Co(II) Complex of Quercetin–Spectral, Anti-/Pro-Oxidant and Cytotoxic Activity in HaCaT Cell Lines

In this study a cobalt(II) complex of quercetin was synthetized in the solid state with the general formula Co(C15H9O7)2∙2H2O. The FT-IR, elemental analysis, and UV/Vis methods were used to study the composition of the complex in a solid state and in a water solution. The anti-/pro-oxidant activity of quercetin and the Co(II) complex was studied by means of spectrophotometric DPPH (2,2-diphenyl-1-picrylhydrazyl), FRAP (ferric reducing antioxidant activity) and Trolox oxidation assays. The cytotoxicity of quercetin and Co(II)-quercetin complex in HaCat cell lines was then established.

Polypodium vulgare L. (Polypodiaceae) as a Source of Bioactive Compounds: Polyphenolic Profile, Cytotoxicity and Cytoprotective Properties in Different Cell Lines

Pteridophytes, represented by ferns and allies, are an important phytogenetic bridge between lower and higher plants. Ferns have evolved independently of any other species in the plant kingdom being its secondary metabolism a reservoir of phytochemicals characteristic of this taxon. The study of the potential uses of Polypodium vulgare L. (Polypodiaceae) as medicinal plant has increased in recent years particularly when in 2008 the European Medicines Agency published a monograph about the rhizome of this species. Our objective is to provide scientific knowledge on the polar constituents extracted from the fronds of P. vulgare, one of the main ferns of European distribution, to contribute to the validation of certain traditional uses. Specifically, we have characterized the methanolic extract of P. vulgare fronds (PVM) by HPLC-DAD and investigated its potential cytotoxicity, phototoxicity, ROS production and protective effects against oxidative stress by using in vitro methods. The 3T3, HaCaT, HeLa, HepG2, MCF-7 and A549 were the cell lines used to evaluate the possible cytotoxic behaviour of the PVM. HPLC-DAD was utilized to validate the polyphenolic profile of the extract. H2O2 and UVA were the prooxidant agents to induce oxidative stress by different conditions in 3T3 and HaCaT cell lines. Antioxidant activity of in vitro PVM in 3T3 and HaCaT cell lines was evaluated by ROS assay. Our results demonstrate that PVM contains significant amounts of shikimic acid together with caffeoylquinic acid derivatives and flavonoids such as epicatechin and catechin; PVM is not cytotoxic at physiological concentrations against the different cell lines, showing cytoprotective and cellular repair activity in 3T3 fibroblast cells. This biological activity could be attributed to the high content of polyphenolic compounds. The fronds of the P. vulgare are a source of polyphenolic compounds, which can be responsible for certain traditional uses like wound healing properties. In the present work, fronds of the common polypody are positioned as a candidate for pharmaceutical applications based on traditional medicine uses but also as potential food ingredients due to lack of toxicity at physiological concentrations.

Improved Efficacy and Stability of Silymarin Loaded Nanocochleates Over Liposomes for the Treatment of Skin Diseases

Aim: Silymarin, a complex polyphenolic component mixture with anti-oxidant, anti-inflammatory, and membrane-stabilizing property is being investigated in several dermatological conditions. Present research aims to evaluate potential of silymarin loaded nanocochleates and liposomal topical application for treating chronic skin diseases. Study Design: Silymarin loaded liposomes and nanocochleates were formulated and optimized using Design Expert software. Different invitro and exvivo tests were performed to compare their performance. Place and Duration of Study: The study was conducted in Smt. Kashibai Navale College of Pharmacy, Pune, India, between January 2019 till February 2020. Methodology: Liposomes were prepared using ethanol injection method and further treated with calcium chloride to form nanocochleates by trapping method. Design of experiments (32 Factorial Design) was used for optimization of nanocochleates. Cell line studies (HaCaT cell lines) and short term stability studies were performed to compare the efficacy and stability respectively. Results: Particle size, entrapment efficiency and drug deposition in Wistar Rat Skin was found to be statistically significant for nanocochleates over liposomes proving superiority of cochleates. Both the carriers sustained release of silymarin for 24h. Antimicrobial efficacy of nanocochleates against E.coli and S.aureus was significant. Inhibition of hyper proliferation of HaCaT cell lines (key mechanism by which most of the antipsoriatic drugs act) demonstrated the superiority of nanocochleates over liposomes.The nanocochleates also displayed better stability compared to liposomes due to decreased entrapment efficacy and leakage of drug. Conclusion: Silymarin loaded Nanocochleates could prove as a promising topical drug delivery system for the treatment of chronic skin diseases like psoriasis.

A New Look at the Purported Health Benefits of Commercial and Natural Clays

Clays attributed to have medicinal properties have been used since prehistoric times and are still used today as complementary medicines, which has given rise to unregulated “bioceutical” clays to treat skin conditions. Recently, clays with antibacterial characteristics have been proposed as alternatives to antibiotics, potentially overcoming modern day antibiotic resistance. Clays with suggested antibacterial properties were examined to establish their effects on common wound-infecting bacteria. Geochemical, microscopical, and toxicological characterization of clay particulates, their suspensions and filtered leachates was performed on THP-1 and HaCaT cell lines. Cytoskeletal toxicity, cell proliferation/viability (MTT assays), and migration (scratch wounds) were further evaluated. Clays were assayed for antibacterial efficacy using minimum inhibitory concentration assays. All clays possessed a mineral content with antibacterial potential; however, clay leachates contained insufficient ions to have any antibacterial effects. All clay leachates displayed toxicity towards THP-1 monocytes, while clay suspensions showed less toxicity, suggesting immunogenicity. Reduced clay cytotoxicity on HaCaTs was shown, as many leachates stimulated wound-healing responses. The “Green” clay exhibited antibacterial effects and only in suspension, which was lost upon neutralization. pH and its interaction with clay particle surface charge is more significant than previously understood to emphasize dangers of unregulated marketing and unsubstantiated bioceutical claims.

Weighted gene coexpression network and experimental analyses identify lncRNA SPRR2C as a regulator of the IL-22-stimulated HaCaT cell phenotype through the miR-330/STAT1/S100A7 axis

Psoriasis is a chronic inflammatory disease of the skin with highly complex pathogenesis. In this study, we identified lncRNA SPRR2C (small proline-rich protein 2C) as a hub gene with a critical effect on the pathogenesis of psoriasis and response to treatment using both weighted gene coexpression network analysis (WGCNA) and differential expression analysis. SPRR2C expression was significantly upregulated in both psoriatic lesion samples and HaCaT cell lines in response to IL-22 treatment. After SPRR2C knockdown, IL-22-induced suppression of HaCaT proliferation, changes in the KRT5/14/1/10 protein levels, and suppression of the IL-1β, IL-6, and TNF-α mRNA levels were dramatically reversed. In the coexpression network with SPRR2C based on GSE114286, miR-330 was significantly negatively correlated with SPRR2C, while STAT1 and S100A7 were positively correlated with SPRR2C. By binding to miR-330, SPRR2C competed with STAT1 and S100A7 to counteract miR-330-mediated suppression of STAT1 and S100A7. MiR-330 overexpression also reversed the IL-22-induced changes in HaCaT cell lines; in response to IL-22 treatment, miR-330 inhibition significantly attenuated the effects of SPRR2C knockdown. STAT1 and S100A7 expression was significantly upregulated in psoriatic lesion samples. The expression of miR-330 had a negative correlation with the expression of SPRR2C, while the expression of SPRR2C had a positive correlation with the expression of STAT1 and S100A7. Thus, SPRR2C modulates the IL-22-stimulated HaCaT cell phenotype through the miR-330/STAT1/S100A7 axis. WGCNA might uncover additional biological pathways that are crucial in the pathogenesis and response to the treatment of psoriasis.

Lactic Acid Modified Natural Rubber–Bacterial Cellulose Composites

Green composite films of natural rubber/bacterial cellulose composites (NRBC) were prepared via a latex aqueous microdispersion process. The acid modified natural rubber/bacterial cellulose composites (ANRBC), in which lactic acid was used, showed significant improvement in mechanical properties, melting temperature, and high resistance to polar and non-polar solvents. The ANRBC films exhibited improved water resistance over that of BC and NRBC films, and possessed a higher resistance to non-polar solvents, such as toluene, than NR and NRBC films. The modification had a slight effect on the degradability of the composite films in soil. The NRBC and ANRBC films were biodegradable; the NRBC80 and ANRBC80 films were degraded completely within 3 months in soil. NRBC and ANRBC showed no antibacterial activity against Escherichia coli and Staphylococcus aureus and did not show cytotoxic effects on the HEK293 and HaCaT cell lines.

DEVELOPMENT AND CHARACTERIZATION OF TOPICAL NANOPARTICULATE ANTIPSORIATIC POLYHERBAL CREAM

Objective: The lack of possible cure and associated disadvantages of allopathic medicines in the topical treatment of psoriasis has led to extensive research for anti-psoriatic activities of plant-based drugs. The present study was aimed to develop topical cream formulation containing the Nanostructured Lipid Carriers of Azadirachta indica leaves extract (AE), Lawsonia inermis leaves extract (LE) and fruit extract of Mallotus philippensis (ME) and assessing the antipsoriatic activity of prepared cream formulation. Methods: The Drug loaded NLCs were prepared via hot homogenization technique and incorporated into the water in oil (w/o) cream base that was prepared by the emulsification method. Evaluation of cream formulation included advanced preclinical trials using Rat Ultraviolet Ray-B photodermatitis and Mousetail method, anti-lipid peroxidation assay, nitric oxide scavenging activity, cytotoxicity and DNA fragmentation on HaCat cell lines to ascertain antipsoriatic potential and in vitro drug diffusion. Results: In spite of the low amount of the drug loading in NLCs their potency was much higher as displayed in Advanced Preclinical studies and cell line studies for psoriasis indicating the suitability of NLCs loaded creams for skin applications due to their various desirable effects on the skin. Occlusive properties of the prepared NLC on the skin provide an increase in drug penetration particularly via skin moisturization. The in vitro drug diffusion studies suggest the prolonged and almost complete release of AE, LE and ME from NLC based cream up to 24 h that was found to be 81.24±0.51%, 74.31±0.25% and 70.43±0.78% respectively. The literature survey supports the novelty of such topical anti-psoriatic polyherbal nanoparticulate cream. Conclusion: It can be concluded that the AE, LE and ME loaded NLC based cream can be used for prolonged topical delivery of drugs for the management of Psoriasis.

TOPICAL ANTI-PSORIATIC NANOPARTICULATE DRUG DELIVERY SYSTEM

Objective: Development of effective drug delivery in the treatment of psoriasis is the major challenge for its successful management. To develop and assess the potential of Nanostructured Lipid Carriers (NLCs) enriched with the powdered leaves extracts of Azadirachta indica (AE), Lawsonia inermis (LE) and fruit extract of Mallotus philippensis (ME) in the management of psoriasis. Methods: Drug loaded NLCs were prepared via hot homogenization technique by adopting 23 factorial design with factors X1 as the concentration of lipids, X2 concentration of surfactants and X3 being the number of homogenization cycle. The responses Y1 and Y2 were particle size and zeta potential. The optimized batch was obtained from Surface response plot and was evaluated for zeta potential, % entrapment efficiency, % drug loading, Scanning Electron Microscopy(SEM), % in vitro diffusion of drugs from the NLCs, anti-lipid peroxidation and nitric oxide scavenging activities, cytotoxicity on HaCat cell lines, Mouse Tail and Rat ultraviolet ray B photodermatitis models for Psoriasis. Results: The optimized batch of NLCs was found within the nanosized range with a relatively low polydispersity index and zeta potential of-20mV. The %EE for an optimized batch of NLCs was found to be 98.97±0.83%, 96.99±0.56% and 99.25±0.55% and the %DL of 21.84±0.15%, 8.55±045%, and 87.91±0.38% respectively for AE, LE and ME. The SEM images showed the spherical vesicular structures of drugs loaded NLCs. The in-vitro diffusion of drugs from the NLCs followed initial burst release thereafter sustained release for 24 h. The AE, LE and ME loaded NLCs proved to possess anti-lipid peroxidation and nitric oxide scavenging activities, cytotoxicity on HaCat cell lines, DNA fragmentation on HaCat cell lines which are biomarkers in the pathogenesis of psoriasis. The results of Mouse Tail and Rat ultraviolet ray B photodermatitis models for Psoriasis supported the anti-psoriatic potential of AE, LE and ME loaded NLCs. Conclusion: AE, LE and ME loaded NLCs can be used for prolonged topical delivery to the psoriatic skin for an effective treatment.

Chemical Composition of Essential Oils from Different Parts of Zingiber kerrii Craib and Their Antibacterial, Antioxidant, and Tyrosinase Inhibitory Activities

The essential oils of the fresh rhizomes; flowers; and leaves of Zingiber kerrii Craib were investigated using different extraction techniques; including solid-phase microextraction (SPME), hydrodistillation (HD), and organic solvent (OS), and characterized by gas chromatography–mass spectrometry (GC–MS). A total of 37 SPME; 19 HD; and 36 OS compounds were identified from the rhizome extract of Z. kerrii; with the major components being α-pinene; β-pinene; and terpinen-4-ol; respectively. From the flower extract; 16 SPME; 2 HD; and 10 OS compounds were identified; (E)-caryophyllene was found as a major compound by these techniques. The leaf extract exhibited 20 SPME; 13 HD; and 14 OS compounds; with α-pinene; (E)-caryophyllene; and n-hexadecanoic acid being the major compounds; respectively. The rhizome extract showed tyrosinase inhibitory activity of 71.60% and a total phenolic content of 22.4 mg gallic acid/g. The IC50 values of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assays were 25.2 µg/mL and 153.6 µg/mL; respectively; and the ferric ion reducing antioxidant power (FRAP) assay value was 318.5 µM ascorbic acid equivalent (AAE)/g extract. The rhizome extract showed weak antibacterial activity. This extract showed no adverse toxicity in human keratinocyte (HaCaT) cell lines at concentrations below 200 µg/mL.

Epidermal Growth Factor and Adenosine Triphosphate Induce Natrium Iodide Symporter Expression in Breast Cancer Cell Lines

AIM: This study aims to investigate the effect of ATP, EGF and combination of those two to the Natrium Iodide Symporter (NIS) expression in MCF7, SKBR3 and HaCaT cell lines. METHODS: MCF7, SKBR3 and HaCaT cell lines were treated with ATP, EGF and combination of those two for 6, 12 and 24 hours. The expression of NIS mRNA was measured through quantitative-reverse transcription-polymerase chain reaction (qRT-PCR). The NIS protein expression was confirmed by immunocytofluorescence. RESULTS: NIS mRNA was expressed in SKBR3 and HaCaT cell lines but not in MCF7. The levels of NIS mRNA expression, after treatment by epidermal growth factor (EGF), adenosine Tri-Phosphate (ATP) or the combination of both for 6 and 12 hours were not significantly different from those of untreated cells. However, the treatment by a combination of ATP and EGF for 24 hours increases the level of NIS mRNA expression by 1.6 fold higher than that of the untreated cells (1.6241 ± 0.3, p < 0.05) and protein NIS expression increase significantly by the treatment than untreated cells (P < 0.05). CONCLUSION: The level of NIS expression varies among the different subtypes of breast cancer cell lines. MCF7 cell line is representing the luminal A subtype of breast cancer does not express NIS. Only SKBR3 cell line express NIS and this subtype might be suitable to receive radioiodine therapy as those cells expressing NIS. A combination treatment of EGF and ATP increases the expression of NIS mRNA and protein at the membrane in SKBR3 cells.