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 Table of Contents  
ORIGINAL ARTICLE
Year : 2022  |  Volume : 6  |  Issue : 3  |  Page : 337-340

Anticancer activity of tin oxide and cerium-doped tin oxide nanoparticles synthesized from Ipomoea carnea flower extract


1 Department of Chemistry, A. V. V. M. Sri Pushpam College, Affiliated to Bharathidasan University, Poondi, Thanjavur, India
2 Department of Physics, A. V. V. M. Sri Pushpam College, Affiliated to Bharathidasan University, Poondi, Thanjavur, India
3 Department of Chemistry, Shrimati Indira Gandhi College, Affiliated to Bharathidasan University, Tiruchirappalli, India
4 P. G. and Research Department of Physics, Sudharsan College of Arts and Science, Affiliated to Bharathidasan University, Pudukkottai, Tamil Nadu, India

Date of Submission27-Apr-2022
Date of Decision18-May-2022
Date of Acceptance20-Jun-2022
Date of Web Publication17-Sep-2022

Correspondence Address:
Gurusamy Muruganandam
Department of Chemistry, A. V. V. M. Sri Pushpam College, Poondi, Thanjavur - 613 503, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/bbrj.bbrj_100_22

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  Abstract 


Background: The aim of the study is to investigate the anticancer potential of tin oxide (SnO2) and different concentrations (2%, 4%, 6%, and 8%) of cerium-doped tin oxide nanoparticles (Ce-SnO2 NPs) using Ipomoea carnea flower extract. The synthesized SnO2 and different concentrations (2%, 4%, 6%, and 8%) of Ce-SnO2 NPs was tested using a colorimetric-based 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay against MCF-7 human breast cancer cell line cells. Methods: Collection and preparation of plant extract is preliminarily carried out followed by the synthesis of Undoped and Cerium doped Tin oxide nanoparticles is achieved by standard protocol along with that its anticancer activity also studied in this research. Results: The anticancer activity increased in direct proportion to the cerium-dopant concentration. Experimental results demonstrated that 8% Ce-SnO2 NPs exhibited a potential anticancer effect compared with SnO2 and other concentrations of Ce-SnO2 NPs. Conclusion: According to the current findings, large-scale manufacturing of Ce-SnO2 NPs might be recommended to have effective anticancer agents against breast cancer cell lines.

Keywords: Anticancer activity, cerium-doped tin oxide nanoparticles, Ipomoea carnea flower


How to cite this article:
Prasanth M, Muruganandam G, Ravichandran K, Dayana Jeyaleela G, Shanthaseelan K, Priyadharshini BP. Anticancer activity of tin oxide and cerium-doped tin oxide nanoparticles synthesized from Ipomoea carnea flower extract. Biomed Biotechnol Res J 2022;6:337-40

How to cite this URL:
Prasanth M, Muruganandam G, Ravichandran K, Dayana Jeyaleela G, Shanthaseelan K, Priyadharshini BP. Anticancer activity of tin oxide and cerium-doped tin oxide nanoparticles synthesized from Ipomoea carnea flower extract. Biomed Biotechnol Res J [serial online] 2022 [cited 2022 Dec 8];6:337-40. Available from: https://www.bmbtrj.org/text.asp?2022/6/3/337/356137




  Introduction Top


Cancer is one of the primary causes of mortality, with an estimated 10.0 million deaths expected in 2020.[1] Furthermore, the World Health Organization (WHO) predicts that by the end of 2040, it will have increased up to thrice.[2] Due to their lifestyle modifications, low- and middle-income nations account for over 70% of these fatalities.[3] Cancer arises when genes that govern the normal cell cycle and cell division are mutated, resulting in uncontrolled cell division and proliferation. Conventional chemotherapeutic medicines, on the other hand, have poor absorption, quick renal clearance, uneven administration, and significant adverse effects.[4] Breast cancer has risen to the top of the list of causes of mortality among women. According to the World Health Organization, these illnesses are estimated to cause 13.1 million deaths by 2030.[5]

Nanotechnology has advanced rapidly in recent years, and it now has a wide range of medical uses. Biomediated nanoparticles (NPs) have grabbed the curiosity of scientists all around the world because they are cost-effective, safe, and biocompatible. These nanoscale materials provide a viable cancer therapy due to their low-risk profile and fewer side effects.[6] As a result, one of the most ardent aims is the creation of robust and effective anticancer medications. Recent advances in nanotechnology have widened the field of study of metal-based silver (Ag) NPs with medicinal uses.

Ag, Au, Ce, Sn, Zn, and Cu are among the most frequent metallic NPs used to treat or diagnose cancer. The formation of reactive oxygen species in cellular compartments is thought to be responsible for these NPs' anticancer properties, which leads to the activation of autophagic, apoptotic, and necrotic death pathways.[7] This study looked at the anticancer properties of SnO2 NPs made from Ipomoea carnea flowers and Ce-doped SnO2 NPs (Ce-SnO2 NPs) in a breast cancer cell line (MCF-7 cell line).


  Methods Top


Collection of materials

The flower of I. carnea was collected in January 2019 from Poondi, Thanjavur District, Tamil Nadu, India. Stannous chloride, NaOH, distilled water, and ethanol were purchased from Merck Chemical Co Private Limited.

Preparation of alcoholic extract

Fresh I. carnea leaves were cut and washed with distilled water. The extraction procedure was as follows: 10 g of leaf was added to 100 mL of ethanol and soaked for 24 h. The obtained extraction was filtered using Whatman No. 1 filter paper, and the filtrate was collected and stored at room temperature for further usage.

Synthesis of cerium-doped tin oxide nanoparticles

For the preparation of Ce-SnO2 NPs, 100 mL of 0.1 M Tin (II) chloride dihydrate (SnCl2.2H2O) solution and different atomic percentage of (NH4) 2Ce (NO3) 6 were added with 100 mL of I carnea flower extract; finally, we get the ash color solution. This solution was stirred constantly at room temperature for 4 h. The colloidal particles dried in a hot air oven at 80°C for 1 h. Further, the precipitate was calcinated at 350°C for 3 h. The synthesized NPs are further used for anticancer activity.


  Determination of Anticancer Activity Top


Cell lines and maintenance

MCF-7 human breast cancer cell line cells used in this study were routinely maintained in Dulbecco's modified Eagle medium (DMEM) containing 10% fetal bovine serum (FBS) at 37°C in a humidified atmosphere with 5% CO2. Penicillin (100 μg/mL) and gentamicin (10 μg/mL) were used as antibiotics. Cell culture work was performed under aseptic conditions inside a laminar airflow hood, and passages were performed once every 3–4 days according to cell densities.

Anticancer activity (3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide assay)

SnO2 with different Ce-SnO2 cytotoxic activity was tested using a colorimetric-based 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay against MCF-7 human breast cancer cell line cells.[8] HeLa cells (1 × 104 cells/mL) were cultured in a 96-well plate containing DMEM with 10% (v/v) FBS and treated at various concentrations of SnO2 with different CE-SnO2 (25, 50, 100, 150, and 200) prepared in 0.4% DMSO, and treatment was separately given for 24 h. After treatment, the MTT assay was performed and the optical density was measured by a microplate reader (VERSAmax, USA) at 595 nm, with 655 nm as reference. All tests were carried out in triplicate, and the percentage of cell viability for treated and untreated cells was determined. In addition, dual labeling with acridine orange/propidium iodide (AO/PI) (Sigma Aldrich, India) revealed a shift in nucleus morphology in HeLa cells. In a Zeiss inverted fluorescent microscope equipped with Indian Institute of Science, the cells were photographed at ×20 using the 485 nm excitation and 535 nm emission filter sets.

Determination of nuclear morphology (acridine orange and propidium iodide)

Fluorescence microscopy was used to analyze morphological changes caused by apoptosis induction using AO/PI dual labeling. In a 12-well plate, 1 × 105 cells were seeded on sterile 18 mm circular coverslips, cultured for 20 h at 37°C in a humidified 5% CO2 incubator, and then treated with the fungal extracts for 24 h. The cells were then rinsed twice with 50 mM PBS before being treated for 2 min with 100 L of a 1:1 combination of Acridine orange/ethidium bromide (AO/EB) (100 g/mL each) solution. In a Zeiss inverted fluorescent microscope equipped with Axiovision® software, cells were photographed at ×20 using the 485 nm excitation and 535 nm emission filter sets. Apoptotic cells stained with both AO and PI had a red-orange–colored appearance with chromatin condensation, whereas viable cells with an unbroken cellular membrane were stained with AO alone and appeared brilliant green. The information offered is based on a minimum of three separate experiments.


  Results and Discussion Top


Breast cancer is the most frequent type of cancer in women all over the world. It is a form of cancer in which breast cells divide and expand out of control. Breast cancer has risen in occurrence during the last 30 years. Between 50% and 75% of breast cancers start in the ducts, 10%–15% start in the lobules, and a few start elsewhere in the breast.[9] Fortunately, the death rate from breast cancer has dropped in recent years as a result of a greater focus on early identification and more effective therapies (http://www.imaginis.com/general-information-about-breast-cancer/what-is-breast-cancer-2). The Cancer Research centres has recognised some regularly used herbs as cancer-preventive agents.[10] As a result, the current research intends to analyze the undoped NP in a low-cost, simple manner.

MCF-7 is a breast cancer cell line derived from a 69-year-old Caucasian woman. It was discovered in 1970. MCF-7 stands for Michigan Cancer Foundation-7, the facility in Detroit where Herbert Soule et al. first created the cell line in 1973.[8] The Barbara Ann Karmanos Cancer Institute has replaced the Michigan Cancer Foundation.[11]

MCF-7 is a human breast cancer cell line that was initially identified in 1970 from a 69-year-old woman's malignant adenocarcinoma breast tissue. Because MCF-7 cells have kept some perfect properties specific to the mammary epithelium, they are suitable for in vitro breast cancer investigations. MCF-7 cells, for example, have the capacity to metabolize estrogen through estrogen receptors. Cytokeratin is likewise toxic to MCF-7 cells. The cell line may form domes, and the epithelial-like cells develop in monolayers when cultured in vitro. Tumor necrosis factor alpha can potentially be used to stop the growth. The human breast cancer cell line MCF-7 is beneficial for in vitro breast cancer investigations because it has kept certain excellent properties specific to the mammary epithelium. MCF-7 cells have the capacity to metabolize estrogen through estrogen receptors.[12]

The cytotoxicity of SnO2 and different concentrations (2%, 4%, 6%, and 8%) of Ce-SnO2 NPs against breast cancer cells is evaluated and summarized in [Table 1] and [Figure 1]. Human breast adenocarcinoma (MCF-7) cell lines were used to evaluate the four different concentrations (25, 50, 100, and 200 μg/mL) of SnO2 and different concentrations (2%, 4%, 6%, and 8%) of Ce-SnO2NPs. It was observed that the SnO2 and different concentrations (2%, 4%, 6%, and 8%) of Ce-SnO2 NPs showed promising anticancer activity toward the cell lines. A significant increase in cell death was recorded with increasing concentrations of SnO2 and different concentrations (2%, 4%, 6%, and 8%) of Ce-SnO2 NPs by MTT assay. The maximum of cell death in SnO2 NPs was 50.72%, 2% of Ce-SnO2 NPs was 69.12%, 4% of Ce-SnO2 NPs was 81.35%, 6% of Ce-SnO2 NPs was 90.82%, and 8% of Ce-SnO2 NPs was 97.1%. Among the various concentrations, 8% of Ce-SnO2 has potential anticancer property observed as compared with other concentrations and SnO2 NPs.
Table 1: Percentage of cell death in different concentrations (25, 50, 100, 150, and 200 μg/mL) tin oxide and cerium-doped tin oxide nanoparticles on cytotoxicity of Michigan Cancer Foundation-7 (breast cancer) cell line as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay

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Figure 1: Percentage of cell death in different concentrations (25, 50, 100, 150, and 200 μg/mL) of SnO2 and cerium-doped SnO2 nanoparticles on cytotoxicity of MCF-7 (breast cancer) cell line as determined by MTT assay. SnO2: Tin oxide, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, CE: Cerium

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Morphological examination

AO/PI staining was performed to determine if exposure to SnO2 and Ce-SnO2 NPs causes cell death by apoptosis in the MCF-7 cell line. In the AO/PI staining, the live cells with intact membranes have a uniform green color in their nuclei. Early apoptotic cells have chromatin condensation with bright green-colored nuclei. Late apoptotic cells have bright red-orange areas of condensed chromatin in the nucleus that distinguish them from necrotic cells, which have a uniform orange color.[13]

[Figure 2] shows the MCF-7 cells stained with AO/PI together. It was found that the untreated control cells were mostly green with an intact nucleus. As shown in [Figure 1], the SnO2 and Ce-SnO2NPs were potent in inducing apoptosis in MCF-7 cells in a time-dependent manner. AO/PI analysis indicated that SnO2 and Ce-SnO2NPs were cytotoxic toward MCF-7 cells through apoptosis when treated with SnO2 and Ce-SnO2 NPs concentration.
Figure 2: Effect of SnO2 and cerium-doped SnO2 nanoparticles (200μg/ml) on cytotoxicity of MCF-7 (breast cancer) cell line as determined by MTT assay and imaged fluorescence microscope (×630). (Live cell: Green color-Cancer; Cancer Dead cells: Orange with red color). MCF: Michigan Cancer Foundation-7, MTT: 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide assay, SnO2: Tin oxide, CE: Cerium

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In the AO/PI analysis, cells treated with SnO2 and Ce-SnO2NPs displayed orange-colored nuclei, which further confirmed the induction of late apoptosis in MCF-7 cells by SnO2 and Ce-SnO2 NPs. This finding was characterized by membrane blebbing, nuclear shrinkage, and significant DNA fragmentation. Necrotic cells were stained red by PI, which penetrated the nuclear matter where the integrity of the cell membrane was compromised. Ce-SnO2 NPs show a higher proportion of apoptotic, necrotic, and viable cells in a population of MCF-7 cells treated with 8% concentration as compared to other concentrations. The apoptotic and necrotic cell populations increased with the increased concentration of Ce-SnO2 NPs when compared with the control cells. This situation may be due to the progression of the apoptotic cells into secondary necrotic cells after a certain period. The term secondary necrosis refers to a process in which late-stage apoptotic cells that failed to be engulfed by phagocytes or neighboring cells undergo necrosis. Secondary necrosis, thus, is a postapoptotic event. The present study agrees with an earlier report by Balaji and Gothandam, [14,15] who reported that cytotoxic effect on cancerous cell lines by biologically synthesized metal NPs. A similar report also observed in the previous studies. Eric et al., studied the cytotoxic effects of novel nanocomposite Ag-doped PEGylated WO3 sheets on MCF-7 human breast cancer cells.


  Conclusion Top


Finally, our findings revealed that SnO2 and Ce-SnO2 NPs exhibit potent cytotoxic action against the MCF-7 human breast cancer cell line. As a result, we believe that they might be potential medicines in cancer treatment, and more research into their specific molecular pathways is required. For the first time, there is a novel design for a probable association between increased band gap energy and improved cytotoxicity as an anticancer therapy.

Limitation of study

Green synthesis of NPs metals has enormous promise; yet, it is hampered by material selection, synthesis circumstances, product quality control, and application. These factors provide obstacles to the adoption of green-synthesized NPs metals for industrial production and large-scale use.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

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Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.  Back to cited text no. 8
    
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    Figures

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    Tables

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