# In Vitro Evaluation and Network Pharmacology Analysis of the Antimicrobial Activity of Pistacia lentiscus

## TL;DR

This study combines network pharmacology and lab tests to show that mastic gum from Pistacia lentiscus has strong antimicrobial effects, potentially useful for treating infections.

## Contribution

The study integrates network pharmacology with in vitro testing to reveal multiple antimicrobial mechanisms of P. lentiscus phytochemicals.

## Key findings

- Four key phytochemicals (alpha-terpineol, linalool, myrcene, verbenone) showed favorable pharmacokinetics and antimicrobial target interactions.
- Ethanolic extract of P. lentiscus achieved ≥5 log reductions in bacterial and fungal strains, comparable to chlorhexidine.
- Antimicrobial mechanisms include membrane disruption, virulence inhibition, and modulation of host inflammation.

## Abstract

Chios mastic gum, a natural resin derived from the mastic tree, has a history of traditional use for its beneficial effects on the gastrointestinal system, its anti‐inflammatory properties, and its antimicrobial activity.

This study aimed to integrate network pharmacology and standardized in vitro analyses to elucidate the antimicrobial mechanisms and therapeutic potential of P. lentiscus phytochemicals.

This study employed a systematic methodology integrating data mining, network construction, and network analysis to explore the intricate relationships between the constituents of mastic gum and their diverse biological targets. The primary active compounds of P. lentiscus were identified through multiple databases, including the Indian Medicinal Plants, Phytochemistry, and Therapeutics database. ADME/T profiling, Lipinski’s rule of five, and target prediction were performed to assess drug likeness and pharmacokinetics. Protein–protein interaction networks and gene ontology (GO) enrichment were analyzed using STRING and Cytoscape to identify biological pathways associated with antimicrobial activity. Moreover, the antibacterial efficacy of these compounds was evaluated following the Polish‐European standard 1040, and antifungal activity was assessed according to the European standard 1650 by enumerating colony‐forming units after a 24‐h incubation period.

Four major phytochemicals — alpha‐terpineol, linalool, myrcene, and verbenone — were identified as key bioactive compounds. These exhibited favorable pharmacokinetic properties and predicted interactions with targets related to inflammation, oxidative stress, and microbial virulence. In vitro, the ethanolic extract of P. lentiscus achieved ≥ 5 log reductions in both bacterial and fungal strains, demonstrating potent antimicrobial activity comparable to chlorhexidine. The hydroalcoholic extract showed moderate yet significant reductions.

The integrative in silico and in vitro analyses indicate that P. lentiscus exerts antimicrobial effects through multiple complementary mechanisms, including membrane disruption, inhibition of virulence and biofilm formation, and modulation of host inflammatory pathways. These findings provide a mechanistic framework supporting the use of P. lentiscus as a natural adjunct for managing periodontal infections and warrant further preclinical and clinical evaluation.