Antibacterial activity of Brazilian red propolis and in vitro evaluation of free radical production

Vital pulp therapy (VPT) preserves the vitality of the remaining pulp tissue in primary and immature permanent teeth (Hanna et al., 2020). Mineral trioxide aggregate (MTA), a calcium silicate-based cement, has demonstrated great potential for pulpotomies and pulp capping procedures, because it is a biocompatible material that can induce dentinal bridge formation, and exhibits regenerative properties, lower cytotoxicity, and higher clinical and radiographic success rates than other pulpotomy agents (Tabarsi et al., 2010).

Evidence-based studies have demonstrated MTA to be the gold standard for pulpotomies, followed by Biodentine™ (Stringhini Junior, dos Santos, Oliveira, & Mercadé, 2019). However, more recent alternatives to VPTs are available, such as a premixed bioceramic material (EndoSequence Root Repair) and a resin-modified tricalcium silicate (TheraCal) (Smaïl-Faugeron et al., 2018), as well as the search for natural products with reduced toxicity and potentially more affordable prices for the population (mainly due to the increasing acceptance of natural medicine), has been undertaken in endodontics (da Silva Barboza, Aitken-Saavedra, Ferreira, Aranha, & Lund, 2021).

Brazilian propolis is a set of resinous materials with high historical, medicinal, and economic value (Sousa, Niege, Furtado, Soares, & Bastos, 2007). Propolis is collected from different plant sources and prepared by bees of the species Apis mellifera (Peter et al., 2017). The chemical composition of propolis is complex and varies according to the flora of the region where it is produced, harvesting season, and genetic characteristics of the bees that produce it, which directly influence the active principles, reflecting the biological and pharmaceutical activities of the different products (Castro et al., 2007).

Brazilian red propolis (BRP), found on the northeastern coast, is derived mainly from the red resinous exudate produced on the surface of Dalbergia ecastophyllum induced by the presence of a parasitic insect (new species of jewel beetle, Agrilus propolis) that feeds on the stem of this plant (Migliore et al., 2022). The pharmacological properties of clinical interest, such as anticaries, antivirus, antifungal, and anti-inflammatory effects, as well as cytotoxic activity against tumor lineages (Freires et al., 2016, Machado et al., 2016) and the antibacterial activity of propolis with satisfactory results against Actinomyces naeslundii, Pseudomys oralis, Prevotella melaninogenica, Porphyromonas gingivalis, Fusobacterium nucleatum, and Veillonella parvula, have already been described (Boeing et al., 2021).

Infected root canals are composed of interacting microbiota that include species of cocci and bacilli, gram-positive and gram-negative, facultative, and strict anaerobes (Aveiro et al., 2020, Gomes et al., 2008, Louzada et al., 2020). Therefore, primary endodontic infections, occur due to the colonization of necrotic pulp tissue by predominantly strict anaerobic bacteria, such as Actinomyces spp., Fusobacterium spp., Porphyromonas spp., Prevotella spp., Parvimonas spp., Pseudoramibacter spp., Tannerella spp., Treponema spp., Dialister spp., Filifactor spp., Olsenella spp., and Pseudoramibacter spp. (Gomes and Herrera, 2018) whereas gram-positive bacteria are strongly associated with secondary/persistent endodontic infection (Barbosa-Ribeiro et al., 2016).

Iranian propolis could promote anti-inflammatory effects in a murine macrophage cell line (RAW 264.7) exposed to lipopolysaccharide (LPS), reducing free radical (nitric oxide – NO and reactive oxygen species – ROS) production, cyclooxygenase-2 (COX-2), interleukin-1β (IL-1β), and interleukin-6 (IL-6) expression (Asgharpour, Moghadamnia, Motallebnejad, & Nouri, 2019). Another study demonstrated that BRP was able to promote anti-inflammatory effects in LPS-activated peritoneal macrophages by decreasing NO levels and reducing the production and expression of proinflammatory cytokines and genes (Bueno-Silva et al., 2017a).

BRP has many biological properties and can provide therapeutic solutions for VPT (da Silva Barboza et al., 2021). First, we investigated the antibacterial and antibiofilm activity of crude BRP extracts against anaerobic bacteria involved in primary endodontic infections. The present study evaluated the viability and free radical production of human dental pulp fibroblasts (HDPF) in direct contact with different concentrations of MTA and BRP. We hypothesized that BRP might show good antibacterial and antibiofilm activity and similar behavior to MTA in terms of free radical production and cell viability.

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