||The study objective was to evaluate the effect of Biofield Treated vitamin D3 and DMEM on bone health. The Test Items (TI) were divided into two parts. One part of each sample was received Consciousness Energy Healing Treatment by Janice Patricia Kinney and those samples were labeled as Biofield Energy Treated (BT) samples, while other parts of each sample were denoted as Untreated Test items (UT). The cell viability showed test samples were found as safe in the tested concentrations. ALP was significantly increased by 212.57% and 134.55% in UT-DMEM + BT-TI and BT-DMEM + BT-TI groups, respectively at 50 µg/mL, while 242.77% increased in BT-DMEM + UT-TI at 10 µg/mL compared to UT-DMEM + UT-TI group. Collagen was significantly increased by 172.04%, 293.55%, and 248.39% in UT-DMEM + BT-TI, BT-DMEM + UT-TI, and BT-DMEM + BT-TI groups, respectively at 0.1 µg/mL compared to untreated group. Moreover, collagen was significantly increased by 99.25% and 172.93% in UT-DMEM + BT-TI and BT-DMEM + BT-TI groups, respectively at 1µg/mL, while increased by 156.38% and 145.64% in UT-DMEM + BT-TI and BT-DMEM + BT-TI groups, respectively at 10 µg/mL than untreated group. Besides, percent of bone mineralization was distinctly increased by 198.62%, 111.72%, and 168.28% in UT-DMEM + BT-TI, BT-DMEM + UT-TI, and BT-DMEM + BT-TI groups, respectively at 0.1µg/mL compared to untreated group. Overall, Biofield Treated vitamin D3 was remarkably improved the bone health parameters and could be beneficial against bone-related disorders (osteoporosis, osteomalacia etc.), stress, inflammatory and autoimmune disorders.