GReen manufactured AntiMicrobial porous matrices for antibiotic-free cell cultures of human primary cells

Antibiotics are molecules that exhibit bactericidal or bacteriostatic properties, controlling bacterial infections and preventing contamination by inducing the death of microbes. These properties make them an effective means of combatting bacteria. However, an excessive use over the years has led to the generation of multidrug-resistant strains that are difficult to treat and to handle. Therefore, antimicrobial resistance has become a global challenge that threatens the future of humanity. Antibiotics are no longer infallible against bacteria, as many bacterial strains can easily develop resistance mechanisms to guarantee their survival. Thus, alternatives to antibiotics, or more effective and sophisticated solutions, are needed. To avoid using antibiotics in human cell cultures, the CNR-ISSMC team produced 2D and 3D matrices based on chitosan and copper ions encapsulated in hydroxyapatite nanoparticles. These matrices are able to reduce the risk of bacterial contamination and confer antimicrobial properties in the absence of antibiotics. At the Centre for Regenerative Medicine (CMR), we tested the viability and the proliferation of T lymphocytes and skin fibroblasts, derived from healthy donors, cultured on 2D and 3D matrices without antibiotic in the medium. In this experimental setting, we assessed also the ability of copper ions to prevent bacterial infections under long-term cell culture conditions. Finally, we investigated the recovery of human fibroblasts and T cells edited by nucleofection of CRISPR/Cas nuclease in a 3D scaffold environment without antibiotics.