TOPICAL ANTI-PSORIATIC NANOPARTICULATE DRUG DELIVERY SYSTEM

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2020v12i2.36536 ◽

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.

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

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2019.v12i6.32594 ◽

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.

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

Frontiers in Pharmacology ◽

10.3389/fphar.2021.727528 ◽

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.

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DESIGN AND CHARACTERIZATION OF NANOSPRAY WITH SELF-NANOEMULSIFYING DRUG DELIVERY SYSTEM USING SINERGISTIC COMBINATION OF MELASTOMA MALABATHRICUM L. FRACTION AND GENTAMICIN

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2021v13i2.40094 ◽

2021 ◽

pp. 254-263

Author(s):

LIZA PRATIWI ◽

RAFIKA SARI ◽

PRATIWI APRIDAMAYANTI

Keyword(s):

Drug Delivery ◽

Particle Size ◽

Zeta Potential ◽

Drug Delivery System ◽

Design Expert ◽

Melastoma Malabathricum ◽

Diffusion Studies ◽

In Vitro Diffusion ◽

Optimal Formula

Objective: This study aimed to design a formula using Design-Expert software to obtain optimal Self-Nanoemulsifying Drug Delivery System (SNEDDS) formulas and to analyze nanospray characteristics of optimal SNEDDS. Methods: The study began with preparing ethanol extract from Melastoma malabathricum. The extract was then fractionated using ethyl acetate. The formulation design stage began with a solubility test of Melastoma malabathricum fraction and gentamicin (MFG) in various surfactants, co-surfactants and oils. Furthermore, the 14 formula of SNEDDS with various compositions of the selected surfactants, co-surfactants and oils were formulated and evaluated with pH response and emulsification time. Analysis was carried out using Design-Expert software with the simplex lattice design method in order to obtain the optimal formula profile. The pH, emulsification time, particle size, and zeta potential of the nanospray from SNEDDS optimal formulas were physically characterized. Stability of SNEDDS and the nanospray was then tested with freeze-thaw cycling and in vitro diffusion studies with Franz diffusion. Results: Based on the study, the ratios of optimal formula SNEDDS composition of Tween 80, propylene glycol, and soybean oil were 2.69: 2.64: 1.67 parts. Nanospray with SNEDDS technology had characteristics of pH 5.61±0.16, emulsification time 7.68±0.18, particle size 270.7 nm, and zeta potential-37.20 mV, and it was stable. Conclusion: Nanospray can be formulated from optimal SNEDDS using Design-Expert software. Nanospray with SNEDDS technology has physical characteristics and is stable. In vitro diffusion studies revealed that the release of Melastoma malabathricum from nanospray was faster than that without preparation.

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Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line.

The Journal of Cell Biology ◽

10.1083/jcb.106.3.761 ◽

1988 ◽

Vol 106 (3) ◽

pp. 761-771 ◽

Cited By ~ 2754

Author(s):

P Boukamp ◽

R T Petrussevska ◽

D Breitkreutz ◽

J Hornung ◽

A Markham ◽

...

Keyword(s):

Epithelial Cell ◽

Cell Line ◽

Human Skin ◽

Cell Lines ◽

Hacat Cell ◽

Epithelial Cell Line ◽

Spontaneous Transformation ◽

Chromosomal Alterations ◽

Hacat Cell Line

In contrast to mouse epidermal cells, human skin keratinocytes are rather resistant to transformation in vitro. Immortalization has been achieved by SV40 but has resulted in cell lines with altered differentiation. We have established a spontaneously transformed human epithelial cell line from adult skin, which maintains full epidermal differentiation capacity. This HaCaT cell line is obviously immortal (greater than 140 passages), has a transformed phenotype in vitro (clonogenic on plastic and in agar) but remains nontumorigenic. Despite the altered and unlimited growth potential, HaCaT cells, similar to normal keratinocytes, reform an orderly structured and differentiated epidermal tissue when transplanted onto nude mice. Differentiation-specific keratins (Nos. 1 and 10) and other markers (involucrin and filaggrin) are expressed and regularly located. Thus, HaCaT is the first permanent epithelial cell line from adult human skin that exhibits normal differentiation and provides a promising tool for studying regulation of keratinization in human cells. On karyotyping this line is aneuploid (initially hypodiploid) with unique stable marker chromosomes indicating monoclonal origin. The identity of the HaCaT line with the tissue of origin was proven by DNA fingerprinting using hypervariable minisatellite probes. This is the first demonstration that the DNA fingerprint pattern is unaffected by long-term cultivation, transformation, and multiple chromosomal alterations, thereby offering a unique possibility for unequivocal identification of human cell lines. The characteristics of the HaCaT cell line clearly document that spontaneous transformation of human adult keratinocytes can occur in vitro and is associated with sequential chromosomal alterations, though not obligatorily linked to major defects in differentiation.

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

Applied Sciences ◽

10.3390/app11199244 ◽

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.

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FORMULATION AND EVALUATION OF ACYCLOVIR LOADED NOVEL NANO-EMULSION GEL FOR TOPICAL TREATMENT OF HERPES SIMPLEX VIRAL INFECTIONS

Journal of Drug Delivery and Therapeutics ◽

10.22270/jddt.v8i5-s.1968 ◽

2018 ◽

Vol 8 (5-s) ◽

pp. 265-270

Author(s):

Swati Patel ◽

Prabhat Jain ◽

Geeta Parkhe

Keyword(s):

Herpes Simplex ◽

Zeta Potential ◽

Viral Infections ◽

In Vitro Release ◽

Stability Study ◽

Oral Drug ◽

Drug Content ◽

Nanoemulsion Gel ◽

In Vitro Diffusion

Acyclovir has low bioavailability mainly due to low solubility. This study aimed to formulate an optimized acyclovir (ACV) nanoemulsion gel for the slow, variable and incomplete oral drug absorption in patient suffering from herpes simplex viral infection. The dispersion solubility of acyclovir was studied in various oils, surfactants and co-surfactants and by constructing pseudo phase ternary diagram nanoemulsion area was identified. The optimized formulations of nanoemulsions were subjected to thermodynamic stability tests. After stability study, stable formulation was characterized for droplet size, pH determination, centrifugation, % drug content in nanoemulsion, Zeta Potential and Vesicle size measurement and than nanoemulsion gel were prepared and characterized for spreadability, measurement of viscosity, drug content, In-vitro diffusion, in-vitro release data. Span 40 was selected as surfactant, PEG 400 as co surfactant and castor oil as oil component based on solubility study. The in vitro drug release from acyclovir nanoemulsion gel was found to be considerably higher in comparison to that of the pure drug. The in-vitro diffusion of nanoemulsion gel was significantly good. Based on this study, it can be concluded the solubility and permeability of acyclovir can be increased by formulating into nanoemulsion gel. Keywords: Acyclovir, Nanoemulsion, In-vitro diffusion, Zeta potential, Stability

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Multivesicular Liposomes for the Sustained Release of Angiotensin I-Converting Enzyme (ACE) Inhibitory Peptides from Peanuts: Design, Characterization, and In Vitro Evaluation

Molecules ◽

10.3390/molecules24091746 ◽

2019 ◽

Vol 24 (9) ◽

pp. 1746 ◽

Cited By ~ 3

Author(s):

Ning Li ◽

Aimin Shi ◽

Qiang Wang ◽

Guoquan Zhang

Keyword(s):

Drug Delivery ◽

Zeta Potential ◽

Sustained Release ◽

Converting Enzyme ◽

Angiotensin I ◽

Ace Inhibitory Activity ◽

Angiotensin I Converting Enzyme ◽

Release Drug ◽

Ace Inhibitory

The multivesicular liposome (MVL) provides a potential delivery approach to avoid the destruction of the structure of drugs by digestive enzymes of the oral cavity and gastrointestinal system. It also serves as a sustained-release drug delivery system. In this study, we aimed to incorporate a water-soluble substance into MVLs to enhance sustained release, prevent the destruction of drugs, and to expound the function of different components and their mechanism. MVLs were prepared using the spherical packing model. The morphology, structure, size distribution, and zeta potential of MVLs were examined using an optical microscope (OM), confocal microscopy (CLSM), transmission electron cryomicroscope (cryo-EM) micrograph, a Master Sizer 2000, and a zeta sizer, respectively. The digestion experiment was conducted using a bionic mouse digestive system model in vitro. An in vitro release and releasing mechanism were investigated using a dialysis method. The average particle size, polydispersity index, zeta potential, and encapsulation efficiency are 47.6 nm, 1.880, −70.5 ± 2.88 mV, and 82.00 ± 0.25%, respectively. The studies on the controlled release in vitro shows that MVLs have excellent controlled release and outstanding thermal stability. The angiotensin I-converting enzyme (ACE) inhibitory activity of ACE-inhibitory peptide (AP)-MVLs decreased only 2.84% after oral administration, and ACE inhibitory activity decreased by 5.03% after passing through the stomach. Therefore, it could serve as a promising sustained-release drug delivery system.

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

Molecules ◽

10.3390/molecules24040699 ◽

2019 ◽

Vol 24 (4) ◽

pp. 699 ◽

Cited By ~ 2

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.

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Soft Cationic Nanoparticles for Drug Delivery: Production and Cytotoxicity of Solid Lipid Nanoparticles (SLNs)

Applied Sciences ◽

10.3390/app9204438 ◽

2019 ◽

Vol 9 (20) ◽

pp. 4438 ◽

Cited By ~ 14

Author(s):

Amélia Silva ◽

Carlos Martins-Gomes ◽

Tiago Coutinho ◽

Joana Fangueiro ◽

Elena Sanchez-Lopez ◽

...

Keyword(s):

Drug Delivery ◽

Surface Properties ◽

Cell Lines ◽

Solid Lipid Nanoparticles ◽

Lipid Nanoparticles ◽

Cationic Lipids ◽

Solid Lipid ◽

Mcf 7

The surface properties of nanoparticles have decisive influence on their interaction with biological barriers (i.e., living cells), being the concentration and type of surfactant factors to have into account. As a result of different molecular structure, charge, and degree of lipophilicity, different surfactants may interact differently with the cell membrane exhibiting different degrees of cytotoxicity. In this work, the cytotoxicity of two cationic solid lipid nanoparticles (SLNs), differing in the cationic lipids used as surfactants CTAB (cetyltrimethylammonium bromide) or DDAB (dimethyldioctadecylammonium bromide), referred as CTAB-SLNs and DDAB-SLNs, respectively, was assessed against five different human cell lines (Caco-2, HepG2, MCF-7, SV-80, and Y-79). Results showed that the cationic lipids used in SLN production highly influenced the cytotoxic profile of the particles, with CTAB-SLNs being highly cytotoxic even at low concentrations (IC50 < 10 µg/mL, expressed as CTAB amount). DDAB-SLNs produced much lower cytotoxicity, even at longer exposure time (IC50 from 284.06 ± 17.01 µg/mL (SV-80) to 869.88 ± 62.45 µg/mL (MCF-7), at 48 h). To the best of our knowledge, this is the first report that compares the cytotoxic profile of CTAB-SLNs and DDAB-SLNs based on the concentration and time of exposure, using different cell lines. In conclusion, the choice of the right surfactant for biological applications influences the biocompatibility of the nanoparticles. Regardless the type of drug delivery system, not only the cytotoxicity of the drug-loaded nanoparticles should be assessed, but also the blank (non-loaded) nanoparticles as their surface properties play a decisive role both in vitro and in vivo.

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