Smaller Copper Oxide Nanoparticles have More Biological Effects Versus Breast Cancer and Nosocomial Infections Bacteria

Abstract

Background and objectives: Despite promising successes in developing new drugs and pharmaceutical biotechnology, infectious diseases and cancer are still the principal causes of mortality and morbidity globally. Therefore, finding effective ways to deal with these pathogens and cancers is critical. Metal nanoparticles are one of the new strategies to combat bacteria and cancers.

Methods: We examined the antimicrobial activity of 30 and 60 nm copper oxide nanoparticles (CuO-NPs) against Acinetobacter baumannii and Staphylococcus epidermidis bacteria responsible for nosocomial infections in standard and clinical strains and anti-cancer activity against 4T1 cell line as malignancy breast cancer cells. Synthesis of CuO-NPs was performed by a one-step reduction method and confirmed by DLS and TEM microscopy at 30 and 60 nm sizes. The antibacterial and anti-cancer activities of the nanoparticles were then investigated against the aforementioned bacteria and breast cancer.

Results: Using disk, well, MIC, MBC methods, and viability/bacterial growth assay, 30 nm CuO NPs were found to have more antibacterial activity on standard and clinical strains than 60 nm CuO NPs. On the other hand, using MTT, apoptosis, and gene expression method, 30 nm nanoparticles were found to have more anti-cancer potential than 60 nm CuO NPs.

Conclusions: Our findings implicate CuO-NPs to possess antimicrobial and anti-cancer effects and more significant potential in smaller sizes, suggesting their pharmaceutical and biomedical capacity.<br />.

Keywords: Acinetobacter baumannii; Copper oxide nanoparticle; Staphylococcus epidermidis; breast cancer; nosocomial infections.

Figure 1

Schematic form of Project Design. A, In the first step, copper oxide nanoparticles are synthesized using the one-step reduction method, then size and morphology characterization of CuO-NPs was performed by DLS and TEM methods. B, In the second step, bacteria resistant to common antibiotics were isolated from the infectious section of the hospital. On the other hand, Standard bacterial strains were prepared. Afterthought antibacterial properties of CuO-NPs on standard and clinical strains of the bacteria of A. baumannii and S. epidermidis were investigated by disk, well diffusion testing, MIC, MBC, and bacterial survival/growth kinetics methods. C, In the third step, the anti-cancer activity of CuO NPs on the 4T1 breast cancer cell line was evaluated by MTT, Apoptosis, and Real-time PCR for gene expression of MMP-2 and VEGF as metastatic and angiogenesis of cancer cell

Figure 2

Hydrodynamic Diameter (Determining the Size) of CuO-NPs via DLS and TEM Results of Synthesized CuO-NPs. A, 30 nm of CuO-NPs. B, 60 nm of CuO-NPs. C, Morphology of CuO-NPs with 30 nm. D: Morphology of CuO-NPs with 60 nm

Figure 3

Antibiotic Resistance Clinical Strain of S. epidermidis and A. baumannii Bacteria by Antibiogram Test. A, Antibiotic disks were used for S. Epidermidis bacteria. B, Antibiotic disks were used for A. Baumannii bacteria

Figure 4

S. epidermidis Standard and Clinical Strains Survival/Growth Curves in the Presence of Different Concentrations (0.1, 0.01, 0.001 mg) and Different Sizes (30 or 60 nm) of CuO NPs (mg/mL). A, Standard strain treated with 30 nm size of CuO NP. B, Clinical strain treated with 30 nm size of CuO NP. C, Standard strain treated with 60 nm size of CuO NP. D, clinical strain treated with 60 nm size of CuO NP. Data shown are means ± SD (n=3) (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 vs similar concentrations in different conditions and each index is represented with assigned color in legend

Figure 5

A. baumannii Standard and Clinical Strains Survival/Growth Curves in the Presence of Different Concentrations (0.1, 0.01, 0.001 mg) and Different Sizes (30 or 60 nm) of CuO NPs (mg/mL). A, Standard strain treated with 30 nm size of CuO NP. B, Clinical strain treated with 30 nm size of CuO NP. C, Standard strain treated with 60 nm size of CuO NP. D, clinical strain treated with 60 nm size of CuO NP. Data shown are means ± SD (n=3) (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 vs similar concentrations in different conditions and each index is represented with assigned color in legend

Figure 6

A, apoptosis morphological change of effect different concentration and the different size of CuO NPs on the 4T1 breast cancer with AO/ PI staining by fluorescence microscope. B, Cancer cell viability in exposing different concentrations of CuO NPs in sizes 30 and 60 nm by MTT assay. C, Determination of apoptosis of breast cancer cells treated with different concentrations of CuO NPs in 30 and 60 nm sizes. Data shown are means ± SD (n=3) (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 vs similar concentrations in different size).

Figure 7

The mRNA Expression Levels of MMP-2 (A) and VEGF (B) in Breast Cancer Cell Treatment with Different Concentrations of CuO NPs in Sizes 30 and 60 nm. HPRT was used for an endogenous control. Data shown are means ± SD (n=3) (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 vs similar concentrations in different size).