eCAM Advance Access originally published online on April 27, 2007
eCAM 2007 4(2):225-232; doi:10.1093/ecam/nem037
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Ki-energy (Life-energy) Stimulates Osteoblastic Cells and Inhibits the Formation of Osteoclast-like Cells in Bone Cell Culture Models
1Philadelphia Biomedical Research Institute, King of Prussia, PA, USA, 2School of Nishino Breathing Method, Shibuya-ku, Tokyo, 3Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, Shizuoka, Japan and 4University of Pennsylvania School of Medicine, Philadelphia, PA, USA
Some practitioners of the Nishino Breathing Method (NBM) were found to have a higher bone density than the average values of age- and gender-matched non-practitioners. Using bone cell culture models, we investigated a possible mechanism behind this observation. For the study of bone mineralization, we performed the following two experiments using cultured osteoblastic MC3T3-E1 cells: (i) Kozo Nishino, a Japanese Ki expert, sent Ki-energy to the cells once for 5 or 10 min after they were seeded in culture dishes in the presence of 10% fetal bovine serum (FBS). They were incubated for 72 h and the cells were counted. The number in the dish with 10-min Ki-exposure was significantly greater than that in the control (P < 0.01 with n = 8). We performed a reverse transcription-polymerase chain reaction (RT–PCR) study using these cells, but the mRNA expressions did not change significantly. (ii) After cells were incubated for 72 h without Ki-exposure (in the presence of FBS), they were further cultured for 48 h (in the absence of FBS) to promote differentiation. At the beginning of the second culture stage, Ki was applied once for 10 min. After 48 h, RT–PCR was performed. The mRNA expressions which are related to bone mineralization, such as Runx2, 
1(I) collagen, alkaline phosphatase and osteocalcin, increased significantly (P < 0.05 and n = 4 for all). For the bone resorption study, we used mouse marrow cultures, which can form osteoclast-like cells in the presence of (1–34) parathyroid hormone (PTH), and stimulate resorption. We exposed these cells to Ki-energy twice for the duration of 5 or 10 min on day 0 and day 4. On day 7, the cells were counted. The number of osteoclast-like cells in dishes with Ki exposure was significantly smaller than those in control dishes (P < 0.05 with n = 5). The difference between 5-min exposure and 10-min exposure was not statistically significant. All of our data suggest that the Ki-effect on osteoporosis should be further explored.
Keywords: bone density – bone mineralization – bone resorption – Ki energy – MC3T3-E1 cells – mouse marrow culture – osteoblastic cells – osteoclast-like cells – osteoporosis – RT–PCR
For reprints and all correspondence: S. Tsuyoshi Ohnishi, Philadelphia Biomedical Research Institute, Suite 250, 100 Ross Road, King of Prussia, PA 19406, USA. Tel: 610-688-6276; Fax: 610-254-9332; E-mail: stohnishi{at}aol.com
Received October 17, 2006; accepted March 13, 2007
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