Evaluation of the toxic potential of Bisphenol-A glycidylmethacrylate (BisGMA) on the third instar larvae of transgenic Drosophila

Abstract

Introduction: In the present study the cytotoxic and genotoxic effects of Bisphenol-A glycidyl methacrylate (BisGMA) was studied on the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg⁹.

Materials and methods: The concentration of BisGMA i.e. 0.005, 0.010, 0.015 and 0.020 M were established in diet and the larvae were allowed to feed on it for 24 h.

Results: A dose dependent significant increase in the activity of β-galactosidase was observed compared to control. A significant dose dependent tissue damage was observed in the larvae exposed to 0.010, 0.015 and 0.020 M of BisGMA compared to control. A dose dependent significant increase in the Oxidative stress markers was observed compared to control. BisGMA also exhibit significant DNA damaged in the third instar larvae of transgenic D. melanogaster (hsp70-lacZ)Bg⁹ at the doses of 0.010, 0.015 and 0.020 M compared to control.

Conclusion: BisGMA at 0.010, 0.015 and 0.020 M was found to be cytotoxic for the third instar larvae of transgenic D. melanogaster (hsp70-lacZ) Bg9.

Keywords: Drosophila; bisphenol-A-glycidylmethacrylate; cytotoxicity; genotoxicity; oxidative stress.

Fig. 1

Quantification of β- galactosidase activity in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lac Z)Bg⁹ after the exposure to various doses of Bisphenol A- glycidyl methacrylate (Bis-GMA) for 24 h. [BisGMA-1 = 0.005 M; BisGMA-2 = 0.010 M; BisGMA-3 = 0.015 M; BisGMA-4 = 0.020 M; positive control (methyl methane sulphonate = 80 μM); C = control;^*significant at P < 0.05 compared to control].

Fig. 2

X-gal staining performed on the third instar larvae of transgenic Drosophila melanogaster (hsp70-lac Z)Bg⁹ after the exposure to various doses of Bisphenol A-glycidyl methacrylate(Bis-GMA) for 24 h [positive control (methyl methane sulphonate = 80 μM (a); control (b); BisGMA-1 = 0.005 M (c); BisGMA-2 = 0.010 M (d); BisGMA-3 = 0.015 M (e); BisGMA-4 = 0.020 M (f); BG-brain ganglia, SG-salivary gland, PV-Proventriculus, FG-foregut, MG-Midgut, HG-hindgut, MT-Malpighian tubule, GC-gastric caeca].

Fig. 3

Trypan staining performed on the third instar larvae of transgenic Drosophila melanogaster (hsp70-lac Z)Bg⁹ after the exposure to various doses of Bisphenol A- glycidyl methacrylate (Bis-GMA) for 24 h [positive control (methyl methane sulphonate = 80 μM) (a); control (b); BisGMA-1 = 0.005 M (c); BisGMA-2 = 0.010 M (d); BisGMA-3 = 0.015 M (e); BisGMA-4 = 0.020 M (f); BG-brain ganglia, SG-salivary gland, PV-Proventriculus, FG-foregut, MG-Midgut, HG-hindgut, MT-Malpighian tubule, GC-gastric caeca].

Fig. 4

Quantification of tissue damage after performing trypan staining on the third instar larvae of transgenic Drosophila melanogaster (hsp70-lac Z)Bg⁹ exposed to various doses of Bisphenol A- glycidyl methacrylate (Bis-GMA) for 24 h [BisGMA-1 = 0.005 M; BisGMA-2 = 0.010 M; BisGMA-3 = 0.015 M; BisGMA-4 = 0.020 M; positive control (methyl methane sulphonate = 80 μM); C = control; ^*significant at P < 0.05 compared to control].

Fig. 5

a) Quantification of glutathione content in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lac Z)Bg⁹ exposed to various doses of Bisphenol A- glycidyl methacrylate(Bis-GMA) for 24 h [BisGMA-1 = 0.005 M; BisGMA-2 = 0.010 M; BisGMA-3 = 0.015 M; BisGMA-4 = 0.020 M; positive control (methyl methane sulphonate = 80 μM); C = control; ^*significant at P < 0.05 compared to control]. b) Quantification of glutathione-S-transferase activity in the third instar larvae of transgenic D. melanogaster (hsp70-lac Z)Bg⁹ exposed to various doses of Bisphenol A- glycidyl methacrylate(Bis-GMA) for 24 h [BisGMA-1 = 0.005 M; BisGMA-2 = 0.010 M; BisGMA-3 = 0.015 M; BisGMA-4 = 0.020 M; positive control (methyl methane sulphonate = 80 μM); C = control; ^*significant at P < 0.05 compared to control]. c) Quantification of lipid peroxidation in the third instar larvae of transgenic D. melanogaster (hsp70-lac Z)Bg⁹ exposed to various doses of Bisphenol A- glycidyl methacrylate (Bis-GMA) for 24 h [BisGMA-1 = 0.005 M; BisGMA-2 = 0.010 M; BisGMA-3 = 0.015 M; BisGMA-4 = 0.020 M; positive control (methyl methane sulphonate =80 μM); C = control; ^*significant at P < 0.05 compared to control].

Fig. 6

a) Quantification of apoptosis after performing acridine orange (AO)/Ethidium bromide (Et-Br) stanning on the midgut cells of third instar larvae of transgenic Drosophila melanogaster (hsp70-lac Z)Bg⁹ exposed to various doses of Bisphenol A- glycidyl methacrylate (Bis-GMA) for 24 h [BisGMA-1 = 0.005 M; BisGMA-2 = 0.010 M; BisGMA-3 = 0.015 M; BisGMA-4 = 0.020 M; positive control (methyl methane sulphonate = 80 μM); C = control; ^*significant at P < 0.05 compared to control]. b) Quantification of Caspase-3 activityin the mid gut cells of the third instar larvae of transgenic D. melanogaster (hsp70-lac Z)Bg⁹ exposed to various doses of Bisphenol A- glycidyl methacrylate (Bis-GMA) for 24 h [BisGMA-1 = 0.005 M; BisGMA-2 = 0.010 M; BisGMA-3 = 0.015 M; BisGMA-4 = 0.020 M; positive control (methyl methane sulphonate = 80 μM); C = control; ^*significant at P < 0.05 compared to control]. c) Quantification of Caspase-9 activityin the mid gut cells of the third instar larvae of transgenic D. melanogaster (hsp70-lac Z)Bg⁹ exposed to various doses of Bisphenol A- glycidyl methacrylate (Bis-GMA) for 24 h [BisGMA-1 = 0.005 M; BisGMA-2 = 0.010 M; BisGMA-3 = 0.015 M; BisGMA-4 = 0.020 M; positive control (methyl methane sulphonate = 80 μM); C = control; ^*significant at P < 0.05 compared to control].

Fig. 7

a) Comet assay performed on the mid gut cells of the third instar larvae of transgenic Drosophila melanogaster (hsp70-lac Z)Bg⁹ exposed to various doses of Bisphenol A- glycidyl methacrylate (Bis-GMA) for 24 h [control (a); BisGMA-1 = 0.005 M (b); BisGMA-2 = 0.010 M (c); BisGMA-3 = 0.015 M (d); BisGMA-4 = 0.020 M (e); positive control (methyl methane sulphonate = 80 μM) (f)]. b) Quantification of DNA damage after performing comet assay on the mid gut cells of the third instar larvae of transgenic D. melanogaster (hsp70-lac Z)Bg⁹ exposed to various doses of Bisphenol A- glycidyl methacrylate(Bis-GMA) for 24 h [BisGMA-1 = 0.005 M; BisGMA-2 = 0.010 M; BisGMA-3 = 0.015 M; BisGMA-4 = 0.020 M; positive control (methyl methane sulphonate = 80 μM); C = control; ^*significant at P < 0.05 compared to control].