DOI: 10.1177/1934578x261500343 ISSN: 1934-578X

In Vitro Evaluation of the Anticancer and Immunomodulatory Activities of Stingless Bee Honey-Loaded Alginate Nanoparticles

Mohammad A. I. Al-Hatamleh, Walhan Alshaer, Ma’mon M. Hatmal, Ali Mussa, Ezaldeen Esawi, Suha Wehaibi, Mohd Zulkifli Mustafa, Rohimah Mohamud
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Objectives

Stingless bee honey (SBH) has reported anticancer activity that is generally attributed to its phenolic acids and flavonoids. We previously described the synthesis, characterization, and wound-healing activity of SBH-loaded alginate nanoparticles (H-ALG NPs). Here, we evaluated the in vitro cytotoxic and anti-migratory activities of H-ALG NPs against cancer cell lines and their effect on the expression of inflammatory and immune-related genes in cancer cells and in macrophage-like cells.

Methods

Cytotoxicity of H-ALG NPs, crude SBH, and free alginate nanoparticles (free ALG NPs) was assessed by MTT assay in four cancer cell lines (MCF-7, MDA-MB-231, A549, U87) and in normal human dermal fibroblasts (HDFa). Cell migration was evaluated by in vitro scratch assay. Transcript levels of inflammatory and immune-related genes were measured by quantitative PCR (qPCR) in cancer cells and in PMA-differentiated, LPS-activated THP-1 macrophage-like cells.

Results

H-ALG NPs showed significantly greater cytotoxicity than crude SBH against the four cancer cell lines (P < 0.01), with IC 50 values of 30.70 ± 3.44 µg/mL (MCF-7), 71.23 ± 5.17 µg/mL (U87), 73.59 ± 7.31 µg/mL (A549), and 84.45 ± 8.81 µg/mL (MDA-MB-231). Viability of HDFa fibroblasts was not reduced below 50% up to 1000 µg/mL. In the scratch assay, H-ALG NPs reduced wound closure more than crude SBH or free ALG NPs. By qPCR, H-ALG NPs increased TNF and IL-6 transcripts in cancer cells but produced a smaller increase in HDFa cells. In LPS-activated THP-1 macrophage-like cells, H-ALG NPs lowered the transcript levels of TNF, IL-6, TLR4, IRAK1, IDO1, FOXP3 , and TFRC , and raised IL-10, PTGES, and CD39 .

Conclusion

H-ALG NPs showed selective in vitro cytotoxicity toward cancer cells relative to a normal fibroblast line, reduced scratch closure, and altered the expression of selected inflammatory and immune-related genes in cancer cells and in macrophage-like cells. Together with our earlier wound-healing findings, the results support further preclinical evaluation of H-ALG NPs.

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