Challenging Applicability of ISO 10993-5 for Calcium Phosphate Biomaterials Evaluation: Towards More Accurate in vitro Cytotoxicity Assessment
Biomaterials Advances 2024
Ilijana Kovrlija, Ksenia Menshikh, Hugo Abreu, Andrea Cochis, Lia Rimondini, Olivier Marsan, Christian Rey, Christele Combes, Jānis Ločs, Dagnija Loča

Research on biomaterials typically starts with cytocompatibility evaluation, using the ISO 10993-5 standard as a reference that relies on extract tests to determine whether the material is safe (cell metabolic activity should exceed 70 %). However, the generalized approach within the standard may not accurately reflect the material's behavior in direct contact with cells, raising concerns about its effectiveness. Calcium phosphates (CaPs) are a group of materials that, despite being highly biocompatible and promoting bone formation, still exhibit inconsistencies in basic cytotoxicity evaluations. Hence, in order to test the cytocompatibility dependence on different experimental setups and material-cell interactions, we used amorphous calcium phosphate, α-tricalcium phosphate, hydroxyapatite, and octacalcium phosphate (0.1 mg/mL to 5 mg/mL) with core cell lines of bone microenvironment: mesenchymal stem cells, osteoblast-like and endothelial cells. All materials have been characterized for their physicochemical properties before and after cellular contact and once in vitro assays were finalized, groups identified as ‘cytotoxic’ were further analyzed using a modified Annexin V apoptosis assay to accurately determine cell death. The obtained results showed that indirect contact following ISO standards had no sensitivity of tested cells to the materials, but direct contact tests at physiological concentrations revealed decreased metabolic activity and viability. In summary, our findings offer valuable guidelines for handling biomaterials, especially in powder form, to better evaluate their biological properties and avoid false negatives commonly associated with the traditional standard approach.


Atslēgas vārdi
Calcium phosphates, Material characterization, Standards, Cytocompatibility guideline, Cytotoxic, Apoptosis, Necrosis, Direct contact, Indirect contact
DOI
10.1016/j.bioadv.2024.213866
Hipersaite
https://www.sciencedirect.com/science/article/pii/S2772950824001092?via%3Dihub

Kovrlija, I., Menshikh, K., Abreu, H., Cochis, A., Rimondini, L., Marsan, O., Rey, C., Combes, C., Ločs, J., Loča, D. Challenging Applicability of ISO 10993-5 for Calcium Phosphate Biomaterials Evaluation: Towards More Accurate in vitro Cytotoxicity Assessment. Biomaterials Advances, 2024, Vol. 160, Article number 213866. ISSN 2772-9516. e-ISSN 2772-9508. Pieejams: doi:10.1016/j.bioadv.2024.213866

Publikācijas valoda
English (en)
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