This study was conducted in order to investigate the reduction activity of red ginseng extract (RGE; Panax ginseng, C. A. Meyer) on hydroxyl radical (·OH) using an electron spin resonance (ESR) spectrometer and spin-trapping techniques. ·OH generated by a Fenton Reaction System was trapped by 5, 5-dimethyl-l-pyrroline-N oxide (DMPO). The decay rate showed approximately pseudo-firs order kinetics over the period of measurement (by 10 min), and the half lifetime of the DMPO/·OH signal was estimated as approximately 8.15 min. However, the half lifetime of RGE/·OH was estimated as approximately 7.5 min, and the half lifetime of RGE was higher than that of DMPO/·OH adduct only. The order of reduction activities was ascorbic acid > N, Nʹ-dimethylthiourea (DMTU) > RGE > trolox > mannitol in the Fenton Reaction System. Thus, these observations indicate that RGE reaction with ·OH has relative reduction activity. The second-order rate constant of RGE/·OH may be 3.5~4.5 × 109 M-¹ ∙ S-¹.

The anti-inflammatory effect of PHBV/Collagen (PHCP) was examined in a mouse model of lipopolysaccharide (LPS)-induced skin inflammation. Vascular permeability on the back skin was measured by the local accumulation of Evan’s blue dye after subcutaneous injection of LPS (30 µg site^-1 ). Dye leakage in the skin showed a significant increase at 2 h after injection of LPS. This LPS-induced dye leakage was also completely inhibited by HO-1 inhibitor, ZnPP, and antioxidants, including methyl gallate, trolox, and mannitol. To study the possible mechanisms underlying the in vivo anti-inflammatory effect of PHCP against LPS-induced inflammation, we also examined the effects of PHCP on malondialdehyde (MDA) and glutathione levels in skin tissues and found that pretreatment with PHCP resulted in inhibited MDA elevation and a remarkable reduction of glutathione level. In addition, similar results were obtained after pretreatment with antioxidants, including trolox and mannitol, and HO-1 inhibitor, ZnPP. Histopathologically, an influx of neutrophils into the skin dermis was detected between 24 h and 72 h after LPS injection (30, 100 µg site^-1), compared to control animals after injection of saline. This increase was greater in mice treated with 100 µg of LPS than in those treated with 30 µg of LPS and was significantly suppressed by pretreatment with PHCP, antioxidants, and HO-1 inhibitor. These results collectively suggest that PHCP has an anti-inflammatory effect against LPS-induced inflammation model in vivo and may be a good candidate for the skin tissue engineering biomedical application primarily through manipulation of the redox state.

In this study, a nanofibrous scaffold was obtained by co-electrospinning poly (3-hydroxybutyrate- co-3-hydroxyvalerate) (PHBV) and collagen in 2,2,2-trifluoroethanol at a ratio of 3/7. The fiber diameters were in the range of 250-600 nm. It was found that PHBV/Collagen (PHCP) nanofibrous scaffold showed greater proliferation than the PHBV nanofibrous scaffold induced by oxidant in NIH3T3 cells. Otherwise, in the early-stage wound-healing mouse model, wound closure was evaluated according to wound size reduction and histology of regenerated skin on the backs of mice. Each of the tissues removed on day 0, 3, 6, 9, 12, 15, and 18 was used for analysis of biochemical and pathological changes. None of the nanofiber-attached mice showed significant difference on the third day, however, from the third day until the ninth day, significantly faster healing was observed in PHCP-attached mice, compared to control wounds in epithelialization, wound contraction, and histopathological examinations. These results strongly support the beneficial effects of biomedical application of PHCP nanofiber in acceleration of the initial phase of wound healing through α-SM actin contraction.

We investigated free radical reactions in lung of living mice using an in vivo electron spin resonance (ESR) spectrometer and nitroxyl radical as a probe. When an aqueous solution of nitroxyl probe was trans-tracheally administered into lung of living mice, a sharp triplet signal was observed at the chest of the mice. The signal showed a gradual decrease with time, obeying first-order kinetics. Signal decay rates of carbamoyl-PROXYL and carboxy-2,2,6,6-tetramethyl-piperidine-N-oxyl were faster than those of CAT-1 and carboxy-PROXYL. The mechanism of signal decay may be attributed to (i) reaction with reactive oxygen species such as ·OH, (ii) transfer into blood circulation, or (iii) reduction by compounds continuously supplied. However, little is known about the clearance mechanism of the nitroxyl probe in lung. To evaluate the disappearance of the ESR signal of the nitroxyl probe in lung, in vivo ESR spectra in chest of mice was recorded after trans-tracheal administration of an aqueous high concentrate solution of nitroxyl probe. A broad signal from the chest was observed immediately after administration due to Heisenberg spin exchange interaction. A sharp triplet signal was superimposed on the broad signal and the appearance of a triplet signal was followed by disappearance of the broad one. Peak-to-peak line width of the sharp signal was almost the same as that after intravenous administration. A distinct signal was detected in blood collected 10 min after trans-tracheal administration of nitroxyl probe. The observations indicate the transfer from lung to blood circulation and its contribution to clearance of probe in lung. Appearance of a sharp signal in blood after trans-tracheal administration was dependent on the kind of nitroxyl probe, showing a different transfer rate from lung to blood.

In vivo redox reaction is involved in the processes of oxidative diseases. Thus, direct and non-invasive measurement of in vivo free radical reactions in living animals should be essential to understanding the roles of free radicals in pathophysiological phenomena. Electron spin resonance (ESR) technique has been utilized in analysis of free radicals, which are generated through imbalance of in vivo redox status. In vivo ESR/spin probe technique using nitroxide radicals as spin probes was developed for estimation of in vivo free radical reactions in whole living animals. This technique using a probe may become a powerful tool for use in clarifying mechanisms of disease and in monitoring pharmaceutical therapy. The application of ESR was introduced and discussed in this article.

Flavonoids are a large family of polyphenolic compounds synthesized by plants that have a common chemical structure. Flavonoids may be further divided into subclasses, Anthocyanidins, Flavan-3- ols, Flavanones, Flavonols, Flavones, Isoflavones. Flavonoids have antioxidant and antiradical activities. The antiradical efficacy of flavonoids have established structure-activity relationships (SAR) of the antioxidant activity. Reactivity of flavonoids is highly dependent on the configuration of OH groups on the flavonoid B and C rings, there being little contribution from A ring to antioxidant effectiveness. This short article presents the current knowledge on structural aspects on their free radical reactions of flavonoids.