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. 2024 Sep 4;16(9):e68653.
doi: 10.7759/cureus.68653. eCollection 2024 Sep.

Modulation of Inflammation in McCoy Cells by Zinc Nanoparticles Conjugated With β-Chitosan

Mampi Payra  1 , Karthik Ganesh Mohanraj  1 , Taniya Mary Martin  1 , Meenakshi Sundaram K  1
Affiliations

Modulation of Inflammation in McCoy Cells by Zinc Nanoparticles Conjugated With β-Chitosan

Mampi Payra et al. Cureus. .

Abstract

Introduction: This study investigated biosynthetically derived β-chitosan-derived zinc nanoparticles (β-Ch-Zn NPs) for their potential anti-inflammatory properties on McCoy cells. β-Ch-Zn NPs were synthesized using a green chemistry approach, and their characterization confirmed successful synthesis, appropriate size, and morphology. The study aimed to evaluate the cytotoxicity of β-Ch-Zn NPs and their effects on inflammatory responses in McCoy cells stimulated with lipopolysaccharide (LPS).

Methods: β-Ch-Zn NPs were synthesized and characterized using Fourier-transform infrared spectroscopy (FTIR), ultraviolet-visible spectroscopy (UV-Vis) spectroscopy, and X-ray diffraction (XRD) to confirm their structural and morphological properties. The cytotoxicity of β-Ch-Zn NPs was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay at various concentrations to determine safe doses for subsequent experiments. To induce inflammation, McCoy cells were pretreated with β-Ch-Zn NPs at different concentrations before LPS stimulations. Gene expression analysis using quantitative real-time polymerase chain reaction was performed to measure the messenger RNA (mRNA) levels of proinflammatory cytokine.

Results: FTIR, UV-Vis spectroscopy, and XRD confirmed the successful synthesis of β-Ch-Zn NPs with the desired size and morphology. The MTT assay demonstrated concentration-dependent cytotoxicity of β-Ch-Zn NPs, indicating safety for cellular studies. Pretreatment with β-Ch-Zn NPs significantly downregulated the mRNA expression of proinflammatory cytokines. The nanoparticles effectively downregulate proinflammatory cytokines and promote anti-inflammatory pathways, as evidenced by the significant reduction in interleukin (IL)-2, IL-6, hypoxia-inducible factor, and nuclear factor kappa B expression in a dose-dependent manner.

Conclusions: This study demonstrated that biosynthetically derived β-Ch-Zn NPs exhibit potent anti-inflammatory effects in McCoy cells. These findings underscore the therapeutic potential of β-Ch-Zn NPs for treating inflammatory conditions and support further investigation into their in vivo efficacy and safety.

Keywords: anti-inflammatory; cytokines; cytotoxicity; green synthesis; mccoy cells; β-chitosan nanoparticles.

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Conflict of interest statement

Human subjects: All authors have confirmed that this study did not involve human participants or tissue. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work.

Figures

Figure 1
Figure 1. UV-Vis absorption spectra of β-Ch-ZnO-NPs
UV-Vis: ultraviolet-visible spectroscopy; β-Ch-ZnO-NPs: β-chitosan-derived zinc oxide nanoparticles
Figure 2
Figure 2. FTIR spectra of β-Ch-ZnO-NPs
FTIR: Fourier-transform infrared spectroscopy; β-Ch-ZnO-NPs: β-chitosan-derived zinc oxide nanoparticles
Figure 3
Figure 3. XRD pattern of the as-prepared and annealed (800°C) β-Ch-ZnO-NPs
XRD: X-ray diffraction; β-Ch-ZnO-NPs: β-chitosan-derived zinc oxide nanoparticles
Figure 4
Figure 4. Cytotoxicity of β-Ch-ZnO-NPs on McCoy cells
The statistical significance was analyzed with the t-test and the Mann-Whitney U test (compared every concentration of the control group to each concentration of the treatment group). A p value of less than 0.05 was considered statistically significant. ***p < 0.001: extremely statistically significant; ns: nonsignificant β-Ch-ZnO-NPs: β-chitosan-derived zinc oxide nanoparticles Image credit: Meenakshi Sundaram K
Figure 5
Figure 5. β-Ch-ZnO-NPs decreased IL-2 expression on McCoy cells in a concentration-dependent manner
The statistical significance was analyzed with the t-test and the Mann-Whitney U test (comparing the control group to each treatment group). A p value of less than 0.05 was considered statistically significant. **p = 0.0079: statistically significant IL-2: interleukin 2; β-Ch-ZnO-NPs: β-chitosan-derived zinc oxide nanoparticles Image credit: Meenakshi Sundaram K
Figure 6
Figure 6. β-Ch-ZnO-NPs decreased IL-6 expression on McCoy cells in a concentration-dependent manner
The values were represented as mean ± standard deviation. The statistical significance was analyzed using the t-test and the Mann-Whitney U test. A p value of less than 0.05 was considered statistically significant. *p = 0.05: statistically significant; **p = 0.01: highly statistically significant IL-6: interleukin 6; β-Ch-ZnO-NPs: β-chitosan-derived zinc oxide nanoparticles Image credit: Meenakshi Sundaram K
Figure 7
Figure 7. β-Ch-ZnO-NPs decreased HIF expression on McCoy cells in a concentration-dependent manner
The values were represented as mean ± standard deviation. The statistical significance was analyzed using the t-test and the Mann-Whitney U test. A p value of less than 0.05 was considered statistically significant. **p = 0.01: highly statistically significant; ***p = 0.008: extremely statistically significant HIF: hypoxia-inducible factor; β-Ch-ZnO-NPs: β-chitosan-derived zinc oxide nanoparticles Image credit: Meenakshi Sundaram K
Figure 8
Figure 8. β-Ch-ZnO-NPs decreased NF-κB expression on McCoy cells in a concentration-dependent manner
The values were represented as mean ± standard deviation. The statistical significance was analyzed using the t-test and the Mann-Whitney U test. A p value of less than 0.05 was considered statistically significant. **p = 0.01: highly statistically significant NF-κB: nuclear factor kappa B; β-Ch-ZnO-NPs: β-chitosan-derived zinc oxide nanoparticles Image credit: Meenakshi Sundaram K
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