Green synthesis of Selenium nanoparticles using Capparis decidua and its anti-in lammatory activity

Janani K1, Preetha S*1, Jeevitha2, Rajeshkumar S3 1Department of Physiology, Saveetha Dental College and Hospitals, Saveetha institute of Medical & Technical Sciences, Saveetha University, Chennai 77, TamilNadu, India 2Department of Periodontics, Pharmacology, Saveetha Dental College and Hospitals, Saveetha institute of Medical & Technical sciences, Saveetha University, Chennai 77, Tamil Nadu, India 3Department of Pharmacology, Saveetha Dental College and Hospitals, Saveetha institute of Medical & Technical Sciences, Saveetha University, Chennai 77, Tamil Nadu, India


INTRODUCTION
In current years, Novel nanotechnologies have seized a lot of attention in research areas as they hold enormous applications in multidisciplinary ields (Cui, 2018). Nanotechnology is the speciality associated with Material science and biology, rather than a speci ic ield. It involves the formulation of particles at nanoscale known as Nanoparticles, where they have control over bulk macroscopic properties of the same material (Fulekar, 2010). Many studies have proved the pharmaceutical and nutraceutical values of nanoparticles, in which they interact at cellular as well as molecu-lar levels with a high degree of speci icity, sensitivity and signalling capability (El-Ghazaly, 2017). These nanoparticles have been utilised in several ields like genomics and proteomics, food industry, cosmetic industry etc.. due to their marvellous ligand binding potential and deep penetrating power (Agarwal et al., 2019). Selenium, being an indispensable dietary trace element, has been recently introduced in biomedicine as a 'Drug nanocarrier'. A lot of research has reported that Selenium possesses strong antioxidant, antibacterial and anti-cancer properties (Song, 2020). Selenium plays a crucial part in boosting immunity, protecting tissues against oxidative stress, Reproduction, growth, development and modulation (Murdock, 2008). Between Selenium's bioactivity and toxicity, there exists a thin margin only. Selenium shows immense biological activity, bioavailability and less toxicity if synthesised biologically rather than physicochemical methods (Wadhwani, 2017).
Capparis decidua is a deciduous bushy shrub, primarily seen in the arid regions of India, Asia, Africa and Saudi Arabia. Capparis decidua is named in versatile languages as Karil (Bengal), Senkam (Tamil), Mumudata (Telugu), Karir (Kannada), Karimulli (Malayalam), Kerdo (Gujarati) etc.. (Plants et al., 1958). It is commonly called 'Caperberry'. This unique arid plant species is resistant to Drought, salinity, soil erosion and tolerates frost to some extent also (Anjum, 2018). Following the arrival/ advent of humanity, This plant has been exploited in Folk medicine (Ayurveda, Unani) and Herbalism and possess enormous medicinal values such as Antibacterial, Anti-diabetic, Anti-fungal, Antirheumatic, Anti-tumour, Antidote Properties (Ayat et al., 2016). This plant has numerous nutraceutical values and is enriched with proteins, carbohydrates, vitamins, ibre, potassium, calcium and used as fodder for livestock (Ozcan, 2005;Romeo, 2007). Capparis decidua contains several alkaloids, Terpenoid, Glycosides and fatty acids (Zhang and Ma, 2018;Reynolds, 2020). The powdered coal of stem of Capparis decidua helps in the healing of the bone fracture, while the paste form of its root is applied for scorpion bite. Apart from the aspects mentioned above, Capparis decidua is an eco-friendly and costeffective biofuel (Nour and El-imam, 2013).
Nanoparticles can modify their physical, chemical and biological characteristics on account of their large surface to volume ratio. 'Green chemistry' plays a vital role in fabricating bioengineered nanoparticles, to attain peculiar composition and function (Darroudi et al., 2010;Sorescu, 2016). The green protocol also eliminates the chances of producing unwanted/ hazardous by-products rather than the conventional physical and chemical methodologies (Gurunathan, 2015;Lee et al., 2014) .
In lammation is part of the body's immune response to remove harmful stimuli and begin the healing process. Chronic in lammation can eventually cause several diseases and conditions, including cancer and rheumatoid arthritis (Byford, 1871). From this study, we are attempting to analyse the combined anti-in lammatory activity of bio-synthesised selenium nanoparticles prepared from Capparis decidua.

Collection and preparation of plant
Fresh fruits of Capparis decidua were obtained, identi ied and authenticated by Botanist and it is double washed with running water and then dried under shade. The dried fruits were thoroughly ground to a ine powder using a blender. The obtained powder of Capparis decidua is stored in an airtight container. One gram of Capparis decidua powder is diluted with 40 ml of distilled water and boiled for 20 mins. The extract is iltered using Whatman ilter paper and allowed to stand undisturbed for 20 mins. 20 ml of iltered extract is obtained and used for green synthesis.

Preparation of Selenium nanoparticle extract
0.01mg of sodium selenite is weighed and mixed with distilled water of 8 ml and mixed with the iltered extract Figure 1. The nanoparticles mixed with the plant extract are permitted to stand in a magnetic stirrer for 1 hour and kept in a shaker for intermixing of the particles to obtain green synthesis. UV spectrometers periodically monitored the reduction of sodium selenite to selenium nanoparticles. The colour change was visually noted and photographed.

Characterisation of Selenium nanoparticles
The synthesised selenium nanoparticles solution is primarily characterised using Ultraviolet (UV)-Visible spectroscopy Figure 2; 3 ml of the solution is taken in the coveted and scanned in double-beam UV-visible spectrophotometer from 300-650 nm wavelength. The results were recorded for the graphical analysis.

Preparation of Nanoparticle powder
Using Lark refrigerated centrifuge, The selenium nanoparticles solution is centrifuged at 8000 rpm for 10 min, and the pellet is collected and washed with distilled water twice. The inal puri ied pellet is collected and dried at 100-150 degree Celsius for 2/24 h, and inally, the nanoparticles powder is collected and stored in airtight Eppendorf tube.

Evaluation of anti-in lammatory activity by Albumin denaturation assay
2 mL of 1% Bovine serum albumin (BSA) was mixed with 400 µL of methanolic crude extract in different concentrations (500-100 µg/mL), and the pH of the reaction mixture was adjusted to 6.8 using 1N HCl. The reaction mixture was incubated at room temperature for 20 min and then heated to 55 • C for 20 min in a water bath. The mixture was cooled to room temperature, and the absorbance value was recorded at 660 nm. A BSA mixture with 30% methanol solution was used as a control. Diclofenac sodium in different concentrations was used as a standard. The experiment was performed in triplicate.
Percentage inhibition was calculated using the following formula:

% inhibition = Absorbance of control -Absorbance of the test x 100
The absorbance of the control

UV -Spectroscopy
The UV-visible analysis of sodium selenite nanoparticles was analysed in the absorbency range of 300-650 nm Figure 5. The peak was found to be maximum at 300 nm. reduction of aqueous metal ions with the Capparis decidua extract indicates the formation and synthesis of the selenium nanoparticles. Selenium nanoparticle synthesised from Capparis decidua was taken in different concentrations of 10 µL, 20 µL, 30 µL, 40 µL and 50 µL Figures 3  and 4. For the concentrations as mentioned above, the following percentage of the zone of inhibition was observed as 50%, 53%, 65%, 70% and 80%. Maximised zone of inhibition, i.e., 80% was noted in the concentration of 50 muL. This depicts that effective anti-in lammatory activity of bio fabricated selenium nanoparticles increased with higher concentrations. Diclofenac, a standard synthetic antiin lammatory drug, exhibits 95% anti-in lammatory activity. This indicates the effectiveness of the extract is close to the standard taken, and its ef icacy can be increased with increasing concentration.  drugs like NSAIDs which are used against in lammation are effective, but they have many side effects like gastrointestinal and renal damage (Pilotto, 2010). In a study done by El-Ghazaly et al., regarding the anti-in lammatory effect of selenium nanoparticles on the in lammation induced on irradiated rats, Nano-Se were administered orally in a dose of 2.55 mg/kg. It has been found that Nano-Se lessened the elevating in lammation in both irradiated and nonirradiated rats (El-Ghazaly, 2017). Melatonin-SeNPs treatment decreased pathological abnormalities of the liver, proin lammatory cytokines and splenocyte proliferation. The combination of silymarin and selenium nanoparticle at Low concentration is an excellent candidate possessing anti-in lammatory as well as antioxidant properties (Khurana, 2019) . To lessen the side effects and toxicity, biologically prepared herbs act as an excellent alternative. Medicinal plants have a wide range of phytochemicals like secondary metabolites which are potent and safe to use. This study has proved the antiin lammatory property of selenium nanoparticles synthesised using Capparis decidua, which as per previous reviews.

CONCLUSION
In this study, Using Capparis decidua, selenium nanoparticles have been synthesised. This bioengineered nanoparticle has proved to exhibit signi icant anti-in lammatory properties with higher concentrations. It is non-toxic, without any side effects as that of steroidal and non -steroidal antiin lammatory drugs. Future studies will be carried out to identify other properties present in selenium nanoparticles synthesised using Capparis decidua.