Cytotoxicity presents one of the required criteria in the biological evaluation of medical devices. In this study, the semi-direct contact test was used to evaluate the potential cytotoxicity of hydrogel wound dressings compared to the conventional extract test. Three types of hydrogel sheets were fabricated from poly(vinyl alcohol) (PVA) by gamma irradiation: Bare sheets, silver (Ag)-coated sheets, and Aloe vera (AV)-coated sheets. In the extract test, L929 cells were cultured in the extract derived from the elution of hydrogel samples. For the semi-direct contact test, the cells were cultured in situ with the hydrogel samples placed inside transwell inserts above the cell monolayers. At the endpoint of both tests, MTT assay was performed, and the cell viability was determined from the absorbance of formazan. Only the bare and AV-coated hydrogel sheets showed cell viability above the 70 % threshold that ensured the non-cytotoxicity by both tests. For Ag-coated sheets, less than 70 % cell viability occurred when Ag coating was ³ 0.2 mg/cm2. Interestingly, the formazan-depleted area underneath the Ag-coated sample could be clearly observed by the semi-direct contact test. The release of Ag in the form of nanoparticles was confirmed by UV-Vis absorption at 420 nm. In conclusion, the semi-direct contact test can serve as a reliable alternative to the conventional extract test in evaluating the potential cytotoxicity of hydrogel wound dressings. HIGHLIGHTS A semi-direct contact test with transwell inserts was investigated as an alternative approach to the conventional extract test in evaluating the cytotoxicity of hydrogel wound dressings Leachable substances from the hydrogel dressings permeated through the porous membrane of transwell inserts to the reach the cell monolayer underneath Cell viability results based on MTT assay obtained from semi-direct contact test were in agreement with the extract test Only the semi-direct contact test provided the visual assessment of localized toxicity caused by the release of silver nanoparticles from hydrogel sheets onto the cell monolayer GRAPHICAL ABSTRACT
Objective: Cannabidiol (CBD), a phytochemical active compound from the Cannabis sativa L., has become a popular ingredient in many industries, especially skincare products. However, the scientific evidence supporting its potential skin benefits and safety concerns are still unclear. Therefore, the aim of this study was to investigate the short-term and long-term cytotoxic effects of CBD and its potential melanin-promoting effect on skin cells in order to deeply evaluate the safety of CBD for use in cosmetics. Material and Methods: HaCaT keratinocytes and B16F10 melanoma cell lines were cultured and investigated in regard to the cytotoxicity of cannabidiol in various concentrations (0-10 μg/ml) in the short term and long term by sulforhodamine B (SRB) assay and clonogenic assay, respectively. Next, the cellular melanin production was measured by melanin content assay. The expression of the related genes was accessed by qPCR. Results: The short-term and long-term cytotoxicity studies revealed that CBD at a low concentration was not toxic to skin cells. In addition, CBD could induce melanogenesis in melanocytes by increasing melanin content and upregulating tyrosinase expression. Also, CBD provoked cell proliferation and enhanced vascular endothelial growth factor (VEGF) mRNA expression in keratinocytes. Conclusion: Our study demonstrated that CBD at a low concentration (0.6 μg/ml) is safe for the skin cells in vitro and should thus also be safe if applied to skin. Additionally, CBD could significantly enhance melanogenesis and cell proliferation, which confirms its potential as a cosmeceutical product.
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