligand (RANKL) for 5 days. RANKL-induced osteoclast differentiation was assessed by staining with tartrate-resistant acid phosphatase. The fracture load and osteoclast number were analyzed using 1-way ANOVA and Tukey's test (α = 0.05). Results: No significant difference in fracture resistance was observed among the groups (p > 0.05). All materials similarly inhibited osteoclastogenesis (p > 0.05), except for BCR, which led to a lower percentage of osteoclasts than did MTA (p < 0.0001). Conclusions: The treatment options for non-vital immature teeth with RRR did not strengthen the teeth and promoted a similar resistance to fractures in all cases. BD, MTA, and BCR showed inhibitory effects on osteoclast differentiation, with BCR yielding improved results compared to the other materials.

• Journal of Periodontal and Implant Science
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• v.53 no.1
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• pp.20-37
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• 2023

Purpose: Our pilot study showed that a 3-dimensional dual drug delivery scaffold (DDDS) loaded with Chinese herbs significantly increased the regenerated bone volume fraction. This study aimed to confirm the synergistic anti-inflammatory and osteogenic preclinical effects of this system. Methods: The targets and pathways of parthenolide and naringin were predicted. Three cell models were used to assess the anti-inflammatory effects of parthenolide and the osteogenic effects of naringin. First, the distance between the cementoenamel junction and alveolar bone crest (CEJ-ABC) and the bone mineral density (BMD) of surgical defects were measured in a rat model of periodontitis with periodontal fenestration defects. Additionally, the mRNA expression levels of matrix metallopeptidase 9 (MMP9) and alkaline phosphatase (ALP) were measured. Furthermore, the number of inflammatory cells and osteoclasts, as well as the protein expression levels of tumor necrosis factor-alpha (TNF-α) and levels of ALP were determined. Results: Target prediction suggested prostaglandin peroxidase synthase (PTGS2) as a potential target of parthenolide, while cytochrome P450 family 19 subfamily A1 (CYP19A1) and taste 2 receptor member 31 (TAS2R31) were potential targets of naringin. Parthenolide mainly targeted inflammation-related pathways, while naringin participated in steroid hormone synthesis and taste transduction. In vitro experiments revealed significant antiinflammatory effects of parthenolide on RAW264.7 cells, and significant osteogenic effects of naringin on bone marrow mesenchymal stem cells and MC3T3-E1 cells. DDDS loaded with parthenolide and naringin decreased the CEJ-ABC distance and increased BMD and ALP levels in a time-dependent manner. Inflammation was significantly alleviated after 14 days of DDDS treatment. Additionally, after 56 days, the DDDS group exhibited the highest BMD and ALP levels. Conclusions: DDDS loaded with parthenolide and naringin in a rat model achieved significant synergistic anti-inflammatory and osteogenic effects, providing powerful preclinical evidence.

• Journal of dental hygiene science
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• v.23 no.2
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• pp.169-175
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• 2023

Background: Although microbial infection is direct cause of periodontal disease, various environmental factors influence the disease severity. Aging is considered a risk factor for oral diseases, with the prevalence of periodontal diseases increasing with age. Moreover, senescence-associated secretory phenotype (SASP) expressed in age-related diseases is a key marker of chronic inflammation and aging phenotypes. Therefore, this study aimed to understand the relevance of senescent cells to periodontal health and disease, investigate the possibility of regulating the expression of aging- and osteolysis-related factors in gingival fibroblasts, and investigate the effect of senescence induction in gingival fibroblasts on osteoclast differentiation in mouse bone marrow-derived macrophages (BMMs). Methods: After stimulation with 400 nM hydrogen peroxidase, human gingival fibroblasts (HGFs) were examined for senescence-associated β-galactosidase. Western blot and enzyme-linked immunosorbent assays were performed to assess the expression of SASP. Osteoclast formation was assessed in BMMs using a conditioned medium (CM) from hydrogen peroxide-stimulated HGFs. Osteoclastic differentiation was investigated using tartrate-resistant acid phosphatase (TRAP) staining and activity. Data analysis was performed using SPSS version 25.0. Results: The expression of senescence-related molecules, including p53, p16, and p21, and the expression of osteolytic factors, including IL-6, IL-8, and IL-17, were found to be significantly higher in the hydrogen peroxide-stimulated HGF than in the control group. Regarding the indirect effects of senescent gingival cells, the number of osteoclasts and TRAP activity increased according to the differentiation of BMM cultured in CM. Conclusion: Our results on the of between osteolytic factors and cellular senescence in gingival fibroblast cells helped to reveal evidence of pathological aging mechanisms. Furthermore, our results suggest that the development of novel therapies that target specific SASP factors could be an effective treatment strategy for periodontal disease.

• BMB Reports
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• v.56 no.10
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• pp.545-550
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• 2023

Osteoporosis is a major public health concern, which requires novel therapeutic strategies to prevent or mitigate bone loss. Natural compounds have attracted attention as potential therapeutic agents due to their safety and efficacy. In this study, we investigated the regulatory activities of boeravinone B (BOB), a natural rotenoid isolated from the medicinal plant Boerhavia diffusa, on the differentiation of osteoclasts and mesenchymal stem cells (MSCs), the two main cell components responsible for bone remodeling. We found that BOB inhibited osteoclast differentiation and function, as determined by TRAP staining and pit formation assay, with no significant cytotoxicity. Furthermore, our results showing that BOB ameliorates ovariectomy-induced bone loss demonstrated that BOB is also effective in vivo. BOB exerted its inhibitory effects on osteoclastogenesis by downregulating the RANKL/RANK signaling pathways, including NF-κB, MAPK, and PI3K/Akt, resulting in the suppression of osteoclast-specific gene expression. Further experiments revealed that, at least phenomenologically, BOB promotes osteoblast differentiation of bone marrow-derived MSCs but inhibits their differentiation into adipocytes. In conclusion, our study demonstrates that BOB inhibits osteoclastogenesis and promotes osteoblastogenesis in vitro by regulating various signaling pathways. These findings suggest that BOB has potential value as a novel therapeutic agent for the prevention and treatment of osteoporosis.

• Journal of Microbiology and Biotechnology
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• v.33 no.9
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• pp.1197-1205
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• 2023

Osteoporosis, Greek for "porous bone," is a bone disease characterized by a decrease in bone strength, microarchitectural changes in the bone tissues, and an increased risk of fracture. An imbalance of bone resorption and bone formation may lead to chronic metabolic diseases such as osteoporosis. Wolfiporia extensa, known as "Bokryung" in Korea, is a fungus belonging to the family Polyporaceae and has been used as a therapeutic food against various diseases. Medicinal mushrooms, mycelium and fungi, possess approximately 130 medicinal functions, including antitumor, immunomodulating, antibacterial, hepatoprotective, and antidiabetic effects, and are therefore used to improve human health. In this study, we used osteoclast and osteoblast cell cultures treated with Wolfiporia extensa mycelium water extract (WEMWE) and investigated the effect of the fungus on bone homeostasis. Subsequently, we assessed its capacity to modulate both osteoblast and osteoclast differentiation by performing osteogenic and anti-osteoclastogenic activity assays. We observed that WEMWE increased BMP-2-stimulated osteogenesis by inducing Smad-Runx2 signal pathway axis. In addition, we found that WEMWE decreased RANKL-induced osteoclastogenesis by blocking c-Fos/NFATc1 via the inhibition of ERK and JNK phosphorylation. Our results show that WEMWE can prevent and treat bone metabolic diseases, including osteoporosis, by a biphasic activity that sustains bone homeostasis. Therefore, we suggest that WEMWE can be used as a preventive and therapeutic drug.

• Journal of Korean Medicine Rehabilitation
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• v.24 no.3
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• pp.29-37
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• 2014

Objectives Osteoporosis is an impending problem in the rapidly aging society. The purpose of this study is to identify the effects of Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) on osteoporosis induced by ovariectomy in rats. Methods 24 rats were randomly assigned to a SHAM group, a control group, and a Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) group (n=8). Ovaries were extracted and for 8 weeks, the rats were given dry feeds and Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) group were given a mixture of dry feeds and Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$). At 8 weeks, their body weight, femur mass, tibia ash per body ratio, overall surface area and thickness of the trabeculae, overall surface area of the osteoblasts, and the number of osteoclasts were measured and levels of albumin, aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total-cholesterol, calcium and estradiol were recorded. Results There was no significant difference in the weight of the femur and the tibia ash per body ratio. The Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) group had significantly thicker trabeculae than the control group and showed a minimal increase of overall surface area of the trabeculae. The overall surface area of the osteoblasts and the number of osteoclasts decreased in the Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) group. There were no statistically significant differences in AST, ALT, ALP, total cholesterol, phosphorus, and estradiol levels. On the contrary the Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) group had significantly higher levels of Albumin, triglyceride, and calcium. Conclusions It can be concluded that oral intake of Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) prevents the thinning process of the trabeculae. Thus, it may have positive effects on the treatment and prevention of osteoporosis in postmenopausal women.