ijdr_987_22_R2_OA

Original Article ‑ In vitro Study (All Types)

1 1
2 2
3 AQ1 Synthesis and Characterisation of Gold Nanoparticles from Acacia Leaf 3
4 and to Evaluate Anticariogenic Activity 4
5 5
6 6
7 Abstract Maher AbdelFattah 7
8 Objective: To synthesise the gold nanoparticles (AuNPs) using Acacia catechu through biogenic Al Shayeb1,2, 8
9 synthesis and evaluate their antimicrobial efficacy against S. mutans and E. coli in vitro. Naresh 9
Methods: Green synthesised AuNPs were characterised using the ultraviolet–visible (UV‑Vis)
10 Yedthare Shetty1,2, 10
spectroscopy, and the size and shape of the synthesised nanoparticles were evaluated using the
11 transmission electron microscopy  (TEM). The antimicrobial efficacy of AuNPs  (30/60/100 µl) Anas Al Jadaa1,2, 11
12 against S. mutans/E. coli was evaluated on the Mueller–Hinton agar by measuring the zone of Syed Kuduruthullah3 12
13 inhibition (ZOI) with ampicillin (15 µl) as a positive control. Results: The synthesised AuNPs were 1
Department of Clinical
13
AQ3
14 confirmed using the UV‑Vis spectroscopy with peaks at 540 nm, and the size of the particle estimated Science, Ajman University, 14
15 using the TEM was between 5 and 15  nm. The antimicrobial efficacy of AuNPs was comparable to Ajman, 2Center of Medical 15
16 that of ampicillin against S. mutans/E. coli, but the difference was not significant. The antimicrobial and Bioallied Health Sciences 16
effects increased in a dose‑dependent fashion but were comparable across all concentrations and Research, Ajman University,
17 Ajman, 3Department of
17
18 ampicillin. Conclusion: Green synthesised AuNPs exhibited significant antibacterial activity against 18
Dentistry, Health Sciences, City
S. mutans and E. coli at par with commercial ampicillin and demonstrated the potential towards
19 University College of Ajman, 19
anticariogenic agent for future use in dentistry. Ajman, UAE
20 20
21 Keywords: Caries, gold nanoparticles, green synthesis, S. mutans 21
22 22
23 23
24 Introduction Acacia catechu also known as Senegalia
24
catechu is a deciduous, thorny tree, which
25 In recent years, nanotechnology has 25
grows up to a height of 15 m. The common
26 attracted great interest due to its expected 26
names for it include kher, catechu, cachou,
27 impact in many areas, such as energy, 27
cutch tree and black cutch.[5] Senegalia
28 medicine and electronics. The synthesis 28
catechu is found in India, China, Japan and
29 of nanoparticles has been carried out 29
the surrounding areas of the Indian Ocean.
30 using many methods, such as sol 30
It contains derivatives of flavonoids, and it
31 process, micelle, chemical precipitation, 31
has acquired its name from the important
32 hydrothermal method, pyrolysis, chemical 32
catechins and catecholamines of chemistry
33 vapour deposition and bio‑based 33
and biology.[6] Flavonoids are a class of Address for correspondence:
34 protocols.[1] To eliminate the use of 34
secondary plant phenolics with significant Dr. Maher AbdelFattah Al
35 traditional toxic substances and reduce the Shayeb, 35
pharmacological activities and possess
36 side effects, the synthesis of nanoparticles Department of Clinical Science, 36
several important functional groups,
37 is carried out using natural products, Ajman University, Ajman, UAE. 37
which aid in reducing the metals into Center of Medical and Bioallied AQ2
38 such as plant extracts.[2] Among all the 38
nanoparticles. Furthermore, the flavonoids Health Sciences Research,
39 nanoparticles, gold nanoparticles (AuNPs) Ajman University, Ajman, UAE. 39
found in plant extract act as both reducing
40 have attracted great interest due to their E‑mail: maheralsahebau@ 40
agents and electrostatic stabilising agents
41 unique and tuneable surface plasmon gmail.com 41
for the synthesised nanoparticles.
42 resonance, and their synthesis using plant 42
43 extract has been proven to be the most Gold is the least active metal, with Received : 29‑12‑2022 43
44 convenient and inexpensive method.[3] excellent biocompatibility and stable Revised : 25‑02‑2023 44
45 chemical properties.[7] AuNPs possess Accepted : 28‑02‑2023 45
Several plant sources, such as gooseberry,
Published : ***
46 aloe vera, coriander, guava, clove buds, excellent antibacterial activity against 46
47 mint, cinnamon and curry leaves, have been gram‑positive and gram‑negative bacteria, Access this article online
47
48 used successfully towards the synthesis of are univalent and have multiple bacterial 48
Website: www.ijdr.in
49 AuNPs.[4] binding sites. Furthermore, because of their 49
50 ability to target a myriad of molecules, drug DOI: 10.4103/ijdr.ijdr_987_22 50
51 Quick Response Code: 51
This is an open access journal, and articles are
52 distributed under the terms of the Creative Commons 52
Attribution‑NonCommercial‑ShareAlike 4.0 License, which allows
53 others to remix, tweak, and build upon the work non‑commercially,
How to cite this article: Shayeb MA, 53
54 as long as appropriate credit is given and the new creations are Shetty NY, Al Jadaa A, Kuduruthullah S. Synthesis 54
licensed under the identical terms. and characterisation of gold nanoparticles from Acacia
55 leaf and to evaluate anticariogenic activity. Indian J 55
56 For reprints contact: WKHLRPMedknow_reprints@wolterskluwer.com
Dent Res 2023;XX:XX-XX. 56

© 2023 Indian Journal of Dental Research | Published by Wolters Kluwer - Medknow 1

 Shayeb, et al.: Anticariogenic potency of biogenic AuNPs

1 resistance is highly unlikely.[8] AuNPs have a high affinity using sterile cotton swabs. The surface of agar plates was 1
2 towards organic species, which could also explain their bored using a gel borer to make wells. Then, different 2
3 effective antibacterial properties.[9] Moreover, gold‑coated concentrations of AuNPs such as 30 µl, 60 µl and 100 µl 3
4 dental floss was found to exhibit antibacterial properties were incorporated in the centre of the plate. Ampicillin (15 4
5 against Streptococcus mutans (S. mutans) and Streptococcus µml) antibiotic was used as a positive control in this 5
6 sobrinus. study. Finally, the plates were incubated at 37°C for 24 6
7 to 48 hours. Triplicate plates were incubated for each 7
Thus, we evaluated the potential of Acacia catechu
8 organism, and the zone of inhibition (ZOI) was evaluated 8
leaves towards a biogenic synthesis of AuNPs and the
9 in millimetres (mm). 9
synthesised nanoparticles were characterised using
10 10
the UV‑Vis spectroscopy and transmission electron Statistical analysis
11 11
microscopy (TEM).[10] Lastly, the AuNPs are investigated
12 The mean difference in the ZOI between various 12
for antimicrobial activity against S. mutans and
13 concentrations of the AuNPs was compared with amipicillin 13
Enterococcus faecalis (E. faecalis).
14 using the Kruskal–Wallis test. The level of significance was 14
15 Materials and Methods set at 5%. 15
16 16
17 Biogenic synthesis of AuNPs Results 17
18 The Acacia catechu extract was obtained from a dedicated 18
Phytochemical analysis of Acacia catechu extract
19 Ayurvedic community pharmacy in Chennai and was 19
20 evaluated for impurities using chromatography. Ethical Qualitative analysis of the triphala extract was found 20
21 clearance was waived for this study as this was an in vitro positive for flavonoids, tannins, phenols, alkaloids, acids, 21
22 study that did not utilise any samples/materials that needed terpenoids, proteins and steroids. 22
23 clearance from the institutional review board. An aqueous 23
Characterisation of AuNPs
24 extract of Acacia catechu (1%) was prepared by boiling 24
25 weighted 10  g of the Acacia catechu powder in 100  ml of The synthesised biogenic AuNPs were confirmed using 25
26 double‑distilled water in a water broth for about 10 minutes the UV‑Vis spectroscopy with changes in the absorption 26
27 at a temperature of about 80°C and then allowing to band beginning at 510  nm until 590  nm. The maximum 27
28 cool down followed by filtration  (Whatman No.  1) of peak was at 540  nm, and the absorption band gradually 28
29 the extract. The gold chloride solution was prepared by increased from 1.6 to 3.1 suggestive of nanoparticle 29
30 dissolving 0.393  mg of AuCl2 in 90  ml of double‑distilled reduction [Figure 1]. The surface morphology of the 30
31 water followed by mixing 10  ml of AuCl2, 10  ml of synthesised AuNPs assessed using TEM was found to be 31
32 aqueous Acacia extract and 80  ml of distilled water. amorphous at 70  nm resolution, and at 20  nm, spherical 32
33 Then, the mixture is placed in the magnetic shaker for the shapes were revealed with the size of synthesised ZnO‑NP 33
34 synthesis of AuNPs and is allowed to mix, and periodic ranging from 5 to 15 nm [Figure 2]. 34
35 UV‑Vis spectrophotometer readings  (Model UV‑D3200) 35
36 Antibacterial activity 36
were recorded to confirm the synthesis of AuNPs.
37 The ZOI was highest for the commercial ampicillin (15 µm) 37
38 Characterisation of ZnO‑NP for both S. mutans and E. coli compared with the other 38
39 UV‑Vis spectrophotometric analysis was conducted on concentrations of AuNPs. However, the difference was not 39
40 2 ml aliquots of the prepared nanoparticle solution at 1, 40
41 12, 18, 24 and 48 hours using a Shimadzu 1700 UV‑Vis 41
42 spectrophotometer at 300–650  nm wavelengths with a 42
43 scanning speed of 1,856  nm/min to confirm the synthesis 43
44 of AuNPs following the change in colour of the solution 44
45 from brown to brick red. The surface morphology and size 45
46 of the synthesised AuNPs were assessed using 200  kV 46
47 high‑resolution TEM. 47
48 48
49 Antibacterial activity of AuNPs 49
50 The antibacterial activities of synthesised AuNPs were 50
51 tested against oral pathogens such as S. mutans and 51
52 E. faecalis (MTCC, India). The antibacterial activity of 52
53 AuNPs was determined using the agar well‑diffusion 53
54 method. This test organism was inoculated in nutrient 54
55 broth agar with a pH of 7 for 24 hours. The isolated test 55
56 organisms were seeded on the Mueller–Hinton agar plates Figure 1: UV‑Vis spectroscopic analysis of gold nanoparticles 56

2 Indian Journal of Dental Research | Volume XX | Issue XX | Month 2023

 Shayeb, et al.: Anticariogenic potency of biogenic AuNPs

1 further research towards safety and toxicity assessments. 1

2 The mechanism of action of AuNPs is not the same against 2
3 gram‑positive and gram‑negative bacteria, and this is 3
4 primarily dependent upon the structure of the membrane 4
5 of the bacteria.[15] The antimicrobial action of AuNPs can 5
6 be attributed primarily to two processes: firstly, they affect 6
7 the metabolism by affecting the membrane potential by 7
8 reducing the activity of adenosine triphosphate synthase; 8
9 secondly, they affect the synthesis of proteins by impacting 9
10 the role of tRNA, thereby causing cell death. 10
11 11
The size of the AuNPs also plays an important role, and
12 12
due to their smaller size, they have higher penetrating
13 13
power and greater surface area to interact with the bacteria.
14 14
Moreover, the efficacy of AuNPs is augmented when the
15 15
overall size of the particles is low.[16] This study evaluated
16 Figure 2: Transmission electron microscopy (TEM) image of gold 16
nanoparticles synthesised using Acacia leaf extract the antimicrobial effect of biogenic AuNPs against
17 17
S. mutans and E. coli and the use of novel techniques
18 18
significant  (P  >  0.05). The ZOI for 30 µl was 16–19  mm; to synthesise AuNPs, and low cost can be considered
19 19
for 60 µl, 17–22  mm; and for 100 µl, 18–23  mm; and an advantage. However, the inability to investigate the
20 20
the control ampicillin had 20–25  mm for S. mutans. With antioxidant potential, material characterisation, and toxicity
21 21
regard to E. coli, 30 µl AuNPs have a ZOI ranging from 12 can be considered potential limitation and it provides scope
22 22
for further research.
23 to 15 mm; 60 µl AuNPs, 13 to 17 mm; and 100 µl AuNPs, 23
24 15 to 19 mm, and ampicillin has 13 to 21 mm. Conclusion 24
25 25
26 Discussion In conclusion, green synthesised AuNPs exhibited 26
27 significant antibacterial activity against S. mutans and 27
AuNPs can be synthesised through several chemical,
28 E. coli at par with commercial ampicillin and demonstrated 28
physical and mechanochemical techniques, but many of
29 the potential of anticariogenic agent for future use in 29
them require the use of chemicals that can produce toxic
30 dentistry. 30
byproducts.[11] The green synthesis of nanoparticles uses
31 the phytochemical contents of plants and other organic Financial support and sponsorship 31
32 sources to produce nanoparticles, which have caused a 32
33 Nil. 33
paradigm shift towards their synthesis. The use of plants to
34 synthesise biogenic nanoparticles provides the advantages Conflicts of interest 34
35 of simplicity, control and mass production without the 35
36 There are no conflicts of interest. 36
added health and environmental impact.[12]
37 37
UV‑Vis spectroscopy is an excellent technique to identify References
38 38
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40 red colour, and we found the peak around 540 nm; previous plants extract mediated synthesis of silver nanoparticles for
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To the best of our knowledge, there are no reports on the
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47 effectiveness of AuNPs on S. mutans; however, gold‑coated 47
herb extract‑mediated biosynthesis of gold nanoparticles and
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commercial ampicillin in our study sample.[14] Based on Puchades R, Maquieira A, et al. Detection of chemical
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33 Author Queries??? 33
34 AQ1: Please suggest whether the phrase “and to Evaluate Anticariogenic Activity” can be changed to “and Evaluation of 34
35 Anticariogenic Activity” in the article title. 35
36 AQ2: Please check corresponding address for correctness 36
37 AQ3: Kindly check all affiliations for correctness 37
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4 Indian Journal of Dental Research | Volume XX | Issue XX | Month 2023