Avaliação de instabilidade genética e citotoxicidade em profissionais expostos ao resíduo de anestésico isoflurano que atuam em centro cirúrgico de hospital veterinário
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Data
2020-08-26
Autores
Figueiredo, Driélle Baptista dos Santos
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Universidade Estadual Paulista (Unesp)
Resumo
Há escassez de dados científicos em relação aos possíveis efeitos tóxicos no material genético e em nível celular frente à exposição ocupacional aos resíduos do anestésico inalatório isoflurano em profissionais atuantes em salas cirúrgicas veterinárias (SCV). Assim, considerando a relevância do tema, o presente estudo objetivou, de forma inédita, monitorar a poluição anestésica em SCV e avaliar os impactos biológicos dessa exposição nos profissionais. Foram recrutados para o estudo do tipo transversal profissionais atuantes em SCV expostos no mínimo há um ano aos resíduos de isoflurano (grupo exposto) e indivíduos sem exposição ocupacional a anestésico (grupo controle), os quais foram pareados com o grupo exposto. Realizou-se a avaliação de danos basais no ácido desoxirribonucleico (DNA) e do sistema de reparo do DNA em células mononucleares pelo teste do cometa e, por citometria de fluxo, foram analisados viabilidade e processo apoptótico em células mononucleares bem como as fases do ciclo celular em mononucleares e polimorfonucleares. Concomitantemente, células esfoliadas orais foram coletadas para realização do buccal micronucleus cytome assay para identificação de marcadores de instabilidade genética (incluindo o micronúcleo - MN), proliferação e morte celular. As concentrações residuais de isoflurano foram mensuradas em SCV sem sistema de exaustão de gases por espectrofotômetro infravermelho portátil e a concentração média de isoflurano foi de 11 partes por milhão, excedendo em 5,5 vezes o limite de recomendação internacional. Não houve diferenças significativas entre os dois grupos para os parâmetros avaliados do teste do cometa, exceto que os profissionais com menor faixa etária apresentaram maior nível de danos basais no DNA que os controles da mesma faixa etária, e em relação ao grupo exposto, os indivíduos mais jovens (com maior carga horária semanal) e indivíduos mais experientes com mais tempo (anos) de exposição apresentaram os maiores níveis de danos no DNA. Também não houve diferenças significativas entre os grupos em relação às análises de citometria de fluxo. Por outro lado, o grupo exposto apresentou maior frequência de MN, broto nuclear, células binucleadas, cariorrexe e cariólise, e menor frequência de células basais que o grupo controle. As mulheres expostas foram mais susceptíveis à instabilidade genética e índice proliferativo enquanto que os homens expostos apresentaram mais alterações de citotoxicidade. Adicionalmente, o grupo exposto com faixa etária mais jovem apresentou maior frequência de MN tanto quando comparado com indivíduos com ≥ 31 anos de idade expostos por até cinco anos como comparado com indivíduos da mesma faixa etária expostos por ≥ 5 anos devido a maior carga horária semanal, portanto com maior exposição residual ao isoflurano. Concluindo, a alta exposição ao isoflurano está associada aos danos no material genético, processo citotóxico e alteração proliferativa na mucosa bucal, mas não altera o sistema de reparo do DNA, o ciclo celular e a frequência de apoptose, quando avaliados em células do sangue periférico. Assim, é necessário mitigar a poluição anestésica no ambiente de trabalho para reduzir a exposição ocupacional em profissionais atuantes em SCV com o intuito de se minimizar o impacto da toxicidade anestésica.
Scarce literature is found about possible toxic effects on genome and cell concerning the occupational exposure to waste anesthetic isoflurane in professionals who work in veterinary operating room (VOR). Because of the relevance of this issue, this is the first study to monitor anesthetic pollution in VOR and assess the biological impact of this exposure on professionals. This cross-sectional study recruited professionals who have been exposed for at least one year to waste anesthetic gas isoflurane (exposed group) and subjects without occupational exposure to anesthetics (control group), who were matched with the exposed group. We evaluated basal deoxyribonucleic acid (DNA) damage and DNA repair system in mononuclear cells by the comet assay, and by flow cytometry, viability and apoptosis were assessed in mononuclear cells and cycle cell phases were detected in both mononuclear and polymorphonuclear cells. Concomitantly, oral exfoliated cells were collected to evaluate the buccal micronucleus cytome assay to detect markers of genetic instability (including micronucleus - MN), cell proliferation and cell death. Concentrations of waste isoflurane were measured in the VOR lacking scavenging system to assess anesthetic pollution by infrared spectrophotometry, and the mean concentration was 11 parts per million, which is 5.5 times higher than the international recommended threshold. There were no significant differences between groups for comet assay parameters, with the exception that younger exposed professionals presented higher basal DNA damage than the matched controls; regarding the exposed group, the younger professionals (with higher week workload) and older professionals with greater time of xiii exposure (years) showed the highest DNA damage levels. No significant differences were also found to analyses obtained by flow cytometry. Differently, the exposed group had higher frequency of MN, nuclear bud, binucleated cells, karyorrhexis and karyolysis, and lower frequency of basal cells than the control group. The exposed women were more vulnerable to genetic instability and proliferative index while the exposed men presented more cytotoxicity. Additionally, the exposed group with lower age range had a higher frequency of MN when compared with both older professionals (≥ 31 years) exposed for up to five years and professionals with the same age range exposed for ≥ 5 years; this might be explained due to higher week workload, therefore they were more exposed to isoflurane. In conclusion, the high exposure to isoflurane is associated with DNA damage, apoptosis and proliferative changes in buccal cells, but does not impair the DNA repair capability, cell cycle or apoptosis rate when analyzed in peripheral blood cells. The findings highlight the need to mitigate anesthetic pollution in the work environment to reduce occupational exposure in professionals who work in VOR to minimize the impact of anesthetic toxicity
Scarce literature is found about possible toxic effects on genome and cell concerning the occupational exposure to waste anesthetic isoflurane in professionals who work in veterinary operating room (VOR). Because of the relevance of this issue, this is the first study to monitor anesthetic pollution in VOR and assess the biological impact of this exposure on professionals. This cross-sectional study recruited professionals who have been exposed for at least one year to waste anesthetic gas isoflurane (exposed group) and subjects without occupational exposure to anesthetics (control group), who were matched with the exposed group. We evaluated basal deoxyribonucleic acid (DNA) damage and DNA repair system in mononuclear cells by the comet assay, and by flow cytometry, viability and apoptosis were assessed in mononuclear cells and cycle cell phases were detected in both mononuclear and polymorphonuclear cells. Concomitantly, oral exfoliated cells were collected to evaluate the buccal micronucleus cytome assay to detect markers of genetic instability (including micronucleus - MN), cell proliferation and cell death. Concentrations of waste isoflurane were measured in the VOR lacking scavenging system to assess anesthetic pollution by infrared spectrophotometry, and the mean concentration was 11 parts per million, which is 5.5 times higher than the international recommended threshold. There were no significant differences between groups for comet assay parameters, with the exception that younger exposed professionals presented higher basal DNA damage than the matched controls; regarding the exposed group, the younger professionals (with higher week workload) and older professionals with greater time of xiii exposure (years) showed the highest DNA damage levels. No significant differences were also found to analyses obtained by flow cytometry. Differently, the exposed group had higher frequency of MN, nuclear bud, binucleated cells, karyorrhexis and karyolysis, and lower frequency of basal cells than the control group. The exposed women were more vulnerable to genetic instability and proliferative index while the exposed men presented more cytotoxicity. Additionally, the exposed group with lower age range had a higher frequency of MN when compared with both older professionals (≥ 31 years) exposed for up to five years and professionals with the same age range exposed for ≥ 5 years; this might be explained due to higher week workload, therefore they were more exposed to isoflurane. In conclusion, the high exposure to isoflurane is associated with DNA damage, apoptosis and proliferative changes in buccal cells, but does not impair the DNA repair capability, cell cycle or apoptosis rate when analyzed in peripheral blood cells. The findings highlight the need to mitigate anesthetic pollution in the work environment to reduce occupational exposure in professionals who work in VOR to minimize the impact of anesthetic toxicity
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Palavras-chave
Exposição ocupacional, Anestésico inalatório, Genotoxicidade, Instabilidade genômica, Apoptose, Occupational exposure, Inhalation anesthetics, Genotoxicity, Genomic instability, Apoptosis