Differential metabolic effects on mitochondria by silica hydride using capillary electrophoresis.

Working as an extension of a newly developed method for a capillary electrophoretic analysis of purine nucleotides, nucleosides, bases, and catabolism, an assay of the differential metabolic properties by a novel organosiliceous anionic hydride compound, silica hydride, was evaluated with Chinese hamster ovary mitochondria using a 50-microm poly(acryloylaminopropanol)-coated, fused-silica capillary. The results of this organellar differential analysis indicate a correlation of increased redox pair of NADH to NAD(+) ratios by two times and an increase in ATP levels in the assayed mitochondria by six times. Glucose levels in the organelles were half of the original values. This study validates the electrophoretic method utilizing live organelle fractions for differential metabolic analysis and additionally illustrates some of the emerging novel properties of silica hydride. As confirmation of the results obtained in this assay, additional methods of standard protocol were used to monitor the mitochondrial metabolic activity.

Antioxidant capacity of silica hydride: a combinational photosensitization and fluorescence detection assay.

Utilizing a novel combinational technique incorporating spectrafluorometry and photosensitization, this analysis determined cell viability and cytotoxicity through the introduction of reactive oxygen species and measurement of plasma membrane integrity. Chinese hamster ovary and mouse hybridoma cells were treated with silica hydride after being photosensitized with singlet oxygen, hydroxyl/superoxide, and hydroxyl reactive oxygen species through the use of rose Bengal diacetate, malachite green, and N,N'-bis(2-hydroperoxy-2-methoxyethyl)-1,4,5,8-naphthaldiimide, respectively. The analysis resulted in an easy and effective method for quantifying reactive oxygen species reduction and characterized the radical reduction efficacy of silica hydride at 97% (+/- 0.68%, sigma = 0.84) against singlet oxygen species and over 87% (+/- 0.56%, sigma = 0.70) for the combination of hydroxyl and superoxide reactive species, and 98% (+/- 0.37%, sigma = 0.47) effective for hydroxyl radical species. Nontreated photosensitized controls showed less than 1% viability under the same conditions.

Evaluation of hydroxyl radical-scavenging abilities of silica hydride, an antioxidant compound, by a Fe2+-EDTA-induced 2-hydroxyterephthalate fluorometric analysis.

The hydroxyl radical scavenging capacity and efficacy of a novel organosiliceous anionic hydride compound, silica hydride, were quantified by a recently developed method. The method measures a direct relationship between the hydroxyl radical scavenging capability of the antioxidant compound and the linear decrease in signal from a fluorescent 2-hydroxyterephthalate product created by reacting an Fe(2+)-EDTA complex in the presence of a potential radical scavenger. A fluorescence signal half-inhibition, IC(50), value of 1.4 +/- 0.1 muM was obtained for silica hydride compounds. The validity of the analysis was verified by electron spin resonance spectroscopy, spectrophotometric analysis of NAD(+)/NADH ratios, mitochondrial membrane potential measurements, and assays of both cytochrome c (Fe(3+)) to cytochrome c (Fe(2+)) and epinephrine to adenochrome reductions.

Antioxidant capability and efficacy of Mega-H silica hydride, an antioxidant dietary supplement, by in vitro cellular analysis using photosensitization and fluorescence detection.

Treatment of Chinese hamster ovary and mouse hybridoma cells with Mega-H brand silica hydride, a marketed antioxidant, after photosensitization with singlet oxygen and hydroxyl/superoxide reactive oxygen species through the use of rose bengal diacetate and malachite green resulted in an effective method of reducing free radical activity by more than 96% against singlet oxygen species and more than 86% for hydroxyl and superoxide free radicals with the dosage recommended by the manufacturer. The analysis used a combinational spectrafluorometric technique to determine cell viability and cytotoxicity through the mechanism of intracellular esterase activity and plasma membrane integrity. Photosensitized controls not treated with silica hydride showed less than 1% viability under the same conditions. The reduction of the introduced free radicals and singlet oxygen species and the consequent high levels of cell viability may be the result of effective and efficient antioxidant and radical scavenging properties of silica hydride.