Assessment of the anticancer and antioxidant potentials of Spinosaurus tooth fossils: Application of energy dispersive X-ray fluorescence and field emission scanning electron microscopy techniques
Yamini MalhotraObjectives:
The study was undertaken to investigate the elemental composition, microstructural characteristics, anticancer activity, and antioxidant-related potential of fossilized Spinosaurus tooth material using field emission scanning electron microscopy (FESEM), energy-dispersive spectroscopy (EDS), energy-dispersive X-ray fluorescence (ED-XRF), gas chromatography–mass spectrometry (GC-MS), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) cytotoxicity assay, and intracellular reactive oxygen species (ROS) analysis. The work aimed to determine whether fossilized Spinosaurus tooth material, beyond its paleontological value, may also represent a mineralized bioactive matrix with potential relevance to cancer-related biological research.
Materials and Methods:
An authenticated fossilized Spinosaurus tooth specimen obtained from the Fossil Era was examined by camera imaging and binocular microscopy and then subjected to FESEM imaging, EDS, and ED-XRF elemental analysis. The fossil was powdered and extracted using ethanol-water (70:30), and the resulting extract was analyzed by GC-MS to identify potentially bioactive compounds. Anticancer activity was evaluated against the MDA-MB-231 human breast cancer cell line using the MTT assay, while intracellular ROS generation was assessed using 2’,7’-dichlorodihydrofluorescein diacetate fluorescence staining. Cell viability, IC 50 values, and ROS fluorescence responses were interpreted in relation to the fossil’s mineral and chemical composition.
Results:
FESEM-EDS and ED-XRF analyses revealed that the fossilized tooth consisted of a highly mineralized matrix enriched with oxygen, calcium, phosphorus, silicon, iron, magnesium, manganese, carbon, and additional trace elements. FESEM–EDS of the upper tooth region showed oxygen (69.16 wt%), carbon (12.96 wt%), iron (7.39 wt%), magnesium (0.49 wt%), and manganese (0.21 wt%), indicating the presence of redox-active and apatite-associated phases. GC-MS profiling identified several compounds, including anthraquinone-related molecules, pyrrole derivatives, aromatic esters, sulfur-containing compounds, and iron-associated organometallic complexes. In vitro cytotoxicity studies demonstrated concentration-dependent inhibition of MDA-MB-231 breast cancer cells, with an IC 50 value of 72.67 µg/mL. Intracellular ROS fluorescence analysis further indicated increased oxidative stress in treated cells.
Conclusion:
The findings suggest that fossilized Spinosaurus tooth material represents a chemically complex mineralized bio-composite with measurable in vitro anticancer and oxidative stress-modulating activity. The observed effects may arise from the synergistic interaction of redox-active trace elements and fossil-associated organic constituents. Although preliminary and requiring further mechanistic, toxicological, and in vivo validation, the present study establishes an interdisciplinary foundation for exploring fossil-derived biomineral materials in anticancer and antioxidant research.