Bone disorders are largely associated with increased morbidity, mortality, and substantial economic and health costs. Vitamin D play an important role for calcium absorption and bone mineralization, which can improve the patients' quality of life. The current study aimed to evaluate the potential of The Trivedi Effect®- Biofield Energy Healing on vitamin D3 and DMEM as test item (TI) on bone cell differentiation using human osteoblast cell line (MG-63, Osteosarcoma). Bone health biomarkers such as alkaline phosphatase enzyme (ALP) activity, collagen levels and bone mineralization were evaluated. The test items were treated with The Trivedi Effect® by William Dean Plikerd and divided as Biofield Energy Treated (BT) and untreated (UT) test items. Cell viability using MTT data showed that the test items were found to be safe. ALP level was significantly increased by 346.4% (at 50 µg/mL), 375.3% (at 100 µg/mL), and 343.2% (at 100 µg/mL) in the UT-DMEM+BT-TI, BT-DMEM+UT-TI, and BT-DMEM+BT-TI groups, respectively as compared to the untreated group. Collagen content was significantly increased by 336.2% and 237.2% in the UT-DMEM+BT-TI at 10 and 50 µg/mL, respectively while 399.3% (10 µg/mL) and 200% (0.1 µg/mL) increased collagen in the BT-DMEM+UT-TI and BT-DMEM+BT-TI groups, respectively. Moreover, the percent of bone mineralization was significantly increased in UT-DMEM+BT-TI, BT-DMEM+UT-TI, and BT-DMEM+BT-TI groups by 416.9% (10 µg/mL), 460.7% (0.1 µg/mL), and 441.7% (10 µg/mL) respectively as compared with the untreated test item and DMEM group. Thus, Biofield Energy Treated vitamin D3 and DMEM would play an important role to control the osteoblast function, improves bone mineralization, and calcium absorption in many bone disorders. Moreover, the bone health parameters such as collagen, calcium and ALP were significantly improved and can be used as supplement to improve bone health. Based on the outstanding results, it is assumed that the Biofield Energy Treated vitamin D3 could be a powerful alternative dietary sources and supplements to fight against various bone related diseases including low bone density and osteoporosis, osteogenesis imperfecta, Paget’s disease of bone, rickets, osteomalacia, bone and/or joint pain, increased frequency of fractures, deformed bones, osteoma, chondrodystrophia fetalis, hormonal imbalance, stress, aging, bone loss and fractures.