In vitro toxicity of various classes of test agents using the neutral red assay on a human three-dimensional physiologic skin model

The objective of this work was to evaluate the biocompatibility of scaffolds of poly(L-lactide) with pure and grafted hydroxyapatite, at various concentrations of reinforcement. The biocompatibility tests were carried out in vivo in Wistar rats by implanting the material into the subcutaneous and muscle tissues from 1 to 14 weeks and evaluating the surrounding tissue stained with hematoxylin-eosin. For in vitro assays, MTT and neutral red assay were used to evaluate any cytotoxicity in Mioblast Muscle C2C12 Cells (ATCC® CRL-1772™) and Bovine Coronary Artery Endothelial Cells (BCAEC); Escherichia coli and Staphylococcus aureus were used to evaluate bacterial adhesion. All variants of scaffolds provoked a mild inflammatory response, without showing necrosis. No evidence of cytotoxicity was presented in cell viability tests and good bacterial cell adhesion was visualized for all of the materials studied.

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Three-dimensional quantitative structure activity relationship (3D-QSAR) analysis for in vitro toxicity of chlorophenols to HepG2 cells

Chemosphere ◽

10.1016/j.chemosphere.2005.04.019 ◽

2005 ◽

Vol 60 (6) ◽

pp. 791-795 ◽

Cited By ~ 6

Author(s):

Y. Liu ◽

J.N. Chen ◽

J.S. Zhao ◽

H.X. Yu ◽

X.D. Wang ◽

...

Keyword(s):

Hepg2 Cells ◽

Three Dimensional ◽

3D Qsar ◽

Quantitative Structure Activity Relationship ◽

Activity Relationship ◽

In Vitro Toxicity ◽

Quantitative Structure ◽

Qsar Analysis ◽

Structure Activity

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Cosmetic efficacy claims in vitro using a three-dimensional human skin model

International Journal of Cosmetic Science ◽

10.1046/j.1467-2494.2001.00098.x ◽

2001 ◽

Vol 23 (5) ◽

pp. 309-318 ◽

Cited By ~ 16

Author(s):

K. Schlotmann ◽

M. Kaeten ◽

A. F. Black ◽

O. Damour ◽

M. Waldmann-Laue ◽

...

Keyword(s):

Human Skin ◽

Three Dimensional ◽

Skin Model

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413. Differences in Inflammatory Mechanisms in Pseudomonas aeruginosa and Staphyloccoccus auereus Infections in Cystic Fibrosis

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz360.486 ◽

2019 ◽

Vol 6 (Supplement_2) ◽

pp. S209-S209

Author(s):

Melissa S Phuong ◽

Rafael E Hernandez ◽

Subash Sad

Keyword(s):

Cystic Fibrosis ◽

Pseudomonas Aeruginosa ◽

Cell Death ◽

Cytokine Production ◽

Neutral Red ◽

Chronic Infections ◽

Neutral Red Assay ◽

The One ◽

The University

Abstract Background Chronic bacterial lung infections are the primary cause of morbidity and mortality in cystic fibrosis (CF). The most common CF pathogens, Pseudomonas aeruginosa (P. aeruginosa) or Staphylococcus aureus (S. aureus), are common commensal or environmental organisms that adapt to the CF lung. We sought to investigate whether adaptation from early lung colonizer to chronic pathogen alters the bacterial effects on host inflammation. Methods P. aeruginosa (n = 25) and S. aureus (n = 25) isolates from CF patients with early and chronic infections were acquired from Seattle Children’s CF. Environmental (n = 8) and clinical, non-CF P. aeruginosa (n = 8) isolates were obtained from the University of Ottawa. P. aeruginosa reference strain PA14 and PA14 transposon mutants for T3SS and flagellin were used to observe the relationship between cell death and cytokine production. We infected THP-1-derived macrophages (PMA differentiated) in vitro for 3 hours with various MOIs. We subsequently measured cell death of THP-1-derived macrophages using neutral red assay and cytokine production using ELISAs. Results Infections with PA14 mutants and non-CF P. aeruginosa isolates demonstrated that rapid cell death of THP-1-derived macrophages caused a reduction in cytokine production relative to strains that did not cause as much cell death. At 10 MOI, early P. aeruginosa isolates from CF patients induced more THP-1-derived macrophage cell death compared with chronic isolates (P < 0.0001). Chronic P. aeruginosa isolates induced greater production of TNF, IL-8, and IL-6 (P < 0.01, P < 0.0001, and P < 0.0001, respectively) compared with early strains. No difference in IL-1β production was observed. When controlling for cell death between the two groups by using heat-killed bacteria, the only difference maintained was in TNF production (P < 0.01). Between early and chronic S. aureus isolates, the one difference observed was greater IL-8 production among early isolates (P < 0.01). Conclusion Chronic P. aeruginosa isolates from CF patients induce less cell death but more TNF, IL-8, and IL-6 production compared with early isolates. This suggests that P. aeruginosa producing chronic infections induce inflammatory signals that may contribute to increased morbidity among CF patients. Disclosures All authors: No reported disclosures.

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Preliminary Study of In Vitro Three-Dimensional Skin Model Using an Ovine Collagen Type I Sponge Seeded with Co-Culture Skin Cells: Submerged versus Air-Liquid Interface Conditions

Polymers ◽

10.3390/polym12122784 ◽

2020 ◽

Vol 12 (12) ◽

pp. 2784

Author(s):

Mh Busra Fauzi ◽

Zahra Rashidbenam ◽

Aminuddin Bin Saim ◽

Ruszymah Binti Hj Idrus

Keyword(s):

Collagen Type ◽

Three Dimensional ◽

Collagen Type I ◽

Liquid Interface ◽

Type I ◽

Skin Model ◽

Skin Cells ◽

Air Liquid Interface ◽

In Vitro Skin Model

Three-dimensional (3D) in vitro skin models have been widely used for cosmeceutical and pharmaceutical applications aiming to reduce animal use in experiment. This study investigate capability of ovine tendon collagen type I (OTC-I) sponge suitable platform for a 3D in vitro skin model using co-cultured skin cells (CC) containing human epidermal keratinocytes (HEK) and human dermal fibroblasts (HDF) under submerged (SM) and air-liquid interface (ALI) conditions. Briefly, the extracted OTC-I was freeze-dried and crosslinked with genipin (OTC-I_GNP) and carbodiimide (OTC-I_EDC). The gross appearance, physico-chemical characteristics, biocompatibility and growth profile of seeded skin cells were assessed. The light brown and white appearance for the OTC-I_GNP scaffold and other groups were observed, respectively. The OTC-I_GNP scaffold demonstrated the highest swelling ratio (~1885%) and water uptake (94.96 ± 0.14%). The Fourier transformation infrared demonstrated amide A, B and I, II and III which represent collagen type I. The microstructure of all fabricated sponges presented a similar surface roughness with the presence of visible collagen fibers and a heterogenous porous structure. The OTC-I_EDC scaffold was more toxic and showed the lowest cell attachment and proliferation as compared to other groups. The micrographic evaluation revealed that CC potentially formed the epidermal- and dermal-like layers in both SM and ALI that prominently observed with OTC-I_GNP compared to others. In conclusion, these results suggest that OTC_GNP could be used as a 3D in vitro skin model under ALI microenvironment.

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Differential toxicities of albendazole and its two main metabolites to Balb/c 3T3, HepG2, and FaO lines and rat hepatocytes

Journal of Veterinary Research ◽

10.1515/jvetres-2016-0073 ◽

2016 ◽

Vol 60 (4) ◽

pp. 495-500 ◽

Cited By ~ 3

Author(s):

Lidia Radko ◽

Maria Minta ◽

Sylwia Stypuła-Trębas

Keyword(s):

Hepatoma Cell ◽

Rat Hepatocytes ◽

Neutral Red ◽

In Vitro Toxicity ◽

Isolated Rat Hepatocytes ◽

Hepatoma Cell Lines ◽

Cellular Models ◽

The Impact ◽

Isolated Rat

Abstract Introduction: The cytotoxicity of anthelmintic agent, albendazole (ABZ) and its two major metabolites, sulfoxide (ABZSO) and sulfone (ABZ-SO2), on non-hepatic Balb/c 3T3 line, two hepatoma cell lines (FaO, HepG2), and isolated rat hepatocytes was investigated. Material and Methods: Cell cultures were exposed for 24, 48, and 72 h to eight concentrations of the compounds ranging from 0.05 to 100 μg/mL (ABZ) and from 0.78 to 100 μg/mL (ABZ-SO and ABZ-SO2). Three different assays were applied in which various biochemical endpoints were assessed: lysosomal activity - neutral red uptake (NRU) assay, proliferation - total protein contents (TPC) assay and lactate dehydrogenase (LDH) leakage assay. Results: The most toxic was albendazole whose EC50 values calculated from the concentration effect curves ranged from 0.2 to 0.5 μg/mL (Balb/c 3T3 ) and from 0.4 to 73.3 μg/mL (HepG2). Rat hepatoma line and isolated rat hepatocytes were less sensitive to the impact of ABZ. Toxic action expressed as EC50 was recorded after 72 h exposure only in LDH release assay at 0.8 μg/mL and 9.7 μg/mL respectively. The toxicity of metabolites was much lower. The most sensitive to ABZ-SO were fibroblasts and EC50-72h values were similar in all three assays used, i.e. NRU (14.1 μg/mL), TPC (15.8 μg/mL), and LDH (20.9 μg/mL). In the case of ABZ-SO2 the mean effective concentrations were the highest, and could be reached only in one LDH assay. These values (μg/mL) were as follows: 65.3 (FaO), 65.4 (HepG2), 75.8 (hepatocytes), and 77.4 (Balb/c 3T3). Conclusion: The differences in in vitro toxicity of albendazole depend on metabolic ability of the cellular models. Primary cultured rat hepatocytes represent a valuable tool to study the impact of biotransformation on the cytotoxicity of drugs.

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