Transdermal nanotherapeutics: Panax quinquefolium polysaccharide nanoparticles attenuate UVB-induced skin cancer.

Ultraviolet (UV) radiation is known to cause an imbalance of the endogenous antioxidant system leading to an increase in skin cancer. Panax quinquefolium (American ginseng) polysaccharides (GPS) can inhibit such an imbalance due to its anti-oxidative and anti-inflammatory properties. The aim of this study was to investigate the therapeutic effects of topical formulations containing GPS nanoparticles (NPs) to inhibit UVB induced oxidative damage and skin cancer. Photoaging was conducted under UVB irradiation with a dose of 300 mJ/cm2 on SKH1 hairless mice. The treatment groups (n = 5) were as follows: sham control, native GPS, GPS NPs and fluorescent labeled GPS NPs. To compare the photoprotective performance, the topical formulations were applied before and after UVB induction (pre-treatment and post-treatment), followed by sacrificing the animals. Then, skin and blood samples were collected, and inflammatory cytokines production was measured using ELISA. Compared to the sham control, GPS NPs pre-treated mice skin and blood samples exhibited a significant lowering in all cytokine production. In addition, skin histology analysis showed that pre-treatment of GPS NPs prevented epidermal damage and proliferation. The results support that topical formulation containing GPS NPs can inhibit UVB induced oxidative damage and skin cancer.

Immunoengineering with Ginseng Polysaccharide Nanobiomaterials through Oral Administration in Mice.

Plant polysaccharides (PS) such as American ginseng polysaccharide (GPS) have drawn immense interest in the field of immunoengineering, as they offer a way to actively control immune cell behavior and stimulation. These pharmacological activities have been limited by PS's inherent physicochemical properties including large molecular size, heterogeneity, and poor solubility. In this work, we hypothesized that by nanosizing and encapsulating GPSs, we could enhance their immunomodulation by increased penetration and absorption through the GI tract. Herein, GPS nanoparticles (NPs) of average size 20 nm (± 4 nm) were prepared using a microfluidic approach, then encapsulated within porous nanospheres (diameter 180 ± 10 nm) of biodegradable gelatin to enhance their oral delivery. To locate the GPS NPs inside the gelatin, we encapsulated fluorescent-labeled GPS in gelatin and analyzed using confocal microscopy. An in vitro investigation on tumor induced macrophage cell lines showed a concentration dependent enhanced immunostimulation with the encapsulated GPS NPs. The immunomodulation was then studied for different formulations of GPS through oral gavage in Swiss albino mice. The results showed that the production of proinflammatory mediators in blood samples was significantly increased for the encapsulated GPS in a dose- and time-dependent manner compared to other GPS treatments. This study shows that GPS and potentially other PS systems' immunomodulation properties can be significantly enhanced for use in simple oral drug delivery.

Fabrication of fluorescent labeled ginseng polysaccharide nanoparticles for bioimaging and their immunomodulatory activity on macrophage cell lines.

Polysaccharides are a major active component of American ginseng root showing various biological activities including anti-carcinogenic, anti-aging, immunostimulatory and antioxidant effects. Although their biological activity has been reported by several groups, no research has explored their cellular uptake and biodistribution, owing to the lack of suitable detection techniques in living cells. This work examines a novel, simple and efficient fluorescent labeling procedure of ginseng polysaccharides (PS), in order to examine their cellular distribution using confocal microscopy. This procedure utilized a one-pot strategy with fluorescein-5-thiosemicarbazide (FTSC) to introduce a thiosemicarbazide group onto the aldehyde group at the reducing saccharide end to form a stable amino derivative through reductive amination. This polysaccharide-FTSC derivative was then characterized by GPC, UV, FTIR, photoluminescence and fluorescence microscopy to confirm attachment and any structural changes. The results demonstrated that the labeled ginseng PS nanostructure showed high fluorescence with minimal changes in PS molecular weight. The labeled PS exhibited almost no cytotoxicity effect against tumor induced macrophage cell lines (RAW 264.7) while retaining high immunostimulating activity similar to the non-labeled ginseng PS. Therefore, the developed approach provides a convenient and highly efficient fluorescent labeling procedure for understanding the mechanism of ginseng PS uptake in macrophage cell lines.

Microfluidic Synthesis of Ginseng Polysaccharide Nanoparticles for Immunostimulating Action on Macrophage Cell Lines.

North American (NA) ginseng root (Panax quinquefolium) has become of increasing scientific interest because of its immune-enhancing properties. Herein, we have developed a novel approach to synthesize ginseng polysaccharide nanoparticles (NPs) from NA ginseng for enhancing their immunostimulation. Nanoparticles of ginseng polysaccharide were prepared using a microfluidic device and compared to other conventional wet chemical processes including nanoprecipitation and reverse microemulsion. The morphology and size of the NPs were characterized by SEM, TEM, DLS and FTIR. Depending on the experimental conditions, microfluidics was found to provide unimodal polysaccharide spheres down to 20 nm (±4 nm) with very narrow particle size distributions. In addition, the immunostimulating effect of the polysaccharide NPs was investigated on Murine macrophage cell lines, with the results revealing an enhanced production of all proinflammatory mediators in a concentration dependent manner. The proposed microfluidic system has the advantages of ease of fabrication, simplicity, and a fast and low-cost process that is capable of producing ginseng polysaccharide NPs with demonstrated enhancement of immunostimulation of macrophage cell lines.