Antibacterial, antibiofilm, and wound healing activities of Piper longum root in multidrug-resistant Staphylococcus aureus infected mice

Abstract

Objective: To explore the effect of a hydrogel of Piper longum (P. longum) root against biofilm-forming multidrug-resistant (MDR) Staphylococcus aureus (S. aureus) through in vitro, in silico, and in vivo studies.

Methods: We isolated the P. longum root ethanolic extract and the compounds using p-HPLC. In vitro antibacterial and antibiofilm activities of P. longum root extract and isolated alkamide compounds against biofilm-forming MDR S. aureus (ATCC 33591) were assessed using agar diffusion and broth microdilution methods, respectively. In silico investigations were conducted to investigate the interaction of alkamide compounds with three target proteins glycogen synthase kinase 3β (GSK3β), matrix metahoproteinases-8 (MMP-8), and inducible nitric oxide synthase (iNOS). In addition, the wound healing effect of P. longum root extract 2% and 5% (w/v)- containing hydrogels was determined in mice.

Results: The ethanolic root extract of P. longum and its compounds exhibited in vitro antibacterial activity with minimum inhibitory concentrations between 50 μg/mL and 700 μg/mL, as well as significantly reduced biofilm formation. Piperdardine isolated from P. longum root extract had the best molecular docking score (−9.7, −9.8, and −9.2 kcal/mol) with target proteins GSK3β, MMP-8, and iNOS. In vivo studies showed that P. longum hydrogels significantly lowered the number of colony-forming units (P < 0.05). The P. longum 5% (w/v) hydrogel-treated group showed enhanced wound healing activity, achieving a wound contraction rate of 99.34% on day 14. Furthermore, histopathological analysis confirmed increased re-epithelialization and reduced inflammation in mice treated with P. longum 5% (w/v) hydrogel.

Conclusions: P. longum root extract has pharmacological potential as an antibacterial and wound-healing agent, and further research is required to confirm its efficacy and clinical application.

Keywords

Piper longum L. / Wound healing / Antibiofilm / Antibacterial / MDR Staphylococcus aureus / In silico docking

Conflict of interest statement

All authors declare that they have no conflict of interest.

Acknowledgments

We thankfully acknowledge the Director, CSIR-NBRI, Lucknow, for providing the necessary analytical and pharmacological facilities.

Funding

The study was supported by NPDF fellowship grants from the Central Council for Research in Ayurvedic Sciences, Ministry of AYUSH, Govt. of India (HQ-ESTT012/60/2022-ESTT/6783).

Data availability statement

The data supporting the findings of this study are available from the corresponding author upon request.

Authors’ contributions

PG carried out biochemical experiments, isolated compounds for p-HPLC analysis, and wrote and edited manuscripts. PS carried out microbial tests for antibacterial and antibiofilm activities. JY carried out both biochemical and histological analyses. CVR was involved in conceptualisation, supervision, data curation, research administration, validation, writing, review, and editing. All authors have approved the final manuscript.

Publisher’s note

The Publisher of the Journal remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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