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Extraction and Analysis of Taiwanese Green Propolis
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A synergistic nanoformulation of propolis and chlorhexidine against Acanthamoeba: encapsulation efficiency, release

Nivetha Marimuthu1, Siriphorn Chimplee2, Shanmuga Sundar Saravanabhavan1

  • 1Department of Biotechnology, Aarupadai Veedu Institute of Technology, Vinayaka Mission's Research Foundation (DU), Chennai, Tamil Nadu, India.

Peerj
|December 24, 2025
PubMed
Abstract

Insights

Chitosan nanoparticles loaded with propolis and chlorhexidine show potent anti-Acanthamoeba activity against resistant cysts with minimal toxicity to mammalian cells, offering a promising biocompatible therapy.

Area of Science:

  • Nanotechnology
  • Microbiology
  • Pharmacology

Background:

  • Acanthamoeba species are opportunistic protozoa causing difficult-to-treat ocular infections due to resistant cysts.
  • Current treatments face challenges in eradicating these resilient parasitic forms.

Purpose of the Study:

  • To evaluate the anti-Acanthamoeba activity and cytotoxicity of chitosan nanoparticles (CS-NPs) loaded with propolis extracts and chlorhexidine (CHX).
  • To investigate the efficacy of these nanoparticles against both trophozoite and cyst stages of Acanthamoeba.

Main Methods:

  • CS-NPs were formulated with propolis extracts from three stingless bee species, alone or combined with CHX.
  • Encapsulation efficiency, drug release kinetics, pH sensitivity, anti-Acanthamoeba activity, and cytotoxicity against Vero cells were assessed.
  • Scanning electron microscopy was used to visualize cyst wall damage.

Main Results:

  • Formulations achieved high encapsulation efficiency (81-92%) and sustained drug release (>80%).
  • Propolis-CHX combinations demonstrated potent cysticidal activity (MICC 1.25%) against Acanthamoeba polyphaga with low cytotoxicity (MCC 2.5%).
  • Drug release followed non-Fickian diffusion, enhanced at acidic pH, and SEM showed cyst wall damage.

Conclusions:

  • Propolis-CHX-loaded CS-NPs are a promising, biocompatible therapeutic strategy against drug-resistant Acanthamoeba cysts.
  • The targeted delivery and enhanced release at acidic pH conditions suggest potential for ocular infection treatment.
Keywords:
Acanthamoeba sppDrug developmentNanoparticlesNatural productPropolis

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