Indirect Cytotoxicity Evaluation of Silver Doped Bioglass Ag-S70C30 on Human Primary Keratinocytes

2005 ◽  
Vol 284-286 ◽  
pp. 431-434 ◽  
Author(s):  
U. Lohbauer ◽  
G. Jell ◽  
Priya Saravanapavan ◽  
Julian R. Jones ◽  
Larry L. Hench
Keyword(s):  
Cell Viability ◽  
Glass Powder ◽  
Culture Medium ◽  
Neutral Red ◽  
Epidermal Keratinocytes ◽  
Mitochondrial Activity ◽  
Human Epidermal Keratinocytes ◽  
Negative Effect ◽  
Healing Agent ◽  
Gel Glasses

Bioactive gel-glasses, such as the silver-doped Ag-S70C30 glass, can be used to modify the inflammatory response in a local body compartment such as in acne lesions and in nonhealing dermal wounds. In this study, the cytotoxicity of soluble silver, calcium and silica ions on human epidermal keratinocytes was investigated by measurements of mitochondrial activity (MTT assay) and neutral red dye uptake (NR assay). Ag-S70C30 extracts were prepared by soaking glass powder in complete culture medium at concentrations of 1 mg/ml and 2 mg/ml (mg of glass powder per ml of culture medium). Silver concentrations for both concentrations of approximately 1 ppm were detected by inductive coupled plasma analysis (ICP). No negative effect on the cell viability was measured for an initial gel-glass concentration of 1 mg/ml and for the two shortest extraction times at a concentration of 2 mg/ml. Based on the results from MTT/ NR assays, a pH rise of approximately one unit had no negative effect on the NHEK-A cell viability. This preliminary study on keratinocyte viability merits future investigations on silver bioglass as a novel antimicrobial wound healing agent.

Antimicrobial Treatment of Dental Osseous Defects with Silver Doped Bioglass: Osteoblast Cell Response

2005 ◽  
Vol 284-286 ◽  
pp. 435-438 ◽  
Author(s):  
U. Lohbauer ◽  
G. Jell ◽  
Priya Saravanapavan ◽  
Julian R. Jones ◽  
Larry L. Hench
Keyword(s):  
Cell Viability ◽  
Culture Medium ◽  
Bactericidal Effect ◽  
Neutral Red ◽  
Antimicrobial Treatment ◽  
Mitochondrial Activity ◽  
Graft Material ◽  
Human Osteoblasts ◽  
Negative Effect ◽  
Silver Dissolution

In dentistry, chronic periodontitis often leads to bone resorption together with an increasing risk of bacteremia. Bioactive glass has found extensive application as dental graft material. A successful antimicrobial bactericidal effect has been shown from the introduction of Ag2O into the glass composition. In this study, the cytotoxicity of soluble silver, calcium and silica ions on primary human osteoblasts was investigated by measurements of mitochondrial activity and neutral red dye uptake. Silver concentrations of 4 - 6 ppm (1 mg/ml conc.) and 6 - 9 ppm (2 mg/ml conc.) have been measured in complete culture medium. It was found that the bioactive gel-glass extract with an initial concentration of 1 mg/ml (1mg glass per ml of culture medium) has no negative effect, whereas increased gel-glass concentration of 2 mg/ml seemed to have a toxic effect on the cell viability of human osteoblasts. It might be concluded that a reduction of the rate of silver dissolution from the bioactive gel-glass might preserve a maximum cell viability.

Biological Interactions of Functionalized Single-Wall Carbon Nanotubes in Human Epidermal Keratinocytes

2007 ◽  
Vol 26 (2) ◽  
pp. 103-113 ◽  
Author(s):  
Leshuai W. Zhang ◽  
Liling Zeng ◽  
Andrew R. Barron ◽  
Nancy A. Monteiro-Riviere
Keyword(s):  
Carbon Nanotubes ◽  
Cell Viability ◽  
Single Wall Carbon Nanotubes ◽  
Epidermal Keratinocytes ◽  
Human Epidermal Keratinocytes ◽  
Single Wall Carbon ◽  
Biological Compatibility ◽  
Tnf Α ◽  
Wide Range ◽  
Significant Difference

Carbon nanotube–based nanovectors, especially functionalized nanotubes, have shown potential for therapeutic drug delivery. 6-Aminohexanoic acid–derivatized single-wall carbon nanotubes (AHA-SWNTs) are soluble in aqueous stock solutions over a wide range of physiologically relevant conditions; however, their interactions with cells and their biological compatibility has not been explored. Human epidermal keratinocytes (HEKs) were dosed with AHA-SWNTs ranging in concentration from 0.00000005 to 0.05 mg/ml. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell viability decreased significantly ( p < .05) from 0.00005 to 0.05 mg/ml after 24 h. The proinflammatory mediators of inflammation cytokines interleukin (IL)-6, IL-8, tumor necrosis factor (TNF)- α, IL-10, and IL-1 β were also assessed. Cytokine analysis did not show a significant increase in IL-6 and IL-8 in the medium containing 0.000005 mg/ml of AHA-SWNTs from 1 to 48 h. IL-6 increased in cells treated with 0.05 mg/ml of AHA-SWNTs from 1 to 48 h, whereas IL-8 showed a significant increase at 24 and 48 h. No significant difference ( p < .05) was noted with TNF- α, IL-10, and IL-1 β expression at any time point. Transmission electron microscopy of HEKs treated with 0.05 mg/ml AHA-SWNTs for 24 h depicted AHA-SWNTs localized within intracytoplasmic vacuoles in HEKs. Treatment with the surfactant 1% Pluronic F127 caused dispersion of the AHA-SWNT aggregates in the culture medium and less toxicity. These data showed that the lower concentration of 0.000005 mg/ml of AHA-SWNTs maintains cell viability and induces a mild cytotoxicity, but 0.05 mg/ml of AHA-SWNTs demonstrated an irritation response by the increase in IL-8.

scholarly journals Skin Barrier-enhancing, Antiwrinkle, and Antimelanogenic Effects of Probiotic Lysates Composed of Nucleotides

2021 ◽  
Vol 27 (6) ◽  
pp. 1343-1350
Author(s):  
Kyung-Min Kim ◽  
Ha-Yeon Kim ◽  
So-Yoon Cha ◽  
Ye-Hyang Kim ◽  
Ji-Won Song ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Cell Viability ◽  
Bifidobacterium Longum ◽  
Radical Scavenging ◽  
Skin Barrier ◽  
Skin Aging ◽  
Epidermal Keratinocytes ◽  
Human Epidermal Keratinocytes ◽  
Plant Materials ◽  
Matrix Metalloproteinase 1

Several previous studies have investigated the skin aging prevention effects of ceramide, hyaluronic acid, and natural or fermented plant materials. Recently, oral administration and dermal application of probiotics or probiotic lysates have shown antiaging effects. The purpose of this study is to optimize the preparation of probiotic lysates with a high concentration of nucleotides and to confirm the effects of probiotic lysates on the skin. Probiotic lysates were prepared by heating at 121°C for various periods with adding of sodium hyaluronic acid. Probiotic lysates of Bifidobacterium longum HDB7072, Lactobacillus paracasei HDB1196, and Lactobacillus acidophilus HDB1014 were applied to normal human epidermal keratinocytes (NHEKs), fibroblast cells, and B16F1 cells, respectively. Cell viability, antioxidant effects, and mRNA expression were evaluated by using MTT assays, DPPH assays, and qRT-PCR. Probiotic lysates prepared by heating the culture medium at 121°C for 2 h with 0.5% sodium hyaluronic acid showed the highest nucleotide concentration. In the three tested skin cells, the cell viability of filtered lysates was similar or higher to that of unfiltered lysates. HDB7072 lysates increased filaggrin expression in NHEKs. HDB1196 lysates showed DPPH radical-scavenging and antiwrinkle effects through the downregulation of matrix metalloproteinase-1 and upregulation of collagen type 1 in fibroblasts. HDB1014 lysates had antioxidant and antimelanogenic effects in B16F1 cells. Cell wall-removed probiotic lysates could be used as novel ingredients to improve skin aging and skin barrier issues.

Demonstration of a Choline Requirement for Optimal Keratinocyte Growth in a Defined Culture Medium

1988 ◽  
Vol 118 (12) ◽  
pp. 1487-1494 ◽  
Author(s):  
Philip R. Gordon ◽  
Liebe K. Gelman ◽  
Barbara A. Gilchrest
Keyword(s):  
Culture Medium ◽  
Basal Medium ◽  
Donor Cell ◽  
Cell Types ◽  
Dermal Fibroblasts ◽  
Cell Yield ◽  
Epidermal Keratinocytes ◽  
Good Growth ◽  
Human Epidermal Keratinocytes ◽  
Defined Culture

Abstract Nutrient requirements for proliferation and differentiated function of individual cell types can be determined using cell culture methodologies. Human epidermal keratinocytes are stimulated to grow by choline supplementation in the presence of myo-inositol when grown in a commercial nutrient medium containing six other defined supplements. The optimal range of choline concentrations varied among donor cell lines, but consistently fell between 36 µM and 180 µM. Addition of 72 µM choline increased cell yield to 250 ± 38% of that produced by myo-inositol supplementation alone and 92 ± 8% of that produced by addition of a highly mitogenic hypothalamic extract, which was previously required for good growth in this culture system. Supplementation of the basal medium with both the extract and choline resulted in 165 ± 13% of the cell yield observed with the extract addition alone. Supplementation with other phospholipid precursors did not further increase keratinocyte growth. Neither dermal fibroblasts nor epidermal melanocytes were stimulated by supplementation with choline, suggesting the high keratinocyte requirement is unusual. This completely defined culture medium for keratinocyte growth should prove useful in analyzing the role of phospholipids and other nutrients in human epidermis.

scholarly journals Ecklonia cava Extract and Dieckol Attenuate Cellular Lipid Peroxidation in Keratinocytes Exposed to PM10

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Jeong-won Lee ◽  
Jin Kyung Seok ◽  
Yong Chool Boo
Keyword(s):  
Lipid Peroxidation ◽  
Particulate Matter ◽  
Cell Viability ◽  
Airborne Particulate Matter ◽  
Ecklonia Cava ◽  
Epidermal Keratinocytes ◽  
Hacat Cells ◽  
Cellular Lipid ◽  
Airborne Particulate ◽  
Human Epidermal Keratinocytes

Airborne particulate matter can cause oxidative stress, inflammation, and premature skin aging. Marine plants such as Ecklonia cava Kjellman contain high amounts of polyphenolic antioxidants. The purpose of this study was to examine the antioxidative effects of E. cava extract in cultured keratinocytes exposed to airborne particulate matter with a diameter of <10 μm (PM10). After the exposure of cultured HaCaT keratinocytes to PM10 in the absence and presence of E. cava extract and its constituents, cell viability and cellular lipid peroxidation were assessed. The effects of eckol and dieckol on cellular lipid peroxidation and cytokine expression were examined in human epidermal keratinocytes exposed to PM10. The total phenolic content of E. cava extract was the highest among the 50 marine plant extracts examined. The exposure of HaCaT cells to PM10 decreased cell viability and increased lipid peroxidation. The PM10-induced cellular lipid peroxidation was attenuated by E. cava extract and its ethyl acetate fraction. Dieckol more effectively attenuated cellular lipid peroxidation than eckol in both HaCaT cells and human epidermal keratinocytes. Dieckol and eckol attenuated the expression of inflammatory cytokines such as tumor necrosis factor- (TNF-) α, interleukin- (IL-) 1β, IL-6, and IL-8 in human epidermal keratinocytes stimulated with PM10. This study suggested that the polyphenolic constituents of E. cava, such as dieckol, attenuated the oxidative and inflammatory reactions in skin cells exposed to airborne particulate matter.

scholarly journals Cytotoxicity assays as tools to assess water quality in the Sinos River basin

2015 ◽  
Vol 75 (2 suppl) ◽  
pp. 75-80 ◽  
Author(s):  
L Trintinaglia ◽  
E Bianchi ◽  
LB Silva ◽  
CA Nascimento ◽  
FR Spilki ◽  
...  
Keyword(s):  
Water Quality ◽  
River Basin ◽  
Water Samples ◽  
Culture Medium ◽  
Culture Media ◽  
Neutral Red ◽  
Mitochondrial Activity ◽  
Preparation Methods ◽  
Cytotoxicity Assays ◽  
Standard Culture

<p>Cytotoxicity assays using cell cultures may be an alternative to assess biological toxicity of surface waters and may help to improve the control of water quality. This study compared two methods to prepare culture media for the exposure of Hep-2 cells to water samples collected from the Rolante River, an important affluent of the Sinos River. The toxicity was evaluated using the MTT and neutral red assays. Two methods were used to prepare culture media. In method 1, the sample was diluted at 1:1, 1:10, 1:100, 1:1000, 1:10.000 (v/v, sample/medium) in a standard culture medium; in method 2, water samples were used as the solvent for the culture medium, which was prepared at concentrations of 100, 80, 60, 40 and 20%. Semi-confluent cultures were then exposed to the media test for 24 hours, and cytotoxicity was determined immediately using the MTT and NR assays. Mitochondrial activity (MTT) was significantly lower at all concentrations in both methods, except at 1:1000 in method 1. However, the lysosome viability (NR) results revealed cytotoxicity only in the 1:1 sample of method 1. Both culture preparation methods were efficient and sensitive to the MTT assay, but method 2 seemed to be more adequate for the NR assay. The Rolante River has cytotoxic contaminants to Hep-2 cells, which may be one of the explanations for the poor water quality of the Sinos River basin.</p>

scholarly journals The Characterization of Scaffolds Based on Dialdehyde Chitosan/Hyaluronic Acid

Materials ◽  
2021 ◽  
Vol 14 (17) ◽  
pp. 4993
Author(s):  
Sylwia Grabska-Zielińska ◽  
Adrianna Sosik ◽  
Anna Małkowska ◽  
Ewa Olewnik-Kruszkowska ◽  
Kerstin Steinbrink ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Physicochemical Properties ◽  
Cell Viability ◽  
Water Content ◽  
Three Dimensional ◽  
Dermal Fibroblasts ◽  
Epidermal Keratinocytes ◽  
Human Epidermal Keratinocytes ◽  
One Step

In this work, two-component dialdehyde chitosan/hyaluronic acid scaffolds were developed and characterized. Dialdehyde chitosan was obtained by one-step synthesis with chitosan and sodium periodate. Three-dimensional scaffolds were prepared by the lyophilization method. Fourier transform infrared spectroscopy (FTIR) was used to observe the chemical structure of scaffolds and scanning electron microscopy (SEM) imaging was done to assess the microstructure of resultant materials. Thermal analysis, mechanical properties measurements, density, porosity and water content measurements were used to characterize physicochemical properties of dialdehyde chitosan/hyaluronic acid 3D materials. Additionally, human epidermal keratinocytes (NHEK), dermal fibroblasts (NHDF) and human melanoma cells (A375 and G-361) were used to evaluate cell viability in the presence of subjected scaffolds. It was found that scaffolds were characterized by a porous structure with interconnected pores. The scaffold composition has an influence on physicochemical properties, such as mechanical strength, thermal resistance, porosity and water content. There were no significant differences between cell viability proliferation of all scaffolds, and this observation was visible for all subjected cell lines.

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