Purpose: The aim of this study was to optimize the immunoresponsive and anti-Candida albicans (C. albicans) properties of guided bone regeneration-resorbable membranes (collagen) using additives based on synergistic mixtures of chlorhexidine with terpene-enriched fractions from Lippia alba essential oils (EO).

Methods: The anti-C. albicans activity of the fractions (individually or combined with chlorhexidine) was evaluated using planktonic and sessile cultures. J774A.1 murine macrophage cells were used to determine the cytotoxicity and immunoresponsive effects of the therapies.

Results: The anti-planktonic and anti-sessile performance of chlorhexidine on C. albicans was improved 2- to 4-fold by supplementation with citral-rich fractions. On macrophages, this fraction also exhibited a potentially cytoprotective action against the toxic effects of chlorhexidine, minimizing damage to the cell membrane, mitochondrial membrane potential, and nuclear integrity. Macrophages growing on collagen-membrane fragments and stimulated with the citral fraction (alone or with chlorhexidine) showed a significant increase in releasing the osteogenic cytokine TNF-α and enhancing the IL-4.

Conclusion: This combined therapy appears as a promising platform for the development of a prophylactic or therapeutic biocidal solution that can optimize the pharmacological characteristics of chlorhexidine (epithelium tolerance and anti-C. albicans consolidation on surfaces), as well as potentiating the immunoresponsive properties of collagen membranes.