https://doi.org/10.5061/dryad.mw6m9062w

[Plain Language Summary]
With the Artemis program, humans will soon return to explore the Moon. However, lunar surface dust has toxic potential that must be assessed in order to clarify short-term and long-term health risks for Artemis astronauts. Numerous studies indicate that Moon dust has chemical and physical properties that may strongly affect dust toxicity. Unlike terrestrial dust, lunar regolith experiences “space weathering” under a vacuum, including the effects of solar wind, which further modifies the bulk and surface properties of this dust. In this work, we used two lunar dust simulant materials that were chemically treated to mimic the effects of space weathering. This treatment strongly increased all the toxic effects of both simulants: cell killing, mitochondrial dysfunction, and damage to DNA. Other experiments point to free radicals as a significant component of these effects. Future work will address whether these radicals arise from the simulants themselves or are generated by cellular activity.

Authors

Jamie Hsing-Ming Chang
Department of Pharmacological Sciences, Renaissance School of Medicine, Stony Brook University, NY 11794, USA
Hsing-Ming.Chang@stonybrook.edu

Zhouyiyuan Xue
Department of Pharmacological Sciences, Renaissance School of Medicine, Stony Brook University, NY 11794, USA
Zhouyiyuan.Xue@stonybrook.edu

Jack Bauer
Department of Pharmacological Sciences, Renaissance School of Medicine, Stony Brook University, NY 11794, USA
Jack.Bauer@stonybrook.edu

Barbara Wehle
Department of Pharmacological Sciences, Renaissance School of Medicine, Stony Brook University, NY 11794, USA
bjm00025@mix.wvu.edu

Donald A. Hendrix
National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL 32310, USA
donh2015@gmail.com

Tristan Catalano
Department of Geosciences, Stony Brook University, NY 11794, USA
Tristan.Catalano@stonybrook.edu

Joel A. Hurowitz
Department of Geosciences, Stony Brook University, NY 11794, USA
Joel.Hurowitz@stonybrook.edu

Hanna Nekvasil
Department of Geosciences, Stony Brook University, NY 11794, USA
Hanna.Nekvasil@stonybrook.edu

Bruce Demple
Departments of Pharmacological Sciences and of Radiation Oncology, Renaissance School of Medicine, Stony Brook University, NY 11794, USA
Bruce.Demple@stonybrook.edu

File Description from Figure 1-6

Fig. 1.zip: In this .zip file, you will see raw files for Figure 1 in the related GeoHealth manuscript with folders of “102821bu” and “102821x” under “SEM images and reports.” The former (folder “102821bu”) includes SEM images of non-reduced LMS-1 (folder “LMS_1”), reduced LMS-1 (folder “LMS_1_R”), LHS-1 (folder “LHS_1”), and reduced LHS-1 (folder “LHS_1_R”), and the later (folder “102821x”) includes spectra files of LHS-1 (folder “LHS_1x”), reduced LHS-1 (folder “LHS_1_Rx”) and reduced LMS-1 (folder “LMS_1_Rx”), and a .xlsx with a table of proving of decrease of size after grinding of lunar dust simulants.

Figure 2, 3, 4 & 6.xlsx: In this .xlsx file with 14 sheets, you will see raw data for Figure 2, 3, 4A, 4B, 6B, and the cell survival for the concentration of N-acetylcysteine (abbreviated as NAC) we used in the related GeoHealth manuscript. For sheets of “NAC survival testing”, “Fig. 2A - untreated LMS”, “Fig. 2A - reduced LMS”, “Fig. 2B - untreated LHS”, “Fig. 2B - reduced LHS”, “Fig. 2C”, and “Fig. 2D”, the UPPER part showed the RAW cell concentration with condition listed upper left (e.g. “A549 1 h LHS-1” stands for A549 cell survival after 1 h-exposure to LHS-1) , and the LOWER part showed the NORMALIZED cell survival after negative control (labeled “Control”) is set to 100% with P values provided. In the sheet of “NAC survival testing”, 70.70 in RED showed that 5 mM NAC is the highest and also the best concentration for pretreating A549 cells before the dust challenge as it is the highest survival among different NAC concentrations. In the sheet of “Fig. 2C” and “Fig. 2D”, the experimental groups pretreated with NAC were compared to their own controls. For sheets of “Fig. 3A - LMS-1”, “Fig. 3A - LHS-1”, “Fig. 3B - LMS-1”, and “Fig. 3B - LHS-1”, RAW fluorescent readings of Long PCR and Short PCR were shown in “Picogreen Readout,” then “Net Value” were calculated after subtracting “PCR Blank” for each group. Later, the ratios were first calculated by dividing the “Net Value” of Long PCR to Short PCR, and then normalizing the controls to 1 with p values provided. In the sheet of “Fig. 3B - LMS-1” and “Fig. 3B - LHS-1”, the experimental groups pretreated with NAC were compared to their own controls. For sheets of “Fig. 4A” and “Fig. 4B”, RAW fluorescent readings of MitoSOX Red were shown under “510 nm>580 nm”, the blanks were then subtracted, and controls are normalized to 1 with p values provided. The experimental groups pretreated with NAC were compared to their own controls. For sheets of “Fig. 6B”, RAW “Tail Moment” values were provided by using ImageJ and OpenComet as described in the manuscript, and then the negative control was normalized to 1 with p values provided.

Figure 6A.zip: In this .zip file, you will see raw image .tif files for Figure 6A in the related GeoHealth manuscript in the designated folder of “(Negative) Control”, “Positive Control (Etoposide)”, “LMS-1”, “Reduced LMS-1”, “LHS-1”, and “Reduced LHS-1”.

Figure 5.pzf: You will need GraphPad Prism to open this file to gain access to raw data points of oxygen consumption continuously detected up to 72 hours after exposure to LMS-1 in A549 cells. Each group usually contains 4 replicates, except for R-0.5 mg/cm² with only 2 replicates and 0.2 mg/cm² with only 3 replicates because of technical issues. Please refer to the captions of Figure 5 in the related GeoHealth manuscript.