The effect of mephedrone on human neuroblastoma and astrocytoma cells.

Mephedrone is an illegal drug that is used recreationally. Few studies have been conducted to investigate the mechanisms by which mephedrone is harming cells. In this research, we investigated the effect of mephedrone using toxicology coupled with LC-MS/MS based metabolomics in the two CNS derived cell lines. Methods of assessment such as neutral red (NR) assay, dimethylthiazolyl diphenyltetrazolium bromide (MTT), lactose dehydrogenase (LDH) measurement, and morphology were performed to identify the effect on cell viability and to identify the best concentration to be used in a metabolomics study. A concentration of 100 µM of mephedrone was used in the metabolomic experiment because at this concentration mephedrone had induced several intracellular changes. Although there no clear indicators of cellular damage caused by mephedrone. In astrocytes there was a clear indication that cell membrane function might be impaired by depletion of ether lipids.

Pulsed electric field treatment as a potential method for microbial inactivation in scaffold materials for tissue engineering: the inactivation of bacteria in collagen gel.

AIMS: To investigate the effectiveness of pulsed electric field (PEF) treatment as a new method for inactivation of micro-organisms in complex biomatrices and to assess this by quantifying the inactivation of Escherichia coli seeded in collagen gels. METHODS AND RESULTS: PEF was applied to E. coli seeded collagen gels in static (nonflowing) chambers. The influence of electric field strength, pulse number and seeded cell densities were investigated. The highest level of inactivation was obtained at the maximum field strength of 45 kV cm(-1). For low levels of E. coli contamination (10(3) CFU ml(-1)), PEF treatment resulted in no viable E. coli being recovered from the gels. However, PEF treatment of gels containing higher cell densities (>or=10(4) CFU ml(-1)) did not achieve complete inactivation of E. coli. CONCLUSIONS: PEF treatment successfully inactivated E. coli seeded in collagen gels by 3 log(10) CFU ml(-1). Complete inactivation was hindered at high cell densities by the tailing effect observed. SIGNIFICANCE AND IMPACT OF THE STUDY: PEF shows potential as a novel, nondestructive method for decontamination of collagen-based matrices. Further investigation is required to ensure its compatibility with other proteins and therapeutic drugs for tissue engineering and drug delivery applications.

Reduced glutathione levels and expression of the enzymes of glutathione synthesis in cryopreserved hepatocyte monolayer cultures.

Cryopreservation of monolayers of hepatocytes in a freezing medium containing 10% (v/v) dimethylsulfoxide, 90% (v/v) foetal calf serum retains cell morphology and viability, but cells lose up to 50% of their intracellular reduced glutathione. This is accompanied by a small increase in glutamate cysteine ligase expression in cryopreserved cultures, but glutathione synthetase expression is undetectable post-cryopreservation. Inclusion of ascorbic acid and alpha-tocopherol in the freezing medium improves maintenance of reduced glutathione content post-cryopreservation at 84% of the levels in non-cryopreserved monolayer cultures, but does not restore glutathione synthetase expression. The inability to synthesise reduced glutathione will mean that cryopreserved hepatocyte monolayers are more susceptible to toxic insults.

Decontamination of collagen biomatrices with combined pulsed electric field and nisin treatment.

Pulsed electric field (PEF) treatment has been proposed as a decontamination method for labile matrices used in tissue engineering applications. Through the application of PEF, a non-thermal treatment that causes bacterial inactivation through the irreversible rupture of microbial cell membranes, inactivation is achieved without loss of scaffold structure and function. However, some microorganisms are less susceptible to PEF treatment. This study shows that treatment with PEF and nisin, a food preservative bacteriocin, has a synergistic effect on the inactivation of Staphylococcus epidermidis in collagen gels. Almost complete inactivation of a 10(3) -10(4) CFU/mL S. epidermidis population was achieved when treated with a combination of PEF and 500 IU/mL nisin, with results demonstrating a 3.4 log(10) reduction, compared with 0.66 log(10) reduction with PEF alone. Nisin, at concentrations up to 3000 IU/mL, had no discernable toxicity to mammalian 3T3 cells when added to the culture medium or incorporated into the collagen gels. This combined decontamination method, involving PEF plus nisin, may provide a non-destructive process for inactivation of PEF-resistant bacteria in labile tissue engineering scaffolds.