## 4 Discussion
Eutropin (0.2 mg/kg), which was used as the positive control in this experiment, is a growth hormone agent proven for its biological efficacy and safety, and it is used to treat growth hormone deficiencies. Seven experimental groups of eight mice were administered AIN-76A feed, and for six of these groups, varying concentrations of each extract were added to the feed. Bone growth and resorption are regulated by osteoblasts and osteoclasts, and bone turnover occurs through repeated bone formation and resorption. Moreover, osteocyte differentiation is closely linked to bone matrix protein expression, and its markers include ALP, pro-collagen I alpha 1, osteocalcin, and calcium. In our study, all the administered aqueous and 30% ethanolic extracts of CB stimulated bone growth, with significantly higher stimulations observed in the mice administered high extract concentrations. However, only a trend of increased bone growth was observed for osteoclastin. Furthermore, the normal group and calcium- and vitamin D-deficient diet groups significantly differed in all markers, with the exception of calcium, indicating that the calcium- and vitamin D-deficient diet induced bone growth suppression. Calcium is generally present in abundance in the bones, and a small amount is present in the blood, body fluids, and muscles. Calcium is regulated by PTH, and PTH increases blood calcium concentration by increasing the amount of calcium released into the blood pool. While calcium is released from the bones to the blood by osteoclasts, the same amount of calcium is simultaneously transported from the blood into the bone matrix by osteoblasts. Although the calcium- and vitamin D-deficient diet groups did not show a significant difference in calcium concentration compared to the normal group, these groups tended to have a lower calcium concentration. These results suggest that blood calcium concentration is influenced by repeated bone remodeling processes involving bone formation and resorption. BMD is the basis of BMC and is a marker of bone health. Compared to the normal group, the calcium- and vitamin D-deficient diet group had significantly reduced BMD and BMC in the femur and tibia, and administering high concentrations of aqueous and ethanolic extracts significantly increased these values. The growth plate is the site of longitudinal bone growth, which occurs through the process of chondrocyte proliferation and enlargement through cell division in soft cartilage tissues and osteogenic differentiation. Both the aqueous and ethanolic extracts of CB significantly lengthened the growth plate in the tibia compared to the control group. According to [PERSON] et al. (2019), CB significantly increased the length of the femur and tibia in a growing rat model. While this study used a different solvent, our results were consistent with those of [8]. GHs are known to induce cellular proliferation by directly acting on osteoblasts. In general, the GH increases IGF-1 production in the liver, but considering that the amount of IGF-1 is separately regulated by the GH, it is believed that the GH is not an absolute determinant of local IGF-1 production in the bones and that various factors, such as estrogen, PTH, and cortisol, are involved in the regulation of IGF-1 [11,23]. Previous studies have reported that the GH induces IGF-1 expression in osteoblasts; however, whether the GH stimulates the IGF-1 expression in human osteoblasts remains unknown. IGF-1 activity in the bone is determined by various insulin-like growth factor binding proteins (IGFBPs) and IGF-1 concentrations, and IGFBP-3, IGFBP-4, and IGFBP-5 are generally involved in this process. In this study, we did not examine the osteogenic mechanisms of CB because [PERSON] et al. (2019) confirmed that _A. fistulosum_ stimulates cell growth through TGF-\(\beta\) and Wnt signaling in osteoblasts [8].