INTRODUCTION:

In the past few years, tremendous interest and substantial research effort were directed to word biomedical evaluation of metallic nanoparticles which are derived from silver and gold, because of their specific and genuine chemical, biological and physical properties [1-3]. In particular, attention was directed towards the biomedicine related assessment of silver nanoparticles (AgNPs), which drew worldwide attention as unconventional anti-microbial agent [4-6]. AgNPs are a class of zero-dimensional materials with distinct morphologies having a size ranging from 1 nm to 100 nm [7].

Plants produce hundreds and thousands different chemical compounds with different biological activities and have been used in the treatment of various human diseases. Herbal medicine has been practicing in rural areas since time immemorial. Implication of the knowledge of those indigenous people into practice helped the researchers to develop the present formulation into a better approach with more therapeutic effect. Medicines that are used today are not definitely the same that were used in ancient times. India is a storehouse of medicinal plants and most of which have been traditionally used in Ayurveda, Unani, Siddha systems of medicine. The medicinal value of plants lies in the phytochemical constituents that produce a definite physiological effect on the human body [8]. Curcuma longa (Linn.) commonly called as “turmeric”, belongs to Zingiberaceae family and it is widely used as an ingredient spice [9,10]. Turmeric has been valued worldwide asa important food because of its health promoting factors [11]. Curcuma species has a dark yellow color, and is a rich source of phenolic compounds, like curcuminoids [12]. Curcuminoids contain three different diarylheptanoids Curcumin (diferuloylmethane), Demethoxycurcumin (p hydroxycinnamoyl methane) and Bisdemethoxycurcumin (di-p-hydroxycinnamoyl methane). Curcumin, which is the major constituent, is reported to be a natural antioxidant with anti-inflammatory and anti-microbial activities [13,14].

MATERIALS AND METHOD:

Turmeric oil was obtained from Synthite Industries Limited, Kerala as a gift sample which was prepared by steam distillation of dried rhizomes.

Preparation of Silver Nanoparticle using Turmeric oil:

1 ml of the turmeric oil was mixed with 10-20 mL of distilled water and kept in boiling water both for 30-60 minutes to dissolve. 90 mL of 1 mM silver nitrate distilled water was added and kept in magnetic stirrer for nanoparticles synthesis.

UV Spectrometric analysis of synthesized nanoparticles:

The color change in the reaction mixture (Silver nitrate solution + turmeric oil) was recorded by visual observation. The synthesized nanoparticles solution was preliminarily characterized by UV-visible spectroscopy. The bio reduced silver nanoparticle solution was filtered through Whatman No.1 filter paper and the absorbance was measured using UV-Visible spectrophotometer. 3ml of the solution was taken in cuvette and scanned in double beam UV-visible spectrometer from 300 nm to 700nm wavelength. The results were recorded for the graphical analysis.

Anti-inflammatory activity of Turmeric oil mediated silver nanoparticle:

Albumin denaturation Inhibitory Assay:

BSA (Bovine Serum albumin) was used as a reagent for the assay. 2mL of 1% Bovine albumin fraction was mixed with 400 micro Litre of the turmeric oil solution in different concentrations (500-100 microgram /mL) and the pH of the reaction mixture was adjusted to 6.8 using 1 N HCl. The reaction mixture was incubated at room temperature for 20 minutes and then heated at 55⁰C for 20 minutes in a water bath. The mixture was cooled to room temperature and the absorbance value was recorded at 660 nm. An equal amount of turmeric oil solution was replaced with DMSO for control. Diclofenac sodium in different concentrations was used as standard. The experiment was performed in triplicate.

% Inhibition was calculated using the following formulae:

% Inhibition= control O.D – sample O.D/ control O.D* 100

RESULT AND DISCUSSION:

Synthesis of silver nanoparticles:

The synthesis of silver nanoparticles through turmeric oil were carried. The aqueous silver ions when exposed to turmeric oil were reduced in solution, there by leading to the formation of silver hydrosol. Change in color varies from plant to plant and is time dependent. The phytochemical present in extract is responsible for the reduction of silver ions. It is well known that silver nanoparticles exhibit yellowish – brown color in aqueous solution due to excitation of yellowish – brown color in the reaction indicate the formation of silver nanoparticles. The color change can be visually observed (Figure 1). Formation of brown colour after addition of AgNO3 indicate synthesis of AgNPs in the process of reduction of Ag+ to Ag nanoparticles and control (AgNO3)

Figure 1: Image of turmeric oil mediated silver nanoparticle

UV-vis spectroscopy:

It is generally recognized that UV–Vis spectroscopy could be used to examine surface Plasmon resonance of the silver nanoparticles in aqueous suspension. Fig. 2 shows the UV-Vis spectra recorded from the reaction medium after 12 hours. The UV–Vis spectra of the reaction mixture of silver nitrate solution with turmeric oil at the peaks observed at 350 nm indicate the presence of silver nanoparticles which is synthesized by turmeric oil.

Fig 2. UV-Vis absorption spectrum of silver nanoparticles synthesized by treating 1 mM aqueous AgNO3 solution with turmeric oil.

Anti-inflammatory activity:

Bovine serum albumin (BSA) makes up around 60% of all the proteins in animal serum. It is commonly used in cell culture, particularly when protein supplementation is necessary and the other components of the serum are unwanted. BSA undergoes denaturation upon heating and starts expressing antigens associated with Type III hypersensitive reaction which are related to diseases such as rheumatoid arthritis, glomerulonephritis, serum sickness and systemic lupus erythematosus [15]. The inflammatory response involves a complex array of enzyme activation, mediator release, fluid extravasations, cell migration, tissue breakdown and repair, which are aimed at host, defense and usually activated in most disease condition [16]. Chronic inflammatory diseases like rheumatoid arthritis are still one of the main health issue of the world's population. At present, although synthetic drugs are dominating the market, element of toxicity that these drugs entail, cannot be ruled out. Their prolonged use may cause severe adverse effects on chronic administration [17]. Currently much interest have been paid in the search of nanoparticle with anti-inflammatory activity which may lead to the discovery of new therapeutic agent that is not only used to suppress the inflammation but also used in diverse disease conditions where the inflammation response amplifies the disease process. There are certain problems in using animals in experimental pharmacological research, such as ethical issues and the lack of rationale for their use when other suitable methods are available or could be investigated. Hence, in the present study the protein denaturation bioassay was selected for in vitro assessment of anti-inflammatory property of turmeric oil and AgNPs. Denaturation of tissue proteins is one of the causes of inflammatory and arthritis. Formation of auto antigens in certain diseases may be due to denaturation of proteins. [18,19]. Agents that can prevent protein denaturation, would be useful for anti-inflammatory drug development.

The increments in absorbance of test samples with respect to control indicated stabilization of protein i.e. inhibition of heat-induced protein denaturation by turmeric oil, AgNPs and reference drug diclofenac sodium. The present findings exhibited a concentration dependent inhibition of protein denaturation by the turmeric oil mediated AgNPs

The greatest effect of AgNPs (150 μg/mL) was found to be more than standard diclofenac sodium. From the present study it can be concluded that turmeric oil mediated AgNPs have good anti-inflammatory effect against the denaturation of protein.

Fig 3: Albumin denaturation inhibitory effect of Turmeric oil mediated silver nanoparticles.

Protein denaturation has been well correlated with the occurrence of the inflammatory response and leads to various inflammatory diseases such as arthritis [20]. Tissue injury can be due to denaturation of the protein constituents of cells or of intercellular substance [21]. Hence, the ability of a substance to inhibit the denaturation of protein signifies apparent potential for anti-inflammatory activity. Plant extracts have capacity to inhibit protein denaturation. capacity of different plant like inflorescence axes, endosperms, leaves, and pericarps of B. Racemosa.to inhibit protein denaturation of albumin which was ranging from 43.33 ± 0.002% to 70.58±0.004% inhibition indicating its anti-inflammatory properties [22].

CONCLUSION:

Silver nanoparticles are intensively explore nano structures for enhanced bio medical application. Silver nanoparticles based nano systems and nano materials are sweat able alternations for drug delivery. A large number of studies have shown that curcumin has wide therapeutic actions such as anti inflammatory, and anti-microbial activities due to inhabitation of AA metabolism, COX, LOX, cytokines and NF-kB.

It may be concluded from this study that turmeric oil mediated silver nanoparticles can be used as potential source of anti inflammatory agent.

ACKNOWLEDGEMENT:

The authors thank Synthite Industries Limited, Kerala for providing turmeric oil as a gift sample

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Received on 27.02.2019 Modified on 23.03.2019

Research J. Pharm. and Tech. 2019; 12(7):3507-3510.

DOI: 10.5958/0974-360X.2019.00596.1