Comparison of enzymatic and thermal synthesis of particulate bone powder derived from cuttlefish
Nozhan Azimi, Shabnam Aghayan, Sadegh HasanniaABSTRACT
Background:
The rising demand for effective bone graft materials stems from the limitations of autografts, such as limited availability and donor site morbidity. Cuttlebone, a natural and abundant calcium carbonate source, exhibits potential as an alternative due to its structural properties. This
Materials and Methods:
In this in vitro study the thermal processing method removed all organic components of the cuttlebone samples, while the enzymatic method retained collagen proteins. The resulting bone powders were characterized using scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, energy-dispersive X-ray spectroscopy, and Brunauer–Emmett–Teller analysis. Cytotoxicity and biocompatibility were evaluated using the MTT assay and cell adhesion test. Data were analyzed using one-way ANOVA test with a significance level of <0.05 and Tukey’s test for pairwise comparisons.
Results:
Enzymatically processed samples retained collagen and showed higher surface area, porosity, and cell adhesion, whereas thermally processed samples were crystalline but cytotoxic. MTT assays demonstrated that the enzymatic group showed over 100% cell viability, indicating excellent biocompatibility, while thermal samples exhibited below 70%.
Conclusion:
Overall, enzymatically processed cuttlebone demonstrated superior structural features that enhanced cell–surface interactions and biocompatibility, suggesting it as a potential alternative for craniofacial bone tissue engineering applications.