Assessment of gallbladder stone – A geological approach through cutting-edge field emission scanning electron microscope and energy-dispersive X-ray fluorescence spectroscopy with anti-cancerous properties examined for hep G2 (liver) cancer cell lines with validation through reactive oxygen species anti-oxidant analysis
Authors: Malhotra Yamini , Subramanian S. R. Singara , Vennila L. , Mukesh M. V. , Kumar N. Senthil 
Published: July 2024

Objectives:
Geology, traditionally focused on the study of Earth, ocean, and planetary rocks, extends to the examination of stones formed within the human body, such as those found in the bladder or kidneys. This research specifically targets the classification and elemental composition of gallbladder stones, with a concentrated analysis on the anionic and cationic constituents. The study employs a combination of advanced imaging and spectroscopic techniques to delve into the intricate details of these stones and evaluates their potential medical applications, particularly their anti-cancer properties.

Material and Methods:
The methodology involved in this research is multifaceted, incorporating several state-ofthe-art techniques. Field emission scanning electron microscopy (FESEM) was utilized to capture high-resolution images of the gallbladder stone samples, providing a detailed look at their surface morphology. Complementing this imaging technique, energy-dispersive X-ray analysis (EDX) was employed to determine the elemental composition of the samples. Additionally, energy-dispersive X-ray fluorescence (EDXRF) spectroscopy was conducted, both with and without chromium coating, to further analyze the elemental makeup of the stones.

Results:
The results from these techniques revealed a comprehensive profile of the elemental composition of gallbladder stones. FESEM provided detailed images, allowing for a thorough examination of the stone’s microstructure. EDX analysis contributed to the identification of various elements present in the samples, highlighting the predominant anions and cations. EDXRF spectroscopy, with its high sensitivity and accuracy, corroborated these findings, ensuring a robust and precise determination of the elemental constituents.Beyond the geological analysis, the study explored the potential biomedical applications of gallbladder stones. Samples were tested for their anti-cancer properties using the MTT assay on Hep G2 liver cancer cells. The MTT assay is a colorimetric assay that measures the metabolic activity of cells, providing an indication of cell viability and proliferation. The gallbladder stones exhibited significant anti-cancerous properties, with an inhibitory concentration (IC50) value of 70.60, indicating their efficacy in inhibiting the growth of liver cancer cells. To further validate these findings, the samples underwent reactive oxygen species (ROS) antioxidant analysis. This analysis assesses the toxicity of the stones and their ability to act as antioxidants. The results confirmed that the gallbladder stones not only possess anti-cancerous properties but also exhibit antioxidant activity. The ROS analysis demonstrated that the stones could effectively neutralize reactive oxygen species, which are known to cause oxidative stress and contribute to the development and progression of cancer.

Conclusion:
The conclusions drawn from this comprehensive study are twofold, providing significant contributions to both geological and biomedical research fields. Geologically, the study offers an in-depth understanding of the nature and classification of gallbladder stones, detailing their elemental composition through advanced imaging and spectroscopic techniques. Biomedically, the research highlights the potential of gallbladder stones as effective anti-cancer agents, supported by rigorous testing and validation through MTT assays and ROS antioxidant analysis. In summary, this study bridges the gap between geology and medicine, unveiling the intricate composition of gallbladder stones and their promising anti-cancer properties. The findings underscore the importance of interdisciplinary research, demonstrating how geological studies can inform and enhance biomedical applications. By providing valuable insights into the composition and medical potential of gallbladder stones, this research opens new avenues for the development of novel anti-cancer therapies and contributes to the broader understanding of both geological and biomedical sciences.