Fabrication and characterization of Agarwood extract-loaded nanocapsules and evaluation of their toxicity and anti-inflammatory activity on RAW 264.7 cells and in zebrafish embryos

Drug Deliv. 2021 Dec;28(1):2618-2633. doi: 10.1080/10717544.2021.2012307.

Abstract

Aquilaria malaccensis has been traditionally used to treat several medical disorders including inflammation. However, the traditional claims of this plant as an anti-inflammatory agent has not been substantially evaluated using modern scientific techniques. The main objective of this study was to evaluate the anti-inflammatory effect of Aquilaria malacensis leaf extract (ALEX-M) and potentiate its activity through nano-encapsulation. The extract-loaded nanocapsules were fabricated using water-in-oil-in-water (w/o/w) emulsion method and characterized via multiple techniques including DLS, TEM, FTIR, and TGA. The toxicity and the anti-inflammatory activity of ALEX-M and the extract-loaded nanocapsules (ALEX-M-PNCs) were evaluated in-vitro on RAW 264.7 macrophages and in-vivo on zebrafish embryos. The nanocapsules demonstrated spherical shape with mean particle diameter of 167.13 ± 1.24 nm, narrow size distribution (PDI = 0.29 ± 0.01), and high encapsulation efficiency (87.36 ± 1.81%). ALEX-M demonstrated high viability at high concentrations in RAW 264.7 cells and zebrafish embryos, however, ALEX-M-PNCs showed relatively higher cytotoxicity. Both free and nanoencapsulated extract expressed anti-inflammatory effects through significant reduction of the pro-inflammatory mediator nitric oxide (NO) production in LPS/IFNγ-stimulated RAW 264.7 macrophages and zebrafish embryos in a concentration-dependent manner. The findings highlight that ALEX-M can be recognized as a potential anti-inflammatory agent, and its anti-inflammatory activity can be potentiated by nano-encapsulation. Further studies are warranted toward investigation of the mechanistic and immunomodulatory roles of ALEX-M.

Keywords: Aquilaria malaccensis; RAW 264.7; anti-inflammatory; polymeric nanocapsules; zebrafish.

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / administration & dosage
  • Anti-Inflammatory Agents / pharmacology*
  • Cell Survival / drug effects
  • Dose-Response Relationship, Drug
  • Drug Liberation
  • Embryo, Nonmammalian
  • Inflammation / pathology*
  • Inflammation Mediators / metabolism
  • Macrophages / drug effects
  • Mice
  • Nanocapsules / chemistry*
  • Nitric Oxide / metabolism
  • Particle Size
  • Plant Extracts / administration & dosage
  • Plant Extracts / pharmacology*
  • Plant Leaves
  • RAW 264.7 Cells
  • Surface Properties
  • Thymelaeaceae*
  • Zebrafish

Substances

  • Anti-Inflammatory Agents
  • Inflammation Mediators
  • Nanocapsules
  • Plant Extracts
  • Nitric Oxide

Grants and funding

This research was funded by the Fundamental Research Grant Scheme (FRGS), grant number [FRGS/1/2019/WAB11/UIAM/02/4] awarded by the Ministry of Higher Education in Malaysia.