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[Study on cytotoxicity of microdoses peracetic acid in vitro]
Mo Zhou1, Youchen Li, Baoming Li
1Shanxi Provincial Tissue Bank, China Institute for Radiation Protection, Taiyuan Shanxi 030006, P R China.
Objective:
To evaluate the cytotoxicity of microdoses peracetic acid (PAA) so as to provide the evidence for making residual limit of PAA sterilization.
Methods:
Mouse fibroblasts (L929 cell line) cultured in vitro were observed to evaluate the influence of microdoses PAA including 1 x 10(-6), 2 x 10(-6), 3 x 10(-6), 4 x 10(-6), 5 x 10(-6), and 10 x 10(-6) (V/V). The proliferation of cells was determined by MTT assay at 2, 4, and 7 days of culture. The growth curve and the relative growth rate (RGR) were obtained. The cytotoxicity of PAA at different concentrations was evaluated according to RGR.
Results:
At 2, 4, and 7 days after culture, fibroblasts of 1 x 10(-6) group grew with normal morphology analogous to control group, while the cell growth of other groups were poor. With the increase of PAA concentration, the absorbance (A) values decreased, which suggested that there was a significant negative correlation between cell proliferation and PAA concentration. And the correlation coefficient was -1.000 at 2 and 4 days, - 0.964 at 7 days. There was no significant difference in A value between 1 x 10(-6) group and the control group (P > 0.05), while there were significant differences in A value between the control group and other concentration groups (P < 0.05). The growth curve of 1 x 10(-6) group was similar to that of the control group, both had obvious phase of exponential growth. The growth curves of other groups had no obvious phase of exponential growth. The cytotoxicity of 1 x 10(-6) group was classified as level 1, 2 x 10(-6) group as level 2, 3 x 10(-6) group as level 3, 4 x 10(-6) group as level 3-4, 5 x 10(-6) group and 10 x 10(-6) group as level 4.
Conclusion:
PAA of 1 x 10(-6) had no obvious cytotoxicity. The residual limit of PAA less than 1 x 10(-6) was recommended.
Insights
Peracetic acid (PAA) at 1 x 10(-6) concentration showed no significant cytotoxicity in mouse fibroblasts. This study recommends a residual limit for PAA sterilization below this concentration to ensure safety.
Area of Science:
- Cell Biology
- Toxicology
- Biochemistry
Context:
- Peracetic acid (PAA) is a potent sterilizing agent used in various industries.
- Determining safe residual limits is crucial for applications involving direct or indirect contact with biological systems.
- Cytotoxicity assessments are essential for evaluating the safety of chemical residues.
Purpose:
- To investigate the cytotoxicity of varying microdoses of peracetic acid (PAA) on mouse fibroblast cells (L929).
- To establish evidence-based recommendations for the residual limit of PAA used in sterilization processes.
- To correlate PAA concentration with cellular proliferation and growth rates.
Summary:
- Mouse fibroblasts (L929) were exposed to PAA concentrations ranging from 1 x 10(-6) to 10 x 10(-6) (V/V).
- Cell proliferation was assessed using MTT assay over 7 days, with growth curves and relative growth rates (RGR) analyzed.
- PAA at 1 x 10(-6) showed no significant cytotoxicity, with normal cell morphology and growth comparable to controls. Higher concentrations exhibited dose-dependent cytotoxicity, significantly inhibiting cell proliferation.
Impact:
- The findings indicate that PAA at 1 x 10(-6) exhibits negligible cytotoxicity.
- A recommended residual limit for PAA sterilization below 1 x 10(-6) is proposed.
- This research provides critical data for regulatory bodies and industries to set safe PAA residue standards.
