INTRODUCTION:

Plants play an important role in the development of new drugs¹. Phytochemicals are the natural compound occur in plants, vegetables and fruits, that work with nutrients and fibres to act specifically against diseases². Glycyrrhiza glabra is one of the most extensively used medicinal herbs. The word Glycyrrhiza is derived from the Greek term glykos (meaning sweet) and rhiza (meaning root). Glycyrrhiza glabra is commonly known as Yashtimadhu.

It is mainly used for the treatment of peptic ulcer, hepatitis C, and pulmonary and skin diseases, although clinical and experimental studies suggest that it has several other useful pharmacological properties such as anti-inflammatory, antiviral, antimicrobial, antioxidative, anticancer, and flavouring agent^3,4. It has been cultivated in Europe since at least the 16^th century ⁵. Nano-technology is the production technology to get the extra high accuracy and ultra fine dimensions, i.e. the preciseness and fineness on the order of 1 nm (nanometer), 10-9 meter in length⁶. The study aimed to Study the antibacterial activity of green synthetic NPs on-one type of Gram-negative (E.coli) and Gram-positive (Staphylococcus aureus) pathogenic bacteria. Investigating the anti-inflammatory (burns healing effects) activity of the green synthetic nanoparticles by investigating the day requiring to heal the burns on the skin of albino male mice ⁶.

MATERIAL AND METHOD:

Study area:

This study was carried out at laboratory of molecular and medical biotechnology department/college of Biotechnology/Al-Nahrain University from September/2019 to September 2020. Green synthetic silver nanoparticles have been prepared previously and supplied⁷ at different green synthetic nanoparticle concentrations (1, 1.5, 1.75, 2mM).

1- Bacterial isolation and identification:

The bacterial sample supplied from the University of Baghdad/biology department/molecular biology laboratory for high graduate.

A- Determination of antibacterial activity:

Muller Hinton agar used to be prepared using dissolving 38g about media within one thousand ml distill cloud (D.W.), then, keyed after ebullition to disappear the middling.

The Muller Hinton agar was once poured in accordance to a deep on 3 to four mm of the glass plate. After solidification, plates were kept at 4°C in conformity with providing a strong surface because wells construction who had been last stuffed along a hundred μL regarding specific concentrations of green synthetic NPs of Glycyrrhiza glabra⁸. In the current study, two different bacterial species were used (Staphylococcus aureus, Escherichia coli). Single colonies from each type of bacteria indicated above text had been grown concerning nutrient agar because of 18-24 hrs yet transferred following reed containing 5ml on Normal saline yet combined nicely using vortex (Griffin/Englan d), below the bacterial boom was compared together with McFarland tube (Baghdad/Iraq). The turbidity regarding norm answer cylinder wide variety was once equal according to a bacterial inoculums attention of 1.5×108 cell/ml. By the use of becoming addicted swab, a touch about the bacterial way of life beside everyday saline was once transferred in conformity with muller Hinton agar prepared above then streaked 3 toughness instances via rotating the pebble approximately 60° concerning the streaking, in imitation of confirming even outgiving touching the inoculums, the inoculated plates have been positioned at automobile temperature because of ten min in conformity with permit absorption about longevity extra sogginess⁹

Then, by means of using sterilized Pasteur pipette (Baghdad/Iraq) building wells (the wells were arranged hence so in imitation of avoiding the improvement about the overlapping of embargo zones) as had been crammed with a hundred μl over of green synthetic NPs of Glycyrrhiza glabra banish along with exceptional concentration [one hundred, two hundred and three hundrerd mg/ml] yet the plates has been incubated at thirty seven degree centigrade for 18-24 hrs. Then incubation, embargo area have been adequate by dictator according to determine their diameters into millimetrs, then the consequences had been recorded¹⁰.

B- Burns skin model in mice:

To assess burns healing activity of green synthetic NPs of Glycyrrhiza glabra , four mice groups were tested. Mice hair were removed by hair removal cream (veet) and a flame was used to induced burns of the skin, the recovery days were calculated by determining the number of days required to heal the wound¹¹.

RESULTS AND DISCUSSIONS:

1- Antibacterial activity of green synthetic silver nanoparticles:

According to the zone of inhibition, the antibacterial activity of green synthetic nanoparticles at (1, 1.5, 1.75, 2mM) against G-ve (E. coli) and G+ve (S. aureus) bacteria were studied. The result showed that green synthetic nanoparticles had effective antibacterial activity increased with increasing concentration of nanoparticles the tested G-ve bacteria with an inhibition zone ranged from 12 to 20mm in diameter. on the other hand, green synthetic NPs had antibacterial activity against G+ve bacteria (Fig 1) , with an inhibition zone ranged from 10 to 22mm, (table 1).

Table (1): Zone of inhibition (mm) produced by green synthetic silver nanoparticles (AgNPs) on Gram-negative (E.coli) and gram-positive ( Staphylococcus aureus).

Concentrations of green synthetic sliver nanoparticles(Mm)	Escherichia coli	Staphylococcus aureus
1	10	17
1.5	22	20
1.75	10	12
2	22	17

(A) (B)

Figure(1 a-b):Antibacterial activity (zone of inhibitions) of green synthetic silver nanoparticles (1, 1.5, 1.75, 2mM) against (A) Gram-negative bacterial isolates (E. coli) and (B)Gram- positive Staphylococcus aureus.

Gram-negative bacteria showed larger zones of inhibition, compared with the Gram-positive bacteria, which may be due to the variation in cell wall composition. The cell wall of Gram-positive bacteria composed of a thick peptidoglycan layer, containing linear polysaccharide chains cross related by short peptides, thus making a more rigid structure leading to hard penetration of the silver nanoparticles, while in Gram negative bacteria the cell wall possesses thinner peptidoglycan layer^12,13. On the other hand, several main mechanisms underlie the biocidal properties of silver nanoparticles against microorganisms. The first one proposed that when silver nanoparticles attach to the negatively charged cell surface alter the physical and chemical properties of the cell membranes, the cell wall and disturb important functions such as permeability, osmoregulation, electron transport and respiration^14,15,16. Another scientist stated that silver nanoparticles can cause further damage to bacterial cells by permeating the cell, where they interact with DNA, proteins and other phosphorus- and sulfur-containing cell constituents; while the third one in silver nanoparticles releases silver ions, generating an amplified biocidal effect, which is size- and dose-dependent^17,18.

However, the same results were recorded by the researchers^19,20 who found that silver nanoparticles have relatively higher antibacterial activity against Gram-negative bacteria than Gram-positive bacteria.

Burns healing effect of green synthetic nanoparticles:

To explore the anti-inflammatory activity of green synthetic nanoparticle, the burns healing effect was assessed. The ability of green synthetic nanoparticle at (1.5mM), silver sulfadiazine (positive control) and negative control to heal the burns by determining days required for recovering was studied.

The results (Table 2) showed that silver green synthetic nanoparticle loaded on Glycyrrhiza glabra could heal burns in 12 days compared with sliver sulfadiazine and negative control which require 16 days and 18 days to heal the burns (figure 2).

Table (2): The recovery of burn healing in mice after different treatments.

Groups	Treatment	Period of recovery
1	Green synthesis of silver nanoparticle of aqueous extract of Glycyrrhiza glabra	Days 12
2	Without any treatment	Days 18
3	Silver sulfadiazine	Days 16

(1) (0day)

(2) (During treatment)

(3) 4 days

(4) 14 days

(A)

(0 day)

(18)days

(B)

(0 day)

(16)days

(C)

Figure (2): Mice healed from burns on (0-18) days after different treatments. (A)Mice treated with green synthetic nanoparticle. (B) Mice without any treatment (negative control). (C) Mice treatment with 1% silver sulfadiazine

It was earlier reported that AgNPs could successfully decrease the infiltration of inflammatory cells, inhibit the production of inflammatory cytokines, and up-regulate the expression of matrix metalloproteinase^21,22. Also, previous reports^23,24 confirmed that administration of AgNPs decreased inflammation by reducing vascular endothelial growth factor and mucous glycoprotein expression in ovalbumin-induced allergy in murine model^25,26. Also, AgNPs effected anti-inflammatory activity in postoperative peritoneal adhesion model. previous studies^27,28indicated that silver-polyvinyl pyrrolidone nanoparticles exhibited anti-inflammatory action by reducing TNF-α^29,30.

CONCLUSION:

It was concluded from the study that green synthetic nanoparticle possessed antibacterial activity against Gram-negative and Gram-positive bacteria and had anti-inflammatory activity in compare to commercially used cream that can be used in medicinal application and scientific field at different degree of activity according to concentration tested.

CONFLICT OF INTEREST:

Last year, there was a widely grow of resistance bacteria which had the ability to cause different disease and inflammation so the requirement to find newly synthesized compound which inhibit these growing increase every year.

AUTHOR'S CONTRIBUTION:

This work was carried out in collaboration between all authors. Author Basma KH. Alani designed the study, performed the statistical analysis, Authors Shahad Basil Ismael and Rasha Abdulelah Ibrahim wrote the protocol, and wrote the first draft of the manuscript. Author Mohanad H.Husain and Zahraa Ahmed Okhti managed the analyses of the study. All authors read and approved the final manuscript.

ACKNOWLEDGEMENT:

This research done by the aid of staff of Biotechnology research center\Al-Nahrain University and especially dr. Jasim Mohammed.

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Received on 27.03.2022 Modified on 14.05.2022

Research J. Pharm. and Tech 2022; 15(12):5571-5575.

DOI: 10.52711/0974-360X.2022.00941