Catarina Satie Takahashi

Effects of high doses of vitamins C and E against doxorubicin-induced chromosomal damage in Wistar rat bone marrow cells

Doxorubicin (DXR) is one of the major antitumoral agents available for clinical use. In addition to intercalating into the DNA molecule, this drug generates free radicals. Vitamins C (VC) and E (VE) can protect normal cells from the damage caused by radicals without interfering with the cytotoxicity of DXR against tumors. The objective of the present study was to investigate the possible protective effect of VC and/or VE on mammalian cells treated with DXR in vivo. Animals treated with the lowest doses of VC and/or VE, alone or in combination, plus a single dose of DXR presented a statistically significant reduction in total number of chromosome aberrations and in number of abnormal metaphases. The highest vitamin doses tested caused no changes in the parameters analyzed when compared with control. Under the present experimental conditions, the efficiency of VC and/or VE in protecting against chromosome damage was dependent on the dose used.

Characterization and in vitro evaluation of bacterial cellulose membranes functionalized with osteogenic growth peptide for bone tissue engineering.

The aim of this study was to characterize the physicochemical properties of bacterial cellulose (BC) membranes functionalized with osteogenic growth peptide (OGP) and its C-terminal pentapeptide OGP[10–14], and to evaluate in vitro osteoinductive potential in early osteogenesis, besides, to evaluate cytotoxic, genotoxic and/or mutagenic effects. Peptide incorporation into the BC membranes did not change the morphology of BC nanofibers and BC crystallinity pattern. The characterization was complemented by Raman scattering, swelling ratio and mechanical tests. In vitro assays demonstrated no cytotoxic, genotoxic or mutagenic effects for any of the studied BC membranes. Culture with osteogenic cells revealed no difference in cell morphology among all the membranes tested. Cell viability/proliferation, total protein content, alkaline phosphatase activity and mineralization assays indicated that BC-OGP membranes enabled the highest development of the osteoblastic phenotype in vitro. In conclusion, the negative results of cytotoxicity, genotoxicity and mutagenicity indicated that all the membranes can be employed for medical supplies, mainly in bone tissue engineering/regeneration, due to their osteoinductive properties.

Potentiation by turmeric and curcumin of γ-radiation-induced chromosome aberrations in Chinese hamster ovary cells

The effect of turmeric and curcumin, two natural antioxidants, on the frequencies of chromosome aberrations induced in Chinese hamster ovary (CHO) cells by gamma-radiation was investigated. Cells were treated with three concentrations of each drug, turmeric (100, 250, and 500 microg/ml) and curcumin (2.5, 5, and 10 microg/ml), and then irradiated (2.5 Gy) during different phases of the cell cycle. Turmeric was not clastogenic by itself, whereas curcumin at 10 microg/ml enhanced the chromosomal damage frequency. Neither of the two antioxidants showed protective effect against the clastogenicity of gamma-radiation. Instead, an obvious increase in the frequencies of chromosome aberrations was observed when turmeric at 500 microg/ml was associated with gamma-radiation during G2/S phase, and curcumin at 10 microg/ml plus gamma-radiation during S and G2/S phases of the cell cycle. The results clearly indicate the exacerbated effect of turmeric and curcumin on radiation-induced clastogenicity, suggesting that these antioxidants are also potentiating agents depending on the experimental conditions.

Protective effects of the amino acid glutamine and of ascorbic acid against chromosomal damage induced by doxorubicin in mammalian cells.

The interaction of antioxidants can provide an essential protection against the damaging effects of free radicals. Beneficial interactions include radioprotection, protection against acute toxicity of chemicals, and antimutagenic and anticarcinogenic activity. The present study was undertaken to evaluate the protective effect of the amino acid glutamine (GLN) and ascorbic acid (AA) on the frequency of chromosomal aberrations induced by the antineoplastic agent doxorubicin (DXR). These micronutrients were tested separately and simultaneously in Wistar rat bone marrow and Chinese hamster ovary (CHO) cells. The treatments with GLN and/or AA significantly decreased the frequency of DXR-induced clastogenic damage in both test systems.

Protection and induction of chromosomal damage by vitamin C in human lymphocyte cultures.

Some chemotherapeutic approaches have proposed the use of antioxidants such as vitamin C (VC) to minimize the cytotoxicity and damage induced in normal tissue by antitumor agents that produce free radicals. Nevertheless, VC can also be cytotoxic, genotoxic, and harmful when combined with antitumor agents in human cells in vitro. The present study was undertaken to investigate the effects of VC (100, 200, 500, and 1,000 microg/ml) on human peripheral blood lymphocytes in vitro and its anticlastogenic effect on chromosomal aberrations induced by doxorubicin (DXR). VC did not show a clastogenic effect by itself, except at 1,000 microg/ml. At the concentration of 100 or 200 microg/ml of VC, administered in pre-, post-, or simultaneous treatment, there was a significant reduction in both chromosome aberrations and number of abnormal metaphases induced by DXR. At the doses of 500 or 1,000 microg/ml, VC did not present the same protective effect and was cytotoxic. Under the present experimental conditions, the efficiency of VC in protecting against chromosome damage was dependent on the doses used.

Mutagenic and cytotoxic activity of an isocoumarin (paepalantine) isolated from Paepalanthus vellozioides

A new isocoumarin with antimicrobial activity was isolated from Paepalanthus vellozioides (a native Brazilian plant) and called paepalantine. This study was carried out to assess the mutagenic activity of this new agent in assays with Salmonella typhimurium TA100, TA98, and TA102 and in Chinese hamster ovary (CHO) cell cultures, as well as cytotoxicity to McCoy cells. paepalantine caused a significant dose-dependent increase in the frequency of revertants in the three strains used in the assay, both with and without S9 mix, in concentrations varying from 2 to 128 micrograms/plate. The mutagenicity was confirmed in assays with CHO cells treated in the G1, S, and G2 phases of the cell cycle. There was an increase in the chromosomal aberration frequency, mainly in the G2 phase. Furthermore, the mitotic index of the treated cultures (40,80, and 160 micrograms/ml) was significantly lower, indicating cytotoxicity. The midpoint cytotoxicity values of McCoy cells by the neutral red (NR) and microculture tetrazolium (MTT) techniques resulted in a NR50 and MTT50 of 30 and 38 micrograms/ml, respectively. Alterations to the paepalantine structure are suggested to reduce its mutagenic and cytotoxic activity in investigations for its antineoplastic potential.

Protective effect of thiourea, a hydroxyl-radical scavenger, on curcumin-induced chromosomal aberrations in an in vitro mammalian cell system.

Natural dietary antioxidants are extensively studied for their ability to protect cells from damage to DNA, protein, and lipids induced by antitumor agents or radiation that leads to the generation of free radical in normal cells in vivo and in vitro. Curcumin is a natural antioxidant known to possess therapeutic properties and has been reported to scavenge free radicals and to inhibit clastogenesis in mammalian cells. However, curcumin has been reported to induce a significant increase in the frequency of chromosomal aberrations in Chinese hamster ovary (CHO) cells. To investigate whether the clastogenic activity of curcumin in CHO cells in culture can be ascribed to a pro-oxidant behavior, mediated by free radical generation, experiments were carried out with the combination of curcumin (15 microg/ml) and thiourea (10, 20, or 40 microg/ml), a potent hydroxyl radical scavenger. The results showed that the clastogenic action of curcumin was statistically decreased in a dose-dependent manner in the presence of thiourea. These data have shown that curcumin-induced chromosomal damage in CHO cells can be mediated by hydroxyl radical generation in the present experimental conditions. Teratogenesis Carcinog. Mutagen. 21:175-180, 2001.

Modulatory effects of curcumin on the chromosomal damage induced by doxorubicin in Chinese hamster ovary cells

Curcumin, a natural phenolic compound, is gaining importance as a free radical scavenger. This study was undertaken to investigate the modulatory effects of curcumin on the chromosomal damage induced by the antitumoral doxorubicin (DXR), a known free radical generator, in Chinese hamster ovary cells in culture. Cells were treated with three concentrations of curcumin (2.5, 5, or 10 microg/ml) and then treated with DXR (1.0 microg/ml) during different phases of the cell cycle. The results show that curcumin induces chromosomal damage in CHO at the highest concentration when compared to the untreated control. Neither treatment with curcumin shows a reduction in the clastogenicity of DXR. Instead, a statistically significant increase in the frequency of chromosomal damage was observed when the middle and the highest concentrations of curcumin were associated with DXR during the G1/S, S, and S/G2 phases of the cell cycle. The results clearly indicate the potentiating effect of curcumin on DXR-induced chromosomal damage.

Basal levels of DNA damage detected by micronuclei and comet assays in untreated breast cancer patients and healthy women

Breast cancer is the second most frequent type of cancer worldwide and is the most common malignant disease among women. Risk factors for breast cancer include early menarche, late menopause, hormonal therapies, exposure to environmental pollutants, smoking and alcohol use. However, increased or prolonged exposure to estrogen is the most important risk factor. It has been suggested that accumulation of DNA damage may contribute to breast carcinogenesis. Epidemiological studies suggest that cytogenetic biomarkers such as micronuclei in peripheral blood lymphocytes may predict cancer risk because they indicate genomic instability in target tissues. The objective of the present study was to evaluate the frequencies of micronuclei and the extent of DNA damage detected by comet assay in peripheral blood lymphocytes of untreated breast cancer patients and healthy women. The study was conducted using peripheral blood lymphocytes from 45 women diagnosed for Ductal “in situ” or invasive breast carcinoma and 85 healthy control women. Micronuclei and comet assays were performed to detect spontaneous DNA damage. The results showed that micronuclei frequencies and tail intensity, detected by comet assay, were significantly higher in the breast cancer group than in controls. The levels of DNA damage were similar in smokers and non-smokers, and aging did not influence the frequencies of micronuclei or tail intensity values observed in either group. In conclusion, the present work demonstrates higher levels of DNA damage in untreated breast cancer patients than in healthy women.

Cytotoxicity and genotoxicity of silver nanoparticles of different sizes in CHO-K1 and CHO-XRS5 cell lines.

Nanoparticles (NPs) have been used in a range of products due to their unique properties. Nevertheless, these NPs can cause adverse biological effects and because of that, there is a great concern about the health and environmental risks related to their use. Recently, silver nanoparticles (Ag NPs) have been used in a variety of cytotoxicity and genotoxicity studies, but there are still controversies regarding the association between the size and the toxicity of these particles. Therefore, in this study, we aimed to evaluate the cytotoxicity and genotoxicity of Ag NPs (10 and 100 nm) in two different cell lines, CHO-K1 and CHO-XRS5, by performing cell viability assay (XTT), clonogenic assay, micronucleus test, comet assay, as well as by investigating the cell cycle kinetics using the flow cytometry. Cell cultures were exposed to different concentrations of AgNPs (0.025-5.0 μg/ml) for 24 h. Our results indicated that cytotoxicity and genotoxicity induced by the 100 nm-Ag NPs were greater than those induced by the 10 nm-Ag NPs for both cell lines, which suggests that the exposure to greater size particles (100 nm) can cause more adverse biological effects than the exposure to the smaller ones (10 nm).