Intake of hydrogen water by tube feeding for patients with DECUBITAL WOUNDSScientific Research
Hydrogen water intake via tube-feeding for patients with pressure ulcer and its reconstructive effects on normal human skin cells in vitro
Abstract
Background
Decubitus ulcers (PU) are common in elderly patients with reduced mobility, and several studies have investigated prevention and treatment methods but have not found sufficient efficacy. The aim of this study was to elucidate the clinical efficacy of hydrogen-dissolved water (HW) ingestion via tube feeding (TF) on wound healing in decubitus patients. Furthermore, normal human dermal fibroblasts OUMS-36 and normal human epidermis-derived cell line HaCaT keratinocytes were studied in vitro to investigate the relevant mechanism of whether hydrogen plays a role in wound healing at the cellular level.
Methods
This study included 22 severely hospitalized elderly Japanese patients with pressure ulcers, aged 71.0 to 101.0 (86.7 ± 8.2) years, 12 males and 10 females, all with eating disorders and bedridden syndrome. Sequelae of various underlying diseases. All patients received usual care for pressure ulcers, combined with 600 mL of daily TF intake of HW rather than partial hydration. On the other hand, HW is produced with a hydrogen bubbler, which produces HW with a dissolved hydrogen concentration (DH) of 0.8-1.3 ppm and a redox range of -602 mV to -583 mV compared to ultrapure reverse osmosis water Potential (ORP) (RW) as reference, DH was < 0.018 ppm, ORP was +184 mV for in vitro experimental studies. In in vitro experiments, OUMS-36 fibroblasts and HaCaT keratinocytes were each cultured in medium prepared with HW and/or RW.Immunostaining was used to detect type I collagen remodeling in OUMS-36 cells. The intracellular reactive oxygen species (ROS) were quantified by NBT method, and the cell viability of HaCaT cells was detected by WST-1 method.
Results
Twenty-two patients were retrospectively divided into the effective group (EG, n=12) and the ineffective group (LG, n=10) according to the endpoint assessment results and healing criteria. The PU hospitalization days were significantly shorter in EG than in LG (113.3 days vs. 155.4 days, p < 0.05), with a reduction rate of approximately 28.1%. In both EG and LG, the reduced change in wound size (EG: 91.4%; LG: 48.6%) was significantly different from before HW ingestion (p<0.05, p<0.001). In vitro data showed that intracellular ROS, quantified by the NBT assay, was decreased by HW but not by RW in ultraviolet A (UVA)-irradiated HaCaT cells. Nuclear condensation and fragmentation occurred in UVA-irradiated HaCaT cells in RW, but hardly in HW, as shown by Hoechst 33342 staining. Furthermore, under UVA irradiation, the mitochondrial reducing capacity of HaCaT cells or the construction of collagen type I in OUMS-36 cells deteriorated in RW-generated medium, as shown by WST-1 assay or immunostaining, but preserved in HW in the resulting medium.
Conclusions
HW uptake via TF has been shown to induce wound size reduction and early recovery in severely hospitalized elderly patients with PU due to type I collagen construction in dermal fibroblasts or by immunostaining or NBT as shown to promote mitochondrial reduction capacity and ROS inhibition or WST-1 detection in epidermal keratinocytes, respectively.
References
Edlich RF, Winters KL, Woodard CR, Buschbacher RM, Long WB, Gebhart JH, Ma EK: Pressure ulcer prevention. J Long Term Eff Med Implants. 2004, 14 (4): 285-304. 10.1615/JLongTermEffMedImplants.v14.i4.20.
Maklebust J: Interrupting the pressure ulcer cycle. Nurs Clin North Am. 1999, 34 (4): 861-871.
European Pressure Ulcer Advisory Panel and National Pressure Ulcer Advisory Panel: Prevention and treatment of pressure ulcers: quick reference guide. 2009, Washington DC: National Pressure Ulcer Advisory Panel
Upton Z, Wallace HJ, Shooter GK, van Lonkhuyzen DR, Yeoh-Ellerton S, Rayment EA, Fleming JM, Broszczak D, Queen D, Sibbald RG, Leavesley DI, Stacey MC: Human pilot studies reveal the potential of a vitronectin: growth factor complex as a treatment for chronic wounds. Int Wound J. 2011, 8 (5): 522-532. 10.1111/j.1742-481X.2011.00859.x.
Lim CP, Phan TT, Lim IJ, Cao X: Cytokine profiling and Stat3 phosphorylation in epithelial-mesenchymal interactions between keloid keratinocytes and fibroblasts. J Invest Dermatol. 2009, 129 (4): 851-861. 10.1038/jid.2008.337.
Kato S, Saitoh Y, Iwai K, Miwa N: Hydrogen-rich electrolyzed warm water represses wrinkle formation against UVA ray together with type-I collagen production and oxidative-stress diminishment in fibroblasts and cell-injury prevention in keratinocytes. J Photochem Photobiol B. 2012, 106: 24-33.
Watanabe S, Saitoh Y, Namba M, Miwa N: Administration with telomeric DNA telomere-like oligonucleotides induces enhancement of telomerase activity and resistance against oxidative stress in telomere reverse transcriptase gene-transfected human fibroblasts. Biomed Pharmacother. 2010, 64 (8): 565-571. 10.1016/j.biopha.2010.02.005.
Yokoo S, Furumoto K, Hiyama E, Miwa N: Slow-down of age-dependent telomere shortening is executed in human skin keratinocytes by hormesis-like-effects of trace hydrogen peroxide or by anti-oxidative effects of pro-vitamin C in common concurrently with reduction of intracellular oxidative stress. J Cell Biochem. 2004, 93 (3): 588-597. 10.1002/jcb.20208.
Qiang S, Kawamura T, Masutani K, Peng X, Qing S, Stolz DB, Pribis JP, Billiar TR, Sun X, Bermudez CA, Toyoda Y, Nakao A: Oral intake of hydrogen-rich water inhibits intimal hyperplasia in arterialized vein grafts in rats. Cardiovasc Res. 2012, 94 (1): 144-153. 10.1093/cvr/cvs024.
Noda K, Tanaka Y, Shigemura N, Kawamura T, Wang Y, Masutani K, Sun X, Toyoda Y, Bermudez CA, Nakao A: Hydrogen-supplemented drinking water protects cardiac allografts from inflammation-associated deterioration. Transpl Int. 2012, 25 (12): 1213-1222. 10.1111/j.1432-2277.2012.01542.x.
Kajiyama S, Hasegawa G, Asano M, Hosoda H, Fukui M, Nakamura N, Kitawaki J, Imai S, Nakano K, Ohta M, Adachi T, Obayashi H, Yoshikawa T: Supplementation of hydrogen-rich water improves lipid and glucose metabolism in patients with type 2 diabetes or impaired glucose tolerance. Nutr Res. 2008, 28 (3): 137-143. 10.1016/j.nutres.2008.01.008.
Ishibashi T, Sato B, Rikitake M, Seo T, Kurokawa R, Hara Y, Naritomi Y, Hara H, Nagao T: Consumption of water containing a high concentration of molecular hydrogen reduces oxidative stress and disease activity in patients with rheumatoid arthritis: an open-label pilot study. Med Gas Res. 2012, 2 (1): 27-10.1186/2045-9912-2-27.
Ohsawa I, Nishimaki K, Yamagata K, Ishikawa M, Ohta S: Consumption of hydrogen water prevents atherosclerosis in apolipoprotein E knockout mice. Biochem Biophys Res Commun. 2008, 377 (4): 1195-1198. 10.1016/j.bbrc.2008.10.156.
Fujita K, Seike T, Yutsudo N, Ohno M, Yamada H, Yamaguchi H, Sakumi K, Yamakawa Y, Kido MA, Takaki A, Katafuchi T, Tanaka Y, Nakabeppu Y, Noda M: Hydrogen in drinking water reduces dopaminergic neuronal loss in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson’s disease. PLoS One. 2009, 4 (9): e7247-10.1371/journal.pone.0007247.
Ohsawa I, Ishikawa M, Takahashi K, Watanabe M, Nishimaki K, Yamagata K, Katsura K, Katayama Y, Asoh S, Ohta S: Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med. 2007, 13 (6): 688-694. 10.1038/nm1577.
Fukuda KI, Asoh S, Ishikawa M, Yamamoto Y, Ohsawa I, Ohta S: Inhalation of hydrogen gas suppresses hepatic injury caused by ischemia/reperfusion through reducing oxidative stress. Biochem Biophys Res Commun. 2007, 361 (3): 670-674. 10.1016/j.bbrc.2007.07.088.
Wood KC, Gladwin MT: The hydrogen highway to reperfusion therapy. Nat Med. 2007, 13 (6): 673-674. 10.1038/nm0607-673.
The actual state investigative committee of 9th Japanese society of pressure ulcers: Investigation of the current clinical practice on pressure ulcers for its prevalence rate. Med Trib. 2007, 10: 11-in Japanese
Saitoh Y, Yoshimura Y, Nakano K, Miwa N: Platinum nanocolloid-supplemented hydrogen-dissolved water inhibits growth of human tongue carcinoma cells preferentially over normal cells. Exp Oncol. 2009, 31 (3): 156-62.
Saitoh Y, Okayasu H, Xiao L, Harata Y, Miwa N: Neutral pH hydrogen-enriched electrolyzed water achieves tumor-preferential clonal growth inhibition over normal cells and tumor invasion inhibition concurrently with intracellular oxidant repression. Oncol Res. 2008, 17 (6): 247-255. 10.3727/096504008786991620.
Saitoh Y, Harata Y, Mizuhashi F, Nakajima M, Miwa N: Biological safety of neutral-pH hydrogen-enriched electrolyzed water upon mutagenicity, genotoxicity and subchronic oral toxicity. Toxicol Ind Health. 2010, 26 (4): 203-216. 10.1177/0748233710362989.
Asada R, Kageyama K, Tanaka H, Matsui H, Kimura M, Saitoh Y, Miwa N: Antitumor effects of nano-bubble hydrogen-dissolved water are enhanced by coexistent platinum colloid and the combined hyperthermia with apoptosis-like cell death. Oncol Rep. 2010, 24 (6): 1463-1470.
Kato S, Hokama R, Okayasu H, Saitoh Y, Iwai K, Miwa N: Colloidal platinum in hydrogen-rich water exhibits radical-scavenging activity and improves blood fluidity. J Nanosci Nanotechnol. 2012, 12 (5): 4019-4027. 10.1166/jnn.2012.6163.
Sanada H: “Pressure Ulcer: Assessment/care guide”. 2009, Nakayama Shoten Co. Ltd, 127-In Japanese, ISBN 978-4-521-73171-12
Wicks G: A guide to the treatment of pressure ulcers from grade 1-grade 4. Wound Essent. 2007, 2: 106-112.
Zeller JL: Pressure ulcers. JAMA. 2006, 296 (8): 23-30.
Matsui Y, Furue M, Sanada H, Tachibana T, Nakayama T, Sugama J, Furuta K, Tachi M, Tokunaga K, Miyachi Y: Development of the DESIGN-R with an observational study: an absolute evaluation tool for monitoring pressure ulcer wound healing. Wound Repair Regen. 2011, 19 (3): 309-315. 10.1111/j.1524-475X.2011.00674.x.
Sanada H, Iizaka S, Matsui Y, Furue M, Tachibana T, Nakayama T, Sugama J, Furuta K, Tachi M, Tokunaga K, Miyachi Y: Clinical wound assessment using DESIGN-R total score can predict pressure ulcer healing: pooled analysis from two multicenter cohort studies. Wound Repair Regen. 2011, 19 (5): 559-567. 10.1111/j.1524-475X.2011.00719.x.
Chuai YH, Sun XJ, Cai J: Biology of hydrogen and its medical applications. Act Biophys Sin. 2012, 28 (9): 705-718. (in Chinese)
Kajiya M, Silva MJ, Sato K, Ouhara K, Kawai T: Hydrogen mediates suppression of colon inflammation induced by dextran sodium sulfate. Biochem Biophys Res Commun. 2009, 386 (1): 11-15. 10.1016/j.bbrc.2009.05.117.
Nakashima-Kamimura N, Mori T, Ohsawa I, Asoh S, Ohta S: Molecular hydrogen alleviates nephrotoxicity induced by an anti-cancer drug cisplatin without compromising anti-tumor activity in mice. Cancer Chemother Pharmacol. 2009, 64 (4): 753-761. 10.1007/s00280-008-0924-2.
Cardinal JS, Zhan JH, Wang YN, Sugimoto R, Tsung A, McCurry KR, Billiar TR, Nakao A: Oral hydrogen water prevents chronic allograft nephropathy in rats: Hydrogen water prevented CAN. Kidney Int. 2010, 77: 101-109. 10.1038/ki.2009.421.
Foschi D, Trabucchi E, Musazzi M, Castoldi L, Di Mattia D, Radaelli E, Marazzi M, Franzini P, Berlusconi A: The effects of oxygen free radicals on wound healing. Int J Tissue React. 1988, 10 (6): 373-379.
Boron WF: Sharpey-Schafer lecture: gas channels. Exp Physiol. 2010, 95 (12): 1107-1130. 10.1113/expphysiol.2010.055244.
Fuchs Y, Steller H: Programmed cell death in animal development and disease. Cell. 2011, 147 (4): 742-758. 10.1016/j.cell.2011.10.033.
Bergmann A, Steller H: Apoptosis, stem cells, and tissue regeneration. Sci Signal. 2010, 3 (145): re8-10.1126/scisignal.3145re8.
Cai JM, Kang ZM, Liu WW, Luo X, Qiang S, Zhang JH, Ohta SG, Sun XJ, Xu WG, Tao HY, Li RP: Hydrogen therapy reduces apoptosis in neonatal hypoxia-ischemia rat model. Neurosci Lett. 2008, 441 (2): 167-172. 10.1016/j.neulet.2008.05.077.
Saitoh Y, Miyanishi A, Mizuno H, Kato S, Aoshima H, Kokubo K, Miwa N: Super-highly hydroxylated fullerene derivative protects human keratinocytes from UV-induced cell injuries together with the decreases in intracellular ROS generation and DNA damages. J Photochem Photobiol B. 2011, 102 (1): 69-76. 10.1016/j.jphotobiol.2010.09.006.
Hoerter JD, Ward CS, Bale KD, Gizachew AN, Graham R, Reynolds J, Ward ME, Choi C, Kagabo JL, Sauer M, Kuipers T, Hotchkiss T, Banner N, Chellson RA, Ohaeri T, Gant L, Vanderhill L: Effect of UVA fluence rate on indicators of oxidative stress in human dermal fibroblasts. Int J Biol Sci. 2008, 4 (2): 63-70.
DOI: 10.1186
Published on: 20131009
Hydrogen water intake via tube-feeding for patients with pressure ulcer and its reconstructive effects on normal human skin cells in vitro
Abstract
Background
Decubitus ulcers (PU) are common in elderly patients with reduced mobility, and several studies have investigated prevention and treatment methods but have not found sufficient efficacy. The aim of this study was to elucidate the clinical efficacy of hydrogen-dissolved water (HW) ingestion via tube feeding (TF) on wound healing in decubitus patients. Furthermore, normal human dermal fibroblasts OUMS-36 and normal human epidermis-derived cell line HaCaT keratinocytes were studied in vitro to investigate the relevant mechanism of whether hydrogen plays a role in wound healing at the cellular level.
Methods
This study included 22 severely hospitalized elderly Japanese patients with pressure ulcers, aged 71.0 to 101.0 (86.7 ± 8.2) years, 12 males and 10 females, all with eating disorders and bedridden syndrome. Sequelae of various underlying diseases. All patients received usual care for pressure ulcers, combined with 600 mL of daily TF intake of HW rather than partial hydration. On the other hand, HW is produced with a hydrogen bubbler, which produces HW with a dissolved hydrogen concentration (DH) of 0.8-1.3 ppm and a redox range of -602 mV to -583 mV compared to ultrapure reverse osmosis water Potential (ORP) (RW) as reference, DH was < 0.018 ppm, ORP was +184 mV for in vitro experimental studies. In in vitro experiments, OUMS-36 fibroblasts and HaCaT keratinocytes were each cultured in medium prepared with HW and/or RW.Immunostaining was used to detect type I collagen remodeling in OUMS-36 cells. The intracellular reactive oxygen species (ROS) were quantified by NBT method, and the cell viability of HaCaT cells was detected by WST-1 method.
Results
Twenty-two patients were retrospectively divided into the effective group (EG, n=12) and the ineffective group (LG, n=10) according to the endpoint assessment results and healing criteria. The PU hospitalization days were significantly shorter in EG than in LG (113.3 days vs. 155.4 days, p < 0.05), with a reduction rate of approximately 28.1%. In both EG and LG, the reduced change in wound size (EG: 91.4%; LG: 48.6%) was significantly different from before HW ingestion (p<0.05, p<0.001). In vitro data showed that intracellular ROS, quantified by the NBT assay, was decreased by HW but not by RW in ultraviolet A (UVA)-irradiated HaCaT cells. Nuclear condensation and fragmentation occurred in UVA-irradiated HaCaT cells in RW, but hardly in HW, as shown by Hoechst 33342 staining. Furthermore, under UVA irradiation, the mitochondrial reducing capacity of HaCaT cells or the construction of collagen type I in OUMS-36 cells deteriorated in RW-generated medium, as shown by WST-1 assay or immunostaining, but preserved in HW in the resulting medium.
Conclusions
HW uptake via TF has been shown to induce wound size reduction and early recovery in severely hospitalized elderly patients with PU due to type I collagen construction in dermal fibroblasts or by immunostaining or NBT as shown to promote mitochondrial reduction capacity and ROS inhibition or WST-1 detection in epidermal keratinocytes, respectively.
References
Edlich RF, Winters KL, Woodard CR, Buschbacher RM, Long WB, Gebhart JH, Ma EK: Pressure ulcer prevention. J Long Term Eff Med Implants. 2004, 14 (4): 285-304. 10.1615/JLongTermEffMedImplants.v14.i4.20.
Maklebust J: Interrupting the pressure ulcer cycle. Nurs Clin North Am. 1999, 34 (4): 861-871.
European Pressure Ulcer Advisory Panel and National Pressure Ulcer Advisory Panel: Prevention and treatment of pressure ulcers: quick reference guide. 2009, Washington DC: National Pressure Ulcer Advisory Panel
Upton Z, Wallace HJ, Shooter GK, van Lonkhuyzen DR, Yeoh-Ellerton S, Rayment EA, Fleming JM, Broszczak D, Queen D, Sibbald RG, Leavesley DI, Stacey MC: Human pilot studies reveal the potential of a vitronectin: growth factor complex as a treatment for chronic wounds. Int Wound J. 2011, 8 (5): 522-532. 10.1111/j.1742-481X.2011.00859.x.
Lim CP, Phan TT, Lim IJ, Cao X: Cytokine profiling and Stat3 phosphorylation in epithelial-mesenchymal interactions between keloid keratinocytes and fibroblasts. J Invest Dermatol. 2009, 129 (4): 851-861. 10.1038/jid.2008.337.
Kato S, Saitoh Y, Iwai K, Miwa N: Hydrogen-rich electrolyzed warm water represses wrinkle formation against UVA ray together with type-I collagen production and oxidative-stress diminishment in fibroblasts and cell-injury prevention in keratinocytes. J Photochem Photobiol B. 2012, 106: 24-33.
Watanabe S, Saitoh Y, Namba M, Miwa N: Administration with telomeric DNA telomere-like oligonucleotides induces enhancement of telomerase activity and resistance against oxidative stress in telomere reverse transcriptase gene-transfected human fibroblasts. Biomed Pharmacother. 2010, 64 (8): 565-571. 10.1016/j.biopha.2010.02.005.
Yokoo S, Furumoto K, Hiyama E, Miwa N: Slow-down of age-dependent telomere shortening is executed in human skin keratinocytes by hormesis-like-effects of trace hydrogen peroxide or by anti-oxidative effects of pro-vitamin C in common concurrently with reduction of intracellular oxidative stress. J Cell Biochem. 2004, 93 (3): 588-597. 10.1002/jcb.20208.
Qiang S, Kawamura T, Masutani K, Peng X, Qing S, Stolz DB, Pribis JP, Billiar TR, Sun X, Bermudez CA, Toyoda Y, Nakao A: Oral intake of hydrogen-rich water inhibits intimal hyperplasia in arterialized vein grafts in rats. Cardiovasc Res. 2012, 94 (1): 144-153. 10.1093/cvr/cvs024.
Noda K, Tanaka Y, Shigemura N, Kawamura T, Wang Y, Masutani K, Sun X, Toyoda Y, Bermudez CA, Nakao A: Hydrogen-supplemented drinking water protects cardiac allografts from inflammation-associated deterioration. Transpl Int. 2012, 25 (12): 1213-1222. 10.1111/j.1432-2277.2012.01542.x.
Kajiyama S, Hasegawa G, Asano M, Hosoda H, Fukui M, Nakamura N, Kitawaki J, Imai S, Nakano K, Ohta M, Adachi T, Obayashi H, Yoshikawa T: Supplementation of hydrogen-rich water improves lipid and glucose metabolism in patients with type 2 diabetes or impaired glucose tolerance. Nutr Res. 2008, 28 (3): 137-143. 10.1016/j.nutres.2008.01.008.
Ishibashi T, Sato B, Rikitake M, Seo T, Kurokawa R, Hara Y, Naritomi Y, Hara H, Nagao T: Consumption of water containing a high concentration of molecular hydrogen reduces oxidative stress and disease activity in patients with rheumatoid arthritis: an open-label pilot study. Med Gas Res. 2012, 2 (1): 27-10.1186/2045-9912-2-27.
Ohsawa I, Nishimaki K, Yamagata K, Ishikawa M, Ohta S: Consumption of hydrogen water prevents atherosclerosis in apolipoprotein E knockout mice. Biochem Biophys Res Commun. 2008, 377 (4): 1195-1198. 10.1016/j.bbrc.2008.10.156.
Fujita K, Seike T, Yutsudo N, Ohno M, Yamada H, Yamaguchi H, Sakumi K, Yamakawa Y, Kido MA, Takaki A, Katafuchi T, Tanaka Y, Nakabeppu Y, Noda M: Hydrogen in drinking water reduces dopaminergic neuronal loss in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson’s disease. PLoS One. 2009, 4 (9): e7247-10.1371/journal.pone.0007247.
Ohsawa I, Ishikawa M, Takahashi K, Watanabe M, Nishimaki K, Yamagata K, Katsura K, Katayama Y, Asoh S, Ohta S: Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med. 2007, 13 (6): 688-694. 10.1038/nm1577.
Fukuda KI, Asoh S, Ishikawa M, Yamamoto Y, Ohsawa I, Ohta S: Inhalation of hydrogen gas suppresses hepatic injury caused by ischemia/reperfusion through reducing oxidative stress. Biochem Biophys Res Commun. 2007, 361 (3): 670-674. 10.1016/j.bbrc.2007.07.088.
Wood KC, Gladwin MT: The hydrogen highway to reperfusion therapy. Nat Med. 2007, 13 (6): 673-674. 10.1038/nm0607-673.
The actual state investigative committee of 9th Japanese society of pressure ulcers: Investigation of the current clinical practice on pressure ulcers for its prevalence rate. Med Trib. 2007, 10: 11-in Japanese
Saitoh Y, Yoshimura Y, Nakano K, Miwa N: Platinum nanocolloid-supplemented hydrogen-dissolved water inhibits growth of human tongue carcinoma cells preferentially over normal cells. Exp Oncol. 2009, 31 (3): 156-62.
Saitoh Y, Okayasu H, Xiao L, Harata Y, Miwa N: Neutral pH hydrogen-enriched electrolyzed water achieves tumor-preferential clonal growth inhibition over normal cells and tumor invasion inhibition concurrently with intracellular oxidant repression. Oncol Res. 2008, 17 (6): 247-255. 10.3727/096504008786991620.
Saitoh Y, Harata Y, Mizuhashi F, Nakajima M, Miwa N: Biological safety of neutral-pH hydrogen-enriched electrolyzed water upon mutagenicity, genotoxicity and subchronic oral toxicity. Toxicol Ind Health. 2010, 26 (4): 203-216. 10.1177/0748233710362989.
Asada R, Kageyama K, Tanaka H, Matsui H, Kimura M, Saitoh Y, Miwa N: Antitumor effects of nano-bubble hydrogen-dissolved water are enhanced by coexistent platinum colloid and the combined hyperthermia with apoptosis-like cell death. Oncol Rep. 2010, 24 (6): 1463-1470.
Kato S, Hokama R, Okayasu H, Saitoh Y, Iwai K, Miwa N: Colloidal platinum in hydrogen-rich water exhibits radical-scavenging activity and improves blood fluidity. J Nanosci Nanotechnol. 2012, 12 (5): 4019-4027. 10.1166/jnn.2012.6163.
Sanada H: “Pressure Ulcer: Assessment/care guide”. 2009, Nakayama Shoten Co. Ltd, 127-In Japanese, ISBN 978-4-521-73171-12
Wicks G: A guide to the treatment of pressure ulcers from grade 1-grade 4. Wound Essent. 2007, 2: 106-112.
Zeller JL: Pressure ulcers. JAMA. 2006, 296 (8): 23-30.
Matsui Y, Furue M, Sanada H, Tachibana T, Nakayama T, Sugama J, Furuta K, Tachi M, Tokunaga K, Miyachi Y: Development of the DESIGN-R with an observational study: an absolute evaluation tool for monitoring pressure ulcer wound healing. Wound Repair Regen. 2011, 19 (3): 309-315. 10.1111/j.1524-475X.2011.00674.x.
Sanada H, Iizaka S, Matsui Y, Furue M, Tachibana T, Nakayama T, Sugama J, Furuta K, Tachi M, Tokunaga K, Miyachi Y: Clinical wound assessment using DESIGN-R total score can predict pressure ulcer healing: pooled analysis from two multicenter cohort studies. Wound Repair Regen. 2011, 19 (5): 559-567. 10.1111/j.1524-475X.2011.00719.x.
Chuai YH, Sun XJ, Cai J: Biology of hydrogen and its medical applications. Act Biophys Sin. 2012, 28 (9): 705-718. (in Chinese)
Kajiya M, Silva MJ, Sato K, Ouhara K, Kawai T: Hydrogen mediates suppression of colon inflammation induced by dextran sodium sulfate. Biochem Biophys Res Commun. 2009, 386 (1): 11-15. 10.1016/j.bbrc.2009.05.117.
Nakashima-Kamimura N, Mori T, Ohsawa I, Asoh S, Ohta S: Molecular hydrogen alleviates nephrotoxicity induced by an anti-cancer drug cisplatin without compromising anti-tumor activity in mice. Cancer Chemother Pharmacol. 2009, 64 (4): 753-761. 10.1007/s00280-008-0924-2.
Cardinal JS, Zhan JH, Wang YN, Sugimoto R, Tsung A, McCurry KR, Billiar TR, Nakao A: Oral hydrogen water prevents chronic allograft nephropathy in rats: Hydrogen water prevented CAN. Kidney Int. 2010, 77: 101-109. 10.1038/ki.2009.421.
Foschi D, Trabucchi E, Musazzi M, Castoldi L, Di Mattia D, Radaelli E, Marazzi M, Franzini P, Berlusconi A: The effects of oxygen free radicals on wound healing. Int J Tissue React. 1988, 10 (6): 373-379.
Boron WF: Sharpey-Schafer lecture: gas channels. Exp Physiol. 2010, 95 (12): 1107-1130. 10.1113/expphysiol.2010.055244.
Fuchs Y, Steller H: Programmed cell death in animal development and disease. Cell. 2011, 147 (4): 742-758. 10.1016/j.cell.2011.10.033.
Bergmann A, Steller H: Apoptosis, stem cells, and tissue regeneration. Sci Signal. 2010, 3 (145): re8-10.1126/scisignal.3145re8.
Cai JM, Kang ZM, Liu WW, Luo X, Qiang S, Zhang JH, Ohta SG, Sun XJ, Xu WG, Tao HY, Li RP: Hydrogen therapy reduces apoptosis in neonatal hypoxia-ischemia rat model. Neurosci Lett. 2008, 441 (2): 167-172. 10.1016/j.neulet.2008.05.077.
Saitoh Y, Miyanishi A, Mizuno H, Kato S, Aoshima H, Kokubo K, Miwa N: Super-highly hydroxylated fullerene derivative protects human keratinocytes from UV-induced cell injuries together with the decreases in intracellular ROS generation and DNA damages. J Photochem Photobiol B. 2011, 102 (1): 69-76. 10.1016/j.jphotobiol.2010.09.006.
Hoerter JD, Ward CS, Bale KD, Gizachew AN, Graham R, Reynolds J, Ward ME, Choi C, Kagabo JL, Sauer M, Kuipers T, Hotchkiss T, Banner N, Chellson RA, Ohaeri T, Gant L, Vanderhill L: Effect of UVA fluence rate on indicators of oxidative stress in human dermal fibroblasts. Int J Biol Sci. 2008, 4 (2): 63-70.
