Identification of Cyclobutane Pyrimidine Dimer-Responsive Genes Using UVB-Irradiated Human Keratinocytes Transfected with In Vitro-Synthesized Photolyase mRNA

Cell culture systems have become increasingly popular as a means of screening toxic agents and studying toxic mechanisms of drugs and other chemicals at the cellular and subcellular levels. These in vitro tests can be conducted rapidly in a broad range of relevant mammalian culture systems; a variety of biological and biochemical cytotoxicity endpoints can be examined. The following study utilized human keratinocytes to evaluate the relative cytotoxicities of nitrofurazone (NF) and silver sulfadiazine (SS), the active ingredients of FURACIN(R) Topical Cream and SILVADENE(R) Cream, respectively. These compounds are anti-infectives used in the treatment of burn patients. Cell ultrastructure and elemental composition were utilized as cytotoxicity endpoints.Normal Human Epidermal Keratinocytes (HK) were prepared from the EpiPackTM culture system (Clonetics Corporation, Boulder, CO). For scanning electron microscopy (SEM) and transmission electron microscopy (TEM), cells were seeded on sterile 35 mm Falcon plastic dishes; for elemental microanalysis, cells were plated on polished pyrolytic carbon discs (E. Fullam, Latham, NY) placed in the culture dishes.

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Cryptotanshinone from Salvia miltiorrhiza Bunge lowers the Cytokeratin 1/10 expression in primary human Keratinocytes in vitro

10.1055/s-0037-1608052 ◽

2017 ◽

Author(s):

S Esch ◽

A Hensel ◽

S Brandt ◽

S König

Keyword(s):

Salvia Miltiorrhiza ◽

Human Keratinocytes ◽

Salvia Miltiorrhiza Bunge ◽

Primary Human Keratinocytes

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Dexamethasone-Loaded Lipomers: Development, Characterization, and Skin Biodistribution Studies

Pharmaceutics ◽

10.3390/pharmaceutics13040533 ◽

2021 ◽

Vol 13 (4) ◽

pp. 533

Author(s):

Eloy Pena-Rodríguez ◽

Maria Lajarin-Reinares ◽

Aida Mata-Ventosa ◽

Sandra Pérez-Torras ◽

Francisco Fernández-Campos

Keyword(s):

In Vitro Release ◽

Human Keratinocytes ◽

Topical Administration ◽

In Vitro Cytotoxicity ◽

Hair Follicles ◽

Vertical Diffusion ◽

Biodistribution Studies ◽

Follicular Targeting ◽

Depot Effect

Follicular targeting has gained more attention in recent decades, due to the possibility of obtaining a depot effect in topical administration and its potential as a tool to treat hair follicle-related diseases. Lipid core ethyl cellulose lipomers were developed and optimized, following which characterization of their physicochemical properties was carried out. Dexamethasone was encapsulated in the lipomers (size, 115 nm; polydispersity, 0.24; zeta-potential (Z-potential), +30 mV) and their in vitro release profiles against dexamethasone in solution were investigated by vertical diffusion Franz cells. The skin biodistribution of the fluorescent-loaded lipomers was observed using confocal microscopy, demonstrating the accumulation of both lipomers and fluorochromes in the hair follicles of pig skin. To confirm this fact, immunofluorescence of the dexamethasone-loaded lipomers was carried out in pig hair follicles. The anti-inflammatory (via TNFα) efficacy of the dexamethasone-loaded lipomers was demonstrated in vitro in an HEK001 human keratinocytes cell culture and the in vitro cytotoxicity of the nanoformulation was investigated.

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Ethosomes and Transethosomes for Mangiferin Transdermal Delivery

Antioxidants ◽

10.3390/antiox10050768 ◽

2021 ◽

Vol 10 (5) ◽

pp. 768

Author(s):

Maddalena Sguizzato ◽

Francesca Ferrara ◽

Supandeep Singh Hallan ◽

Anna Baldisserotto ◽

Markus Drechsler ◽

...

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Radical Scavenging ◽

Human Keratinocytes ◽

Antioxidant Response ◽

Correlation Spectroscopy ◽

Inflammatory Status ◽

Transmission Electron ◽

Anti Inflammatory

Mangiferin is a natural glucosyl xanthone with antioxidant and anti-inflammatory activity, making it suitable for protection against cutaneous diseases. In this study ethosomes and transethosomes were designed as topical delivery systems for mangiferin. A preformulation study was conducted using different surfactants in association with phosphatidylcholine. Vesicle dimensional distribution was monitored by photon correlation spectroscopy, while antioxidant capacity and cytotoxicity were respectively assessed by free radical scavenging analysis and MTT on HaCaT keratinocytes. Selected nanosystems were further investigated by cryogenic transmission electron microscopy, while mangiferin entrapment capacity was evaluated by ultracentrifugation and HPLC. The diffusion kinetics of mangiferin from ethosomes and transethosomes evaluated by Franz cell was faster in the case of transethosomes. The suitability of mangiferin-containing nanovesicles in the treatment of skin disorders related to pollutants was investigated, evaluating, in vitro, the antioxidant and anti-inflammatory effect of ethosomes and transethosomes on human keratinocytes exposed to cigarette smoke as an oxidative and inflammatory challenger. The ability to induce an antioxidant response (HO-1) and anti-inflammatory status (IL-6 and NF-kB) was determined by RT-PCR and immunofluorescence. The data demonstrated the effectiveness of mangiferin loaded in nanosystems to protect cells from damage. Finally, to gain insight into the keratinocytes’ uptake of ethosome and transethosome, transmission electron microscopy analyses were conducted, showing that both nanosystems were able to pass intact within the cells.

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