褥瘡患者の栄養チューブからの水素水摂取と 水素水の正常ヒト皮膚細胞に対する復元作用(in vitro) | Hydrogen water intake via tube-feeding for patients with pressure ulcer and its reconstructive effects on normal human skin cells in vitro.

Med Gas Res. 2013 Sep 10;3(1):20.

Hydrogen water intake via tube-feeding for patients with pressure ulcer and its reconstructive effects on normal human skin cells in vitro.

Li Q, Kato S, Matsuoka D, Tanaka H, Miwa N.



Pressure ulcer (PU) is common in immobile elderly patients, and there are some research works to investigate a preventive and curative method, but not to find sufficient effectiveness. The aim of this study is to clarify the clinical effectiveness on wound healing in patients with PU by hydrogen-dissolved water (HW) intake via tube-feeding (TF). Furthermore, normal human dermal fibroblasts OUMS-36 and normal human epidermis-derived cell line HaCaT keratinocytes were examined in vitro to explore the mechanisms relating to whether hydrogen plays a role in wound-healing at the cellular level.


Twenty-two severely hospitalized elderly Japanese patients with PU were recruited in the present study, and their ages ranged from 71.0 to 101.0 (86.7 +/- 8.2) years old, 12 male and 10 female patients, all suffering from eating disorder and bedridden syndrome as the secondary results of various underlying diseases. All patients received routine care treatments for PU in combination with HW intake via TF for 600 mL per day, in place of partial moisture replenishment. On the other hand, HW was prepared with a hydrogen-bubbling apparatus which produces HW with 0.8-1.3 ppm of dissolved hydrogen concentration (DH) and -602 mV to -583 mV of oxidation-reduction potential (ORP), in contrast to reversed osmotic ultra-pure water (RW), as the reference, with DH of < 0.018 ppm and ORP of +184 mV for use in the in vitro experimental research. In in vitro experiments, OUMS-36 fibroblasts and HaCaT keratinocytes were respectively cultured in medium prepared with HW and/or RW. Immunostain was used for detecting type-I collagen reconstruction in OUMS-36 cells. And intracellular reactive oxygen species (ROS) were quantified by NBT assay, and cell viability of HaCaT cells was examined by WST-1 assay, respectively.


Twenty-two patients were retrospectively divided into an effective group (EG, n = 12) and a less effective group (LG, n = 10) according to the outcomes of endpoint evaluation and the healing criteria. PU hospitalized days in EG were significantly shorter than in LG (113.3 days vs. 155.4 days, p < 0.05), and the shortening rate was approximately 28.1%. Either in EG or in LG, the reducing changes (EG: 91.4%; LG: 48.6%) of wound size represented statistically significant difference versus before HW intake (p < 0.05, p < 0.001). The in vitro data demonstrate that intracellular ROS as quantified by NBT assay was diminished by HW, but not by RW, in ultraviolet-A (UVA)-irradiated HaCaT cells. Nuclear condensation and fragmentation had occurred for UVA-irradiated HaCaT cells in RW, but scarcely occurred in HW as demonstrated by Hoechst 33342 staining. Besides, under UVA-irradiation, either the mitochondrial reducing ability of HaCaT cells or the type-I collagen construction in OUMS-36 cells deteriorated in RW-prepared culture medium, but was retained in HW-prepared culture medium as shown by WST-1 assay or immunostain, respectively.


HW intake via TF was demonstrated, for severely hospitalized elderly patients with PU, to execute wound size reduction and early recovery, which potently ensue from either type-I collagen construction in dermal fibroblasts or the promoted mitochondrial reducing ability and ROS repression in epidermal keratinocytes as shown by immunostain or NBT and WST-1 assays, respectively.


Med Gas Res. 2013 Sep 10;3(1):20.

水素水の正常ヒト皮膚細胞に対する復元作用(in vitro)





さらに水素に細胞レベルで傷を癒す働きがあるかどうかのメカニズムを調べるため、in vitroで正常ヒト線維芽細胞OUMS-36と正常ヒト表皮由来角化細胞株HaCaTを用いた研究を行いました。




0.8~1.3ppmの水素濃度と-602mV~-583mVの酸化還元電位を持つ水素水を作り出す水素バブリング装置を使って水素水を供給する一方で、in vitro試験では対照水として逆浸透膜を使った純水を用い、この水素濃度は0.018ppmで酸化還元電位は+184mVでした。

in vitro試験では、OUMS-36線維芽細胞とHaCaT角化細胞を、水素水あるいは純水を使った培養液でそれぞれ培養しました。OUMS-36細胞中の1型コラーゲン合成を検出するために免疫染色を用いました。また、細胞間質の活性酸素種はNBTアッセイで、HaCaTの生存能力はWST-1アッセイでそれぞれ定量しました。



EGではLGに比べて有意に褥瘡による入院日数が短く(113.3日 vs 155.4日,p<0.05)、短縮比はおおよそ28.1%でした。また、EGとLGのどちらにおいても、水素水を摂取する前と比べて、褥瘡サイズが統計学的に有意差をもって減少しました(EG:91.4%,p<0.05; LG:48.6%,p<0.001)。

一方in vitroのデータによると、UVA照射HaCaT細胞間質の活性酸素量を定量(NBTアッセイ)しましたが、水素水で培養すると活性酸素は減り、純水では減りませんでした。