The effects of eggshell calcium (Biomin H® ) and its combinations with alfacalcidol (1α-hydroxyvitamin D3) and menaquinone-7 (vitamin K2) on ovariectomy-induced bone loss in a rat model of osteoporosis

Abstract

The purpose of this study was to investigate the impact of eggshell calcium (Biomin H^® dietary supplement) and its combinations with alfacalcidol (1α-hydroxyvitamin D₃ ) and menaquinone-7 (vitamin K₂ ) on ovariectomy-induced bone loss in rats. Adult female rats (n = 48) were divided into 6 groups of 8 individuals each: sham-operated rats (SHAM); ovariectomized (OVX) rats untreated; OVX rats treated with Biomin H^® (BIO); OVX rats simultaneously receiving Biomin H^® , vitamin D₃ (BIO + D₃ ); OVX rats simultaneously treated with Biomin H^® , vitamin K₂ (BIO + K₂ ) and OVX rats treated with Biomin H^® , vitamin D₃ , vitamin K₂ (BIO + D₃ + K₂ ) during 8 weeks. Biochemical parameters, bone mineral density (BMD), bone mineral content (BMC) and femoral bone microstructure were determined. Plasma calcium and phosphate were increased in BIO + D₃ and BIO + D₃ + K₂ groups as compared to OVX. Alkaline phosphatase was elevated in OVX, BIO versus SHAM, BIO + D₃ + K2 groups. When compared to OVX group, decreased urine deoxypyridinoline was observed in all treated groups and femoral BMD, BMC were higher in BIO, BIO + D₃ , BIO + D₃ + K₂ groups. The BIO + K₂ rats had similar densitometrical values than OVX individuals. Microcomputed tomography revealed increased trabecular relative bone volume (due to an increase in trabecular number) in BIO + D₃ , BIO + D₃ + K₂ as compared to OVX. The higher relative bone volume in BIO + D₃ , BIO + D₃ + K₂ groups was also accompanied by an increase in bone surface. In the cortical bone, an enhanced periosteal bone apposition was identified in BIO, BIO + D₃ , BIO + K₂ , BIO + D₃ + K₂ groups. The rats from BIO + D₃ + K₂ group had a higher area of primary osteon's vascular canals. In BIO + D₃ , BIO + K₂ , BIO + D₃ + K₂ groups, an increased area of secondary osteons was determined in comparison with OVX. Our results indicate the beneficial effect of triple application of Biomin H^® , vitamin D₃ , vitamin K₂ , as well as simultaneous administration of Biomin H^® , vitamin D₃ on the inhibition of ovariectomy-induced bone loss in a rat model of osteoporosis.

Keywords: Biomin H®; eggshell calcium; osteoporosis; ovariectomized rats; vitamin D3; vitamin K2.