Antioxidant Activity of Lawsonia alba Mediated Silver Nano-Particles

Abirami Arthanari^1*, Kandhal Yazhini² and S Rajeshkumar^3
*Correspondence: Abirami Arthanari, Department of Forensic Odontology, Saveetha Institute of Medical and Technical Sciences (SIMATS), Saveetha University, Chennai, Tamil Nadu, India, Email:

Keywords

Antioxidant activity • Lawsonia alba • Silver nano particle • DPPH assay • Innovative technique herbal

Introduction

Nanotechnology is one of the foremost active research areas within modern material science. Nanotechnology is one of the fastest developing sciences over the past few years [1]. This is an emerging field of modern research dealing with the synthesis and designing of particle structures ranging from approximately 1-100 nm [2]. Nanotechnology being a nascent innovation has a lot of potential for medical and dental applications. It continues to influence several new developments and future advancements of orthodontics and dentistry [3]. Nanoparticles are known to decrease toxicity, increase bioactivity and also improve cell targeting. Nanoparticles    of metal and metal oxides such as silver, zinc oxide, zirconium oxide, copper oxide, gold, selenium, hydroxyapatite, titanium oxide, and copper sulfide, have been used in many medicinal applications, in particular for cancer detection, screening  purposes,  drug  delivery  systems,  antisense  and gene therapy applications, and tissue engineering [4]. Silver nanoparticles (AgNPs) have a wide range of  medicinal  and  diagnostic  uses.  Silver  is one of the most common metal nanoparticles,  owing  to  its  antimicrobial and pharmaceutical properties [5–7]. New and innovative strategies are of potential interest for the synthesis of silver nanoparticles (AgNPs), which    are used in a huge range of consumer products [8]. The utilization of nanoparticles as a medicine in the treatment of diabetes using the mediated cerium oxide nanoparticles (HMCeO₂  NPs),  herbal  mediated  silver nanoparticles (HMAg NPs) and Lawsonia inermis extract have been attempted [9]. Lawsonia alba is cultivated for the medicinal and cosmetic value of its leaves. The shrub’s stem bark, roots, flowers, and seeds have also been used in traditional medicine [10]. Henna is the common name   for this herb, which is widely distributed in tropical and subtropical regions. This plant is a worldwide known cosmetic agent used to stain hair, skin, and nails [11]. It is a small tree or a multi-branched shrub belonging to the family Lythraceae. It is typically 2-6 m in height [12]. Since antiquity, the Lawsonia alba plant's leaf paste has been used for dying blood, skin, and  nails. Besides cosmaceutical usages, the plant also harbors a well-documented folklore history for treating  convulsion,  jaundice,  and  malignant  ulcers  [13]. Lawsonia alba was evaluated to have antimicrobial, antioxidant, anti- complementary activity, anti-sticking activity, and catalytic properties [14]. Studies on this important medicinal plant have resulted in the isolation and structure elucidation of more than 80 compounds up to 2011 and evaluation of biological activities of some of them. Lawsonia inermis (Lythraceae) commonly known as Henna is a well-known plant used in Indian [15]. Traditional Indian medicine has used different parts of this herb. The plant has a wide range of phytochemicals [16]. Antioxidants are compounds that may shield the cells from free radicals, which may cause heart disease, cancer, and other  illnesses  [17,18].  When  your  body  breaks  down  food or when you're exposed to cigarette smoke or radiation, free radicals are created [19]. Our team has extensive knowledge and research experience that has translate into high quality publications [20-39]. The research is novel and hence not much reference material available. Very few studies have been done on the use of Lawsonia alba as a source of silver nanoparticles.

The synthesis of metal nanoparticles from herbal sources is also tedious   and time-consuming. Previous literature majorly focused on Lawsonia  inermis mediated silver nanoparticles and their antimicrobial activity, the present study aims to evaluate the antioxidant activity of Lawsonia alba mediated silver nanoparticles [40].

Materials and Methods

Extract preparation

1 gm of Lawsonia Alba was added in 100 ml of distilled water and boiled for 10-15 minutes at 70 degrees celsius. After boiling, the plant extract was filtered by Whatman No 1 filter paper. 90 ml of 1 millimolar silver nitrate is prepared in 250 ml of a  conical  flask;  40  ml  of  filtered  plant  extract was mixed to it and kept in a magnetic stirrer for nanoparticle synthesis (Figure 1). The synthesized nanoparticle was preliminarily analyzed using  UV visible spectroscopy. Before the final step, the nanoparticle solution was centrifuged at 8000 rpm to prepare nanoparticle pellet powder; it was dried in a hot air oven at 80 degrees Celsius. The dried powder was sent     for characterization. Finally, the leftover solution was taken to calculate antioxidant activity.

Antioxidant activity

DPPH method: A DPPH assay was used to test the antioxidant activity of biogenic synthesized silver nanoparticles. Diverse concentrations (2-10 μg/ml) of Lawsonia alba extract mediated silver nanoparticle  were  mixed with 1 ml of 0.1 mM DPPH in methanol and 450 μl of 50 mM Tris HCl buffer (pH 7.4) and incubated for 30 minutes (Figure 2). Later, the reduction in the quantity of DPPH free radicals was assessed dependent on the absorbance at 517 nm. Vitamin C was employed as control. The percentage of inhibition was determined from the following equation.

%inhibition=(Absorbance of control-Absorbance of test sample × 100)/ Absorbance of control

Result and Discussion

DPPH has been widely used as a stable free radical to test reducing substances as well as a useful reagent for investigating the component's   free radical scavenging behavior. From the results obtained it has been inferred that there is  a  consistent  increase  in  the  antioxidants  activity  with Lawsonia Alba mediated silver nanoparticles. At 10 μl concentration 54.8% is observed by silver nanoparticle and 76.56% by standard, 67.8% antioxidant activity was observed by AgNPs and 78.52% by the standard      at 20 μl concentration, at 30 μl concentration AgNPs showed 71.9% antioxidant property and 85.63% by standard, at 40 μl concentration 75.7% was observed by AgNPs and 88.68% by standard, at 50 μl concentration 79.2% and 93.15% is observed by AgNPs and standard respectively. Hence maximum antioxidant activity is observed at 50 μl concentration. The present study shows that the antioxidant activity of Lawsonia alba mediated silver nanoparticles was seen  to  be  increased  as  the  concentration  increased in a dose-dependent manner, hence can act as good antioxidant control.   The graph represents a comparison of antioxidant activity of Lawsonia alba mediated silver nanoparticles at different concentrations. From the above graph it has been observed that the antioxidant activity of Lawsonia alba mediated silver nanoparticles was seen to be increased as the concentration increased in a dose-dependent manner, hence can act as good antioxidant control (Graph 1).

DPPH radical scavenging  assay  was  used  to  determine  AgNPs' major antioxidant capacity. The BHT (butylated hydroxytoluene) has been used as the  standard.  Kharat  (2016)  analyzed  the  antioxidant  behavior  of the synthesized  nanoparticles  using  the  DPPH  assay  and  detected  the antioxidant ability of photosynthesized nanoparticles [41]. They  suggested that photosynthesized NPs could be used as  possible  free  radical scavengers. The color difference was caused by the  reduction  of Ag+ into silver nanoparticles when exposed to the Lawsonia alba extract.  The extract was yellow when it was freshly prepared. The extract turned  dark brown after being treated with AgNO3 and incubated. The surface plasmon resonance effect results in color variations in aqueous solutions  [42]. Plants including Cymbopogon citratus, Garcinia mangostana bark extract, Mucuna pruriens seed, Kalanchoe pinnata leaf,  Acorus  calamus root and Chrysanthemum Indicum have previously  been  reported  to produce AgNPs [43–48]. The prior studies indicated that the antioxidant property may be due to the existence of flavonoids and tannins in leaves   and phytochemicals such as phytol, sterols, and fatty acids [49]. Both the amounts of oxidants and antioxidants in tissues should  be  controlled.  If ROS oxidants are present at high levels, oxidative stress is produced in      the body by destroying DNA, carbohydrate, and protein repair, thereby disrupting the regular metabolism of the body  [50].  Yet  it  also  tends  to stop aging. Antioxidants play an important role in the neutralization of free radical species generated as end products of natural biochemical processes in the body, thereby regulating the aging process and other degenerative diseases [51]. In a previous study done by Anand findings indicated that, relative to vitamin C, AgNPs have greater antioxidant activity [51]. In terms of DPPH radical scavenging, Patra in his study showed high antioxidant activity (IC50 385.87 g/mL). The findings suggest that AgNPs can be used   as natural antioxidants to protect against various types of oxidative stress linked to degenerative diseases. In particular, AgNPs must be tested for antioxidants before being used in vivo models or human applications [52]. The Ag-NPs coating is used to secure the NPs by causing electrostatic and electrostatic repulsions between them. The coating  also  defends  against the cytotoxicity of Ag-NPs, according to several studies. Nanoparticles may help with vascular changes, particularly endothelial dysfunction caused by oxidative stress [53]. This syndrome can cause a decrease in Nitric Oxide (NO) bioavailability, which can affect vascular tone control and endothelial dysfunction, the first stage of cardiovascular disease. Thus, the antioxidant nanoparticles produced in this study may be used to treat vascular disease caused by hypertension, diabetes, or atherosclerosis (Figures 3 and 4) [54].

Conclusion

The current study was limited to the use of DPPH assay for evaluating antioxidant activity. Other conclusive methodologies like  ABTS  assay,  FRAP assay, FOX assay and  FTC  assay  need  to  be  employed  for  further evaluation. Future research  into  silver  nanoparticles  examining  their biological properties such as ant diabetic, anti-inflammatory, and antimicrobial activities both in vitro and  in vivo will lead to the production      of Nano-formulations as therapeutics in various diseases. Lawsonia Alba mediated  silver  nanoparticles  were  proved  to  possess  a  strong  level    of antioxidant activity. The results of the study reinforce the opinion that medicinal plants are promising sources of potent antioxidants that may be useful for therapy.

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