Possible contribution of sialic acid to the enhanced tumor targeting efficiency of nanoparticles engineered with doxorubicin.

Doxorubicin (DOX)-engineered poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) including phloretin (PHL) were designed and the feasible contribution of sialic acid (SA) to the improved tumor targeting and penetration capabilities was elucidated in lung adenocarcinoma models. DOX has been clinically used as liposomal formulations after its introduction to the inner side of vehicles, however DOX is anchored in the outer surface of PLGA NPs for improved tumor penetration by interactions with SA in this study. DOX (positively charged at physiological pH) was adsorbed onto the negatively charged PLGA NPs via electrostatic interactions and consequent binding of SA (negatively charged at physiological pH) to DOX located in NPs was also elucidated. DOX layer in DOX@PLGA NPs rendered improved endocytosis and partial contribution of SA (expressed in cancer cells) to that endocytosis was demonstrated. DOX@PLGA/PHL NPs provided enhanced antiproliferation potentials in A549 cells rather than single agent (DOX or PHL)-installed NPs. In addition, DOX-SA interactions seemed to play critical roles in tumor infiltration and accumulation of DOX@PLGA NPs in A549 tumor-xenografted mouse model. All these findings support the novel use of DOX which is used for the surface engineering of NPs for improved tumor targeting and penetration.

Development of iron(II) sulfate nanoparticles produced by hot-melt extrusion and their therapeutic potentials for colon cancer.

Particle size reduction of FeSO4 (iron(II) sulfate, IS) from micron to nano size was achieved by a combination of hot-melt extrusion (HME) processing and the input of Span 80, Tween 80, and poly(ethylene glycol) (PEG) 6000. Conveying, kneading, and extruding steps of the HME process and a decrease in the surface tension by surfactants were introduced to produce FeSO4 nanoparticles (NPs) in an aqueous environment. The FeSO4-based NPs (ISNPs) in the dispersion were composed of FeSO4, Span 80, Tween 80, and PEG 6000 and displayed a hydrodynamic size of 350-400 nm (5-50 mg/mL ISNPs concentration range) and a spherical shape. Considering the feeding ratio of FeSO4 (20%, w/w) used for preparing the ISNPs, FeSO4 appears to be wrapped by Span 80, Tween 80, and PEG 6000 according to the results of X-ray photoelectron spectroscopy (XPS) analysis. ISNPs exhibited different thermodynamic properties from those of FeSO4 itself. In colon adenocarcinoma (Caco-2) cells, the ISNPs group exhibited enhanced antiproliferation and apoptosis potentials compared to the FeSO4 group (p < 0.05). Histological staining data of a dissected intestine after oral administration of ISNPs suggest the absence of severe intestinal toxicities compared to the control (no treatment) group. All of these results imply the feasibility of the use of the developed ISNPs for the treatment of colon cancers with oral administration.

Therapeutic Efficacies of Artemisia capillaris Extract Cream Formulation in Imiquimod-Induced Psoriasis Models.

A cream formulation containing Artemisia capillaris (AC) extract (ACE) was developed for psoriasis therapy. Although ACE can be dissolved in organic solvents, its topical application is restricted because of toxicities. Therefore, a cream formulation was developed for the convenient and safe local application of ACE on skin lesions. The antipsoriatic properties of the ACE cream were evaluated using an imiquimod- (IMQ-) induced psoriasis-like mouse model. In psoriasis-like mouse models, the cumulative score (redness, thickness, and scaling) of the IMQ + ACE cream group was significantly lower than those of the other groups on day 4 (p < 0.05). The results of the hematoxylin and eosin staining of skin tissues revealed that the epidermal thickness value of the IMQ + ACE cream group was significantly lower than those of the other experimental groups (p < 0.05). The expression level of intracellular adhesion molecule-1 (ICAM-1), which indicates the leukocyte infiltration into the skin and subsequent interactions with keratinocytes, was also lower in the IMQ + ACE cream group than in the IMQ group. These results indicate that ACE cream formulation could be used safely and conveniently for psoriasis treatment.

Preparation of cupric sulfate-based self-emulsifiable nanocomposites and their application to the photothermal therapy of colon adenocarcinoma.

Nanocomposites (NCs) of cupric sulfate monohydrate (CuSO4) were fabricated by hot-melt extrusion (HME) system equipped with twin screws. Micron-sized bulk powder of CuSO4 was dispersed in the mixture of surfactants (Span 80 and Tween 80) and hydrophilic polymer (polyethylene glycol (PEG) 6000) by HME process. Reduction of surface tension by surfactants and homogeneous dispersion in hydrophilic polymer along with HME technique were introduced to prepare CuSO4 NCs. Dispersion of CuSO4 NCs exhibited approximately 204 nm hydrodynamic size, unimodal size distribution, and positive zeta potential values. Encapsulation of CuSO4 in CuSO4 NCs and the physicochemical interactions between CuSO4 and pharmaceutical excipients were investigated by solid-state studies. Of note, CuSO4 NCs group exhibited higher antiproliferation efficacies, compared with bulk CuSO4, in Caco-2 (human adenocarcinoma) cells at 75 and 100 µg/mL CuSO4 concentrations (p < 0.05). Also, near-infrared laser irradiation to CuSO4 NCs group elevated the antiproliferation efficacies, compared with non-irradiation group, in Caco-2 cells. After intravenous injection in mice, CuSO4 NCs did not show severe in vivo toxicities. Developed CuSO4 NCs can be one of promising candidates of photothermal therapeutic agents for colon cancers.

Development of Resveratrol-Loaded Herbal Extract-Based Nanocomposites and Their Application to the Therapy of Ovarian Cancer.

Resveratrol (RSV) and the ethanol extract of Angelica gigas Nakai (AGN Ex)-based nanoparticles (NPs) were prepared using the nanocrystal concept. AGN/RSV NPs with 224 nm hydrodynamic size, unimodal size distribution, and negative zeta potential values were developed with the emulsification and solvent evaporation techniques. The crystalline properties of AGN Ex and RSV were transformed during the emulsification and solvent evaporation processes, thus, AGN NPs and AGN/RSV NPs exhibited amorphous states. AGN/RSV NPs held up their initial hydrodynamic size after 24 h of incubation in serum-included media. Sustained release profiles (for 5 days) of decursin (D) and decursinol angelate (DA) (the representative markers of AGN Ex) and RSV were observed at normal physiological pH (pH 7.4). In ovarian cancer (SKOV-3) cells, although AGN/RSV NPs showed a lower cellular entry rate rather than AGN NPs, the cellular accumulated amount of AGN/RSV NPs was similar with that of AGN NPs after 4 h of incubation. The antiproliferation efficiency of AGN/RSV NPs group was significantly higher than the AGN Ex, AGN NPs, and AGN NPs + RSV groups in SKOV-3 cells. AGN/RSV NPs can be one of the promising candidates for therapeutic nanoplatforms against ovarian cancers.

Polydopamine-coated nanocomposites of Angelica gigas Nakai extract and their therapeutic potential for triple-negative breast cancer cells.

Polydopamine (PD)-coated nanocomposites (NCs) based on the ethanol extract of Angelica gigas Nakai (AGN EtOH ext) were fabricated and evaluated for breast cancer therapy. AGN NCs were prepared using a modified emulsification-solvent evaporation method and were further incubated in dopamine solution (at pH 8.6) to be covered with the PD layer. PD-AGN NCs with a 213-nm mean diameter, narrow size distribution, and negative zeta potential values were fabricated in this study. Less negative (close to zero) zeta potential value of PD-AGN NCs than that of AGN NCs implied the existence of the PD layer in the outer surface of NCs. The PD layer in PD-AGN NCs was also identified by X-ray photoelectron spectroscopy (XPS) and ultraviolet (UV)/visible absorption analyses. The sustained release of decursin (D) and decursinol angelate (DA), as major active pharmacological components of AGN, was observed in both AGN NCs and PD-AGN NCs. Enhanced cellular binding property of PD-AGN NCs, compared to AGN NCs, in MDA-MB-231 (human breast adenocarcinoma; triple-negative breast cancer) cells was observed. Improved anticancer activities of PD-AGN NCs compared with those of AGN EtOH ext and AGN NCs were also shown in MDA-MB-231 cells. The developed PD-AGN NCs may be used as remarkable platform nanocarriers for efficient breast cancer therapy.

Antiproliferation of keratinocytes and alleviation of psoriasis by the ethanol extract of Artemisia capillaris.

The therapeutic potentials of the ethanol extract of Artemisia capillaris (ACE) for psoriasis were verified in HaCaT cells (as an immortalized human keratinocyte cell line) and imiquimod (IMQ)-induced psoriasis-like mouse models. In HaCaT cells, IC50 value of ACE was 37.5 µg/ml after incubating for 72 hr. The antiproliferation activity of ACE in HaCaT cells was further verified by apoptosis assays. The percentage of apoptotic population in ACE-treated group was significantly higher than that of control group (p < .05). The result of cell cycle arrest assay also supported the observed antiproliferation efficacy of ACE in HaCaT cells. In IMQ-induced psoriasis-like mouse models, the Psoriasis Area and Severity Index score of ACE (50 mg/ml; ACE50)-treated group was significantly lower than that of IMQ group on Day 4 (p < .05). After topical application of ACE on psoriasis-like lesion for 4 days, the epidermal thickness of (IMQ + ACE50) group was significantly lower than that of IMQ group (p < .05). The expression levels of Ki-67 and intracellular adhesion molecule-1 in excised skin tissues of (IMQ + ACE50) group were also lower than those of IMQ group. All these findings suggest that ACE can be used as a promising antipsoriatic agent.

Fabrication of polymer matrix-free nanocomposites based on Angelica gigas Nakai extract and their application to breast cancer therapy.

Nanocomposites (NCs) based on the ethanol extract of Angelica gigas Nakai (AGN EtOH ext) were developed for breast cancer therapy. Polymer matrix-free nano-sized particles based on the extract of natural product were fabricated using a modified emulsification-solvent evaporation method. Without the use of polymer matrix, toxicity can be minimized and the clinical application may be assured. AGN NCs with approximately 200nm mean diameter, narrow size distribution, and negative zeta potential were prepared in this study. Sustained release of decursin (D) and decursinol angelate (DA) (as major components of AGN) from AGN NCs was observed at pH 7.4. Cellular accumulation efficiency and intracellular distribution of AGN NCs were evaluated in MCF-7 (human breast adenocarcinoma) cells. According to the results of antiproliferation assay in MCF-7 cells, IC50 value of AGN NCs group (27.4±4.0µg/mL) was lower than that of AGN EtOH ext group (75.3±13.7µg/mL) (p<0.05). Also, the percentage of apoptotic events of AGN NCs group was significantly higher than that of AGN EtOH ext group (p<0.05). All these findings suggest that developed AGN NCs can be used as one of promising nanosystems for the therapy of breast cancers.

Phloretin-loaded fast dissolving nanofibers for the locoregional therapy of oral squamous cell carcinoma.

Fast dissolving nanofiber (NF) composed of poly(vinyl alcohol) (PVA) and d-α-tocopheryl polyethylene glycol succinate (TPGS) was developed for locoregional delivery of phloretin to oral cancers. PVA/TPGS/phloretin NF with 321nm mean diameter and >90% drug entrapment efficiency was fabricated by an electrospinning method. Transformation of drug from crystalline to amorphous state was identified by solid-state studies. NF structure was changed to nanoparticles after its dispersing in the aqueous medium. PVA/TPGS/phloretin NF exhibited fast wetting property and smaller hydrodynamic size of dispersion, compared with PVA/phloretin NF. The amphiphilic property of TPGS also contributed to the improved drug release from PVA/TPGS/phloretin NF. The anticancer activities of phloretin, via the inhibition of glucose uptake into the cancer cells, in NFs were assessed in YD-9 cells (oral squamous cell carcinoma from buccal cheek). The antiproliferation efficacy of PVA/TPGS/phloretin NF was significantly higher than that of phloretin solution and PVA/phloretin NF (p<0.05). Higher apoptotic events were also observed in PVA/TPGS/phloretin NF group rather than phloretin solution and PVA/phloretin NF groups (p<0.05). All these results support that PVA/TPGS/phloretin NF can be a promising fast dissolving formulation for the treatment of oral cancers.

Angelica gigas Nakai extract-loaded fast-dissolving nanofiber based on poly(vinyl alcohol) and Soluplus for oral cancer therapy.

A poly(vinyl alcohol) (PVA) and Soluplus (SP)-based nanofiber (NF) mat was fabricated using an electrospinning method for the delivery of Angelica gigas Nakai (AGN) extract (ext) to oral cancers. AGN/SP NF (mean diameter: 75±26nm; entrapment efficiency: 84.6±18.6%) and AGN/PVA/SP NF (mean diameter: 170±35nm; entrapment efficiency: 81.0±10.1%) were fabricated using an electrospinning method. Amorphization of AGN EtOH ext was verified by X-ray diffractometry (XRD) analysis during the electrospinning process for the fabrication of NF structures. The AGN/PVA/SP NF group exhibited instant wetting (within 2s) and rapid disintegration (within 3min) properties compared with those in the AGN/PVA NF group, assuring the successful and conventional application of AGN/PVA/SP NF film in the oral cavity without the intake of beverages. After the spontaneous dispersion of NF in the aqueous media, it was converted to nanoparticles with a narrow size distribution. In YD-9 cells (oral squamous cell carcinoma from buccal cheek), the anti-proliferation activity was ordered as follows: AGN EtOH ext suspension < AGN/PVA NF < AGN/PVA/SP NF. All of these findings indicated that AGN/PVA/SP NF can be used as a fast-dissolving mat formulation for the therapy of oral cancers.

Nanocomposites based on Soluplus and Angelica gigas Nakai extract fabricated by an electrohydrodynamic method for oral administration.

Nanocomposites (NCs) based on Soluplus (SP) were fabricated by an electrohydrodynamic (EHD) method for the oral delivery of Angelica gigas Nakai (AGN). Nano-sized particles were obtained after dispersing the resultant, produced by the EHD technique, in the aqueous environment. AGN/SP2 (AGN:SP=1:2, w/w) NC dispersion in aqueous media exhibited a 130nm mean diameter, narrow size distribution, and robust stability in the tested concentration range of the ethanol extract of AGN (AGN EtOH ext) and at pH 1.2 and 6.8. Amorphization of the components of AGN and their interactions with SP in the AGN/SP2 NC formulation were demonstrated by X-ray diffractometry (XRD) analysis. The released amounts of decursin (D) and decursinol angelate (DA), major components of AGN, from NCs were improved compared with those from the AGN EtOH ext group at both pH 1.2 and 6.8. As D and DA can be metabolized into decursinol (DOH) in the liver after oral administration, the DOH concentrations in plasma were quantitatively determined to evaluate the oral absorption of AGN. In a pharmacokinetic study in rats, higher oral absorption and the maximum concentration in plasma (Cmax) were presented in the AGN/SP2 NC group compared with the AGN EtOH ext and AGN NC groups. These findings indicate the successful application of developed SP-based NCs for the oral delivery of AGN.