Local delivery of cardiac stem cells overexpressing HIF-1α promotes angiogenesis and muscular tissue repair in a hind limb ischemia model.

Critical limb ischemia (CLI) is the most advanced stage of peripheral artery disease, associated with significant risk of limb loss, morbidity and mortality; however, there remains unmet therapeutic needs for arterial revascularization and ischemic tissue repair. Stem cell therapies have emerged as compelling candidates due to beneficial proangiogenic and immunosuppressive function. Nevertheless, in vivo efficacy was insufficient in proliferation, differentiation and survival/engraftment rate. Cardiac stem cells (CSCs) was firstly attempted for CLI as a novel therapeutic modality to provide superior angiogenic potency to bone marrow-derived stem cells (BMSCs). It was noted that CSCs demonstrated 3.2-fold in HGF, 2.9-fold in VEGF and 8.7-fold in PDGF-B higher gene expressions compared to BMSCs. To enhance the hypoxia-induced proangiogenic effect, CSCs were transfected with hypoxia-inducible factor-1 alpha (HIF-1α) by using electroporation method, specifically optimized for CSCs yielding 45.77% of transfection efficiency and 89.75% of viability. HIF-1α overexpression significantly increased CSC survival in hypoxia, proangiogenic factors production and endothelial differentiation. In mouse hind limb ischemia model, local intramuscular delivery of CSC overexpressing HIF-1α (HIF-CSC) significantly improved the blood flow recovery. Histological analysis revealed that muscle degeneration and fibrosis in the ischemic limb were attenuated. Local delivery of HIF-CSC might be a promising option for ischemic tissue restoration.

The effect of antioxidant and whitening action on Plantago asiatica L. leaf ethanol extract for health care.

BACKGROUND: The Plantago asiatica L. is easy to cultivate and has been used as a folk remedy since ancient times because of various pharmacological actions such as anti-inflammation and antioxidation. It also contains a variety of flavonoids such as aucubin, which is thought to be excellent for whitening, antioxidant and anti-inflammatory action. OBJECTIVE: We investigated the effect of P. asiatica L. leaf ethanol extracts containing various active ingredients on antioxidative, anti-inflammation and whitening action and investigated its potential as a health care material. P. asiatica L. has been widely used in folk remedies. RESULTS: The cell toxicity test using RAW264.7 cells showed a high cell survival rate of over 75%, thus demonstrating the safety of the sample. In order to study the antioxidant activity of P. asiatica L. leaf ethanol extracts, we studied a sample which showed radical scavenging activity in a dose-dependent manner. To observe the antioxidant activity at the cell level, RAW 264.7 cells were used and inhibition of ROS production was measured. The ROS production was suppressed in a dose-dependent manner and the scavenging activity was stronger than the sample's own radical scavenging ability. To observe the anti-inflammatory effect of P. asiatica L. leaf ethanol extracts, inhibition of NO generation was observed using LPS-induced RAW 264.7 cells. NO generation was inhibited in a dose-dependent manner and was strongly inhibited by 31% at 100 µg/mL. In vitro, L-DOPA and L-tyrosine were used to inhibit tyrosinase action in a dose-dependent manner. The concentration of melanin at 1, 10, and 100 µg/mL was suppressed in B16 F10 melanin cells supplemented with α-MSH in the cells, and the inhibition was suppressed to 29% at 100 µg/mL. In the B16 F10 melanin cell stimulated with MSH, the P. asiatica L. leaf ethanol extracts inhibited melanin formation in a dose-dependent manner. CONCLUSION: P. asiatica L. leaf ethanol extracts are expected to be developed as whitening cosmeceutical ingredients and as health care ingredients with antioxidant and anti-inflammatory properties.