The objective was to investigate the potential of Biofield Treated vitamin D3 and DMEM on bone markers. The test items (TIs) were separated into two parts. One part of each sample was received Consciousness Energy Treatment by Alice Branton and termed as Biofield Treated (BT) samples, while other parts of each sample denoted as untreated TI (UT). MTT data found TIs were safe and nontoxic at tested concentrations. ALP was significantly elevated by 390% and 200% in UT-DMEM + BT-TI at 0.1 and 1µg/mL, respectively; while, increased by 135.29% in BT-DMEM + BT-TI at 0.1µg/mL than UT-DMEM + UT-TI. ALP was significantly increased by 106.34%, 138.47%, and 212.68% in UT-DMEM + BT-TI, BT-DMEM + UT-TI, and BT-DMEM + BT-TI, respectively at 10µg/mL than untreated. Collagen was significantly increased by 288.68% and 106.96% in UT-DMEM + BT-TI and BT-DMEM + BT-TI, respectively at 10µg/mL than untreated. Further, collagen was significantly increased by 435.78% and 139.72% in UT-DMEM + BT-TI and BT-DMEM + UT-TI, respectively at 50µg/mL than untreated. Percent of bone mineralization was significantly increased by 299.13%, 239.91%, and 218.77% in BT-DMEM + BT-TI at 0.1, 1, and 10µg/mL, respectively than untreated. Besides, percent of bone mineralization was distinctly increased by 192.51% in BT-DMEM + UT-TI at 10µg/mL than untreated. Overall, Biofield Treated vitamin D3 remarkably improved bone parameters in MG-63 cells and could be an alternative to vitamin D3 deficiency and used various bone disorders viz. low bone density, osteoporosis, autoimmune and inflammatory diseases, stress management and prevention, and anti-aging by improving overall health.