Cytotoxicity Evaluation of Green-synthesized MgO Nanoparticles with the Combination of Curcuma, Cardamom, and Pippali for Potential Cardioprotective Therapy in Diabetes-associated CVDs

Diabetes mellitus increases the risk of Cardiovascular Diseases (CVDs), highlighting the need for new treatments. Herbal remedies like turmeric (Curcuma longa), with its bioactive compound curcumin, are known for their anticoagulant effects. Cardamom (Elettaria cardamomum) helps lower diastolic blood pressure and inflammatory markers linked to CVDs. Pippali (Piper longum) acts as an anti-inflammatory, analgesic, and bioenhancer. In the current study, acknowledging their properties, these three herbs were combined (Curc+Card+Pip) and conjugated with green-synthesized MgO nanoparticles from curry leaves (Murraya koenigii). The aim was to evaluate the cytotoxicity of the herbal combination with and without MgONPs before employing it as a potential novel therapeutic approach for CVD-related research in diabetes.

This case-control study included two groups: a) normoglycemic individuals and b) those with Type 2 Diabetes Mellitus (T2DM). Hydroethanolic extracts were prepared and characterized. Cytotoxicity of Curc+Card+Pip and Curc+Card+Pip+MgONps was assessed using the MTT assay over 24 and 48 hours at concentrations of 100-500 µg/ml. Peripheral Blood Mononuclear Cells (PBMCs) from both groups were used as the primary cell lines. Statistical analysis was conducted using SPSS 26.0 and Excel 2021.

The MTT assay on PBMCs from Normoglycemic (NG) and T2DM groups with i) Curc+Card+Pip and ii) Curc+Card+Pip+MgONps showed minimal cytotoxicity at concentrations of 100-500 µg/ml. The CC50 order was NG 24 hours >; T2DM 24 hours >; NG 48 hours >; T2DM 48 hours. A significant difference in cell viability was observed between 24 hours (96.45% ± 0.012) and 48 hours (73.62% ± 0.48) at 100 µg/ml in NG (p >; 0.01), while T2DM showed 93.63% ± 0.012 and 68.41% ± 0.048 at 24 and 48 hours, respectively. For NG, Curc+Card+Pip+MgONps exhibited viabilities of 92.75% ± 0.026 and 62.02% ± 0.046 at 24 and 48 hours, respectively, while T2DM showed 90.82% ± 0.018 and 59.44% ± 0.044.

Cytotoxicity increased from 100-500 µg/ml, reducing viability. Cytotoxicity increased at 400-500 µg/ml at 48 hours (p >; 0.001) in both groups and treatments, suggesting that cell tolerance to MgO NPs was similar to that without NPs. The herbal mixture with MgONPs displayed a cytotoxicity pattern similar to that of the mixture without NPs in both groups.

The CC50 for Cur+Card+Pip without and with NPs was 452 and 410 µg/ml, respectively, within 24 hours, whereas in T2DM patients, the same treatments were found to have CC50 values of 300 and 330 µg/ml, respectively, with and without NPs. The values were the same in the case of T2DM (48 hours), whether it was with or without NPs (CC50 = 240 µg/ml), allowing for future exploration of its anti-inflammatory and antioxidant effects on PBMCs, which could suggest its potential as an alternative therapy for managing CVD in diabetic patients.