1 Background

Nanotechnology has resulted in various scientific advancements, which functions as a fundamental unit [1]. Many agencies such as proteins, medicines, dendrites, DNA, RNA [2], and ligand improve nanoparticle application in delivery of drugs [3, 4] for the treatment of many conditions such as wound infection, gene therapy, malignancies, and many more [5, 6]. The cobalt, titanium, and silver nanoparticles have emerged as promising tool in drug delivery [7, 8]. Factors such as desiccation, infection [9], maceration, necrosis, pressure, trauma, and edema [10] can all impede wound healing [11]. Syzygium aromaticum (cloves) are aromatic flower buds [12] that belong to Myrtaceae family [13]. It has been known to be a rich source of phenolic chemicals including eugenol, eugenol acetate, and gallic acid [14]. Eugenol extracted from clove buds has been reported to contain different phytochemicals like flavones, aldehyde, and phenol.

The aggrandized prospects of nanoparticles in different biomedical application have led to the quest of their synthesis using different methodology like chemical, physical, and biological [15]. However, the biological method has been mostly recognized in this decade due to their eco-friendly preparation method and the use of non-hazardous chemicals. These methodologies have been called as green methods of synthesis [16]. Green synthesis of the nanoparticle is mainly used due to its advantages like eco-friendly approach, easily availability of the biological materials, and rapid process [17,18,19].

The biosynthesis of nanoparticles by plant extract gives an advancement over other green methods as it is simple, environment friendly, cost-effective, and relatively reproducible [20]. Plant extracts have the potential to act as both reducing agent and stabilizing agent in the nanoparticle synthesis [13, 14], and it affects the features of the synthesized nanoparticles also [21, 22]. The synthesis has been successfully reported to form nanoparticles because of the presence of several concentrations and amalgamations of organic reducing agents in variety of plant extracts [23, 24]. The various phases included in the process of nanoparticle synthesis involve the activation phase, which includes metal ion reduction, and then their nucleation [25]. The primary phase is nanoparticle synthesis using plant extracts, the second phase (growth phase) is tiny nanoparticles and coalesce, and the final phase is the termination phase, during which the nanoparticles take on their final shape [26].

Evaluation of drug release is crucial in the biomedical field, as it is important for the analysis of drug release type [19]. The importance of kinetic models is often helpful in elucidating release mechanisms and that can be of use in the control of release mechanism [20, 27]. Further advantage of the kinetics is to signify various release data with one or two parameters. Results obtained from such a different standard model can be employed for evaluating different delivery systems of a drug [28]. Drug release studies have a fundamental role in analyzing the release of drug in different dosage [29]. A theoretical justification of the best models is evaluated by different mathematical models [22, 23].

The aim of the study is to determine the antimicrobial activity of eugenol-loaded nanocomposite against wound-associated pathogens, evaluation of minimum inhibitory concentration (MIC), and minimum bactericidal concentration test (MBC). This study is of the opinion that eugenol-loaded nanocomposite with significant antimicrobial activity may enhance the wound healing with special reference to impaired wound healing in diabetes.

2 Materials and Methods

2.1 Materials

Silver nitrate (AgNO3), cobalt nitrate (CoNO3), and titanium dioxide (TiO2) were purchased from Merck. Polyvinylpyrrolidone (PVP), 1,1-diphenyl-2-picrylhydrazyl (DPPH), and other components were purchased from Sigma-Aldrich.

2.2 Syzygium aromaticum Extract Preparation

Dried clove buds were collected and powdered using mortar and pestle, and 1 gm of the powder was dissolved in 100 ml of a 40% butanol solution and continuously shaken in a rotary shaker for 3–4 h. The extract was then filtered through Whatman filter paper, and a crude extract was prepared [30].

2.3 Nanocomposite Synthesis and Characterization

The synthesis of Ag-Ti-Co NC was done was done by using eugenol extracts obtained from Syzygium aromaticum (S. aromaticum). Plant extract was added dropwise into a solution containing AgNO3 (0.001 M) contained with Titanium dioxide (TiO2) (0.001 M) and Cobalt Nitrate (CoNO3) (0.01 M) stirred at 350 rpm for 2 h at 60 °C [31]. PVP (polyvinyl pyrrolidone) (0.001 mM; 0.5 ml) was added into the solution. The formation of color change indicated the synthesis of nanocomposite. The synthesized Ag-Ti-Co nanocomposite was characterized by UV–Vis spectroscopy, X-ray diffraction (XRD) powder method, Fourier transform infrared (FTIR) spectroscopy, Energy dispersive X-ray (EDAX), ZETA Sizer and scanning electron microscope (SEM) (Model JSM6100) analysis. Different parameters of the nanocomposite such as shape, nature, size of the particles, crystallinity, surface area, charge and stability are obtained by these techniques [32].

2.4 Loading of Eugenol on Nanocomposite

The loading of eugenol on prepared nanocomposite was done through sonication technique. Five hundred microgram/milliliter of nanocomposite was dissolved in eugenol and bath sonicated for 15 min [33]. Allowed to be kept on dissolved in distilled water through a sonicator and in a different vial, 500 μg/ml of eugenol was prepared. Then, the sample was centrifuged at 8000 rpm for 10 min and filtered through a cellulosic white membrane filter. Followed by the filtration, the supernatant was collected to determine the loading amount of eugenol [28]. The loading capacity was calculated based on the following formula:

$$\mathrm{\%Eugenol\;loading}=\frac{\mathrm{Amount\;of\;eugenol\;added\;initially}-\mathrm{amount\;of\;eugenol\;in\; supernatant}}{\mathrm{Amount\;of\;eugenol\;added\;initially}}\times 100$$

The eugenol-loaded nanocomposite (Eu@NC) (collected on the pellet was dried under 30–40 °C and washed with 40% ethanol [34].

2.5 Drug Release Kinetics

Drug release of eugenol-loaded nanocomposite was analyzed separately by two methods given with some modification, i.e., direct method and dialysis bag method [28, 35].

2.5.1 Direct Addition Method

Five hundred microgram of eugenol-loaded nanocomposite (Eu@NC) was added in a tube containing 2 ml of buffer with pH 7 and shaked continuously. A small aliquot of dissolution fluid was withdrawn at different time intervals and maintained with an equal volume of the buffer after per withdrawal. The collected samples were centrifuged for 10 min at 10,000 rpm, and supernatant was analyzed by UV–visible spectrophotometer at 543 nm [36].

2.5.2 Dialysis Method

Eu@NC was dialyzed against 40-ml buffer (pH 7) at 25 °C with constant stirring at 100 rpm. At different time intervals, sample was pipetted out (2 ml) and was analyzed spectrophotometrically. Simultaneously, add an equal amount of buffer (2 ml) during every cycle of sample collection [35].

Different types of mathematical models are used for evaluating drug dissolution [28] to determine the mechanism of drug release.

Zero-order equation:

$$Q_{t} = Q_{0} + K_{0}^{t}$$

Here, Qt is the quantity of drug dissolved in time t, K0 is the zero-order release (concentration/time) constant, and Q0 is the initial quantity of drug in the solution, mostly Q0 = 0.

First-order equation:

$${\text{log }}C = {\text{log }}C_{0} - K_{t} /{2}.{3}0{3}$$

Here, C0 is the initial concentration of drug and K is the first-order constant.

Higuchi’s equation:

$$Q_{t} = K_{H} t^{{1}}_{{/{2}}}$$

Here, KH is the release rate constant (Higuchi model) and Qt is the amount of drug released in time t.

$$Mt/M\overline{\alpha } = Kt^{n}$$

where Mt/ M ᾱ is a fraction of drug released at time t. K is the release rate of constant, and n is the release exponent.

2.6 Evaluation of Antimicrobial Activity

Antimicrobial activity of Eu@NC (Eu @Ag-Ti-Co NC) was estimated using well diffusion method against four wound-associated microorganisms E. coli (MTCC-40), Proteus mirabilis (MTCC-3310), Pseudomonas aeruginosa (MTCC-424), and Staphylococcus aureus (MTCC-9760). 0.5 McFarland standard (108 CFU per ml) was used for estimation of antimicrobial activity.

2.7 Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)

MIC of Eu@NC for Escherichia coli (MTCC-40), Proteus mirabilis (MTCC-3310), Pseudomonas aeruginosa (MTCC-424), and Staphylococcus aureus (MTCC-9760). The Minimum inhibitory concentration value of loaded nanocomposite conjugate and others was estimated according to the standard method [37]. All experiments were conducted in triplicate. Five hundred microgram/milliliter stock solution was prepared, and two-fold serial dilutions of the eugenol-loaded nanocomposite were carried out. 96-well microtiter plate was used for the MIC test by standard broth microdilution processes [38]. After 24 h, measurements were taken using ELISA reader. The lowest concentration of nanocomposites that inhibits the growth of bacteria was taken as a MIC [39]. MBC analysis was performed by plating the suspension [40]. The lowest concentration that shows no visible growths is taken as the MBC value [41].

2.8 Antioxidant Activity

The Eu@NC DPPH assay [42] was assessed for eugenol, eugenol-loaded nanocomposite, and unloaded nanocomposite using the standard technique [43]. The obtained solution was kept in the dark for 30 min and measured at 517 nm using by UV–vis spectrophotometer. The percentage of scavenging activity was calculated as follows:

$$\mathrm D\mathrm P\mathrm P\mathrm H\;\mathrm f\mathrm r\mathrm e\mathrm e\;\mathrm r\mathrm a\mathrm d\mathrm i\mathrm c\mathrm a\mathrm l\;\mathrm s\mathrm c\mathrm a\mathrm v\mathrm e\mathrm n\mathrm g\mathrm i\mathrm n\mathrm g\;(\%)=\frac{\mathrm{control}-\mathrm{sample}}{\mathrm{control}}\times100$$

3 Results and Discussion

3.1 Nanocomposite Synthesis and Their Characterization

In nanoparticle synthesis, there are three fundamental phases of metal nanoparticle synthesis with plant extracts [44]: firstly, the activation phase in which metal ions are reduced and metal atoms are formed; during the second phase, which is the growth phase, the small nanoparticles spontaneously coalesce into large particle size; and the third phase called termination phase gives the final shape of the nanoparticles [45]. The synthesis of Eu@NC is achieved by reducing silver nitrate, titanium dioxide, and cobalt nitrate (0.001 M, 0.001 M, 0.01 M, respectively), with eugenol as a reducing agent and PVP as a capping agent. The changes in the color of the solution after the addition of plant extract, from pink to deep brown precipitate, confirmed the formation of the nanocomposite. The color change of the precipitate can be reasoned to the shift of the surface plasmon band because of the size changes of the particle [46, 47].

Further, the biosynthesized Eu@NC was characterized by FTIR spectroscopy, SEM, XRD, EDAX, ZETA, size distribution pattern, and UV–Vis spectroscopy to determine their physiochemical properties.

3.1.1 UV–Vis Spectroscopy Analysis

Surface plasmon resonance (SPR), also known as surface plasmon excitation, is a phenomenon that arises when applied electromagnetic fields excite surface plasmons on the surfaces of nanoparticles [36, 37]. Sarekha et al. previously discussed that eugenol-loaded nanoparticle range comes between 250 and 300 nm range. The UV–visible absorbance spectrum of the synthesized Eu@NC displayed a broad peak at 258 nm. The absorption peak can be attributed to the surface plasmon resonance of tri-metallic nanocomposite [48]. The data indicated the loading of eugenol into tri-metallic nanocomposite.

3.1.2 FTIR Spectroscopy

FTIR is a technique for the characterization of nanoparticles that can be used for the analysis of the vibration frequencies of bonds present between the molecules [38, 40]. FTIR analysis determined the phytochemicals likely to be responsible for the reduction and capping of reduced silver, titanium, and cobalt nanoparticles with surface adsorbent characteristics.

As shown in Fig. 1, the FTIR results of eugenol gave characteristic peak at 3551 cm−1, 3476 cm−1, 3413 cm−1, 3236 cm−1, 2929 cm−1, 2030 cm−1, 1637 cm−1, 1617 cm−1, 1381 cm−1, 1362 cm−1, 1204 cm−1, 1043 cm−1, 620 cm−1, 480 cm−1, and 408 cm−1 which corresponds to O–H stretch vibration of phenol, O–H stretch vibration of carboxylic acid, N–H stretch vibration of amines, C-H stretch vibration of alkane, C-H stretch vibration of alkanes, C≡C stretch vibration of alkyne, C–C stretch vibration of aromatic ring, C–C stretch vibration of allyl group, C-H stretch vibration of alkane, C-H stretch vibration of alkyl methyl, -C–O–C- stretch vibration of alcohol, -CH-CH2 stretch vibration of vinyl group, C-S stretch vibration of halo compound, and C-S stretch vibration of halo compound, respectively [49].

Fig. 1
figure 1

Physiochemical characterization of nanomaterials determined by FTIR spectra of eugenol and Eu@NC

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The FTIR spectrum of the Eu@NC is shown in Fig. 1. The biosynthesis of tri-metallic nanocomposite loaded with eugenol gave characteristic peaks. The peaks at 3551 cm−1, 3476 cm−1, 3413 cm−1, 3236 cm−1, 2929 cm−1, 2030 cm−1, 1637 cm−1, 1618 cm−1, 1497 cm−1,1444 cm−1, 1382 cm−1, 1195 cm−1, 1047 cm−1, 1082 cm−1, 23 cm−1, and 488 cm−1 were N–H stretch vibration of amines, C-H stretch vibration of alkane, C-H stretch vibration of alkane, C≡C stretch vibration of alkyne C–C stretch vibration of allyl, C–C stretch vibration of allyl, C = O stretch vibration of ketone, C = O stretch vibration of carbonyl group, C-H stretch vibration of alkane, -C–O–C- stretch of alcohol, C–O–C stretch vibration of ether, C–O–C stretch vibration of ether, C-S stretch vibration of halogen compound, respectively. The presence of similar peaks was considered for the presence of eugenol which has been reported previously [49].

3.1.3 XRD Analysis

The XRD spectra were utilized to verify the crystalline nature of the tri-metallic nanocomposite. The XRD spectrum of Eu@NC exhibited strong peaks at 2θ values: 32.1512, 38.079, 46.1373, 47.9564, and 54.7137 corresponding to (111), (200), (211), (220), and (221) planes, respectively (Fig. 2). The different peaks corresponded to the standard peaks of silver, titanium, and cobalt nanocomposite reported in JCPDS 21–1272 and 80–1538. As calculated, the lattice parameter of eugenol-loaded Ag-Ti-Co nanocomposite was found to be 4.6344 Å. The average nanocomposite size is calculated by the Debye Scherrer equation: D = Kλ/β cosθ where D is the average particle size, λ (1.5406 ˚ A); β is the full width at half maximum of the peak (FWHM), k (constant 0.9); and θ is the diffraction angle. The average calculated crystal size of Eu@NC was found as 41 nm, corresponding to orthombic structures [41, 42].

Fig. 2
figure 2

Physiochemical characterization of nanomaterials determined by XRD spectra of Eu@NC

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3.1.4 SEM and EDAX Analysis

The surface morphology and size of the Eu@NC were determined and validated by SEM [43, 44] equipped with EDS, and SEM results indicated the size of the nanocomposite in the nanorange (10–100 nm) shown in Fig. 3. The EDS analysis of Eu@NC is shown in Fig. 4. The result demonstrates the clear elemental composition profile of the biosynthesized eugenol-loaded Ag-Ti-Co nanocomposite. The validation of the presence of silver, titanium, and cobalt in synthesized nanocomposite confirmed the formation of a tri-metallic nanocomposite, and the presence of oxygen and carbon was also found which may be due to the loading of eugenol (C10H12O2).

Fig. 3
figure 3

Physiochemical characterization of nanomaterials determined by SEM image of the Eu@NC

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Fig. 4
figure 4

Physiochemical characterization of nanomaterials determined by EDS analysis of Eu@NC

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3.1.5 Stability Analysis

The Zeta potential is a characterization technique to measure the magnitude of the electrical charge present on the lipid bilayer, which is used to evaluate the charge stability of a dispersed system [50, 51]. An electric charge is applied over the sample for measuring the Zeta potential of the nanoparticle in a folded capillary flow cell [52]. It is significant to examine the stability of nanocomposite in a medium to consider its complementary effect. Accumulation and stability depend on the charge of the nanoparticles [53].

The Zeta potential of synthesized Eu@NC was found to be − 34.3 mV with PDI value 1.000 (Fig. 5) which indicated the net negative charge at the surface of the synthesized nanoparticles. PDI value is basically a representation of the distribution of size populations within the sample according to the PDI value of the sample that shows the highly polydisperse sample. Zeta potential results confirmed the stability of synthesized EU@NC. Zeta potential’s absolute value indicates a high electric charge on the surface of the eugenol-loaded nanocomposite, which has potent repulsive effects between particles to stop the nanocomposite in the solution from aggregating [54].

Fig. 5
figure 5

Physiochemical characterization of nanomaterials determined by Zeta potential analysis of Eu@NC

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3.2 Antimicrobial Activity of Eu@NC

3.2.1 Well Diffusion Assay

As shown in Fig. 6, the antimicrobial activity of the Eu@NC showed significant antimicrobial activity against wound-causing pathogens as compared with eugenol, the antimicrobial activity of eugenol-loaded nanocomposite against E. coli, S. aureus, P. aeruginosa, and Proteus mirabilis (i.e., 16 mm, 17 mm, 14 mm, and 21 mm, respectively). The antimicrobial activity of this eugenol-loaded Ag-Ti-Co tri-metallic nanocomposite is not yet performed in any previous research work. This unique investigation was performed in our research work.

Fig. 6
figure 6

Antimicrobial activity of of eugenol and eugenol-loaded nanocomposite (Eu@NC) in wound-associated pathogens

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3.2.2 MIC and MBC value of Eu@NC

The MIC and MBC value of eugenol-loaded nanocomposite against E. coli, P. aeruginosa, Proteus mirabilis, and S. aureus. The MIC (minimum inhibitory concentration) of eugenol-loaded nanocomposite was analyzed in microtiter plate by ELISA reader. The MBC value was observed by the agar plate method. The MIC value of Eu@NC is shown in Table 1, the MBC value is shown in Table 2, and Fig. 7 shows the 96-well microtiter plate. The minimum bacterial concentration of nanocomposite was analyzed in microtiter plate. Wells of the microtiter plate contain nanocomposite ranging from 3.125 to 0.098 μg/ml showed bacterial growth, whereas no growth was seen in wells containing nanocomposite 6.25 to 50 μg/ml. But in the case of Proteus mirabilis, no growth observed at 3.125 μg/ml concentration. One hundred microliter of the sample was poured on an agar plate, and after 24 h of incubation, no bacterial growth was observed (Table 2) at particular concentration of nanocomposite. The result determined the MIC and MBC value of the nanocomposite and confirmed its antimicrobial activity. The phenomenon can be reasoned to the three possible ways for the antimicrobial activity of nanocomposite: (a) eugenol-loaded nanocomposite may bind to the bacterial cell wall and degrade the membrane integrity and cell respiratory functions also; (b) eugenol-loaded nanocomposite may penetrate inside the bacterial cell and generate cell death; and (c) nanocomposite may release nanoforms of metal ion which cause bactericidal effect [55].

Table 1 Minimum inhibitory concentration (MIC) value of eugenol-loaded nanocomposite analyzed by ELISA reader against different wound-causing pathogens: (A) Proteus, (B) Staphylococcus aureus, (C) P. aeruginosa, and (D) E. coli. Positive ( +), indicating growth; Negative ( −), indicating absence of growth
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Table 2 Minimum bactericidal concentrations (MBC) of eugenol-loaded nanocomposite after 24 h. E.coli (12.5 μg), S. aureus (25 μg), P. aureginosa (25 μg), and Proteus (12.5 μg). Positive ( +), indicating growth; Negative ( −), indicating no growth
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Fig. 7
figure 7

Pictorial presentation showing 96-well plate set up for the finding of the MIC value of Eu@NC using the broth microdilution method. Nutrient broth media was used in the microdilution assay. Each test well possesses 100 μl of defined Eu@NC dilution and 100 μl of 0.5 McFarland solution of the bacterial suspension. Negative control wells have media only to provide turbidity control. The positive control contained 100 μl of nutrient broth and 100 μl of 0.5 McFarland bacterial culture. 1 24 h incubation without staining; 2 24 h incubation with staining. Wells contained different bacterial cultures: (A) E. coli, (B) Proteus, (C) P. aeruginosa, and (D) S. aureus

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3.3 DPPH Radical Scavenging Activity

The antimicrobial activity of the nanocomposite was hypothesized to be mediated through the antioxidant property of the nanocomposite [48, 56]. The antioxidant activity was investigated using the radical scavenging (DPPH) experiment. A stable free radical called DPPH reacts with a hydrogen donor. After reacting, the DPPH radical produces the reduced form of DPPH that is hydrazine form, which changes from purple to yellow color. The loss of color is due to the antioxidant content [57]. Eugenol extract shows 86.8% inhibition, eugenol-loaded nanocomposite 67.1% inhibition, and unloaded nanocomposite 36.3% inhibition which could be related to crosslinking. A graph is shown in Fig. 8, and different samples are shown in Fig. 8a. Pure eugenol shows high inhibition compared to other samples, and the loaded nanocomposite shows high inhibition compared to the unloaded nanocomposite.

Fig. 8
figure 8

a DPPH-RSA activity of three different samples: (A) eugenol, (B) nanocomposite (NC), and (C) loaded nanocomposite (@NC). b DPPH-RSA inhibition percentage for eugenol, loaded nanocomposite, and unloaded nanocomposite

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3.4 Drug Release Kinetics

Different kinetic models were used to perform analysis of in vitro drug release (Fig. 9) [49, 58]. Kinetics analysis of the dialysis method and the direct method is shown in Fig. 10 and Fig. 11. The maximum degree of correlation coefficient determined the proper mathematical model that follows release kinetics [59]. In the dialysis method, the results obtained for drug release follow Kosermayer Peppas model, with an R2 value of 0.9487. As per Kosermayer Peppas model, R2 value < 0.89 follows non-Fickian release, which describes drug release type, diffusion, and dissolution. In the direct method, drug release data were best fitted with the Higuchi drug release model, with an R2 value of 0.9511. Higuchi release model describes the diffusion type of drug release (Fig. 11). Thus, in vitro drug release performed by both the methods, direct and dialysis bag method, is approximately equal to the burst release of eugenol from the nanocomposite.

Fig. 9
figure 9

In vitro eugenol release from nanocomposite at pH 7: direct method and dialysis method

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Fig. 10
figure 10

Kinetics analysis of drug release by dialysis method: 1 zero-order kinetics, 2 first-order kinetics, 3 Higuchi plot, and 4 Kosermayer-Peppas model

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Fig. 11
figure 11

Kinetics analysis of drug release by direct method: 1 zero-order kinetics, 2 first-order kinetics, 3 Higuchi plot, and (4) Kosermayer-Peppas model

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The study showed a novel way to synthesize eugenol loaded tri-metallic nanocomposite (Ag-Ti-Co) using green methodology. Synthesized nanocomposite was well characterized using different techniques. UV–Vis determined the SPR of the nanocomposite, and the core optical properties as well as stability were determined. The nanocomposites showed a distinct antimicrobial activity with a display of antioxidant activity. With reference to the experimental data and the literature, it can be intrigued that the synthesized nanocomposite showed antibacterial activity against different microbes through the phenomenon of antioxidant activity which further causes imbalance in the physiology of the bacterial strains, thereby leading to the cell death of the microbes.

4 Future Prospects

Multidrug-resistant bacteria which is an ever-increasing problem these days could be resolved by using this biosynthesized eugenol-loaded nanocomposite. The major challenge is to discover the effective techniques for isolating and purifying newer and safer naturally occurring antimicrobials against MDR pathogenic microorganisms. A better understanding of the structure, function, and action mechanism of existing and newly identified antibiotics will help researchers to fine-tune proper design of the drug to work against MDR pathogenic microorganisms. The current research focuses on most common approach for antimicrobial therapy and checks on excessive use of antibiotics and the spread of antibiotic resistant bacteria. Furthermore, the basic technique for assessing antibiotic sensitivity is time consuming; thus, this therapy may help preventing unnecessary use of antibiotics and the spread of antibiotic resistance bacteria. The future of antibiotics requires advancement in the areas that are dependent on highly traditional methods. Thus, there is an urge to perform substantial changes in policy, an understanding of the mode of action of these drugs, and investment in alternatives to traditional antibiotics. The current study may provide new insight in the development of accelerated wound healing phenomena/slows skin infection and allowing patients to take a more active role in daily wound/skin infection care while potentially lowering healthcare costs, in numerous forms such as bandages, antiseptics, or antiseptics which may be beneficial for patients to apply with ease and comfort.

5 Conclusion

The present work focuses on and highlights the biosynthesis of eugenol-loaded nanocomposite for enhancement of wound healing. Due to the existing major problems in wound healing, there is a need for easy, low costing, eco-friendly, and non-toxic biosynthesized nanocomposite for biomedical applications like wound healing. Eugenol was loaded successfully on the nanocomposite using the sonication technique. The loaded nanocomposite showed good antibacterial activity against wound-causing pathogens. Thus, a eugenol-loaded Ag-Ti-Co nanocomposite will be a promising alternative to develop an antibacterial agent against wound-causing microorganisms, which are multidrug-resistant strains of microbes. Accordingly, this research may accelerate wound healing by this biosynthesized Eu@NC.