Regulation of Human Beta-Defensin 3(hBD-3) in Human Keratinocyte(HaCaT) Cell Lines

2003 ◽  
Vol 15 (1) ◽  
pp. 1 ◽  
Author(s):  
Yu-Jin Kim ◽  
Chang-Kwun Hong ◽  
Seong-Jun Seo
Keyword(s):  
Cell Lines ◽  
Hacat Cell ◽  
Human Keratinocyte ◽  
Hacat Cell Lines

scholarly journals Bioactivity Determination of a Therapeutic Recombinant Human Keratinocyte Growth Factor by a Validated Cell-based Bioassay

Molecules ◽  
2019 ◽  
Vol 24 (4) ◽  
pp. 699 ◽  
Author(s):  
Wenrong Yao ◽  
Ying Guo ◽  
Xi Qin ◽  
Lei Yu ◽  
Xinchang Shi ◽  
...  
Keyword(s):  
Growth Factor ◽  
Cell Lines ◽  
Hacat Cell ◽  
Keratinocyte Growth Factor ◽  
Growth Factor Receptor ◽  
Luciferase Reporter ◽  
Hematopoietic Stem ◽  
Human Keratinocyte ◽  
Serum Response ◽  
Hacat Cell Lines

The therapeutic recombinant human keratinocyte growth factor 1 (rhKGF-1) was approved by the FDA for oral mucositis resulting from hematopoietic stem cell transplantation for hematological malignancies in 2004. However, no recommended bioassay for rhKGF-1 bioactivity has been recorded in the U.S. Pharmacopoeia. In this study, we developed an rhKGF-1-dependent bioassay for determining rhKGF-1 bioactivity based on HEK293 and HaCat cell lines that stably expressed the luciferase reporter driven by the serum response element (SRE) and human fibroblast growth factor receptor (FGFR2) IIIb. A good responsiveness to rhKGF-1 and rhKGF-2 shared by target HEK293/HaCat cell lines was demonstrated. Our stringent validation was completely focused on specificity, linearity, accuracy, precision, and robustness according to the International Council for Harmonization (ICH) Q2 (R1) guidelines, AAPS/FDA Bioanalytical Workshop and the Chinese Pharmacopoeia. We confirmed the reliability of the method in determining rhKGF bioactivity. The validated method is highly timesaving, sensitive, and simple, and is especially valuable for providing information for quality control during the manufacture, research, and development of therapeutic rhKGF.

scholarly journals EVALUATION OF ANTIPROLIFERATIVE ACTIVITY OF SIDDHA ANTI-PSORIATIC FORMULATION PANCHAMUGA CHENDHURAM USING CULTURED HUMAN KERATINOCYTE CELL LINES

2019 ◽  
pp. 280-283
Author(s):  
RAJALAKSHMI S ◽  
RAMYA VT ◽  
SAMRAJ K
Keyword(s):  
Cell Lines ◽  
Antiproliferative Activity ◽  
Hacat Cell ◽  
Scientific Evidence ◽  
Positive Control ◽  
Traditional Use ◽  
Human Keratinocyte ◽  
Ic50 Values ◽  
Hacat Cell Lines

Objectives: This study was aimed at scientifically evaluating the in vitro antipsoriatic activity of Siddha drug Panchamuga Chendhuram (PMC) in human keratinocyte (HacaT) cell lines. Methods: The Siddha drug PMC tested for antipsoriatic activity on HacaT cell lines was morphologically examined by phase contrast microscopy, and the cell viability was determined by 3- (4, 5 dimethyl thiazole-2 yl) -2.5-diphenyl tetrazolium bromide assay. About 100 μl of different concentrations (2, 6, 10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 μg/ml) of the test samples were prepared in the cell culture medium and incubated for 24 h and 48 h to determine the viable cells. Results: The results revealed that Siddha drug PMC showed hopeful antiproliferative activity. In vitro studies showed that after 24 h and 48 h incubation, the inhibitory concentration 50 (IC50) values of PMC (IC50 20 μg/ml) were 72.08±27.56 μg/ml and 43.91±17.71 μg/ml, respectively, as compared with Asiaticoside as a positive control with an IC50 value of 20.13 μg/ml. Conclusion: Thus, this study provides scientific evidence about the efficacy of the Siddha drug PMC against the HacaT cell lines confirming its traditional use in psoriasis treatment and also emphasizes the need for antipsoriatic evaluation in animal models.

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

2021 ◽  
Vol 11 (19) ◽  
pp. 9244
Author(s):  
Monika Kalinowska ◽  
Hanna Lewandowska ◽  
Marek Pruszyński ◽  
Grzegorz Świderski ◽  
Ewelina Gołębiewska ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Solid State ◽  
Cytotoxic Activity ◽  
General Formula ◽  
Cell Lines ◽  
Elemental Analysis ◽  
Hacat Cell ◽  
Water Solution ◽  
Ft Ir ◽  
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.

scholarly journals 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

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Meijunzi Luo ◽  
Pan Huang ◽  
Yi Pan ◽  
Zhu Zhu ◽  
Rong Zhou ◽  
...  
Keyword(s):  
Cell Lines ◽  
Hacat Cell ◽  
Differential Expression Analysis ◽  
Coexpression Network ◽  
Cell Phenotype ◽  
Mrna Levels ◽  
Psoriatic Lesion ◽  
Gene Coexpression Network ◽  
Gene Coexpression ◽  
Hacat Cell Lines

AbstractPsoriasis 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.

TOPICAL ANTI-PSORIATIC NANOPARTICULATE DRUG DELIVERY SYSTEM

2020 ◽  
pp. 76-85
Author(s):  
KIRAN BHISE ◽  
SHADAB KHAN ◽  
GHAZALA MULLA
Keyword(s):  
Nitric Oxide ◽  
Drug Delivery ◽  
Lipid Peroxidation ◽  
Zeta Potential ◽  
Cell Lines ◽  
Hacat Cell ◽  
In Vitro Diffusion ◽  
Ultraviolet Ray ◽  
Hacat Cell Lines

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.

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

2021 ◽  
Vol 12 ◽  
Author(s):  
Adrià Farràs ◽  
Montserrat Mitjans ◽  
Filippo Maggi ◽  
Giovanni Caprioli ◽  
María Pilar Vinardell ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Lines ◽  
Hacat Cell ◽  
Polyphenolic Compounds ◽  
European Medicines Agency ◽  
Food Ingredients ◽  
Traditional Uses ◽  
Polyphenolic Profile ◽  
Hacat 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.

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

2021 ◽  
pp. 163-176
Author(s):  
Rukhsana Rub ◽  
Neha Munot ◽  
Akshay Wadate
Keyword(s):  
Cell Lines ◽  
Skin Diseases ◽  
Hacat Cell ◽  
Entrapment Efficiency ◽  
Significant Inhibition ◽  
Component Mixture ◽  
Ethanol Injection ◽  
Chronic Skin ◽  
Chronic Skin Diseases ◽  
Hacat Cell Lines

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.

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

2019 ◽  
Vol 7 (13) ◽  
pp. 2088-2092 ◽  
Author(s):  
Aisyah Elliyanti ◽  
Andani Eka Putra ◽  
Yunia Sribudiani ◽  
Noormartany Noormartany ◽  
Johan S. Masjhur ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Mrna Expression ◽  
Cell Line ◽  
Cell Lines ◽  
Hacat Cell ◽  
Breast Cancer Cell Lines ◽  
Epidermal Growth ◽  
Hacat 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.

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