The aim of the present study was to evaluate the significant role of Consciousness Energy Healing based vitamin D3 and DMEM medium on bone health parameters in vitro using MG-63 cells such as alkaline phosphatase enzyme (ALP) activity, collagen levels and bone mineralization. The test items (TI) i.e. vitamin D3 and DMEM medium were divided into two parts. The test samples received Consciousness Energy Healing Treatment by Laura Nelson Streicher and samples were defined as the Biofield Energy Treated (BT) samples, while the other parts of each sample were denoted as the untreated test items (UT). MTT assay for cell viability showed that increased range 76% to 139% cell viability with safe and nontoxic profile of test samples on MG-63 cell line. ALP was significantly increased by 232.7% (1 µg/mL), 156.4% (100 µg/mL) and 262.5% (100 µg/mL) in the UT-DMEM+BT-TI, BT-DMEM+UT-TI, and BT-DMEM+BT-TI groups, respectively as compared with the untreated group. The level of collagen was significantly increased by 145.8% and 151.6% at 10 and 50 µg/mL, respectively in the UT-DMEM+BT-TI group, while 177.1%, 235.6%, and 165.3% at 1, 10, and 50 µg/mL, respectively in the BT-DMEM+UT-TI group as compared with the untreated group. In addition, BT-DMEM+BT-TI group showed a significant increased collagen level by 252.1% at 1 µg/mL as compared with the untreated test item and DMEM group.The percent of bone mineralization was significantly increased by 92.8%, 65.8%, and 32.2% at 0.1, 1, and 10 µg/mL, respectively in the UT-DMEM+BT-TI group, while 76.8%, 132.5%, and 7.6% at 0.1, 1, and 10 µg/mL, respectively in the BT-DMEM+UT-TI group as compared with the untreated group. In addition, BT-DMEM+BT-TI group showed a significant increased bone mineralization by 92.8%, 38.6%, and 12.3% at 0.1, 1, and 10 µg/mL, respectively as compared with the untreated group. Biofield Energy Treatment might be vital in promotion and maintenance of strong and healthy bones and quality of life by assisting them in maintaining optimal vitamin D levels. It regulates the osteoblast function, improvesbone mineralization, and calcium absorption in wide range of bone disordersalong with wide range of adverse health conditions, comprising cancer and certain autoimmune diseases. https://www.trivedieffect.com/science/a-prospective-study-on-bone-health-effect-of-biofield-energy-treated-vitamin-d3-using-mg-63-cell-line http://www.springjournals.net/gjfst/springjournals.netgjfstarticlesindex=2streicheretal