# Therapeutic Potential of Chrysin in Improving Bone Health
## 4 Discussion
In addition, the diminished availability of serum biochemical estimations of essential minerals such as calcium, magnesium, and phosphorus witnessed in animals with insufficient vitamin D (group II) can also be analogous to the decreased hardness in the 4 th lumbar and 8 th thoracic vertebrae, suggesting a deterioration in bone mass that corroborates previous studies. Comparatively, other therapeutic groups showed amplified bone strength concerning increased 4 th lumbar and 8 th thoracic vertebral hardness. A comparable effect in bone strength was observed with increased serum calcium, magnesium, and phosphorus in earlier reports [42]. Previous research revealed the imperative link between bone mineralization and bone ash content in bone-related diseases. In this regard, the vitamin-D-deficient diet control group showed decreased bone ash content, i.e., calcium, phosphorus, and magnesium, which confirms decreased bone mineral deposits [43]. Nevertheless, all the treatment groups showed enhanced calcium content in bone ash, which can be attributed to the increased serum calcium levels. None of the therapeutic interventions caused any increase in magnesium deposits in the bones. However, chrysin, when given in combination with vitamin D\({}_{3}\) and CaCO\({}_{3}\) caused a prominent increase in the phosphorus content in bone ash. The role of CaCO\({}_{3}\) and vitamin D in increasing bone mineralization has been established in earlier studies [8]. Femur bone mineral density (BMD) is one of the gold standard tests for the determination of the mineral strength of bone and was assessed by performing DEXA [30]. The animals treated with a vitamin-D-deficient diet showed decreased bone mineral density compared with the normal group, which indicates decreased bone calcification, mineralization, and bone mass. Significantly, animals in different treatment groups showed intense bone mineral density, which supports the results obtained from biochemical and bone ash estimations [30]. To estimate the inhibitory effect of chrysin on vitamin D metabolism and its effect on low-dose vitamin D\({}_{3}\) (40 IU) supplementation, the estimation of serum 25-OH-D\({}_{3}\) levels was considered a reliable marker for vitamin D status [44]. From our experimental analysis, the diet control group showed decreased serum 25-OH-D\({}_{3}\). In contrast, increased serum 25-OH-D\({}_{3}\) levels were obtained in the animals treated with different therapeutic interventions, reinforcing its physiological effect on bone associated with the serum biochemical parameters discussed above. Chrysin, when given alone, effectively elevated 25-OH-D\({}_{3}\) levels, but the combination of chrysin with CaCO\({}_{3}\) and vitamin D\({}_{3}\) has shown a major impact in increasing the serum 25-OH-D\({}_{3}\) levels, which partly confirms the metabolic interactions of chrysin on vitamin D catabolism, even at a low doses of vitamin D\({}_{3}\) (40 IU). The histopathological data also revealed a greater number of bone-forming cells in the epiphyseal plate of the animals treated with different therapeutic interventions, which also favors the data obtained from other biochemical parameters. However, an uneven distribution of osteocytes in the bone matrix was spotted in animals along with a smaller number of bone-forming cells in the epiphyseal plate among the diet control group (group II). Bone specimens resemble decreased bone formation and can be attributed to hypovitaminosis D, as mentioned earlier [45].