Abstract

This study explores the use of Curecetin nanoparticles, synthesized and characterized through X-ray Diffraction (XRD), Fourier-Transform Infrared Spectroscopy (FTIR), and measured using Scanning Electron Microscopy (SEM) with sizes ranging from 13.40 to 44.66 nm. We conducted in vivo experiments on laboratory animals, applying nanoparticle doses of 10 mg, 50 mg, and 100 mg to 30 mm wounds. Skin wounds pose significant medical challenges due to potential complications like rupture and delayed healing. Results indicated that Curecetin nanoparticles significantly accelerated wound healing, with wound size reduction observed as early as the second day and substantial healing by the eighth day, suggesting the potential of nanoparticles in therapeutic applications for skin wounds.

Highlights:

• Precise Characterization: Curecetin nanoparticles were precisely synthesized and characterized using XRD, FTIR, and SEM.
• Rapid Healing: The study showed that Curecetin nanoparticles significantly accelerated wound healing in laboratory animals.
• Clinical Potential: Results indicate a promising future for nanoparticle applications in treating skin wounds clinically.

Keywords: Nanoparticles, Curecetin, Wound Healing, Nanotechnology, Organism

Introduction

When the wound does not heal on time, it may take a very long time to heal [1]. Skin wounds must first heal to close the wound, restore damaged tissue and apply treatments that speed up the healing process Infection can develop when different bacteria penetrate damaged skin or mucous membranes, a condition known as wound [2]. Globally, wound infections cause illness and mortality due to the susceptibility of open wounds to many types of microorganisms [3]. The main difficulty is infections Wounds in high antibiotic resistance among bacteria, fungi and viruses. Tissue damage can result from multiple bacterial infections or from colonization of areas affected by more than one type of microbial [4]. Researchers around the world have tried to use many products and medicines to treat acute and chronic wounds, but none has been able to cure the problem of wounds not healing [5]. Many preclinical and experimental studies have proven the effectiveness of using nanomaterials and natural materials to treat wounds [6]. Because they are inexpensive, effective, and easy to use, these materials are great alternatives to costly chemotherapy [7].

When refined, quercetin has a yellowish color, is slightly soluble in water, and forms solid crystals [8]. However, its solubility in alcohol and alkaline water solutions is enhanced. Quercetin is found in fruits and vegetables ( pear, oats , soybeans, beans and coffee, tea, medicinal herbs, spinach, garlic, and onions [9]. Quercetin is a secondary metabolite and pigment found in plants and is a powerful flavonoid found in plants. Chemical studies have shown that it has antioxidant properties [10]. Curcetin is a natural multifunctional chemical. Pharmaceutical effects include anti-inflammatory drugs and a wide range of biological effects, such as antiviral ones, neuroprotective factors, cardiovascular disease, microbes, diabetes, hypertension, obesity, and liver disease. The creation of drug-carrying hybrid molecules is facilitated by this antimalarial agent [11].

The different physical and chemical properties of nanomaterials such as size, shape, composition, surface charge, surface halo or aggregation affect their biomedical applications and the size of the nanomaterial is mainly dominant while other properties are controlled because reducing the size of the nanomaterial provides an opportunity to improve absorption and possibilities of interaction with biological tissues to a greater extent [12]. Hybrid nanomaterials to control sensory functions are an important topic in the pharmaceutical and nanomedicine industries. The mechanical strength, chemical stability, and flexibility of hybrid nanomaterials make them ideal for improving human health. Among the many important applications are drug delivery, antimicrobial effects, nutrition, dentistry, orthopedics, the creation of new antifungals, antioxidants, wound healing enhancers, and antibiotics [13].

Objectives of the Study

a. Preparation of a nanomaterial from quercetin.

b. Study the effect of nano-quercetin on wound healing and make assessments on vivo.

Methods

A. Experimental Work

1. Study the Effect of Curcetin Nanoparticles on Wound Healing and in Vivo Tests

a. Preparation of Q uercetin N anomaterials

The curcetin nanoparticles were ground in the laboratories of the Faculty of Science and the Department of Physics at the University of Diyala. Quercetin was crushed for 30 minutes in an electric mortar at room temperature after which the said substance was transferred to a sterile ivory mortar using ethanol and dry heat. A small amount of 1-2 grams of quercetin was ground again in the mortar. This process was repeated until the entire amount of quercetin was ground and finally an opaque glass container was used to protect the material . of light. The next step was to confirm the presence of the nanomaterial by sending it for testing.

b. X-ray diffraction (XRD)

Quercetin nanoparticles were diagnosed by X-ray diffraction ( XRD) examination at Kashan University – Iran.

c. Scanning Electron Microscopy (SEM)

The samples were examined at Kashan University in Iran with scanning electron microscopy ( SEM) analysis. When the focused beam interacts with the samples, the electrons collide with the specimen atoms and transmit the decomposition signals of the surface structure and detectors, allowing the device to scan the samples with extremely high resolution and image them with a magnification power of 25-250,000 times. Tests were carried out on quercetin nanoparticles to show their properties and structural appearance by converting electron density into stored digital voltage and generating three-dimensional images with a size range of 1-5 nm [14].

d. Transmission Electron Microscopy (TEM)

The sample at Kashan Pal University was examined by a transmission electron microscope ( TEM) and found that the quercetin nanoparticles manufactured by physical method , specifically the grinding method. The electron beam from the microscope, which has a voltage of 200 kV, was directed at the thin sample to observe its size, shape, density and crystal structure through atomic processes [15].

e. Industry of the Material to be Applied to Live Wounds

Quercetin nanopowder mixed with local butter was prepared in concentrations of 10 mg, 50 mg and 100 mg, and each concentration was placed in Petri dishes as shown in Figure 1 [15].

Figure 1. S hows the preparation of curocetin nanoparticles powder mixed with local butter in concentrations of 10 mg, 50 mg, 100 mg

f. Experiment of Wounding and Infecting Laboratory Mice and Applying Nano-Curetin

The laboratory mice were equipped and hair was remov ed from them by a hair remover using coarse gauze in order to facilitate the removal process, after that a machine was used for a skin mask . Artery and wounds were made in the back area for mice with a length of mm 30.00 by a sharp blade for surgeries after using an anesthetic as shown in Figure (2 ), then the pre-prepared compounds are used in this competition.

Figure 2.A: The hair removal of the dorsal area of mice using a depilator, B: The use of an arterial grab machine to make a wound in the dorsal region of the mouse, C: Skin grasping artery grabber, D: Making a wound with a surgical blade.

g. Statistical Analysis

The data of the current study were statistically analyzed using the statistical software SPSS version 25 . The Least Significant Difference (LSD) test was used to compare the arithmetic averages of the areas affected by the wounds [16].

Results and Discussion

A. Results of Infrared Spectrometry (FTIR ) for Quercetin

Instant infrared spectroscopy proved the obtained quercetin ; the hydroxyl group OH is shown with a strip at a wavelength of (3407.04 cm-1), as shown in Figure 3. In accordance with the results [17], we note the presence of water and the absorption range at (399.193 ^cm-1 ).

Figure 3.Quercetin Infrared Spectrometry (FTIR)

B. X-ray Diffraction Tests

Quercetin patterns were detected by analyzing the results of X-ray diffraction and understanding the locations of the peaks. The polycrystalline membrane confirms its findings [18] on the optimal development directions of crystalline granules, namely 200, 220 and 330. The directions (140) and (440) are also identical , as shown in Figure 4.

Figure 4. Shows the X-ray diffraction of quercetin

C. Scanning Electron Microscopy ( SEM) Examinations

Figure 5 shows the results of the scanning electron microscope (SEM) used to distinguish the nanostructures of q uercetin . At 150 KX magnification , the image clearly reveal s that quercetin is primarily dense and its shapes are not completely round. Their sizes range consistently between (13.40-44.66) nm, and this is confirmed by the results that were consistent with the results [19].

Figure 5.SEM image showing the morphological appearance of quercetin nanoparticles

D. Transmission Electron Microscope (TEM) T ests

Electron micrographs of quercetin nanoparticles mixed with local butter are shown as in Figure 6 . The image is q uercetin nanoparticles ranging in size from 4-60 nm. The results agreed with the researchers' findings [20].

Figure 6. TEM images of curetin nanomaterial, which was mixed with local butter

1. The Results of the Effect of Curestine Nano Mixed with Local Butter on Wounds in the Body of Living Organisms that Have Been Subjected to the Following Concentrations

a. The M ice were Exposed to a Concentration of 10 mg of Nano Q uercetin

As shown in Figure 7, the average length of wounds on the first, second, third and fourth day was 30.00 mm, 28.51 mm, 26.32 mm, 23.78 mm, 21.14 mm, 16.68 mm, 9.48 mm, and 4.33 mm respectively.

Figure 7.Shows the effect of exposing mice to the nano-quercetin plant preparation at a concentration of 10 mg

b. When Exposed to a Concentration of 50 mg C

The mice were exposed to n ano - quercetin at a concentration of mg50 and the average length of the wounds on the first day was 30.00 mm , the second day was 27.33 mm and the third day 25. 47 mm , the fourth day 19.22 mm , the fifth day 14.38 mm, the sixth day 10.25 mm, the seventh day 7.73 mm and the eighth day 2.11 mm as shown in Figure (8).

Figure 8. Shows the effect of exposing wounds to nano- quercetin at a concentration of 50 mg

c. When Exposed to a Concentration of 100 mg

The mice were exposed to a concentration of 100 mg of nano- quercetin . As shown in Figure 9. , the average length of wounds on the first, second, third and fourth day was 30.00 mm, 27.04 mm, 22.2 mm, 16.26 mm, 11.14 mm, 08.14 mm, 05.74 mm, and 01.09 mm respectively.

Figure 9.The effect of exposing wounds to nano-curetine at a concentration of 100 mg

Experimental skin wounds exposed with different doses of nanomaterials (10 mg, 50 mg, 100 mg) showed efficacy and effect on the vivo. On the eighth day of the trial, the wounds healed significantly ( P<0.05). On the second day of the experiment,

The average and varied recovery at doses was 10 mg, 50 mg and 100 mg when reaching the length of the wound on the second day ( Average ± Standard error (Mean ± Std ): ( 28.517± 0.50 1 , 27.337±0.574 , 27.040 ± 0.061 ) . It is compared with the length of the wounds o n the eighth and second days respectively ( 4.333 ± 0.577 , 4.333± 0.577 , 1.090 ± 0.020 ) . The healing of skin wounds significantly improved in mice. R at skin wounds heal faster after applying the formula Nano- Topical quercetin , as shown in the table ( 1 ) because the quercetin is hydrophobic and does not penetrate the skin . T his limits its use as a topical healing agent. So , t his study used n anoparticles in w ound h ealing d ue to the change of chemical and physical properties and the improvement of the process of absorption and solubility of nano-kerosene and conductivity to the target . Also, t he skin of laboratory mice was cured from the second to the eighth day g radually w ith reduced deposition of inflammatory cells, proper regulation of collagen fibers, and remodeling of epithelial tissue . T he results were consistent with the [21] .

Conclusion

Nanomaterials with high specifications and unique sizes ranging from 13.40 to 44.66 nm were produced by grinding the quercetin nanoparticles from top to bottom. Multiple concentrations (10, 50 and 100 μg/mg) have been shown to be effective in wound healing rate on the skin in vivo experiments .‏

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