Synthesis and Characterization of Citrus limonum Copper Nanoparticles and its in vitro Evaluation of Cytotoxicity against JURKAT Cells

Biosynthesized copper nanoparticles (CuNPs) are eco-friendly, cost-effective, and biocompatible agents reported for extensive biomedical and bioengineering applications. Different chemical synthesis approaches have been established recently, with challenges of higher toxicity and high cost involved in the synthesis process. Green synthesized nanoparticles emerged and was extensively reported to address the challenges faced by traditional chemical synthesis processes. However, the high toxicity remains a significant challenge for translating the green synthesized nanoparticles into clinically valuable products.

To synthesize, characterize, and evaluate the citrus extract-based CuNps cytotoxicity against JURKAT cell lines.

An aqueous extract of the citrus fruit was used as a reducing agent, and the CuNps were synthesized. Fourier Transform Infra-Red (FTIR), scanning electron microscopy (SEM), dynamic light scattering (DLS), Ultraviolet-visible (UV-vis) spectrophotometry, and X-ray diffraction (XRD) were used to confirm the synthesis of CuNp and its structure. Furthermore, the effect of CuNPs on cell viability and toxicity was evaluated by mitochondrial toxicity tests (MTT) and LDH assays against the JURKAT cell lines.

The synthesized nanoparticle’s size ranged from 40 to 70 nm, as confirmed through nanoparticle tracking analysis (NTA) and SEM. The synthesized nanoparticles were confirmed to be anti-proliferative with a high percent of cytotoxicity as found from MTT and LDH leakage assays. The size and shape of the synthesized CuNPs as studied by SEM were found to be 30–70 nm and more or less spherical. MTT reported 64.87% inhibition at 320 µg/mL with an IC₅₀ value of 80.78 µg/mL (p < 0.05). Cytotoxicity as measured by the LDH assay was found to be 53.12 ± 0.89% at 320 µg/mL with an IC₅₀ of 23.12 ± 0.39 when compared to the positive control (11.21 ± 0.15 µg/mL). Green-synthesized CuNPs exhibited potential anti-cancerous activity in JURKAT cell lines, as evidenced by the LDH and MTT assays.