‌Hydrogen(H2) in Skin and Aging

One of the most visible signs of aging is a change in the appearance of skin.
Some of the hallmarks of aging skin include increased fragility and diminished collagen production, resulting in loss of elasticity and wrinkles.
These negative characteristics are caused primarily by exposure to ROS/RNS that damage cellular proteins, membranes and DNA .

The ROS concentrations in skin are among the highest of any other organ because of exposure to extrinsic environmental factors, such as ultraviolet light, ionizing radiation and pollutants.
Cosmetic interventions to improve skin appearance―including pharmaceutical, surgical, and topical approaches―are considered temporary solutions, unless they deliver antioxidants to skin tissue and prevent ROS/RNS damage. Antioxidants effective in reducing ROS/RNS have been proposed to hold promise in improving skin structure and appearance.

Antioxidants have been delivered to skin in lotions, creams and oils, and by bathing . For example, hydrogen(H2) is considered a novel antioxidant for combating oxidative damage in skin and promoting a youthful appearance, and it has been used in water for bathing. By bathing daily for 3 months in H2-water (0.2 - 0.4 ppm H2), Japanese subjects showed significant improvements in neck wrinkles at the end of the 90-day bathing sessions.

This same publication examined the ability of H2-enriched water to stimulate production of type-1 collagen in skin fibroblasts and keratinocytes after UVA-exposure. They found that type-1 collagen synthesis was increased over 2-fold after 3 - 5 days in the H2-enriched water samples compared to controls .
Another approach has been to drink H2-water. Using healthy four month-old rats fed H2-enriched or control water the effects of H2-water have been examined in aging periodontal tissues. The animals fed H2-water and control water were examined after 16 months. At this time, the animals were examined for the expression of inflammation-associated genes. Although the expression of interleukin-1β was not different between the two groups of animals, the H2-water fed group was found to have activated Nod-like receptor protein 3 inflammasomes in periodontal tissue. In addition, oxidative damage was determined in periodontal tissue by measuring the levels of 8-hydroxydeoxyguanosine (8-OHdG) as a marker for DNA damage. Over time 8-OHdG levels increased in the control group (p <  0.05), but in the H2-water fed group the levels of 8-OHdG were significantly lower than the control animals (p <  0.05).
Also, the serum levels of 8-OHdG were examined. In the control group the serum levels of 8-OHdG increased in an age-dependent manner, whereas in the H2-water fed group the serum levels of 8-OHdG did not change during aging.

When periodontal tissues were examined histologically in the H2-water and control water animals, the linear distances between the cemento-enamel junction and the alveolar bone crest were significantly lower in the H2-water fed group than in the control water group (p <  0.05).
These authors also examined the level of alveolar bone loss for the medial root region of the first molar, but significant differences were not found. In addition, the numbers of TRAP-positive osteoclasts were lower in the experimental H2-water group than in the control group (p <  0.05), but there were no significant differences in the ratios of interleukin-1β-positive cells to total cells between the two groups.
Interestingly, examination of gene expression in H2-water and control animals revealed that the expression of inflammasome NLRP3-associated caspase-1, ASC and interleukin-1β in periodontal tissues was higher in the H2-water group (p <  0.05), whereas expression of NF-κB was significantly lower in the H2-water group (p <  0.05). Thus although drinking H2-water decreased oxidative damage to DNA, it did not suppress inflammatory reactions in aging periodontal tissue .

The protective effects of hydrogen have also been examined in animals exposed to cutaneous burns.
Rats were divided into sham, burn plus saline, and burn plus H2-enriched saline groups and analyzed at various times (6, 24 and 48 hours) after burning by contact with a hot metal comb for 20 seconds.

Indexes of oxidative stress, apoptosis and autophagy were measured in each group, and the zone of stasis was evaluated using immuno-fluorescence staining, ELISA, and Western blot analysis.
H2-enriched saline, but not control saline, attenuated the increases in apoptosis and autophagy seen in burn wounds, as measured by the expression of TUNEL staining and the expression of Bax, Bcl-2, caspase-3, Beclin-1, and Atg-5 proteins. Additionally, H2-saline treat- ment lowered the level of myeloperoxidase and expression of inflammation markers tumor necrosis factor-α, interleukin-1β, and -6 in the zone of stasis while augmenting interleukin-10. The elevated levels of Akt phosphorylation and NF-κB p65 expression post-burn were also down-regulated by H2-saline treatment.
The results indicated that H2-enriched saline treatment reduces the inflammation associated with cutaneous burns.

When skin is burned, there are typically changes in the epidermis and dermis tissue.
Sections of the skin from H2-saline and control saline treated animals were examined.
The interspaces between two burn wounds in saline control animals gradually narrowed and had a tendency to merge following the burn, whereas the interspaces remained relatively stable at various time points in the H2-saline treated skin. Certain characteristics, such as severe epidermis layer thinning, epithelium nuclei elongation, and dermis layer swelling with collagen alterations, could be observed in the normal saline-treated animals, whereas in the H2-enriched saline-treated animals these changes were alleviated over time.

Lipid peroxidation was also examined in the animals after burning their skin.
Skin tissue homogenates from the burn wounds reacted with thiobarbituric acid-reactive species (TBARS), a method that has been used to determine the malondialdehyde (MDA) levels.
Tissue superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) activities were also evaluated to determine the oxidative stress status in the skin tissues of burn wounds. The burn-induced increases in MDA were reduced in the animals treated with H2-saline, while the activities of endogenous antioxidant enzymes were significantly increased.

The results indicated that H2-enriched saline treatment attenuates burn-induced oxidative damage in the burn-wounded tissue of rats by inhibiting oxidative stress and increasing the activities of endogenous antioxidant enzymes.

Pressure ulcers are a routine problem in long-term hospitalization of aged patients. Li et al. examined the effects of H2-water in 22 elderly Japanese patients (mean = 86.7 ± 8.2 years) with pressure ulcers. The purpose of this study was to clarify the clinical effectiveness of H2-water given by tube feeding.

All patients received routine care treatments for pressure ulcers in combination with H2-water (600 mL per day) for partial moisture replenishment.
Routine care included: ointment, gauze dressing, wrapping, and bed-pad use after washing by the acidic water disinfection. Pressure relief modalities and nutritional support were also employed.
The 22 patients were divided into two groups: an effective care group (EG, n = 12) and a less effective group (LG, n = 10) according to the outcomes of endpoint evaluation and healing criteria. Pressure ulcer hospitalization days in EG patients were significantly shorter than in LG (113.3 days versus 155.4 days, p <  0.05), and the reduced rate was approximately 28.1% less.
In both the EG and LG groups the wound size reductions (91.4% and 48.6%, respectively) were statistically significant with the intake of H2-water (p <  0.05). The results demonstrated that H2-water intake via tube feeding reduced wound size in hospitalized elderly patients with pressure ulcers.

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