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Patent
Neochemir Inc., Kobe University and Co2Be Medical Engineering K.K. | Date: 2012-07-13

The present invention provides a prophylactic or therapeutic agent for fracture and bone diseases as well as a bone growth promoting agent containing carbon dioxide as an active ingredient. Further, according to the present invention, an additive or synergistic effect for preventing or treating fracture and bone diseases as well as for promoting bone growth by combining the therapeutic agent of the present invention and a bone absorption inhibitor and/or a bone formation promoter is obtained.


Patent
Neochemir Inc., Kobe University and Co2Be Medical Engineering K.K. | Date: 2013-09-25

A liquid medicine of the present invention comprising liquid having carbon dioxide dissolved therein, said liquid being administered by using means of liquid injection, and a method of treatment of the present invention using a liquid medicine comprising liquid having carbon dioxide dissolved therein, said liquid being administered by using means of liquid injection can reduce or eliminate tumors with few side effects. When the liquid medicine of the present invention comprising liquid having carbon dioxide dissolved therein, said liquid being administered by using means of liquid injection, and the method of treatment of the present invention using a liquid medicine comprising liquid having carbon dioxide dissolved therein, said liquid being administered by using means of liquid injection are used in combination with surgical therapy, chemotherapy, radiotherapy, or immunotherapy of tumors, effects can be enhanced or side effects can be reduced compared with monotherapy or multidisciplinary therapy thereof.


Onishi Y.,Kobe University | Ueha T.,NeoChemir Inc. | Kawamoto T.,Kobe University | Hara H.,Kobe University | And 5 more authors.
Scientific Reports | Year: 2014

A number of studies have reported that decreased mitochondrial numbers are linked with neoplastic transformation and/or tumor progression, including resistance to apoptosis. Peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) is a multi-functional transcriptional coactivator that regulates the activities of multiple nuclear receptors and transcriptional factors involved in mitochondrial biogenesis. In this study, we observed that the number of mitochondria in sarcoma tissues, such as osteosarcoma and malignant fibrous histiocytoma, is significantly lower than that in normal muscle tissue or benign tumors, and that increasing the number of mitochondria by PGC-1α overexpression induces mitochondrial apoptosis in human sarcoma cell lines. The findings suggest that decreased mitochondrial numbers may contribute to musculoskeletal tumor progression, and that regulation of mitochondrial numbers by PGC-1α could be a potent therapeutic tool for human malignancies. © 2014 Macmillan Publishers Limited. Source


Onishi Y.,Kobe University | Kawamoto T.,Kobe University | Ueha T.,NeoChemir Inc. | Kishimoto K.,Kobe University | And 9 more authors.
PLoS ONE | Year: 2012

Mitochondria play an essential role in cellular energy metabolism and apoptosis. Previous studies have demonstrated that decreased mitochondrial biogenesis is associated with cancer progression. In mitochondrial biogenesis, peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) regulates the activities of multiple nuclear receptors and transcription factors involved in mitochondrial proliferation. Previously, we showed that overexpression of PGC-1α leads to mitochondrial proliferation and induces apoptosis in human malignant fibrous histiocytoma (MFH) cells in vitro. We also demonstrated that transcutaneous application of carbon dioxide (CO2) to rat skeletal muscle induces PGC-1α expression and causes an increase in mitochondrial proliferation. In this study, we utilized a murine model of human MFH to determine the effect of transcutaneous CO2 exposure on PGC-1α expression, mitochondrial proliferation and cellular apoptosis. PGC-1α expression was evaluated by quantitative real-time PCR, while mitochondrial proliferation was assessed by immunofluorescence staining and the relative copy number of mitochondrial DNA (mtDNA) was assessed by real-time PCR. Immunofluorescence staining and DNA fragmentation assays were used to examine mitochondrial apoptosis. We also evaluated the expression of mitochondrial apoptosis related proteins, such as caspases, cytochorome c and Bax, by immunoblot analysis. We show that transcutaneous application of CO2 induces PGC-1α expression, and increases mitochondrial proliferation and apoptosis of tumor cells, significantly reducing tumor volume. Proteins involved in the mitochondrial apoptotic cascade, including caspase 3 and caspase 9, were elevated in CO2 treated tumors compared to control. We also observed an enrichment of cytochrome c in the cytoplasmic fraction and Bax protein in the mitochondrial fraction of CO2 treated tumors, highlighting the involvement of mitochondria in apoptosis. These data indicate that transcutaneous application of CO2 may represent a novel therapeutic tool in the treatment of human MFH. © 2012 Onishi et al. Source


Uefuji A.,Kobe University | Matsumoto T.,Kobe University | Matsushita T.,Kobe University | Ueha T.,NeoChemir Inc. | And 3 more authors.
American Journal of Sports Medicine | Year: 2014

Background: The anterior cruciate ligament (ACL) does not heal spontaneously after injury, and patients of different ages respond differently to treatment. CD34+ stem/progenitor cells derived from the ACL remnant and associated tissues contribute to tendon-bone healing, but the relationship between age and the ACL's healing potential has not been clarified. Hypothesis: The ACL remnant and associated tissues from adolescent patients have more CD34+ cells, and this population of cells from younger patients exhibits a higher potential for proliferation and differentiation in vitro. Study Design: Descriptive laboratory study. Methods: Ruptured ACL remnants and associated tissues were harvested from 28 patients (mean age, 24.6 ± 1.6 years) who had undergone primary arthroscopic ACL reconstruction. Patients were divided into 3 patient groups by age: 10-19 years (teens group; n = 10), 20-29 years (20s group; n = 10), and ≥30 years (30s group; n = 8). The ACL remnant cells were characterized using fluorescence-activated cell sorting (FACS). Expansion potential was evaluated using population doubling (PD), and multi-lineage differentiation potential was assessed and compared. Results: The FACS analysis showed numerous CD34+ cells in the teens group compared with the 30s group (mean, 25.4% ± 7.9% vs 16.9% ± 3.9%, respectively; P = .044). The PD results indicated that the teens group had a significantly higher expansion potential than the 30s group at passage 3 (mean, 3.3 ± 0.2 vs 2.8 ± 0.2, respectively; P = .039). Young ACL remnant cells had a higher potential for osteogenic differentiation according to alkaline phosphatase activity (teens group, 169.5 ± 37.9 × 10 ng/mL vs 30s group, 64.9 ± 14.6 × 10 ng/mL; P = .029) and osteocalcin gene expression (teens group, 1.0 ± 0.25 vs 30s group, 0.39 ± 0.01; P = .01). In addition, the teens group displayed a higher differentiation potential to angiogenic lineages (acetylated low-density lipoprotein/Ulex europaeus lectin-stained cell counts) than other groups (teens group, 15.9 ± 1.9 vs 20s group, 8.9 ± 1.3 [P = .04]; teens group, 15.9 ± 1.9 vs 30s group, 7.2 ± 1.5 [P = .008]) and also tube length (teens group, 6939 ± 470 |xm vs 30s group, 4119 ± 507 |xm; P = .009). Conclusion: The ACL remnants of adolescent patients had more CD34+ cells, and those cells had a higher potential for proliferation and multilineage differentiation in vitro. Clinical Relevance: During remnant-preserving or remnant-transplanted ACL reconstruction, surgeons should consider the patient's age when predicting the healing potential. © 2014 The Author(s). Source

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