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. 2017 Sep 25;4:543–553. doi: 10.1016/j.toxrep.2017.09.005

Table 3.

Summary of the genotoxicity & reproductive toxicology studies with bacterial cellulose.

Type of study Cell line/animal model Dosages Main results Ref.
In vitro
Comet assay Chinese hamster ovary (CHO) cells Assay: 0.1, 0.5 or 1 mg BC/ml DNA damages in the presence of BC fibres are similar to the negative control for each BC concentration; Moreira et al. [31]
Positive control: hydrogen peroxide (100 mM) Around 95% of cells showed none or insignificant DNA damage (comet class 0 and 1)
Negative control: water
Ames test Salmonella typhimurium (TA 89, TA 100, TA 1535, TA 1537, TA 1538) with and without metabolic activation Assay: 0, 66.7, 100, 333, 667, 1000, and 2500 μg/plate BC did not cause an increase in the number of histidine revertants (mutations) per plate in any bacterial strain, either in the presence or absence of S9 microsomal enzymes Schmitt et al. [28]
Positive controls used without metabolic activation:
  • –

    2-nitrofluorene (TA 98, TA 1538)

  • –

    sodium azide (TA 100, TA 1535)

  • –

    ICR-191 with TA 1537

  • –

    Positive controls with metabolic activation:

  • –

    2- aminoanthracene was used with all strains
Salmonella tryphimurium (TA97a, TA98, TA100 and TA102) Assay, with and without S9 mixture: 0.1, 0.5 or 1.0 mg BC/ml The results obtained, in the presence of BC without S9 mixture, correspond to spontaneous reversion for each strain and are similar to those obtained to negative control; Moreira et al. [31]
Negative control: distilled water In the presence of S9 mixture, an increase of revertant colonies per plate, for the TA98 and TA100 strains, was detected as compared with control; however, the increases were in each case <2-fold and did not appear to be dose-related
Positive controls:
  • –

    0.1 μg/plate 4-nitroquinoline 1-oxide (TA97a, TA98)

  • –

    5.0 μg/plate sodium azide (TA100)

  • –

    0.5 μg/plate mytomicyn C (TA102)
S. typhimurium (TA97, TA98, TA100, TA102) Assay: 8, 40, 200, 1000 and 5000 μg CB/dish The numbers of colonies of each group at any BC dose, with or without S9 did, did not exceed twice of those of spontaneous reverse mutation group; Hagiwara et al. [30]
Positive controls without S9: 9-fluorenone, sodium azide, mitomycin C; Reversion mutation colonies did not grow with increasing dosages of BC, when compared to the solvent plates, indicating that no dose-response relationship was reflected.
Positive controls with S9: 1,8-dihydroxy anthraquinone, 2-amino fluorine
Cytogenetic Assay CHO cells Assay: 0.333 μg/ml to 10,000 μg/ml Cellulon in McCoy's Sa culture medium No significant increase in cells with chromosomal aberrations was observed at the concentrations analysed; [28]
Positive controls: mitomycin C, nonactivation series; cyclophosphamide, metabolic activation series BC was considered negative for inducing chromosomal aberrations in CHO cells under both non-activation and metabolic activation conditions
UDSa Assay Rat primary hepatocytes Assay: replacement of the culture media with
  • –

    2,5 mL WMEI with 10 μCi/ml 3H-thymidine (50 Ci/mmol), BC (501, 1000, 2000, 3010, 4010, and 5010 μg/ml)

 BC did not to induce significant changes in the nuclear labelling of rat primary hepatocytes within the range of tested concentrations;
Positive controls: (2- acetylaminofluorene) None of the criteria used to indicate UDS were approached by any of the analysed treatments and no dose-related response was observed
Negative control:
  • –

    WMEI with 10 pCi/ml 3H-TdR,

  • –

    WMEI with sucrose
CHO/HGPRTb Forward Mutation Assay CHO-KI-BH4 cells Assay with and without S9 metabolic activation: BC at 0.098-5.0 mg/ml, in F12 culture medium BC was considered negative for inducing forward mutations at the HGPRT locus in CHO cells under both nonactivation and S9 metabolic activation conditions
Negative control: Sucrose
Positive control: (nonactivation assay, 5-bromo-2′ deoxyuridine (BrdU)
Metabolic activation: 3-methylcholanthrene
LALc assay Assay: BC (0.5% Cellulon fibre, 99.5% water) BC was negative for the presence of gram-negative bacterial endotoxin (<0,25 EU/ml)
Negative: sterile water and endotoxin
Mouse sperm abnormality test Kunming male mice Assay: BC meals with 1.3, 2.5, 5.0 g/Kg bw, through oral gavage; No significant differences in the rate of sperm abnormality between each BC dosage group and the solvent control group (CMC); Li-ming et al. [29]
Negative control: 1% CMC There was a significant difference between the positive control group (cyclophosphamide) and the solvent control group
Positive control: cyclophosphamide, (40 mg/kg bw)
Teratogenicity test Fertilized SD rats Assay: during gestation (7–16 days), dosage groups received oral gavage of 1.0, 2.0, 4.0 g BC/kg; No deaths and no gross anatomical abnormalities were observed to any pregnant rat in all groups;
Control group: 10.0 mL/kg bw of 1% CMC No abnormalities in the anatomy of the rats in each dose group;
No significant differences in the:
  • –

    weight and weight gain of pregnant rats,

  • –

    placental weight,

  • –

    incidence of foetal and stillbirth in pregnant rats,

  • –

    foetal body length and tail length,

  • –

    absorption rate (0–5.8%), rate of stillbirth (0%), rate of malformation (0%), rate of visceral deformity (0%),

  • –

    litter size and skeletal deformities,

between each dose group and the control group



In vivo
Mouse bone marrow micronucleus assay Kunming mice Assay: oral gavage of 1.3, 2.5, 5.0 g BC/Kg bw No significant differences in the incidence of micronucleus in the bone marrow of female and male mice in each dose group, as compared to the solvent control group (CMC); Hagiwara et al. [30]
Negative control: 1% CMC The micronucleus rate in the positive control group (cyclophosphamide) was significantly higher than that of the CMC group
Positive control: cyclophosphamide, at 40 mg/kg bw
a

Unscheduled DNA Synthesis.

b

Chinese hamster ovary cell/hypoxanthine-guanine phosphoribosyl-transferase.

c

Limulus amebocyte Iysate.