# Enhancing anti-inflammatory activity of Eucalyptus camaldulensis by upregulating secondary metabolites using suspension cultures techniques

## TL;DR

This study shows that growing Eucalyptus camaldulensis in tissue culture increases its anti-inflammatory compounds and makes them more effective than natural leaves.

## Contribution

The study demonstrates that tissue culture enhances secondary metabolite production and anti-inflammatory activity in Eucalyptus camaldulensis.

## Key findings

- Callus extracts contain more phytoconstituents (54) than leaf extracts (32).
- Callus volatile oil has higher levels of bioactive compounds like 1,8-cineole and α-terpineol.
- Callus volatile oil shows greater anti-inflammatory activity than leaf oil and indomethacin.

## Abstract

Eucalyptus camaldulensis Dehn (Family Myrtaceae) is among the most prominent Eucalyptus species, extensively exploited for its anti-inflammatory efficacy. The present study undertakes a comparative phytochemical and pharmacological evaluation of leaf and tissue culture (callus) extracts and their corresponding volatile oils. Methanolic extracts from both plant sources were subjected to liquid chromatography–mass spectrometry (LC/MS), revealing 32 phytoconstituents in leaf extracts and 54 in callus extracts. Identified chemical classes included flavonoids, tannins, coumarins, and phenolic acids, with higher relative abundance in callus-derived extracts. Volatile oil analysis using gas chromatography–mass spectrometry (GC/MS) identified 58 compounds in leaf oil and 52 in callus oil. Quantitative profiling demonstrated a significant elevation in bioactive volatiles within callus oil: 1,8-cineole content showed a 2.1-fold increase compared to leaf oil, and both α-terpineol and sabinene exhibited approximately fourfold increases relative to leaf oil. Bioassays further indicated that callus extracts and volatile oils possess enhanced anti-inflammatory and antioxidant activities, exhibiting higher potency than leaf-derived counterparts. Remarkably, the anti-inflammatory activity of callus volatile oil exceeded that of the standard pharmaceutical agent indomethacin. These findings underscore the potential of in vitro tissue culture as a biotechnological tool for amplifying the yield and bioactivity of secondary metabolites in E. camaldulensis.

The online version contains supplementary material available at 10.1038/s41598-025-34963-8.