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Biosynthesis of Chitosan Nanocomposite with
Myrrh-Mediated Nanosilver for Controlling Skin
Pathogenic Microbes
Hager A. Saad
1
, Mona Assas
1
, Asmaa Abdella
2
, Hend A. Gad
1
,
and Ahmed A. Tayel
1
Abstract
Background: The usages of biosynthesized nanomaterials for microbial pathogens’fighting have numerous rationales and effec-
tiveness. Skin microbes could acquire drug-resistance that needs innovative approaches for overcoming. Objectives:
Phytosynthesized silver nanoparticles (AgNPs) with Commiphora myrrh resin extract (MR) and their nanoconjugates with chitosan
nanoparticles (Cht) were fabricated and assessed as potential antimicrobial agents for controlling antibiotic-resistant microbial
skin pathogens, Materials and methods: AgNPs biosynthesis was achieved within MR solution and they were composited with
Cht. The syntheses of nanomaterials were validated using infrared spectroscopy and electron microscopy and they were loaded
onto cotton textiles, then all fabricated nanomaterials/textiles were assessed for inhibiting skin pathogens Staphylococcus aureus and
Candida albicans. Results: Nanomaterials’characterization appointed the mean size of MR-synthesized AgNPs to be 22.58 nm,
whereas the mean diameter of Cht/MR/AgNPs nanocomposites was 130.34 nm and carry +25.9 mV charges. The infrared assess-
ment validated the interactions between the employed materials. The loaded cotton textiles with MR/AgNPs and Cht/MR/AgNPs
could effectively inhibit the growth of Staphylococcus aureus and Candida albicans, Cht/MR/AgNPs was the most powerful. The scan-
ning microscopy confirmed the antimicrobial action of Cht/MR/AgNPs toward the skin pathogens; the microbes mostly lysed and
deformed within 12 h of exposure to nanocomposites. Conclusions: The Cht/MR/AgNPs nanocomposite provided potent anti-
microbial actions toward skin microbial pathogens.
Keywords
antimicrobial, biopolymers, green synthesis, hygienic textiles, nanomaterials
Received: August 2nd, 2024; Accepted: October 21st, 2024.
Introduction
The skin is vital for shielding the body from infections and
external harm. Normal skin microbiota, which is primarily com-
mensal in nature, is colonized on the skin. But these microbiota
act like opportunistic pathogens in aberrant or disturbed condi-
tions.
1
A common method for pathogens to bypass the skin
barrier is through a wound or tear, which enables them to
enter the body, cause an infection, and evade the body’s
natural defenses. Candida albicans and Staphylococcus aureus are
prime examples of such pathogens.
2
Folliculitis and hidradenitis can develop from the growth of
pathogenic microbes in skin appendages, which promptly trig-
gers nearby keratinocytes to release inflammatory cytokines
and chemokines. This, in turn, attracts neutrophils and mono-
cytes to the affected area.
3
Severe injuries can impact appear-
ance and health, increasing the risk of infection. Antimicrobial
resistance exacerbates treatment difficulties, leading to higher
morbidity, mortality, and healthcare costs.
4,5
Nanotechnology has been recognized as one of the most
crucial research endeavors of the twentieth century, primarily
due to its use of the unique properties of atomic and molecular
structures at the nanoscale. It provides the scientific and indus-
trial foundation for creating and utilizing nanomaterials.
Nanotechnology enables the manipulation of materials at the
nanoscale (1-100 nm), driving significant advancements across
various sectors, including energy, electronics, and medicine.
1
Department of Fish Processing and Biotechnology, Faculty of Aquatic and
Fisheries Sciences, Kafrelsheikh University, Kafrelsheikh, Egypt
2
Department of Industrial Biotechnology, Genetic Engineering and
Biotechnology Research Institute, University of Sadat City, El-Sadat City, Egypt
Corresponding Author:
Ahmed A. Tayel, Department of Fish Processing and Biotechnology, Faculty of
Aquatic and Fisheries Sciences, Kafrelsheikh University, Kafrelsheikh 33516,
Egypt.
Email: ahmed_tayel@fsh.kfs.edu.eg
Creative Commons Non Commercial CC BY-NC: This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License
(https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission
provided the original work is attributed as specified on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage).
Natural Products-Based Nanomaterials: Biosynthesis and Biomedical Applications –Original Research Article
Natural Product Communications
Volume 19(11): 1–11
© The Author(s) 2024
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DOI: 10.1177/1934578X241297994
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This scale of material engineering allows for unprecedented
improvements in functionality and performance.
6
A nanometer
(nm) is a billionth of a meter. By working at the atomic and
molecular levels, scientists and engineers can develop new fea-
tures and applications. However, one of the main challenges in
nanotechnology is producing nanoparticles with the appropriate
characteristics for specific applications. Achieving desirable out-
comes requires comprehensive knowledge of nanoparticle
properties, synthesis techniques, and operating conditions.
7
The antibacterial efficacy of metallic nanoparticles, or nano-
metals, has been shown in a variety of biological and pharma-
ceutical applications.
8
This is mostly because of the metals’
cytotoxicity against microbial infections and interactions with
their membranes and internal routes.
6
Nanotechnology explores
nanoscale materials, like silver nanoparticles (AgNPs), for
medical applications. AgNPs, along with polymer nanocomposites
(NCs), improve treatments like targeted delivery.
5
The antimicro-
bial activities of silver nanoparticles (AgNPs) are well-documented.
These activities include the release of Ag +ions, the production of
reactive oxygen species (ROS) within the inner and outer microbial
membranes, interference with the cellular membrane, disruption of
the ribosome-mitochondrial complex, and disturbances in nucleic
acids.
9
Conventional synthesis methods face cost and environmen-
tal issues, prompting interest in eco-friendly alternatives like green
synthesis, using plant metabolites. Plants, known for medicinal
properties, contain phenolic compounds with antimicrobial, anti-
inflammatory, and antioxidant benefits for treating skin infections.
5
The genus Commiphora (Burseraceae) encompasses approxi-
mately 200 species, including Commiphora myrrh, from which
the myrrh resin is derived. With a rich historical background,
myrrh extract (MR) has been extensively utilized as a traditional
remedy for various skin conditions such as wounds, infections,
acne, and boils.
10
Myrrh, along with several related species of Commiphora,is
defined by the United States Pharmacopeia (USP 34) as the gum-
resin extracted from the stems and branches of C. molmol.Ethiopia
and Somalia are the primary producers of this resin. It is designated
as Generally Recognized as Safe (GRAS) by the Food and Drug
Administration (FDA) for use as a food additive.
11
Additionally,
Commission E has approved myrrh extract for its astringent
and antibacterial properties.
12
As a functional food with antioxi-
dant activity and potential to help prevent human colon cancer,
it was suggested adding it to yogurt or milk.
13
Myrrh is used in complementary medicine for its antiviral,
expectorant, antifungal, and antibacterial properties.
14
Numerous
clinical and experimental studies have been conducted locally to
investigate the effects of myrrh extracts on various microorgan-
isms. While the results have been mixed, several studies have sup-
ported the effectiveness of myrrh, especially when used in the
recommended daily dosages.
12
Chemically, myrrh is an oleo gum
resin. The gum component, which constitutes 40%–60% of the
total, is primarily made up of polysaccharides, proteins, and
oxidase enzymes and is water-soluble. In contrast, the resin, com-
prising 20%–40%, along with the volatile oil fractions, is
alcohol-soluble.
12,14
Chitosan (Cht), a versatile polymer, exhibits biocompatibility,
non-toxicity, and ease of extraction, making it a promising can-
didate for biomedical applications.
15
Chitosan nanoparticles
have shown potential in enhancing the bioactivities of metal
nanoparticles, including their antimicrobial properties.
16
Combining chitosan with AgNPs enhances the stability and
dispersibility of the nanoparticles, thereby boosting their thera-
peutic potential for combating cancer and microbial infec-
tions.
17
Chitosan’s pH responsiveness and mucoadhesive
properties enable controlled drug release and targeted delivery
to mucosal areas. Additionally, its film-forming capability
makes it ideal for applications requiring thin protective coatings.
The ease of modifying chitosan also allows for the addition of
new functions to AgNPs-chitosan composites.
18
Despite the potential benefits, there is currently a dearth of
published studies on the biosynthesis of the Cht/MR/AgNPs
nanocomposite. Therefore, our study aimed to bridge this gap
by synthesizing the nanocomposite, characterizing its physico-
chemical and structural properties, evaluating its antimicrobial
efficacy, and assessing its suitability for textile coatings.
Materials and Methods
Chemicals and Reagent
The experiment’s materials and reactants were all obtained
from Sigma-Aldrich Co. (St. Louis, MO, USA) and approved
for use in analysis.
Preparation of Myrrh Extract
Dried MR underwent pulverization to a 60-mesh size using a
mixing grinder, followed by mixing 200 g of MR powder in
1000 mL (w/v) of 70% ethanol (Sigma-Aldrich, MO). The
mixture was stirred at a speed of 160 xg for 22 h at room tem-
perature (25 ±2 °C). Upon filtration by filter paper (Whatman
no. 41), after removing the plant remnants, the MR extract
was dried by rotary evaporation (IKA, RV 10, Germany) at
44 °C. For the purposes of the next investigations, the dried
MR was then dissolved in a 2% Tween 80 solution in deionized
water (DW) to produce a concentration of 1 mg/mL.
19
Green Synthesis of Silver Nanoparticles
The green synthesis of AgNPs involved the use of myrrh
extract; 5 mL of the MR extract solution (concentration of
1 mg/mL) was applied dropwise to 50 ml of AgNO
3
(1.0 mM) at 45 °C for 15 min. After that, the resultant
AgNPs were heated to 60 °C and stirred with a magnetic
stirrer and electric heater until a brown-yellow color that is sug-
gestive of AgNPs production was seen. Since research has
shown that this suspension is stable in this environment, it
was then kept at ambient temperature in a conical glass flask
covered with foil until it was needed.
20
2Natural Product Communications
Chitosan Extraction
Erugosquilla massavensis (mantis shrimp) shell debris served as the
primary source for chitosan extraction. These remnants were
gathered from the “Seafoods processing plants, Kafrelsheikh
University, Egypt,”then cleaned, washed with deionized water
(DW), and subsequently dried at 45 °C for 42 h.
21
After grind-
ing the shells, chitosan extraction followed a series of steps:
demineralization (using 16 volumes of 1 M HCl, 12 h, at
room temperature); deproteinization (using 16 volumes of
1 M NaOH, 12 h, at room temperature); and deacetylation
(using 18 volumes of 55% NaOH solution, 95 min, at 115 °
C). Each stage was followed by DW rinsing and drying, and
the resultant powder was stored for further analysis.
16
Preparation of Cht/ MR /Ag Nanocomposites
The following solutions had to be made in order to synthesize
Cht-NPs and load them with MR/AgNPs: TPP (0.5 mg/ml in
DW), Cht (1 mg/ml in a 1% acetic acid solution), and MR
extract (1 mg/ml in a 2% Tween 80 solution in DA).
16
Using
a syringe needle, the TPP solution was gradually added to the
Cht solution at a rate of 0.35 ml/min until the volumes were
equal after the pH of the Cht solution was adjusted to 5.2.
An equal amount of MR/AgNPs was added to the Cht solution
prior to the addition of TPP in order to produce Cht/MR/
AgNPs.
Characterization of Synthesized Nanoparticles
NPs’Optical Analysis. In order to verify the creation of metal
nanoparticles through the identification of their surface
Plasmon resonance, which is linked to unbound electrons on
the surfaces of the nanoparticles, the AgNPs spectrum was exam-
ined utilizing a UV-Vis spectrophotometer (UV-2450, Shimadzu,
Japan) operating in the 300–1000 nm wavelength range.
FTIR Analysis. In transmission mode (at a wavenumber range
of 450-4000 cm1), the infrared characteristics of synthesized
MR, Cht, MR/AgNPs, and Cht/MR/AgNPs were examined
using Fourier transform infrared spectroscopy (FTIR; Perkin
ElmerTM FTIR-V. 10.03.08, Germany).
22
Zetasizer Analysis. Using Zetasizer (Zeta plus, Brookhaven,
USA), the AgNPs zeta potential (ζ) was assessed using the
dynamic light scattering approach.
Transmission Electron Microscopy (TEM). TEM (Leica-Leo 0430;
Cambridge, UK) were used to screen the Ps and define the mor-
phology and dispersion of Cht/MR/AgNPs.
Antimicrobial Textiles Preparation
The textile was immersed in SPSs for 90 min at 25 °C with con-
tinuous stirring, padded, and compressed to 100% wet pickup.
This process was modified from former work.
23
The textile was
then forced air dried for 120 min and cured for 15 min at 100 °
C. After being cut into 2 cm
2
squares, the SPS-treated textiles
were tested for microbiological resistance utilizing the inhibition
zone (IZ) appearance method on the microorganisms under
investigation. Textiles treated with SPS were placed on the
infected surface after microbial strains were streaked onto the
suitable solid media. The diameters of the IZs were measured,
and their mean values were computed, in triplicate throughout
the experiments. Antimicrobial textiles’resilience was assessed
following several cycles of washing in a domestic laundry
machine with neutral water at 41 ±3 °C.
23
Antimicrobial Activity of Treated Cotton Textiles. A sterile skin pro-
tectant solution (SPS) comprising 1% of Cht, MR, MR/AgNPs,
and Cht/MR/AgNPs was applied to scoured cotton plain
weave (108 g/m2) obtained from Misr Weaving and Spinning
Co., Egypt. This procedure was carried out utilizing a modified
pad-dry-cure method.
23
Evaluation of NPs Antibacterial/Anticandidal Activity
Cultures of Bacteria and Fungi. For antimicrobial screening,
Staphylococcus aureus and Candida albicans were used as common
skin pathogens. The screened strains included C. albicans-S
(ATCC-10231), C. albicans-R (resistant isolate to fluconazole,
from skin lesion), S. aureus-S (ATCC-25923) and resistant S.
aureus-R isolate (resist methicillin, from skin wound infection).
They were achieved from the microbial culture collection, Ain
Shams University, Egypt. On nutrient agar and broth (NA
and NB, Difco Laboratories, Detroit, MI), the whole bacterial
and fungal strains were kept alive and subcultured aerobically
at 37 °C.
24
Inhibition Zone (IZ) Assay. After the disc diffusion test, the inhi-
bition zones (IZ) that emerged were taken into consideration as
indicators of the antibacterial/antifungal bioactivity of the pro-
duced agents. Clean Whatman The 2% solutions of MR/
AgNPs, MRand Cht/MR/AgNPs were impregnated into No.
4 paper discs (6 mm in diameter) and then placed onto
freshly infected NA plates with each bacterial culture. After
measuring the IZ diameters with a precision calliper and incu-
bating the plates at 37 °C for 18 to 24 h, they were turned
upside down and the standard deviation of their triplicate
means was computed.
23
SEM “Scanning Electron Microscopy”Imaging. SEM imaging was
utilized to observe the morphological changes in Staphylococcus
aureus cell surfaces after exposure to a 2.0% (w/v) solution of
Cht/MR/AgNPs. These changes were monitored over an incu-
bation period ranging from 0 to 12 h at 37°C. SEM images
were captured at 20 kV and a magnification of x10,000, docu-
menting the distortions in bacterial and fungal cell structures.
22
Saad et al. 3
Cytotoxicity Assay
The normal HDF “Human Dermal Fibroblasts; C-12302,
Sigma-Aldrich”were used for assessing the cytotoxicity of
Cht/MR/AgNPs nanocomposite. In 96 well plate, cells
(10.000 cells/well) were seeded in DMEM medium
“Dulbecco’s Modified Eagle Media”supplemented with 10%
FBS “fetal calf serum”and Antibiotic-Antimycotic mixture
(1%), followed by their incubation in CO
2
incubator at 95%
humidity and 37 °C temperature. Wells were amended with
gradual concentrations of Cht/MR/AgNPs and further incubated
then cell proliferation/viability were assessed using MTT
“3.[4.5-dimethylthiazol-2-yl]-2.5-diphenyltetrazolium bromide”assay.
Statistical Analysis
In triplicate studies, each analysis was conducted three times.
Standard deviations were calculated using SPSS. Statistical anal-
ysis to determine data significance (at p < 0.05) was perfor med
using the “t-test and ANOVA”tests from the SPSS package
(V-11.5, Chicago, IL).
Results
NPs’Optical Analysis
Direct observation and UV-vis spectrophotometric analysis of
MR/AgNPs provide optical evidence that NP production
follows MR interaction. Direct examination of the MR/
AgNPs solutions revealed a progressive color change within
30 min of the reaction, going from clear to deep blackish-brown
(Figure 1A); no more color changes were seen after that. MR/
AgNPs had the largest NPs absorption peaks (λmax) at 412 nm
for that. MR/AgNPs according to UV-vis analysis (Figure 1B).
FTIR Analysis
The molecules produced, including MR and its composites with
AgNPs and Ch/AgNPs, were analyzed using FTIR to clarify bio-
chemical bonding and interactions between the composited agents
and the possible My groups in responsible for NP formation.
FTIR analysis of Cht (Figure 2-Cht) showed a band at
3426 cm
−1
, indicating strained intramolecular hydrogen bonds
(between O–H and N–H). Polysaccharide bands at
2921 cm
−1
and 2872 cm
−1
showed asymmetric and symmetric
C–H stretching. Chitosan biological bonding can be identified by
the following bands: 1153 cm
−1
(asymmetric C–O–Cbridge
stretching), 1067 cm
−1
and 1024 cm
−1
(C–Ostretching),
1654 cm
−1
(amide I C =O stretching), 1321 cm
−1
(C–Nstretching
vibration), 1412 cm
−1
and 1357 cm
−1
(CH2 bending/CH3 sym-
metrical deformations). The peaks at 1153 cm
−1
and 1066 cm
−1
indicate C–O overlapping and the formation of chitosan due to
the interaction with NH
4
groups in chitosan molecules.
The MR extract FTIR analysis (Figure 2-MR) showed bio-
molecules that involved in AgNPs synthesis, with absorption
peaks for O–H groups of phenols and C–H aromatic stretch
at around 3350 and 1480 cm
−1
, respectively. MR/AgNPs dis-
played absorption at 3464.32, 2205.12, 2346.38, 2032.89,
2020.91, 2001.85, 2160.98, 1963.83, 1637.98, 851.64, and
630.54 cm
−1
(Figure 2-MR/AgNPs), that are emerged after
interaction of MR and AgNPs during biosynthesis, which indi-
cate the new bonds between the two molecules. In
(Figure 2-MR), the peak at 2185.12 cm
−1
disappeared in the
AgNPs absorption bands; therefore, this peak, which corre-
sponds to (C =C) alkyne, might have helped C. myrrha to
produce nanoparticle.
The Cht/MR/AgNPs nanocomposites’FTIR analysis
(Figure 2-Cht/MR/AgNPs) showed numerous functional
groups from MR/AgNPs’interaction with Cht, with a broad
Figure 1. Evidences of AgNPs synthesis using myrrh extract, including visual appearance (A), UV-vis spectral analysis (B).
4Natural Product Communications
band at 3236 cm
−1
(carboxylic acids), a narrow band at
2877 cm
−1
(C–H stretch alkanes), and bands at 1397 cm
−1
(aromatic amines) and 1021 cm
−1
(aliphatic amines). The spec-
trum had many transferred bands from either Cht or MR/
AgNPs spectra, which appoint the electrostatic interactions
between both compounds.
Zetasizer Analysis
The nanoparticle size average diameter of the AgNPs was deter-
mined to have a mean Z-average size of 4.43 nm and negatively
charged (−23.7 mV). The polydispersity index (PDI) of the
AgNPs was 0.503, indicating minor size variation and agglom-
eration. The mean diameter of Cht/MR/AgNPs nanocompo-
sites was 130.34 nm and carry +25.9 mV charges, indicating
efficient MR/AgNPs encapsulation inside Cht.
Transmission Electron Microscopy (TEM)
The TEM imaging was used to examine. The images revealed
relatively circular nanoparticles with an average diameter
ranging from 0.52 to 8.28 nm., with mean dimeter of
4.81 nm. Synthesized AgNPs exhibited a spherical shape,
minimal aggregation, and varying sizes, as depicted in Figure 3.
Evaluation of NPs Antibacterial/Anticandidal Activity
From Table 1, it was observed that Cht/MR/AgNPs composite
had the most effective inhibitory action against S. aureus S, with
an inhibition zone diameter of 36.5 mm, while it possessed less
effect against C. albicans R, with an inhibition zone diameter of
29.2 mm; and the inhibition zone diameter against S. aureus R
and C. albicans S was 30.7 mm and 32.0 mm respectively. In
Figure 2. FTIR of chitosan(Cht), Myrrh Extract(MR), Myrrh/AgNPs (MR/AgNPs), and (Cht/MR/AgNPs) nanocomposites.
Saad et al. 5
the meantime, Ag-NPs biosynthesized by Myrrh extract showed
high inhibitory action against S. aureus S with an inhibition zone
diameter of 31.1 mm and moderate inhibitory action against S.
aureus R and C. albicans S with an inhibition zone diameter of
27.5 mm and 28.5 mm, respectively, while it possessed less
effect against C. albicans R, with an inhibition zone diameter
of 26.2 mm. Compared with lower effect of commercial
Myrrh extract, recording inhibition zone diameters of
22.7 mm against C. albicans S, 19.1 mm against C. albicans R,
23.2 mm against S. aureus S and 20.8 mm against S. aureus R;
the synergistic effect of combined materials were evidenced.
All agents were synthesized in accordance with the approved
concentrations for use as conventional antibacterial and anti-
fungal treatments.
Antimicrobial Activity of Treated Cotton Textiles
In general, fabrics treated with Cht/MR/AgNPs demonstrated
greater antimicrobial activity against all strains compared to
those treated with MR extract and MR/AgNPs alone.
Moreover, fabrics treated with the composite Cht/MR/
AgNPs showed a significant enhancement in antimicrobial effi-
cacy against all tested strains compared to fabrics treated with
each individual agent. Among the strains tested, S. aureus
showed greater sensitivity compared to C. albicans, and
antibiotic-sensitive isolates exhibited higher susceptibility than
antibiotic-resistant strains to all treated fabrics, as illustrated in
Figure 4. The appeared dark spots on some textiles’surfaces
(eg C-2, S-2, C-3 and S-3) are typically attributed to the
loaded AgNPs, and not from growth of microbes onto textiles.
Scanning Electron Microscopy (SEM)
SEM imaging (Figure 5) was utilized to observe the morpholog-
ical changes in S. aureus and C. albicans following exposure to
minimum inhibitory concentrations (MICs) of Cht/MR/
AgNPs. Initially (Figure 5- 0), bacterial cells exhibited well-
preserved and intact cell walls. However, after 3 to 6 h of expo-
sure, signs of wall softening and notable distortions became
evident, accompanied by leakage of internal constituents
(Figure 5- 1,2). By the ninth hour mark, some treated S.
aureus and C. albicans cells were lysed (Figure 5- 3). After 12 h
of exposure, nearly all treated cells were lysed, with residual
cell walls appearing in conjunction with leaked internal compo-
nents and nanomaterials (Figure 5 - 4).
The cytotoxicity assay of Cht/MR/AgNPs nanocomposite
toward normal HDF (C-12302) cells using MTT method illus-
trated minimal biotoxicity of nanocomposites toward human
dermal fibroblasts, up to concentration of 50 mg/mL.
Discussion
The biosynthesis of Cht/MR/AgNPs nanocomposite presents
a promising strategy for effectively controlling skin pathogenic
microbes. The synthesized nanocomposite exhibited significant
antimicrobial activity against a range of microbial strains,
including S. aureus and C. albicans using a simple, economical,
and environmentally outgoing nanobiotechnological approach.
The usage of use nanosilver, despite its potential cytotoxicity
and carcinogenicity based on several reasons; their potent anti-
bacterial actions, their synthesis with biogenic materials (MR),
and their conjugation with biopolymers (Cht), which are
believed to greatly minimize their biotoxicity toward mamma-
lian cells,
18,20,25
and this was verified in current study. The
TEM imaging of MR-synthesized AgNPs appointed some
agglomeration, which occurred due to the constituents of the
extract aggregating and adhering to the surface of AgNPs.
25
The MR-synthsized AgNPs apperaed with well-despersion
and compositesd with MR residues (Figure 3). However the
biosynthesis process is suggested to be obtimized via incorpo-
rating of further stabilizing materials (eg biopolymers, thickning
agents, …), and by managing the zeta potential of synthsizing
nanometals through adjusting process pH and other conditions,
to diminsh the potential agglumeration of nanomaterials
17,20,25
SEM imaging revealed notable morphological changes in
bacterial and fungal cells following exposure to the
Table 1. Antimicrobial Performance of Myrrh (MR), Synthesized
Silver Nanoparticles Using Myrrh Extract (MR/AgNPs), and
Cht/MR/AgNPs Composites, Using Zone of Inhibition (ZOI; in mm)
Assays.
Treatment*
Inhibition zone (mm)**
Candida
albicans S
Candida
albicans R
Staphylococcus
aureus S
Staphylococcus
aureus R
MR 22.7 ±1.1 19.1 ±0.7 23.2 ±1.2 20.8 ±1.1
MR/AgNPs 28.5 ±1.2 26.2 ±1.4 31.1 ±1.5 27.5 ±1.3
Cht/MR/AgNPs 32.0 ±1.8 29.2 ±1.4 36.5 ±1.6 30.7 ±1.7
*Textiles were treated by 1% MR in addition to active compounds.
**Inhibition zones are triplicates means (including textile width of 20 mm)
standard deviation.
Figure 3. Transmission microscope screening of synthesized agNPs.
6Natural Product Communications
nanocomposite, indicative of its disruptive effects on cell walls
and internal constituents.
26
Moreover, the enhanced antimicro-
bial potential of the composite fabric highlights its potential for
applications in textile coatings aimed at preventing microbial
infections. Overall, the findings suggest that the Cht/MR/
AgNPs nanocomposite holds considerable promise as a thera-
peutic agent for combating skin pathogenic microbes.
The FTIR analysis demonstrated the effectiveness of bio-
organics derived from myrrh extract as capping or reducing
agents for AgNPs. These bio-organics included germacrene
B, limonene, curzerene, beta selinene, myrcenol, isocerice-
nine, and spathulenol.
25,27
The phytosynthesis of metals
nanoparticles was effectually gathered due to the reducing
powers of phyto-constituents and their capping potentialities
that generates homogenous NPs sizes with minimum
agglomeration.
28,29
Cht/MR/AgNPs nanocomposite could burst, destroy, or
break down bacterial and candidal cells. More research is
required to fully understand the mechanisms of such types of
Cht/MR/AgNPs nanocomposites’antibacterial and anticandi-
dal capabilities, additionally to its use as an antibacterial and
anticandidal agent.
16
The aim of the current study was to
extract and apply a variety of bioactive natural agents, such as
Cht, MR, and AgNPs, for the possible prevention of skin
Figure 4. Antimicrobial activity of treated cotton textiles with 1% from myrrh extract (1), and with nano silver synthesized from Myrrh extract (2),
and Cht/MR/AgNPs (3) against antibiotic resistant strains from Candida albicans (C) and Staphylococcus aureus (S).
Saad et al. 7
infections. This was advised because natural products have a
wide range of antimicrobial activities, are expected to be
biosafe, and are compatible with the human body.
23
The antimicrobial actions of fabricated nanocomposites are
suggested to involve many factors. The ratio between Cht and
MR/AgNPs imparts high importance for the net zeta potential
of nanocomposites; the higher Cht ratio derives nanocompo-
sites to positivity, whereas the higher MR/AgNPs ratio enforces
nanocomposites negativity. The composites charges (especially
the positive charges) trigger their attachment and interactions
with microbial cells that are negatively charged.
9,16,18
Additionally, the high ratio of MR-mediated AgNPs is the
Figure 5. SEM of treated Staphylococcus aureus (S) and Candida albicans (C) with cht/mr/agNPs nanocomposite for 0, 3, 6, 9 and 12 h (form 0 to 4 in
figure, respectively).
8Natural Product Communications
suggested main responsible for antimicrobial actions, due to syner-
gistic microbicidal actions of MR and AgNPs (including cellular
penetration, disruptions and disturbing their biosystems).
7,9,20,25
However, the suggested specific mechanisms that are
involved in the destruction of microbial cells by the Cht/
MR/AgNPs nanocomposite include the attachment of
Cht-based capsules onto cells surfaces (due to their opposite
charges) to disrupt cellular permeability, the release of MR/
AgNPs from the nanocomposite to enter the cells, and the
destructive interactions between each component (Cht, MR
and AgNPs) with the cellular segments (eg membranes,
enzymes, DNA, RNA, proteins, …).
9,16,18
AgNPs that have been produced and their antimicrobial
action in human skin cells may be key components of novel
anti-inflammatory and sore treatment medicines. The antibacte-
rial properties of green synthesized AgNPs made with myrrh
extract show more promise against a variety of harmful micro-
organisms.
30
For eco-friendly AgNP synthesis, myrrh extract
can be utilized as a green reducing and covering agent. Even
after four months, there were no discernible alterations to the
synthesized AgNPs, demonstrating their high stability.
Furthermore, when compared to myrrh extract, AgNPs exhib-
ited enhanced antifungal activity and high bacterial and fungal
activity against both S. aureus and C. albicans. AgNPs are adapt-
able and can be employed in a variety of biological and biomed-
ical applications as antimicrobial agents.
25
Regarding the promising attained antimicrobial activity of
MR-mediated AgNPs and their nanoconjugates with Cht, it
can be suggested to potentially improve the AgNPs antibacterial
actions to make them more effective against more resistant
microorganisms or emerging strain via biosynthsizing with
more effectual natural derivatives (contained higher amunts of
bioactive compouns), and compositing with aditional biopoly-
mers to diminsh their potential biotoxicity toward mammalian
cells.
31,32
MR was selected due to its well-established benefits; it has
astringent, antimicrobial, and anti-inflammatory properties. It
is a common ingredient in many skin care products due to its
unique characteristics and therapeutic advantages. It has mois-
turizing and soothing properties and can be used to relieve acne,
blisters, dryness, and inflammation.
33
Within this context, the
Cht/MR/AgNPs matrix has demonstrated numerous benefits
in terms of having favorable biological, chemical, and physical
properties for the defense of skin tissues. Because of their bio-
activities, compatibility, and structural enhancement of the pro-
duced composites, Cht/MR/AgNPs were recommended as
ideal nanocomposites for skin protection and were therefore
encouraged to be applied in tissue engineering and other skin
regeneration sectors.
34
The elevated biosafety of Cht/MR/AgNPs nanocomposites
toward normal HDF cells, after validating with MTT method,
could advocate their practical usage for human skin protection
and disinfection using these innovative natural materials.
1,23,34-37
The outstanding properties of Cht/MR/AgNPs components
can additionally promote their prospective assessment as
candidates for skin regeneration, especially after wounds/burns
injuries. Because of its amazing features, such as its anti-
inflammatory, biocompatible, antibacterial, and wound-healing
properties (such as cytokine production, fibroblast activation,
and promotion of My synthesis), Cht was also recommended
for skin protection.
35
Cotton fiber coated with Cht has the poten-
tial to accelerate wound healing by stimulating PMN (polymor-
phonuclear) cells to infiltrate the injured skin.
36
One important consideration for the bioactivity of treated
textiles’potential uses in the medical field is the longevity of
their antibacterial action.
6
Treated textiles containing Cht/
MR/AgNPs maintained much higher antimicrobial potentiali-
ties than those reported in other pertinent studies involving tex-
tiles loaded with AgNPs or other individual antimicrobial
agents.
36-38
This is likely due to their synergistic composition,
which strengthened the cross linkage with cotton fibers and pre-
vented their rapid loss.
39-41
The main limitations of this study
may include the need for assessing nanomaterials biosafety
using animal models, the evaluation of antimicrobial durability
of fabricated textiles after several washing cycles, the probable
inflammation consequence when applying coated textiles with
nanomaterials, and the coast effectiveness evaluation of pro-
duced materials.
Conclusion
The AgNPs was perfectly biosynthesized using MR with minute
size and homogenous dispersal. The Cht/MR/AgNPs nano-
composite was innovatively fabricated and characterized; the
antimicrobial actions of this nanocomposite were evidenced
toward different bacterial and mycotic skin pathogens. The
loaded textiles with Cht/MR/AgNPs nanocomposite exhibited
remarkable antimicrobial actions, where the nanocomposite
could destruct microbial cells and prevent their development.
The fabricated natural nanocomposites and their loaded
hygienic textiles are promisingly suggested for controlling path-
ogenic skin microbes. The prospective role of fabricated nano-
composites in tissue regeneration is recommended to be
investigated using in vivo, in situ and molecular studies. It is
also suggested to evaluate the stability of the nanocomposite
under different environmental conditions, such as prolonged
exposure to sunlight or extreme temperatures.
Acknowledgements
The authors declare their appreciation and foremost gratefulness to
ALLAH for the merciful and generous guiding throughout this work.
Author Contributions
HAS: data collection, methodology, prepared tables and figures, study
administration, writing the original manuscript. MA: conceptualiza-
tion, supervision, resources, written the original manuscript and cor-
rection of the original manuscript. AA: supervision, study
registration, edition. HAG: methodology, data analysis, writing the
original manuscript. AAT: methodology, data analysis
Saad et al. 9
conceptualization, supervision, resources, written the original manu-
script and correction of the original manuscript. All authors agreed
to submit the manuscript.
Data Availability
“The datasets used and analyzed during the current study are all pro-
vided in the manuscript.”
Declaration of Conflicting Interests
The authors declared no potential conflicts of interest with respect to
the research, authorship, and/or publication of this article.
Ethical Approval
“Ethical Approval is not applicable for this article.”
Funding
The authors received no financial support for the research, authorship,
and/or publication of this article.
Statement of Human and Animal Rights
“This article does not contain any studies with human or animal
subjects.”
Statement of Informed Consent
“There are no human subjects in this article and informed consent is
not applicable.”
ORCID iD
Ahmed A. Tayel https://orcid.org/0000-0001-9411-134X
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