processes

Review
Copper and Copper Nanoparticles Applications and Their Role
against Infections: A Minireview
Iliana A. Ivanova 1 , Dragomira S. Daskalova 1 , Lilia P. Yordanova 1 and Elitsa L. Pavlova 2,3, *

1 Faculty of Biology, Sofia University “St. Kliment Ohridski”, 8 Dragan Tsankov Blvd., 1164 Sofia, Bulgaria
2 Faculty of Physics, Sofia University “St. Kliment Ohridski”, 5 James Boucher Blvd., 1164 Sofia, Bulgaria
3 Center of Competence “Clean Technologies for Sustainable Environment—Water, Waste, Energy for Circular
Economy”, 1000 Sofia, Bulgaria
* Correspondence: [email protected]

Abstract: The focus of this review article is to present a retrospective analysis of copper applications
focusing on ions and nanoparticles as broad-spectrum antimicrobials. Copper nanoparticles are
presented as an alternative to rising antibiotic resistance. The basic mechanisms of bacterial, fungal,
and viral inactivation, which explain their potential, are presented. The green biosynthesis of copper
nanoparticles using biomaterials is also presented and considered a very promising trend for future
biotechnology and medical applications.

Keywords: antibacterial; antifungal; antiviral; copper; nanoparticles; coatings

1. Introduction and History of Copper Applications in the Prevention and Treatment

of Infections
The Smith Papyrus, the oldest known medical document in history, presents the first
documented use of copper as an anti-infection agent. The information there is attributed to
an Egyptian physician around 1700 BC, but is based on information even since 3200 BC. In
hieroglyphics, copper was denoted by the ankh symbol, which was used by the Egyptians
Citation: Ivanova, I.A.; Daskalova, and Indians to symbolize endless life. They used various forms of copper and copper
D.S.; Yordanova, L.P.; Pavlova, E.L. compounds to maintain hygiene for many ailments. In ancient times, the Egyptians used
Copper and Copper Nanoparticles to sterilize wounds and drink water by applying metals (silver, copper, etc.) [1].
Applications and Their Role against As early as 1600 BC, the Chinese utilized copper coins as medicine to cure bladder
Infections: A Minireview. Processes problems and stomach and heart aches. To keep battle wounds from infection, Phoenician
2024, 12, 352. https://doi.org/ sailors sprinkled shavings from their bronze swords inside.
10.3390/pr12020352 Women have known for thousands of years that storing water in copper vessels
Academic Editor: Fabio Carniato
protected their children from diarrhea, and kept this knowledge for the next generations.
The Romans described various medicinal applications of copper. Copper compounds were
Received: 14 December 2023 also used by the Aztecs to heal sore throats; in Persia and India, copper was applied to treat
Revised: 27 January 2024 venereal ulcers, boils, and eye infections [2].
Accepted: 5 February 2024 The power of copper’s antimicrobial effect is extraordinary. A team of scientists
Published: 7 February 2024
checked the old railings of the Grand Central Terminal in New York and found that the
copper still works just as well as the day it was installed more than a hundred years ago,
and the antimicrobial effect still remains.
Copper kills the bacteria that cause Legionnaires’ disease, as well as drug-resistant
Copyright: © 2024 by the authors.
Licensee MDPI, Basel, Switzerland.
microorganisms such as methicillin-resistant Staphylococcus aureus (MRSA). Viruses concern-
This article is an open access article
ing the global health community, such as the Middle East Respiratory Syndrome (MERS),
distributed under the terms and the 2009 swine flu pandemic (H1N1), and the 2019 pandemic (SARS-CoV-2) have been
conditions of the Creative Commons shown to die from contact with pure copper in minutes [3–8]. Ahmadi et al. revealed
Attribution (CC BY) license (https:// the antiviral efficacy of copper nanoparticles biosynthesized by Juglans regia green husk
creativecommons.org/licenses/by/ aqueous extracts and the synergistic action of copper and iron nanoparticles on viruses [9].
4.0/).

Processes 2024, 12, 352. https://doi.org/10.3390/pr12020352 https://www.mdpi.com/journal/processes

Processes 2024, 12, 352 2 of 13

Patoo et al. and Rai et al. [10,11] also contributed with proofs to the synergistic effects of
metal and metal oxide nanoparticles on SARS-CoV-2 and other viruses.

2. Methods for the Synthesis of Copper Nanoparticles

There are basically three groups of methods for nanoparticle synthesis: physical, chem-
ical, and biological [12]. One of the physical methods that is most preferred and popular
among authors is radio frequency magnetron sputtering. This is one of the most promising
methods for applying thin layers due to the possibility of obtaining a homogeneous coating
over a large area under controllable conditions [13]. The main disadvantage of this outdated
method is related to the high cost of nanoparticle synthesis [14]. This method also allows
the co-pulverization of the newly synthesized nanomaterials, which leads to an important
characteristic: the antimicrobial activity [15].
According to Hachem et al. [16], the synthesis of nanoparticles using chemical methods
is based on fundamental chemical reactions. These reactions are precipitation, oxidation,
hydrolysis, thermolysis, polymerization, and condensation [17]. Depending on the reac-
tions, several preferred chemical methods for synthesis stand out. One of them is plasma
chemical vapor deposition, which is used to deposit thin films from a gaseous to a solid
state over a substrate [18]. Another widely used technique is the “sol-gel” method. This
technique is applied to obtain metal oxides, mostly copper nanoparticles [19,20]. Chemical
deposition is one of the most economical techniques for acquiring nanoparticles because it
does not depend on expensive machinery and equipment [21]. “Green” chemical synthesis
is an innovative and relatively new method for the synthesis of metallic nanomateri-
als, where water, which contains environmentally friendly reducing agents, is used as a
chemical solvent [22].

3. Green Synthesis of Copper Nanoparticles

“Green synthesis” controls the size and shape as well as the production of stable
nanomaterials without harmful reagents. Biological synthesis is superior to chemical and
physical methods in terms of cost, environmental friendliness, and ease of synthesis screen-
ing. “Green synthesis” is ecological, economical, and safe and uses biological materials
such as cell cultures of bacteria, fungi, and plants [23]. Several factors affect this method,
including the temperature, pH, reaction time, volume of reagents added, and volume of the
biological extract. The interaction of these factors is critical for determining the shape and
size of the synthesized nanoparticles. Various plants and their parts are excellent producers
of copper nanoparticles, such as the leaf extract of Hagenia abyssinica L., the juice of Citrus
medica Linn., and Syzygium guineense [24–26]. The plants are excellent producers of copper
nanoparticles because they are widely available, easy to process, and a source of many
metabolites. They are rich in pharmacological constituents that act as both reducing and
capping agents in the process of synthesis [24–26].
Over the past few decades, chemical and physical synthesis approaches have been
widely used to produce nanoparticles [27]. However, they have been proven to be dan-
gerous to both the environment and human health because of the harsh chemicals used.
Biosynthesis technologies have been identified as potential and optimal alternatives. These
approaches hold promise in a variety of contexts, including green chemistry. For example,
copper nanoparticles can be produced in large quantities using Rhatany root extract as a
“renewable resource” [26]. The literature review reveals that biologically synthesized cop-
per nanoparticles using microbial substances instead of plant extracts are sparsely explored.
Future research can be focused in this direction because “green synthesis” appears to be
the future of nanotechnology synthesis.

4. Copper Ions and Nanoparticles

The majority of research on metal-based nano-antimicrobials has demonstrated that
the nanomaterial’s biological efficacy is much greater and longer than the metal salt’s bioac-
tivity. Numerous factors can affect this effect, including the size of the nanoparticles, the
 Processes 2024, 12, x FOR PEER REVIEW 3 of 14

4. Copper Ions and Nanoparticles

Processes 2024, 12, 352 3 of 13
The majority of research on metal-based nano-antimicrobials has demonstrated that
the nanomaterial’s biological efficacy is much greater and longer than the metal salt’s
bioactivity. Numerous factors can affect this effect, including the size of the nanoparti-
high
cles,surface-to-volume ratio of ultrafine
the high surface-to-volume ratio ofparticles,
ultrafinethe massivethe
particles, release of ions,
massive as well
release as the
of ions, as
unusual
well as features connected
the unusual to theconnected
features existence oftosurface stabilizers.
the existence of Anti-agglomeration
surface stabilizers.com- An-
pounds, in fact, have
ti-agglomeration frequentlyin
compounds, been
fact,shown
have to modify the
frequently ionshown
been releasetoofmodify
the nanoparticles
the ion re-
and, consequently, their antibacterial characteristics [23,27–30].
lease of the nanoparticles and, consequently, their antibacterial characteristics [23,27–30].
ItIthas
hasbeen
beendemonstrated
demonstrated that soluble
that soluble copper
coppercompounds
compounds exhibit superior
exhibit antibacterial
superior antibac-
activity against a variety of pathogens, such as viruses, bacteria, fungi,
terial activity against a variety of pathogens, such as viruses, bacteria, fungi, and and algae [5,7–9],
algae
all of which
[5,7–9], all ofare generally
which safe forsafe
are generally medical implants
for medical and human
implants and humanconsumption
consumption[31–33].
[31–
Copper primarily enters microbial cells in the form of ions [28–30]. It alters
33]. Copper primarily enters microbial cells in the form of ions [28–30]. It alters the pH the pH level
and
levelfunction of theofmembrane
and function the membranechannels, destroys
channels, the functional
destroys groups
the functional and and
groups structure
struc-
of the cell and inner membranes, and produces reactive oxygen
ture of the cell and inner membranes, and produces reactive oxygen species (ROS) species (ROS) that can
that
cause irreversible
can cause damages
irreversible like protein
damages oxidation,
like protein the rupture
oxidation, theofrupture
crucial internal molecules
of crucial internal
like enzymes, DNA, and RNA, and the destruction of the entire cell membrane due to
molecules like enzymes, DNA, and RNA, and the destruction of the entire cell membrane
free-radical lipid peroxidation (Figure 1).
due to free-radical lipid peroxidation (Figure 1).

Figure1.1.Effect
Figure Effectof
ofcopper
coppernanoparticles
nanoparticlesand
andions
ionson
onaamicrobial
microbialcell.
cell.

Longano
Longanoetetal.al.[34]
[34]have
haveoutlined
outlined several methods
several methods forfor
producing
producing copper nanoparticles
copper nanoparti-
with
cles varying sizes, shapes,
with varying and coatings
sizes, shapes, attachedattached
and coatings to variousto polymers. Torras andTorras
various polymers. Roig [35]
and
present contemporary techniques for using microwaves to manufacture
Roig [35] present contemporary techniques for using microwaves to manufacture copper copper and alloyed
copper particles
and alloyed and trace
copper their and
particles photocatalytic
trace theiractivity. One of the
photocatalytic most prevalent
activity. One of the uses of
most
this metal in contemporary healthcare facilities is the control of Legionella
prevalent uses of this metal in contemporary healthcare facilities is the control of Le- sp. in hospital
water distribution
gionella systems
sp. in hospital by the
water ionization method
distribution systemsofbycopper and silvermethod
the ionization [36–40].ofA copper
broad
spectrum of bacteria, including spore-forming Bacillus subtilis, Salmonella enterica,
and silver [36–40]. A broad spectrum of bacteria, including spore-forming Bacillus subtilis, Campi-
lobacter jejuni,
Salmonella Escherichia
enterica, coli, and Staphylococcus
Campilobacter aureus,coli,
jejuni, Escherichia are susceptible to the antimicrobial
and Staphylococcus aureus, are
activity of copper ions originating from nanoparticles. Moreover,
susceptible to the antimicrobial activity of copper ions originating from 99.9% of most bacteria
nanoparticles.
are killed within two hours of contact [37–39,41,42]. Additionally, this
Moreover, 99.9% of most bacteria are killed within two hours of contact [37–39,41,42]. metal occasionally
exhibits superior
Additionally, thisqualities in comparison
metal occasionally to other
exhibits precious
superior metalsinwith
qualities antibacterial
comparison ac-
to other
tivity, like platinum and gold [40,43]. By direct contact, bacteria, yeasts, and
precious metals with antibacterial activity, like platinum and gold [40,43]. By direct con- viruses can
be eliminated from copper surfaces. According to reports, contact death happens over a
tact, bacteria, yeasts, and viruses can be eliminated from copper surfaces. According to
prolonged incubation period and at a pace of at least seven to eight orders of magnitude
reports, contact death happens over a prolonged incubation period and at a pace of at
per hour. There were no live microbes found on copper surfaces. This gives rise to the
least seven to eight orders of magnitude per hour. There were no live microbes found on
concept of self-cleaning materials made of copper [36]. Additionally, copper is a vital
copper surfaces. This gives rise to the concept of self-cleaning materials made of copper
trace element for human health since it is regarded as a metal that is not too harmful to
[36]. Additionally, copper is a vital trace element for human health since it is regarded as
humans and is utilized in intrauterine and other devices [37,38]. Some authors claim that
copper nanoparticles cause severe toxicological effects and serious damage to the kidneys,
liver, and spleen of experimental mice, but micro-copper particles do not do so in the
majority [40]. There is a relationship between the characteristics of nanomaterials and their
Processes 2024, 12, 352 4 of 13

nanotoxicity [43]. Several factors, such as the specific surface area of the nanoparticles,
their high reactivity, the overexpression of H+ , etc., are predicted to probably cause the
severe nanotoxicity observed in vivo [43]. It should be experimented further in vivo by
green synthesized copper nanoparticles to check their toxicity completely.
Synthesized copper nanoparticles against Escherichia coli, Pseudomonas aeruginosa,
Staphylococcus aureus, and Bacillus subtilis have exceptional antibacterial capabilities, thanks
to an extract from the leaves of the herb Hagenia abyssinica [44]. According to Ganga’s re-
search, gram-positive bacteria like Staphylococcus aureus and Bacillus subtilis, as well as gram-
negative bacteria like Escherichia coli and Klebsiella pneumoniae, were effectively neutralized
by green-synthesized copper nanoparticles derived from Riccia fluitans bryophytes [45].
From all the combinations tested against the targeted bacterial strains, copper nanoparticles
demonstrated the strongest antibacterial efficacy. Similar outcomes were observed by
Malaikozhundan et al. [46] when copper nanoparticles were synthesized using Mentha
spicata leaf extract to inhibit Klebsiella pneumoniae, Escherichia coli, and the gram-positive
Streptococcus pyogenes and Staphylococcus aureus.

5. Gene Expression after Nanoparticle Interaction

Reverse transcriptase polymerase chain reaction about gene expression induced the
activation of copper oxidase, converting the toxic first-valence copper ions into less toxic
second-valence ions. The activation was observed in the fifth minute of the interaction of
bacterial cells with copper and silver ions. A statistically significant activation of copper
oxidase was found in E. coli ATCC 10536 and in the presence of titanium dioxide thin
films embedded with copper and silver nanoparticles by RT-qPCR, despite the significantly
lower concentration of ions released from the thin film.
The representativeness of these results is ensured by the use of two types of negative
controls: the so-called clear glass plate (uncoated but cleaned as for coating) and the
titanium dioxide-coated one. The gene expression was similar when tested against the
control, pure glass, or TiO2 . This allows us to conclude that TiO2 does not act as a stress-
determining factor and that the occurrence of stress responses is entirely due to the copper
and silver nanoparticles entering the component composition of the thin films’ coatings.
This confirms that titanium nanoparticles or dissolved ions do not have a suppressive effect
on the bacterium. It proves that the dissolution of metal ions from the atomic clusters of
copper and silver nanoparticles is responsible for their combined toxic effect [47].
When analyzing the data for 30 and 60 min, the results are comparable. The effect
is weaker with time, i.e., the expression of copper oxidase CuO in the first minutes of
the treatment is most noticeable. The tendency for a decrease in the expression of copper
oxidase with time is clear. The result in the 5th minute shows the rapid reaction of the
bacteria to the nanomaterials and the stress response. Nine-fold activation is achieved at
the 5th minute, and seven-fold activation is achieved at the 30th minute [47].

6. Combinations between Copper, Copper Oxide, and Other Nanoparticles

Copper nanoparticles could be applied to critical points in some key industries to im-
prove the quality of protective clothing [48]. Several reported studies have given promising
results. The Cu-SiO2 nanocomposite, copper-doped hydroxyapatite nanopowders, alginate
nano-copper cotton cellulose fibers, and copper nanoparticle-treated fibers were all tested
for antibacterial activity using the agar diffusion test [34]. The antibacterial performance
results of the fibers loaded with nano-copper demonstrated a distinct zone of inhibition
surrounding the visible copper nanoparticle fiber bundle and a reduced number of micro-
biological colonies. Grace et al. also reported another clear concentration effect [49]. The
same scientists demonstrated in a different work that cotton-cellulose fibers treated with
nano-copper alginates efficiently prevented the growth of bacteria [31]. CuO nanocompos-
ites completely inhibited the development of E. coli cells when applied to cotton fibers [50].
The addition of CuO nanoparticles at doses of 0.01–0.5% and 1% to adhesive composite
trans bond XT increased its antibacterial properties against Streptococcus mutans. The shear
Processes 2024, 12, 352 5 of 13

bond strength of the metal brackets linked to the human premolars did not exhibit any
negative consequences [51].
Copper alloys can be created by combining copper nanoparticle coatings with other
metals. The US Environmental Protection Agency has registered over 450 alloys that are
resistant to six different kinds of bacteria, including MRSA, VRE (Vancomycin-resistant
Enterococcus faecium), P. aeruginosa, S. aureus, and E. coli. Copper-containing coatings can
be applied to any surface that is in contact with pathogens and come in a variety of
colors and shapes. They do not need expensive extra cleaning techniques and are robust,
recyclable, and sustainable. These surfaces work incredibly well; MRSA at 106 CFU/mL
was eliminated in 90 min at 25 ◦ C on a C197 copper surface when it was contaminated
by sneezing and splashing. For clean copper surfaces with 106 CFU at 22 ◦ C, a 45-min
elimination period in a dry touch simulation yields a faster effect. At 4 ◦ C, complete
disinfection is accomplished in 6 h [52].
At 20 ◦ C and 50% relative humidity, copper surfaces were more effective in eradicating
MRSA than silver and iron-containing materials and stainless-steel surfaces, with almost
no live bacteria remaining 75 min after the microbial application to the studied surface [53].
Within 10 min, an even quicker bactericidal response was seen for vancomycin-resistant
Enterococcus (VRE) [54]. The increased effectiveness of contact bacterial inactivation on
copper surfaces evaluated at higher temperatures and humidity levels is demonstrated by
this and other research. Copper nanomaterials generally exhibit quick and wide-ranging
biocidal action against a variety of microorganisms, including viruses (Influenza A and
Norovirus), fungi (Candida albicans, Penicillium sp.), bacteria (Legionella pneumophila, E. coli,
and Clostridium difficile—both vegetative cells and spores), as well as resistant strains of
these organisms (MRSA, VRE) [2,55]. Due to their multiple non-specific modes of action
and quick bactericidal effect, copper-containing contact surfaces are preferred in the fight
against resistance in comparison to chemical antibacterial treatments [56,57].
The agar diffusion of the Cu-SiO2 nanocomposite was determined by placing a paper
disk saturated with different nanocomposite aqueous suspensions on an agar surface in-
oculated with E. coli (ATCC 25922). Our result showed that the filter paper prevented the
nanoparticles’ diffusion into the nutrient medium, but the nanocomposite had a stronger
antibacterial effect than the copper nanoparticles [57]. The antibacterial activity of Cu-SiO2
nanocomposites depends on the good distribution and lack of aggregation of the copper
nanoparticles on the surface of the SiO2 nanoparticles [58]. The effectiveness of nanocom-
posites is also confirmed by the work of Maniprasad and Santra [59], who investigated the
antibacterial activity of copper nanoparticles deposited on SiO2 particles. They estimated
a minimum inhibitory concentration (MIC) of 2.4 µg of Cu metal/mL against both E. coli
and B. subtilis. Antibacterial studies clearly present that Cu/SiO2 nanocomposites are more
effective compared to insoluble Cu(OH)2 at equivalent concentrations of copper, indicating
the higher bioavailability of ions (i.e., more soluble Cu) in Cu/SiO2 nanocomposites, deter-
mined by the “core-shell” structure. The antimicrobial test of copper-doped hydroxyapatite
nanopowders against E. coli ATCC 25922 by the agar diffusion method and in liquid media
showed that all metal-doped hydroxyapatite samples caused the death of viable bacterial
cells [60]. Another study by Yaseen et al. [61] reported that the CuO-SiO2 -nanocomposite
had good photocatalytic activity, anti-leishmanial activity, and antioxidant activity, i.e., had
promising biological attributes.
Compared to silver, copper coatings have been the subject of less investigated antibac-
terial activity; however, a Cu/TiO2 thin film was investigated under low UV light using
E. coli that is resistant to copper ions. Sunada [62] showed that the synergistic action of the
photocatalytic activity of TiO2 and that of copper ions produced a bactericidal impact of the
Cu/TiO2 combination even on copper-resistant E. coli cells under very low UV irradiation.
Cu-doped TiO2 showed superior antibacterial activity against E. coli when compared to
undoped TiO2 , as reported by Mingmongkol et al. [63]. However, the high Cu doping
concentrations of 1.0 wt.% and Cu-doped TiO2 exhibited detrimental effects on the charge
transfer, ·OH generation, and surface charge of the nanoparticles.
Processes 2024, 12, 352 6 of 13

Several authors reported about various formulations and combinations of silver and
copper nanoparticles and their synergistic effects on different bacteria [64,65]. Other authors
reported on the immunomodulatory effects of copper on some proteins, influencing innate
host defenses. Zinc, copper, and silver appear to be the metals with the greatest activity
in regard to the increased concentration of nitric oxide (NO·), a kind of reactive nitrogen
species that has synergistic effects on host immune systems against pathogens [66–68].
Nitric oxide can oxidize copper too. It is present in protective proteins. It may cause the
release of copper ions that boost the toxicity against microbial and tumor cells [66,69].

7. Combination of Copper with Polymers for Antimicrobial Effect

Using electro-synthesized copper nanoparticles dissolved in various polymers, as well
as ion-deposited composites (Ion Beam Sputtering technique) and copper fluoropolymer
nanocomposite films, Cioffi et al. [27] conducted biological studies on E. coli. Copper
nanoparticles stabilized with tetra-butyl-ammonium perchlorate were found to have clear
biostatic activity on Saccharomyces cerevisiae, Escherichia coli, Staphylococcus aureus, and Lyste-
ria sp. when they were embedded in polymer matrices like polyvinyl-methyl-ketone, PVC
(polyvinyl chloride), and polyvinylidene fluoride. Furthermore, it was consistently noted
that copper nanoparticle-polyvinylidene fluoride films had the least effective biostatic ac-
tion, while copper nanoparticle-polyvinyl-methyl-ketone films exhibited the best biostatic
effect [27]. On the other hand, the ammonium salt had a substantial growth inhibition
impact and significantly reduced the growth of bacteria in the cases of copper nanoparti-
cles stabilized by tetra-octyl-ammonium chloride and embedded in polymer matrices like
polyvinyl-methyl-ketone. It is interesting to note that the simultaneous action of implanted
metal nanoparticles and tetra-octyl-ammonium chloride resulted in increased biological
activity for these nanocomposites. Furthermore, because of the exceptionally high concen-
tration of unattached copper atoms, copper nanopaticle-polyvinylidene fluoride coatings
demonstrated even greater disinfection efficacy against E. coli [27].
The antibacterial properties of copper-encrusted multi-walled carbon nanotubes (Cu-
MWCNT) were also evaluated in compliance with ISO 14729:2001 (EU) regulations [70].
At a dose of 21 µg/mL, pure MWCNTs showed a comparatively poor impact against E.
coli (20 ± 2.5%). When compared to pure copper nanoparticles, combined Cu-MWCNTs
demonstrated a good level of antibacterial efficiency (~75%) and around 52% bactericidal
activity [71]. Copper nanoparticles were functionalized with polyethyleneimine (PEI) and
4-aminobutyric acid (GABA) by Jardón-Maximino et al. [32]. These composites with 2, 5,
and 5.0 wt.% of Cu showed outstanding antibacterial action against Staphylococcus aureus
and Pseudomonas aeruginosa. By employing bile juice to create a hemocompatible composite
Cu-MWCNT, Mondal and Mondal [33] demonstrated potent antibacterial activity against
S. aureus and P. aeruginosa biofilms, with minimum inhibitory concentrations (MIC) of 24
and 16 µg/mL.
At a lower dosage of approximately 3 µg of Cu per gram composite gel, agarose-Cu
nanoparticle composites at two distinct concentrations (3 and 6 µg of Cu per milliliter of
LB broth) demonstrated full suppression against E. coli [72].

8. Copper Oxide’s Biological Effects

According to Perelshtein et al. [50], copper oxide has a very high activity (1.4 wt.%),
completely inactivating S. aureus and E. coli in just 3 h. The toxicity of zinc oxide and
titanium oxide was compared with that of nano-sized and powdered copper oxide on the
bacteria Vibrio fischeri, the water flea Daphnia magna, and the freshwater crustacean Tham-
nocephalus platyurus. Compared to the other nanoparticles, the copper nanoparticles were
more harmful [73]. The antibacterial properties of cuprous oxide nanoparticles produced
with linear low-density polyethylene by co-extrusion, thermal adhesion, and attachment
to epoxy resin, three-methoxy-vinyl-silane, and ethyl cyanoacrylate were examined by
Gurianov et al. [74]. The impacts of the nanocomposites were ranked as follows: thermal
adhesion > ethyl cyanoacrylate > trimethoxyvinylsilane > epoxy resin > extrusion > epoxy
Processes 2024, 12, 352 7 of 13

resin > produced by extrusion. The composites that exhibit the highest activity may find
application as materials for the disinfection of waste-water and tap water.
An enhanced antimicrobial effect was obtained with a combination of TiO2 with CuO
and two-layer structures of Ag and CuO in combination with TiO2 . [57,75–77]. Structures
where Ag and CuO are inlaid on TiO2 have higher biocidal activity.
Fan et al. [68] reported the synergistic antimicrobial effect of copper and silver nanopar-
ticles when mixed and alloyed. The antimicrobial behavior is even better enhanced in
Ag–Cu nanoparticles.

9. Mechanisms of Antimicrobial Activity

The antimicrobial properties are attributed to the release of ions from the surface of
the metal copper, copper oxide, and nanoparticles, the rate of which mainly depends not
only on the chemical composition of the material, but also on the reactive oxygen and
nitrogen species (ROS and NS) that are formed. Several mechanisms of action could be
outlined [28,42,78].
Copper ions hinder the development of bacterial resistance through many mechanisms
simultaneously. The bacterial membranes become damaged when they come into contact
with copper surfaces. Copper ions have the ability to directly harm bacterial proteins
and, via a Fenton-like reaction, generate extremely reactive hydroxyl radicals (·OH) that
interact with other proteins, DNA, and enzymes to cause lipid peroxidation and the
destruction of membrane structures [42,79–81]. CuO nanoparticles have been shown to
promote advantageous outcomes, including enhanced angiogenesis and wound healing.
Through surface functionalization, the surface charge of the nanoparticle can be altered,
which contributes to the inactivation of bacteria. Positively charged nanoparticles may
bind faster and easily kill bacteria because the bacterial cell wall is, in general, negatively
charged [17,28].
An even stronger antimicrobial effect was observed when combining chitosan with cop-
per nanoparticles, and their size depends on the concentration of chitosan [82]. Vanti et al. [83]
established 98% fungal growth inhibition at a 0.1% concentration of economically feasible
chitosan-coupled copper nanoparticles. The study emphasizes the benefits of synthesized
chitosan-copper nanoparticles on agricultural crops as fungicides and growth promot-
ers. It can be a safe alternative to pesticides in order to avoid hazardous effects on the
environment [55].
Copper nanoparticles have the potential to be used as antibacterial agents; however,
when administered as an aerosol, they cause severe inflammatory reactions and cellular
damage. This toxicity may be decreased by coating metal nanoparticles with polysac-
charides, such as the antibacterial and biocompatible chitosan. In vitro experiments have
demonstrated a considerable reduction in the ROS production and the toxicity of copper
nanoparticles after 24 and 52 h when coated with chitosan. In contrast, mice exposed to
chitosan-coated as opposed to uncoated copper nanoparticles showed an increase in the
inflammatory response presented by the total protein, cytokines/chemokines, and white
blood cell count in their broncho-alveolar fluid. These findings imply that covering metal
nanoparticles with muco-adhesive polysaccharides (chitosan, for example) may expand
their potential for use in the regulated release of copper ions; nevertheless, if inhaled, this
will lead to a greater inflammatory response [84,85]. Other studies on the copper-chitosan
nanocomposite showed that it had no cytotoxicity on the normal cell line, but exhibited
very low cell viability against bladder cancer (UM-UC-3 (transitional cell carcinoma)),
squamous cell carcinoma, and transitional cell carcinoma Grade IV cell lines [86].
Copper nanoparticles are mainly used in antiseptic technical materials such as var-
nishes, paints, and other coatings; for ointments with antimicrobial action in medicine;
and also for antiseptic materials in wound healing [28]. In agriculture, the nano-crystalline
powders of iron, cobalt, and copper in extremely low concentrations increase the germina-
tion rate of seeds as well as the chlorophyll index, the number of nodules, and the yield of
the crop [87].
Processes 2024, 12, 352 8 of 13

In vitro studies in mice proved that a dose of 430 mg/kg live weight of 23.5 nm copper
nanoparticles is lethal. The targets of the attack were the liver, spleen, and kidneys [88].
Exposure to concentrations of 10, 25, and 50 µg/mL of CuO nanoparticles in human pul-
monary epithelial cells (A549) resulted in the depletion of glutathione and the stimulation
of catalase, lipid peroxidation, and superoxide dismutase activities. The CuO nanoparticles
reduced cell viability to 75, 66, and 48%, respectively, according to the MIT assay [89].
The inclusion of copper atoms in the layers of TiO2 nanoparticles shifts the light
absorption to visible light, and therefore irradiation with ultraviolet light is not necessary.
The antimicrobial effect is preserved with irradiation with visible light, achieving high
bacterial inactivation in 30 min-99.9999% [90]. Qin et al. [91] conducted experiments
comparing the antibacterial action of three oxides. The results show that copper oxide is
most effective, followed by zinc and finally titanium. Copper oxide nanoparticles work in
low concentrations, and it is believed that their action is due to the released copper ions,
the generation of ROS, membrane cell disruption, protein and mitochondrial damage, and
DNA dysfunction [42].
According to Sandle [92], the control of surfaces as vectors for pathogen transmission
increasingly forms part of the overall infection control strategy. In a real hospital setting,
various studies have shown that in rooms equipped with copper surfaces, after proper
pre-positioning, a reduction in the bacterial load and infection rate is observed [93]. In
some studies, it was found that the contact inactivation of bacteria was observed at a rate
of 7–8 log/h, with no surviving microorganisms after only a few hours of contact [94].
In 2010, a similar study was conducted at a hospital in Birmingham, UK, where some of
the surfaces were replaced with copper alloys. These have been used for an experimental
composite toilet seat as well as other key risk items such as door locks, etc.
All the data collected prove that bacteria on copper surfaces are 90–100% less compared
to similar plastic and aluminum surfaces. Based on this evidence, many hospital facilities
are beginning to use copper or copper alloys for touch surfaces such as door handles,
switches and control buttons, tables, bed boards, and hospital cabinets. Furthermore, the
use of copper nanoparticles is not limited to hard surfaces; some manufacturers are targeting
CuO nanoparticle-impregnated textiles for beds, pillowcases, and hospital clothing for
patients and healthcare workers. A clinical trial in intensive care units at Calama Hospital
in Chile reported a similar reduction. An ambulatory study confirmed the reduction in
the microbial load, resulting in reduced contamination in close proximity to the copper
surfaces. The estimate shows a 40% reduction in ICU-acquired infections, with the potential
for a reduction of up to 70%. This also leads to reduced care costs and improved patient
outcomes [95,96]. After 6 h of exposure, Cu-containing ZnO films led to a strong reduction
in the number of viable E. coli bacteria in a hospital environment [96–98].
Airborne pathogens are also risky, as are surfaces. Copper nanoparticles are used in
heating, ventilation, and air conditioning systems in hospitals. Typically, these systems
use aluminum components. Depending on the design and outdoor air pollution, these
components can promote the development of resistant biofilms of bacteria and fungi on
heat exchanger coils, fins, condensate drains, air ducts, etc. [55,99,100].

10. Conclusions
There is a strong connection between the past applications of copper as a metal and the
continuing interest in this material in the following new forms: nanoparticles, alloys with
other metals, and combinations with other materials such as polymers, carbon nanotubes,
etc. Nanocomposites accumulate promising results, especially in combined activity and
synergistic effects with silver, zinc, silica, and titanium dioxide, including ferrite nanoparti-
cles. The nanocomposites with chitosan and other polymers have the advantage of causing
less toxicity to mammals but present strong antibacterial, antiviral, and antifungal effects.
Copper is improving the immune system’s defense against pathogens. Copper will become
one of the most precious metals for human well-being. Green synthesized nanoparticles and
Processes 2024, 12, 352 9 of 13

nanocomposites could be nontoxic for humans as antimicrobials, bioremediation agents,

and growth factors for agricultural plants.

11. Future Directions

The future application of green synthesized copper nanoparticles could improve the
antimicrobial effect of combined copper nanocomposites and diminish their toxic effect on
mammalian cells [34,101,102]. The copper nanoparticles obtained by green synthesis using
tea leaf extracts could be used not only for biomedical applications, but also for water biore-
mediation [103]. Authors obtain copper oxide nanoparticles by “green synthesis” using,
for instance, the leaf tissue from Camellia sinensis L. and lavender (Lavandula anguistifolia).
It is proven that they have a successful effect at the lowest concentrations (4 µg mL−1 )
in the root and shoot development of lettuce and tomato seedlings [104,105]. So, these
nanomaterials could be used in the future for bioreclamation and as new fertilizers. The
literature review reveals that biologically synthesized copper nanoparticles using plant
extract are sparsely explored, and much future research can be conducted in this direction
as “green synthesis” is the future of nanotechnology.

Author Contributions: I.A.I., D.S.D. and L.P.Y. have gathered the information and written the text;
E.L.P. has reviewed and edited the whole text and supported its publication financially. All authors
have read and agreed to the published version of the manuscript.
Funding: The authors acknowledge the financial support of the following projects: “Clean technolo-
gies for sustainable environment-waters, waste, energy for circular economy”, Ministry of Education
and Science, Bulgaria, Contract Number: BG05M2OP001-1.002-0019, “Study of the qualities of
borehole waters in Sofia and small settlements around the capital”, Scientific Research Fund, Sofia
University “St. Kliment Ohridski”, Bulgaria, Contract Number: 80-10-174/16.05.2023.
Data Availability Statement: The data presented in this study are available on request from the
corresponding author.
Conflicts of Interest: The authors declare no conflict of interest.

References
1. Wade, K. The sword and the knife: A comparison of ancient Egyptian treatment of sword injuries and present day knife trauma.
Res Med. J. R. Med. Soc. 1957, 24, 47–56. [CrossRef]
2. Grass, G.; Rensing, C.; Solioz, M. Metallic copper as an antimicrobial surface. Appl. Environ. Microbiol. 2011, 77, 1541–1547.
[CrossRef]
3. Vincent, M.; Duval, R.E.; Hartemann, P.; Engels-Deutsch, M. Contact killing and antimicrobial properties of copper. J. Appl.
Microbiol. 2018, 124, 1032–1046. [CrossRef]
4. Hutasoit, N.; Kennedy, B.; Hamilton, S.; Luttick, A.; Rashid, R.A.R.; Palanisamy, S. Sars-CoV-2 (COVID-19) inactivation capability
of copper-coated touch surface fabricated by cold-spray technology. Manuf. Lett. 2020, 25, 93–97. [CrossRef]
5. Merkl, P.; Long, S.; McInerney, G.M.; Sotiriou, G.A. Antiviral activity of silver, copper oxide and zinc oxide nanoparticle coatings
against SARS-CoV-2. Nanomaterials 2021, 11, 1312. [CrossRef]
6. Jagaran, K.; Singh, M. Nanomedicine for COVID-19: Potential of copper nanoparticles. Biointerface Res. Appl. Chem. 2021, 11,
10716–10728. [CrossRef]
7. Poggio, C.; Colombo, M.; Arciola, C.R.; Greggi, T.; Scribante, A.; Dagna, A. Copper-alloy surfaces and cleaning regimens against
the spread of SARS-CoV-2 in dentistry and orthopedics. From fomites to anti-infective nanocoatings. Materials 2020, 13, 3244.
[CrossRef]
8. Ramos-Zúñiga, J.; Bruna, N.; Pérez-Donoso, J.M. Toxicity mechanisms of copper nanoparticles and copper surfaces on bacterial
cells and viruses. Int. J. Mol. Sci. 2023, 24, 10503. [CrossRef] [PubMed]
9. Ahmadi, M.; Elikaei, A.; Ghadam, P. Antiviral activity of biosynthesized copper nanoparticle by Juglans regia green husk aqueous
extract and iron nanoparticle: Molecular docking and in-vitro studies. Iran J. Microbiol. 2023, 15, 138–148. [CrossRef] [PubMed]
10. Patoo, T.S.; Khanday, F.; Qurashi, A. Prospectus of advanced nanomaterials for antiviral properties. Mater. Adv. 2022, 3, 2960–2970.
[CrossRef]
11. Rai, M.; Deshmukh, S.D.; Ingle, A.P.; Gupta, I.R.; Galdiero, M.; Galdiero, S. Metal nanoparticles: The protective nanoshield against
virus infection. Crit. Rev. Microbiol. 2016, 42, 46–56. [CrossRef]
12. Nam, N.H.; Luong, N.H. Nanoparticles: Synthesis and applications. Mater. Biomed. Eng. 2019, 211–240. [CrossRef]
13. Al-Ghamdi, A.A.; Khedr, M.H.; Ansari, M.S.; Hasan, P.M.Z.; Abdel-Wahab, M.S.; Farghali, A.A. RF Sputtered CuO thin films:
Structural, optical and photo-catalytic behavior. Phys. E Low-Dimens. Syst. Nanostructures 2016, 81, 83–90. [CrossRef]
Processes 2024, 12, 352 10 of 13

14. Kim, D.Y.; Kwon, J.H.; Jang, G.S.; Hwang, N.M. Effect of pressure on the film deposition during RF magnetron sputtering
considering charged nanoparticles. Coatings 2021, 11, 132. [CrossRef]
15. Pakpum, C. Preparation of copper thin film mask by sputtering technique assisted by polymer mask photolithography. IOP Conf.
Ser. Mater. Sci. Eng. 2017, 213, 012023. [CrossRef]
16. Hachem, K.; Ansari, M.J.; Saleh, R.O.; Kzar, H.H.; Al-Gazally, M.E.; Altimari, U.S.; Hussein, S.A.; Mohammed, H.T.; Hammid,
A.T.; Kianfar, E. Methods of chemical synthesis in the synthesis of nanomaterial and nanoparticles by the chemical deposition
method: A review. BioNanoScience 2022, 12, 1032–1057. [CrossRef]
17. Gawande, M.B.; Goswami, A.; Felpin, F.X.; Asefa, T.; Huang, X.; Silva, R.; Zou, X.; Zboril, R.; Varma, R.S. Cu and Cu-based
nanoparticles: Synthesis and applications in catalysis. Chem. Rev. 2016, 116, 3722–3811. [CrossRef] [PubMed]
18. Dörner, L.; Cancellieri, C.; Rheingans, B.; Walter, M.; Kägi, R.; Schmutz, P.; Kovalenko, M.V.; Jeurgens, L.P. Cost-effective sol-gel
synthesis of porous CuO nanoparticle aggregates with tunable specific surface area. Sci. Rep. 2019, 9, 11758. [CrossRef]
19. Etefagh, R.; Azhir, E.; Shahtahmasebi, N. Synthesis of CuO nanoparticles and fabrication of nanostructural layer biosensors for
detecting Aspergillus niger fungi. Sci. Iran. 2013, 20, 1055–1058. [CrossRef]
20. Patel, M.; Mishra, S.; Verma, R.; Shikha, D. Synthesis of ZnO and CuO nanoparticles via sol gel method and its characterization
by using various technique. Discov. Mater. 2022, 2, 1. [CrossRef]
21. Carlsson, J.-O.; Martin, P.M. Chemical vapor deposition. In Handbook of Deposition Technologies for Films and Coatings, 3rd ed.;
Martin, P.M., Ed.; Elsevier: Amsterdam, The Netherlands, 2010; Chapter 7; pp. 314–363. ISBN 9780815520313. [CrossRef]
22. Bhattacharya, D.; Gupta, R.K. Nanotechnology and potential of microorganisms. Crit. Rev. Biotechnol. 2005, 25, 199–204.
[CrossRef]
23. Singh, A.; Gautam, P.K.; Verma, A.; Singh, V.; Shivapriya, P.M.; Shivalkar, S.; Sahoo, A.K.; Samanta, S.K. Green synthesis of
metallic nanoparticles as effective alternatives to treat antibiotics resistant bacterial infections: A review. Biotechnol. Rep. 2020,
25, e00427. [CrossRef]
24. Ben Mosbah, M.; Alsukaibi, A.K.D.; Mechi, L.; Alimi, F.; Moussaoui, Y. Ecological synthesis of CuO nanoparticles using Punica
granatum L. peel extract for the retention of methyl green. Water 2022, 14, 1509. [CrossRef]
25. Shende, S.; Ingle, A.P.; Gade, A.; Rai, M. Green synthesis of copper nanoparticles by Citrus medica Linn. (Idilimbu) juice and its
antimicrobial activity. World J. Microbiol. Biotechnol. 2015, 6, 865–873. [CrossRef]
26. Jahan, I.; Erci, F.; Isildak, I. Facile microwave-mediated green synthesis of non-toxic copper nanoparticles using Citrus sinensis
aqueous fruit extract and their antibacterial potentials. J. Drug Deliv. Sci. Technol. 2021, 61, 102172. [CrossRef]
27. Cioffi, N.; Ditaranto, N.; Torsi, L.; Picca, R.A.; Sabbatini, L.; Valentini, A.; Novello, L.; Tantillo, G.; Bleve-Zacheo, T.; Zambonin,
P.G. Analytical characterization of bioactive fluoropolymer ultra-thin coatings modified by copper nanoparticles. Anal. Bioanal.
Chem. 2005, 381, 607–616. [CrossRef] [PubMed]
28. Vasiliev, G.; Kubo, A.-L.; Vija, H.; Kahru, A.; Bondar, D.; Karpichev, Y.; Bondarenko, O. Synergistic antibacterial effect of copper
and silver nanoparticles and their mechanism of action. Sci. Rep. 2023, 13, 9202. [CrossRef]
29. Zhang, E.; Zhao, X.; Hu, J.; Wang, R.; Fu, S.; Qin, G. Antibacterial metals and alloys for potential biomedical implants. Bioact.
Mater. 2021, 6, 2569–2612. [CrossRef] [PubMed]
30. Yin, S.; Liu, J.; Kang, Y.; Lin, Y.; Li, D.; Shao, L. Interactions of nanomaterials with ion channels and related mechanisms. Br. J.
Pharmacol. 2019, 176, 3754–3774. [CrossRef] [PubMed]
31. Grace, M.; Chand, N.; Bajpai, S.K. Copper alginate-cotton cellulose (CACC) fibers with excellent antibacterial properties. J. Eng.
Fibers Fabr. 2009, 4, 24–35. [CrossRef]
32. Jardón-Maximino, N.; Cadenas-Pliego, G.; Ávila-Orta, C.A.; Comparán-Padilla, V.E.; Lugo-Uribe, L.E.; Pérez-Alvarez, M.; Tavizón,
S.F.; Santillán, G.d.J.S. Antimicrobial property of polypropylene composites and functionalized copper nanoparticles. Polymers
2021, 13, 1694. [CrossRef]
33. Mondal, A.; Mondal, N.K. Efficacy of bile juice mediated Cu MWCNT nanostructure, with respect to antibacterial and antibiofilm
activity. Mater. Lett. 2023, 335, 133828. [CrossRef]
34. Longano, D.; Ditaranto, N.; Sabbatini, L.; Torsi, L.; Cioffi, N. Synthesis and antimicrobial activity of copper nanomaterials.
Nano-Antimicrob. 2011, 26, 85–117. [CrossRef]
35. Torras, M.; Roig, A. Copper oxide nanocubes wrapping metals by microwave synthesis. Cryst. Growth Des. 2021, 21, 5027–5035.
[CrossRef]
36. Osonga, F.J.; Eshun, G.; Kalra, S.; Yazgan, I.; Sakhaee, L.; Ontman, R.; Jiang, S.; Sadik, O.A. Influence of particle size and shapes
on the antifungal activities of greener nanostructured copper against Penicillium italicum. ACS Agric. Sci. Technol. 2022, 2, 42–56.
[CrossRef]
37. Mahmoodi, S.; Elmi, A.; Hallaj-Nezhadi, S. Copper nanoparticles as antibacterial agents. J. Mol. Pharm. Org. Process. Res. 2018,
6, 140. [CrossRef]
38. Chen, Z.; Meng, H.; Xing, G.; Chen, C.; Zhao, Y.; Jia, G.; Wang, T.; Yuan, H.; Ye, C.; Zhao, F.; et al. Acute toxicological effects of
copper nanoparticles in vivo. Toxicol. Lett. 2006, 163, 109–120. [CrossRef] [PubMed]
39. Mehtar, S.; Wiid, I.; Todorov, S.D. The antimicrobial activity of copper and copper alloys against nosocomial pathogens and
Mycobacterium tuberculosis isolated from healthcare facilities in the Western Cape: An in-vitro study. J. Hosp. Infect. 2008, 68, 45–51.
[CrossRef] [PubMed]
Processes 2024, 12, 352 11 of 13

40. Karakoti, A.S.; Hench, L.L.; Seal, S. The potential toxicity of nanomaterials—The role of surfaces. J. Miner. 2006, 58, 77–82.
[CrossRef]
41. Aillón-García, P.; Parga-Landa, B.; Guillén-Grima, F. Effectiveness of copper as a preventive tool in health care facilities. A
systematic review. Am. J. Infect. Control 2023, 51, 1038–1048. [CrossRef] [PubMed]
42. Ma, X.; Zhou, S.; Xu, X.; Du, Q. Copper-containing nanoparticles: Mechanism of antimicrobial effect and application in dentistry-a
narrative review. Front. Surg. 2022, 9, 905892. [CrossRef] [PubMed]
43. Freedman, J.H.; Ciriolo, M.R.; Peisach, J. The role of glutathione in copper metabolism and toxicity. J. Biol. Chem. 1989, 264,
5598–5605. [CrossRef]
44. Ruparelia, J.P.; Chatterjee, A.K.; Duttagupta, S.P.; Mukherji, S. Strain specificity in antimicrobial activity of silver and copper
nanoparticles. Acta Biomat. 2008, 4, 707–716. [CrossRef]
45. Hans, M.; Erbe, A.; Mathews, S.; Chen, Y.; Solioz, M.; Mücklich, F. Role of copper oxides in contact killing of bacteria. Langmuir
2013, 29, 16160–16166. [CrossRef]
46. Malaikozhundan, B.; Lakshmi, V.N.; Krishnamoorthi, R. Copper oxide nanoparticles using Mentha spicata leaves as antibacterial,
antibiofilm, free radical scavenging agent and efficient photocatalyst to degrade methylene blue dyes. Mater. Today Commun.
2022, 33, 104348. [CrossRef]
47. Stoyanova, D.S. Antimicrobial Activity of Nanomaterials. Ph.D. Thesis, Sofia University St. Kliment Ohridski, Sofia, Bulgaria, 2018.
48. Fijan, S.; Cencič, A.; Turk, S.S. Hygiene monitoring of textiles used in the food industry. Braz. J. Microbiol 2006, 37, 356–361.
[CrossRef]
49. Grace, M.; Bajpai, S.K.; Chand, N. Copper (II) ions and copper nanoparticles-loaded chemically modified cotton cellulose fibers
with fair antibacterial properties. J. Appl. Polym. Sci. 2009, 113, 757–766. [CrossRef]
50. Perelshtein, I.; Applerot, G.; Perkas, N.; Wehrschuetz-Sigl, E.; Hasmann, A.; Gübitz, G.; Gedanken, A. CuO–cotton nanocomposite:
Formation, morphology, and antibacterial activity. Surf. Coat. Technol. 2009, 204, 54–57. [CrossRef]
51. Toodehzaeim, M.H.; Zandi, H.; Meshkani, H.; Firouzabadi, A.H. The effect of CuO nanoparticles on antimicrobial effects and
shear bond strength of orthodontic adhesives. J. Dent. 2018, 19, 1–5.
52. Noyce, J.O.; Michels, H.; Keevil, C.W. Potential use of copper surfaces to reduce survival of epidemic methicillin-resistant
Staphylococcus aureus in the healthcare environment. J. Hosp. Infect. 2006, 63, 289–297. [CrossRef] [PubMed]
53. Michels, H.T.; Noyce, J.O.; Keevil, C.W. Effects of temperature and humidity on the efficacy of methicillin-resistant Staphylococcus
aureus challenged antimicrobial materials containing silver and copper. Appl. Microbiol. 2009, 49, 191–195. [CrossRef] [PubMed]
54. Warnes, S.L.; Green, S.; Michels, H.T.; Keevil, C.W. Biocidal efficacy of copper alloys against pathogenic enterococci involves
degradation of genomic and plasmid DNAs. Appl. Environ. Microbiol. 2010, 76, 5390–5401. [CrossRef]
55. Chalandar, H.E.; Ghorbani, H.R.; Attar, H.; Abolhasan, S. Antifungal effect of copper and copper oxide nanoparticles against
Penicillium on orange fruit. Alavi Biosci. Biotech. Res. Asia 2017, 14, 279–284. [CrossRef]
56. Borkow, G.; Gabbay, J. Copper as a biocidal tool. Curr. Med. Chem. 2005, 12, 2163–2175. [CrossRef]
57. Angelov, O.; Stoyanova, D.; Ivanova, I.A. Antimicrobial effect of TiO2 doped with Ag and Cu on Escherichia coli and Pseudomonas
putida. In Proceedings of the INERA Conference “Vapor Phase Technologies for Metal Oxide and Carbon Nanostructures”, Velin-
grad, Bulgaria, 6–8 July 2016. Available online: https://ui.adsabs.harvard.edu/abs/2016JPhCS.764a2014A/abstract (accessed on
10 December 2023).
58. Kim, K.H.; Yang, M.; Song, Y.; Kim, C.H.; Jung, Y.M.; Bae, N.H.; Chang, S.J.; Lee, S.J.; Kim, Y.T.; Choi, B.G.; et al. Touchable 3D
hierarchically structured polyaniline nanoweb for capture and detection of pathogenic bacteria. Nano Converg. 2021, 8, 30–41.
[CrossRef]
59. Maniprasad, P.; Santra, S. Novel copper (Cu) loaded core-shell silica nanoparticles with improved Cu bioavailability: Synthesis,
characterization and study of antibacterial properties. J. Biomed. Nanotechnol. 2012, 8, 558–566. [CrossRef]
60. Stanić, V.; Dimitrijević, S.; Antić-Stanković, J.; Mitrić, M.; Jokić, B.; Plećaš, I.B.; Raičević, S. Synthesis, characterization and
antimicrobial activity of copper and zinc doped hydroxyapatite nanopowders. Appl. Surf. Sci. 2010, 256, 6083–6089. [CrossRef]
61. Yaseen, M.; Farooq, S.; Khan, A.; Shah, N.; Shah, L.A.; Bibi, S.; Khan, I.U.; Ahmad, S. CuO-SiO2 based nanocomposites: Synthesis,
characterization, photocatalytic, antileishmanial, and antioxidant studies. J. Chin. Chem. Soc. 2022, 69, 1637–1653. [CrossRef]
62. Sunada, K.; Kikuchi, Y.; Hashimoto, K.; Fujishima, A. Bactericidal and detoxification effects of TiO2 thin film photocatalysts.
Environ. Sci. Technol. 1998, 32, 726–728. [CrossRef]
63. Mingmongkol, Y.; Trinh, D.T.T.; Phuinthiang, P.; Channei, D.; Ratananikom, K.; Nakaruk, A.; Khanitchaidecha, W. Enhanced
photocatalytic and photokilling activities of Cu-doped TiO2 nanoparticles. Nanomaterials 2022, 12, 1198. [CrossRef] [PubMed]
64. Paszkiewicz, M.; Gołabiewska,
˛ A.; Rajski, L.; Kowal, E.; Sajdak, A.; Zaleska-Medynska, A. Synthesis and characterization
of monometallic (Ag, Cu) and bimetallic Ag-Cu particles for antibacterial and antifungal applications. J. Nanomater. 2016,
2016, 2187940. [CrossRef]
65. Ameh, T.; Gibb, M.; Stevens, D.; Pradhan, S.H.; Braswell, E.; Sayes, C.M. Silver and copper nanoparticles induce oxidative stress
in bacteria and mammalian cells. Nanomaterials 2022, 12, 2402. [CrossRef] [PubMed]
66. Pant, J.; Goudie, M.J.; Hopkins, S.P.; Brisbois, E.J.; Handa, H. Tunable nitric oxide release from S-nitroso-N-acetylpenicillamine via
catalytic copper nanoparticles for biomedical applications. ACS Appl. Mater. Interfaces 2017, 9, 15254–15264. [CrossRef] [PubMed]
67. Crane, J.K. Metal nanoparticles in infection and immunity. Immunol. Investig. 2020, 49, 794–807. [CrossRef]
68. Fan, X.; Yahia, L.; Sacher, E. Antimicrobial properties of the Ag, Cu nanoparticle system. Biology 2021, 10, 137. [CrossRef]
Processes 2024, 12, 352 12 of 13

69. Rolim, W.R.; Pieretti, J.C.; Renó, D.L.; Lima, B.A.; Nascimento, M.H.; Ambrosio, F.N.; Lombello, C.B.; Brocchi, M.; de Souza,
A.C.S.; Seabra, A.B. Antimicrobial activity and cytotoxicity to tumor cells of nitric oxide donor and silver nanoparticles containing
PVA/PEG films for topical applications. ACS Appl. Mater. Interfaces 2019, 11, 6589–6604. [CrossRef]
70. EN ISO 14729:2001; Ophthalmic Optics—Contact Lens Care Products—Microbiological Requirements and Test Methods for
Products and Regimens for Hygienic Management of Contact Lenses. ISO: Geneva, Switzerland, 2001.
71. Mohan, R.; Shanmugharaj, A.M.; Sung Hun, R. An efficient growth of silver and copper nanoparticles on multiwalled carbon
nanotube with enhanced antimicrobial activity. J. Biomed. Mater. Res. B Appl. Biomater. 2011, 96, 119–126. [CrossRef]
72. Datta, K.K.R.; Srinivasan, B.; Balaram, H.; Eswaramoorthy, M. Synthesis of agarose–metal/semiconductor nanoparticles having
superior bacteriocidal activity and their simple conversion to metal–carbon composites. J. Chem. Sci. 2008, 120, 579–586. [CrossRef]
73. Heinlaan, M.; Ivask, A.; Blinova, I.; Dubourguier, H.C.; Kahru, A. Toxicity of nanosized and bulk ZnO, CuO and TiO2 to bacteria
Vibrio fischeri and crustaceans Daphnia magna and Thamnocephalus platyurus. Chemosphere 2008, 71, 1308–1316. [CrossRef]
74. Gurianov, Y.; Nakonechny, F.; Albo, Y.; Nisnevitch, M. Antibacterial composites of cuprous oxide nanoparticles and polyethylene.
Int. J. Mol. Sci. 2019, 20, 439. [CrossRef]
75. Ditta, B.; Steele, A.; Liptrott, C.; Tobin, J.; Tyler, H.; Yates, M.; Sheel, W.; Foster, A. Photocatalytic antimicrobial activity of thin
surface films of TiO2 , CuO, and TiO2 /CuO dual layers on Escherichia coli and bacteriophage T4. Appl. Microbiol. Biotechnol. 2008,
79, 127–133. [CrossRef]
76. Ivanova, I.A.; Pavlova, E.L.; Stoyanova, D.S.; Angelov, O.I. Antibacterial effect of TiO2 : Cu: Ag thin coatings on Pseudomonas
strain measured by microbiological and ATP assays. J. Basic Microbiol. 2019, 59, 1165–1172. [CrossRef]
77. Shimabukuro, M. Antibacterial property and biocompatibility of silver, copper, and zinc in titanium dioxide layers incorporated
by one-step micro-arc oxidation: A review. Antibiotics 2020, 9, 716. [CrossRef]
78. Ivanova, I.; Stoyanova, D.; Nenova, E.; Staneva, A.; Kostadinova, A. Antimicrobial and cytotoxic properties of metal and graphene
nanomaterials (review). J. Chem. Technol. Metall. 2020, 55, 239–250.
79. Villapún, V.; Dover, L.; Cross, A.; González, S. Antibacterial metallic touch surfaces. Materials 2016, 9, 736. [CrossRef] [PubMed]
80. Leyland, N.S.; Podporska-Carroll, J.; Browne, J.; Hinder, S.J.; Quilty, B.; Pillai, S.C. Highly efficient F, Cu doped TiO2 anti-bacterial
visible light active photocatalytic coatings to combat hospital-acquired infections. Sci. Rep. 2016, 6, 24770. [CrossRef] [PubMed]
81. Rtimi, S.; Giannakis, S.; Sanjines, R.; Pulgarin, C.; Bensimon, M.; Kiwi, J. Insight on the photocatalytic bacterial inactivation by
co-sputtered TiO2 -Cu in aerobic and anaerobic conditions. Appl. Catal. B Environ. 2016, 182, 277–285. [CrossRef]
82. Usman, M.S.; El Zowalaty, M.E.; Shameli, K.; Zainuddin, N.; Salama, M.; Ibrahim, N.A. Synthesis, characterization, and
antimicrobial properties of copper nanoparticles. Int. J. Nanomed. 2013, 8, 4467–4479. [CrossRef]
83. Vanti, G.L.; Masaphy, S.; Kurjogi, M.; Chakrasali, S.; Nargund, V.B. Synthesis and application of chitosan-copper nanoparticles on
damping off causing plant pathogenic fungi. Int. J Biol. Macromol. 2020, 156, 1387–1395. [CrossRef] [PubMed]
84. Qi, L.; Xu, Z.; Jiang, X.; Wang, M. Cytotoxic activities of chitosan nanoparticles and copper-loaded nanoparticles. Bioorg. Med.
Chem. Lett. 2005, 15, 1397–1399. [CrossRef]
85. Li, X.; Xu, H.; Zhe-Sheng, C.; Chen, G. Biosynthesis of nanoparticles by microorganisms and their applications. J. Nanomater. 2011,
2011, 270974. [CrossRef]
86. Hongfeng, Z.; El-Kott, A.; Ahmed, A.E.; Khames, A. Synthesis of chitosan-stabilized copper nanoparticles (CS-Cu NPs): Its
catalytic activity for C-N and C-O cross-coupling reactions and treatment of bladder cancer. Arab. J. Chem. 2021, 14, 103259.
[CrossRef]
87. De La Torre-Roche, R.; Cantu, J.; Tamez, C.; Zuverza-Mena, N.; Hamdi, H.; Adisa, I.O.; Elmer, W.; Gardea-Torresdey, J.; White,
J.C. Seed biofortification by engineered nanomaterials: A pathway to alleviate malnutrition? J. Agric. Food Chem. 2020, 68,
12189–12202. [CrossRef] [PubMed]
88. Hejazy, M.; Koohi, M.; Bassiri, M.P.A.; Najafi, D. Toxicity of manufactured copper nanoparticles—A review. Nanomed. Res. J. 2018,
3, 1–9. [CrossRef]
89. Naz, S.; Gul, A.; Zia, M. Toxicity of copper oxide nanoparticles: A review study. IET Nanobiotechnology 2020, 14, 1–13. [CrossRef]
[PubMed]
90. Mathew, S.; Ganguly, P.; Rhatigan, S.; Kumaravel, V.; Byrne, C.; Hinder, S.J.; Bartlett, J.; Nolan, M.; Pillai, S.C. Cu-doped TiO2 :
Visible light assisted photocatalytic antimicrobial activity. Appl. Sci. 2018, 8, 2067. [CrossRef]
91. Qin, Q.; Li, J.; Wang, J. Antibacterial activity comparison of three metal oxide nanoparticles and their dissolved metal ions. Water
Environ. Res. 2017, 89, 378–383. [CrossRef]
92. Sandle, T. Risk Management Library Volume 4: Practical Approaches to Risk Assessment and Management Problem Solving: Tips and Case
Studies; DHI Publishing, LLC: River Grove, IL, USA, 2018.
93. Salgado, C.D.; Sepkowitz, K.A.; John, J.F.; Cantey, J.R.; Attaway, H.H.; Freeman, K.D.; Sharpe, P.A.; Michels, H.T.; Schmidt, M.G.
Copper surfaces reduce the rate of healthcare-acquired infections in the intensive care unit. Infect. Control Hosp. Epidemiol. 2013,
34, 479–486. [CrossRef] [PubMed]
94. Santo, C.E.; Lam, E.W.; Elowsky, C.G.; Quaranta, D.; Domaille, D.W.; Chang, C.J.; Grass, G. Bacterial killing by dry metallic
copper surfaces. Appl. Environ. Microbiol. 2011, 77, 794–802. [CrossRef]
95. Efstathiou, G.; Papastavrou, E.; Raftopoulos, V.; Merkouris, A. Factors influencing nurses’ compliance with standard precautions
in order to avoid occupational exposure to microorganisms: A focus group study. BMC Nurs. 2011, 10, 1. [CrossRef]
Processes 2024, 12, 352 13 of 13

96. Khan, H.A.; Baig, F.K.; Mehboob, R. Nosocomial infections: Epidemiology, prevention, control and surveillance. Asian Pac. J. Trop.
Biomed. 2017, 7, 478–482. [CrossRef]
97. Abraham, J.; Dowling, K.; Florentine, S. Can copper products and surfaces reduce the spread of infectious microorganisms and
hospital-acquired infections? Materials 2021, 14, 3444. [CrossRef] [PubMed]
98. Montero, D.A.; Arellano, C.; Pardo, M.; Vera, R.; Gálvez, R.; Cifuentes, M.; Berasain, M.A.; Gómez, M.; Ramírez, C.; Vidal, R.M.
Antimicrobial properties of a novel copper-based composite coating with potential for use in healthcare facilities. Antimicrob.
Resist. Infect. Control 2019, 8, 3. [CrossRef] [PubMed]
99. Schmidt, M.; Bréchot, N.; Hariri, S.; Guiguet, M.; Luyt, C.E.; Makri, R.; Leprince, P.; Trouillet, J.L.; Pavie, A.; Chastre, J.; et al.
Nosocomial infections in adult cardiogenic shock patients supported by venoarterial extracorporeal membrane oxygenation. Clin.
Infect. Dis. 2012, 55, 1633–1641. [CrossRef] [PubMed]
100. Pontin, K.P.; Borges, K.A.; Furian, T.Q.; Carvalho, D.; Wilsmann, D.E.; Cardoso, H.R.P.; Alves, A.K.; Chitolina, G.Z.; Salle, C.T.P.;
de Souza Moraes, H.L.; et al. Antimicrobial activity of copper surfaces against biofilm formation by Salmonella enteritidis and its
potential application in the poultry industry. Food Microbiol. 2021, 94, 103645. [CrossRef] [PubMed]
101. DeAlba-Montero, I.; Guajardo-Pacheco, J.; Morales-Sánchez, E.; Araujo-Martínez, R.; Loredo-Becerra, G.M.; Martínez-Castañón,
G.A.; Ruiz, F.; Compeán Jasso, M.E. Antimicrobial properties of copper nanoparticles and amino acid chelated copper nanoparti-
cles produced by using a soya extract. Bioinorg. Chem. Appl. 2017, 2017, 1064918. [CrossRef]
102. Mohindru, J.; Garg, U. Green synthesis of copper nanoparticles using tea leaf extract. Int. J. Eng. Sci. Res. Technol. 2017, 6, 307–311.
[CrossRef]
103. Atiya, M.A.; Hassan, A.K.; Kadhim, F.Q. Green Synthesis of copper nanoparticles using tea leaves extract to remove ciprofloxacin
(CIP) from aqueous media. Iraqi J. Sci. 2021, 62, 2832–2854. [CrossRef]
104. Khaldari, I.; Naghavi, M.R.; Motamedi, E. Synthesis of green and pure copper oxide nanoparticles using two plant resources via
solid-state route and their phytotoxicity assessment. RSC Adv. 2021, 11, 3346–3353. [CrossRef]
105. Sutradhar, P.; Saha, M.; Maiti, D. Microwave synthesis of copper oxide nanoparticles using tea leaf and coffee powder extracts
and its antibacterial activity. J. Nanostruct. Chem. 2014, 4, 86. [CrossRef]

Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual
author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to
people or property resulting from any ideas, methods, instructions or products referred to in the content.