Antitumor effects of nano-bubble hydrogen-dissolved water are enhanced by coexistent platinum colloid and the combined hyperthermia with apoptosis-like cell death

Abstract

In order to erase reactive oxygen species (ROS) related with the proliferation of tumor cells by reducing activity of hydrogen, we developed functional water containing nano-bubbles (diameters: <900 nm for 71%/ population) hydrogen of 1.1-1.5 ppm (the theoretical maximum: 1.6 ppm) with a reducing ability (an oxidation- reduction potential -650 mV, normal water: +100-200 mV) using a microporous-filter hydrogen-jetting device. We showed that hydrogen water erased ROS indispensable for tumor cell growth by ESR/spin trap, the redox indicator CDCFH-DA assay, and was cytotoxic to Ehrlich ascites tumor cells as assessed by WST-8 assay, crystal violet dye stain and scanning electron microscopy, after 24-h or 48-h incubation sequent to warming at 37˚C or 42˚C. Hydrogen water supplemented with platinum colloid (0.3 ppm Pt in 4% polyvinylpyrrolidone) had more antitumor activity than hydrogen water alone, mineral water alone (15.6%), hydrogen water plus mineral water, or platinum colloid alone as observed by decreased cell numbers, cell shrinkage and pycnosis (nuclear condensation)/karyorrhexis (nuclear fragmentation) indicative of apoptosis, together with cell deformation and disappearance of microvilli on the membrane surface. These antitumor effects were promoted by combination with hyperthermia at 42˚C. Thus, the nano-bubble hydrogen water with platinum colloid is potent as an anti- tumor agent.

Study details

Article type

Original Research

Disease / injury model

Ehrlich ascites tumor (EAT) cells (mouse cancer cell line)

Dose comparison

Different concentrations and combinations (hydrogen water, platinum colloid, mineral water) were compared for their effects on tumor cell viability.

Dose-dependent effect

Yes

Outcome

Positive

Findings

Nano-bubble hydrogen-dissolved water exhibits antitumor effects on Ehrlich ascites tumor cells, which are further enhanced by the addition of platinum colloid and by hyperthermia (42°C). The combination leads to increased apoptosis-like cell death, cell shrinkage, and loss of microvilli, indicating potent anti-tumor activity.