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A novel particulate matter sampling and cell exposure strategy based on agar membrane for cytotoxicity study
Si-Si Chen1, Tian-Qi Wang1, Wan-Chen Song2
1State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry & Chemical Engineering and Center of Materials Analysis, Nanjing University, Nanjing, 210023, China.
Abstract:
Laboratories use different strategies to sample and extract atmospheric particulate matter (PM), some of which can be very complicated. Due to the absence of a standard protocol, it is difficult to compare the results of PM toxicity assessment across different laboratories. Here, we proposed a novel PM sampling and cell exposure strategy based on agar membrane. The agar membrane, prepared by a simple freeze-drying method, has a relatively flat surface and porous interior. We demonstrated that the agar membrane was a reliable substitute material for PM sampling. Then the PM on the agar membranes was directly extracted with the culture medium by vortex method, and the PM on the polytetrafluoroethylene (PTFE) filters was extracted with water by the traditional ultrasonic method for comparison. The extraction efficiency was evaluated and in vitro cytotoxicity assays were carried out to investigate the toxic effects of PM extracted with two strategies on macrophage cells. The results showed that the PM extracted from agar membranes induced higher cytotoxicity and more differentially expressed proteins. Overall, the novel PM sampling-cell exposure strategy based on the agar membrane is easy to operate, biocompatible and comparable, and has low disturbance, could be an alternative sampling and extraction method for PM toxicity assessment.
Insights
A new agar membrane method simplifies atmospheric particulate matter (PM) sampling and extraction for toxicity studies. This approach offers a standardized, low-disturbance alternative for reliable PM toxicity assessment.
Area of Science:
- Environmental Science
- Toxicology
- Materials Science
Background:
- Standardization of atmospheric particulate matter (PM) sampling and extraction is lacking, hindering comparative toxicity assessments.
- Current methods can be complex and may affect the integrity of PM for toxicological analysis.
Purpose of the Study:
- To develop and evaluate a novel, simplified strategy for PM sampling and cell exposure using an agar membrane.
- To compare the efficiency and biological impact of PM extraction from agar membranes versus traditional polytetrafluoroethylene (PTFE) filters.
Main Methods:
- A novel agar membrane was prepared using a simple freeze-drying technique for PM sampling.
- PM was extracted from agar membranes via vortexing with culture medium and from PTFE filters using sonication in water.
- In vitro cytotoxicity assays were performed on macrophage cells exposed to PM extracted by both methods.
Main Results:
- The agar membrane proved to be a reliable material for PM sampling.
- PM extracted from agar membranes exhibited higher in vitro cytotoxicity compared to PM from PTFE filters.
- Analysis revealed more differentially expressed proteins in macrophage cells exposed to PM from agar membranes.
Conclusions:
- The proposed agar membrane-based strategy offers a simple, biocompatible, and low-disturbance method for PM sampling and extraction.
- This novel approach could serve as a valuable alternative for standardized PM toxicity assessment.
- The method facilitates direct cell exposure and enhances the detection of PM's toxicological effects.
