M Cellulon was not cytotoxic in either mutation assay (with or without the need of S9 metabolic activation) TNF-alpha Protein Bovine within the selection of tested concentrations. The mutant frequencies of treated cultures varied randomly with Cellulon dose, inside the range acceptable for background mutant frequencies that is significantly less than 15 10-6. In the 14 cultures treated with Cellulon, only one culture, inside the activation mutation assay, had a mutant frequency that was statistically elevated over the mutant frequencies of the concurrent automobile control cultures. This observation is constant with regular variation in background mutant frequency in independent cultures. As a result, BC was thought of unfavorable for inducing forward mutations in the HGPRT locus in CHO cells below each nonactivation and S9 metabolic activation situations. 5.six. Limulus IMPA1 Protein E. coli amebocyte iysate (LAL) assay Schmitt et al. [28] assayed the pyrogenicity of BC in Cellulon by using the Limulus amebocyte Iysate (LAL) assay (Table 3). As negativeTable three Summary from the genotoxicity reproductive toxicology research with bacterial cellulose. Dosages Key results Ref.F. Dourado et al.Form of studyCell line/animal modelIn vitro Comet assay Assay: 0.1, 0.5 or 1 mg BC/ml Optimistic manage: hydrogen peroxide (100 mM) DNA damages in the presence of BC fibres are equivalent to the negative manage for each BC concentration; About 95 of cells showed none or insignificant DNA harm (comet class 0 and 1) BC didn’t bring about an increase within the quantity of histidine revertants (mutations) per plate in any bacterial strain, either inside the presence or absence of S9 microsomal enzymesChinese hamster ovary (CHO) cellsMoreira et al. [31]Ames testSalmonella typhimurium (TA 89, TA 100, TA 1535, TA 1537, TA 1538) with and with no metabolic activationNegative manage: water Assay: 0, 66.7, one hundred, 333, 667, 1000, and 2500 g/plateSchmitt et al. [28]Salmonella tryphimurium (TA97a, TA98, TA100 and TA102) Adverse control: distilled waterPositive controls made use of with out metabolic activation: 2-nitrofluorene (TA 98, TA 1538) sodium azide (TA one hundred, TA 1535) ICR-191 with TA 1537 Good controls with metabolic activation: 2- aminoanthracene was utilized with all strains Assay, with and devoid of S9 mixture: 0.1, 0.five or 1.0 mg BC/mlThe outcomes obtained, inside the presence of BC without the need of S9 mixture, correspond to spontaneous reversion for each strain and are similar to these obtained to negative manage; Within the presence of S9 mixture, an increase of revertant colonies per plate, for the TA98 and TA100 strains, was detected as compared with control; nonetheless, the increases have been in every case 2-fold and didn’t appear to become dose-relatedMoreira et al. [31]Positive controls with out S9: 9-fluorenone, sodium azide, mitomycin C; Constructive controls with S9: 1,8-dihydroxy anthraquinone, 2-amino fluorine Assay: 0.333 g/ml to ten,000 g/ml Cellulon in McCoy’s Sa culture medium Constructive controls: mitomycin C, nonactivation series; cyclophosphamide, metabolic activation series Assay: replacement from the culture media with two,5 mL WMEI with 10 Ci/ml 3H-thymidine (50 Ci/ mmol), BC (501, 1000, 2000, 3010, 4010, and 5010 g/ml) Positive controls: (2- acetylaminofluorene) Unfavorable control: WMEI with ten pCi/ml 3H-TdR, WMEI with sucrose Assay with and devoid of S9 metabolic activation: BC at 0.098-5.0 mg/ml, in F12 culture medium Negative control: Sucrose Good manage: (nonactivation assay, 5-bromo-2 deoxyuridine (BrdU) Metabolic activation: 3-methylcholanthreneS. typhimurium (TA97, TA98, TA10.