SILVER NANOPARTICLE-SUPPORTED, NATURAL, BIOCOMPATIBLE, ANTIMICROBIAL, HEMOSTATIC, AND WOUND-HEALING COMPOSITE WOUND DRESSING.

Abstract

The invention relates to a multifunctional composite wound dressing containing silver nanoparticles, natural biopolymers, and plant extracts, characterized by its inclusion of sodium alginate, aloe vera, Hypericum perforatum extract, Achillea millefolium extract, Equisetum arvense extract, and silver nanoparticles.

Description

1 TARIFF SILVER NANOPARTICULAR-SUPPORTED, NATURAL, BIOCOMPATIBLE PRODUCT. ANTIMICROBIAL, HEMOSTATIC AND WOUND HEALING COMPOSITE WOUND DRESSING 5 Technological Field: The invention involves a multi-layered mixture containing silver nanoparticles, natural biopolymers, and plant extracts. It relates to functional composite wound dressings. 10 State of the Art: A significant portion of the traditional wound dressings used in current techniques cover the wound surface. 15 It performs basic functions. However, it is particularly useful in treating chronic wounds and burns. For surgical wounds and open wounds with a high risk of infection, only physical protection is recommended. This is insufficient; limiting microbial growth and stopping bleeding quickly is not enough. additional requirements such as controlling and supporting tissue regeneration This is why classic dressings, which have a single function in wound care, are 20 The effectiveness of these materials remains limited in many cases. Advanced wound dressings incorporate antibacterial agents, hydrogel or biopolymer structures, Metal nanoparticles and various biologically active substances can be used. However, this Cell compatibility is 25 during the enhancement of antibacterial efficacy in such systems. protection, retention of active ingredients on the wound surface for a sufficient period of time and uncontrolled release Prevention presents significant technical problems. Especially silver-based antibacterial the amount of silver used while ensuring sufficient antimicrobial efficacy in the systems It needs to be maintained at an appropriate level in terms of biocompatibility. Similarly Materials exhibiting hemostatic properties do not always promote wound healing or have antibacterial properties. 30 It cannot provide the same level of protection. 2 On the other hand, plant extracts and natural biopolymers are used in wound care products. While it can offer advantages in terms of biocompatibility and biological activity, this The process involves homogeneously and permanently fixing the components onto a textile carrier. during which time it maintains its functions and remains stable together with other active ingredients. Creating a composite structure is not easy. Therefore, 5 within a single wound dressing. Antibacterial activity, rapid hemostasis, biocompatibility, and wound healing support. Providing both of these features simultaneously also makes the structure in question feasible and viable. The fundamental principle in the technical field is that it can be obtained through a repeatable production method. It remains one of the needs. Description of the invention: The primary purpose of the invention is not limited to merely physically closing the wound surface. not remaining, reducing the risk of infection, controlling bleeding quickly 15 multifunctional devices that contribute to the removal and support of the wound healing process. The goal is to develop a composite wound dressing. In this context, the different types of wound dressings used in classic wound dressings will be considered separately. antibacterial effect, hemostatic properties, biocompatibility and tissue compatibility were observed. needs such as supporting regeneration are brought together within a single structure. The aim was to bring it about. The invention involves sodium alginate, aloe vera, on a cotton-based carrier surface. With extracts of Hypericum perforatum, Achillea millefolium and Equisetum arvense Silver nanoparticles are used together. Thanks to this structure, it can be applied to wound dressings. A composite material with natural ingredients and functional properties is being created using sodium alginate. While contributing to liquid absorption and biocompatible matrix formation, the plant-based components 25 It supports wound healing processes, while silver nanoparticles are antibacterial. It contributes to ensuring the effectiveness of the event. One of the key advantages of the invention is that the composite wound dressing is Gram-negative. High resistance to Escherichia coli and Gram-positive Staphylococcus aureus bacteria (30%). It exhibits antibacterial activity. In quantitative analyses performed, it showed antibacterial activity against E. coli. a reduction of approximately 4.88 log₁₀ and over 99.99%, while for S. aureus it was approximately 2.00 log₁₀. 3 A reduction and approximately 99% antibacterial efficacy were achieved. Thus, the wound an effective structure for limiting bacterial contamination in the region is provided. Another advantage of the invention is its rapid hemostatic effect. The composite wound dressing has 5 After application to the bleeding area, the bleeding stops in approximately 3.70 seconds. It has been determined that it stops bleeding. This feature is particularly important where rapid bleeding control is crucial. It provides advantages in wound care and emergency intervention applications. The wound dressing that is the subject of the invention is also suitable in terms of biocompatibility. 10 It has been determined that no cytotoxic effect was observed in MTT analyses, and in scratch analyses... Cell proliferation and wound healing processes have been shown to be supported. In vivo Assessments revealed a decrease in inflammation and edema, and the formation of granulation tissue. re-epithelialization and angiogenesis are supported; also VEGF-A and TGF-β It has been determined that their expressions have increased. These results indicate that the invention only covers the wound surface 15 It is not a passive material that closes the wound, but rather participates in wound healing and tissue regeneration processes. This ensures that it serves as a functional wound dressing that contributes to the wound care process. The invention also involves the use of a cotton-based carrier and natural ingredients. Its durability and applicability in the textile industry through impregnation-drying-curing processes 20 Its adaptability to industrial production is due to its ease of manufacturing. In this respect, it is low-cost. developing a cost-effective, sustainable and domestically produced wound care product It provides an opportunity. Explanation of the Figures: 25 The invention will be described by referring to the attached figures, so that the features of the invention can be explained. It will be understood and appreciated more clearly, but the purpose of this invention is this obvious It is not about limiting it with regulations. On the contrary, the invention is defined by the accompanying claims. all alternatives, modifications and 30 that can be included within the defined area The aim is to cover their equivalences. The details shown are only for the present invention. It is shown to illustrate the preferred arrangements and both the methods 4 shaping, as well as the most useful and conceptual features of the invention's rules and principles It should be understood that these drawings are presented to provide an easily understandable definition. In these drawings; Figure 1. FTIR analysis of the composite wound dressing made of cotton fabric, the subject of this invention. Figure 2 XRD spectrum of the composite wound dressing subject to the invention 5 Figure 3a SEM image of cotton. Figure 3b SEM image of the composite wound dressing that is the subject of the invention. Figure 4 EDS spectrum (a) Cotton (b) Composite product subject to the invention Figure 5. Cell viability analysis results for the groups used in the experimental study. (*: p<0.05) 10 Figure 6a Control group (commercial DMEM-control fabric extract), 2-32 applications (commercial DMEM-2-32 fabric extract) wound test images Figure 6b % wound closure data (**; p< 0.01) Figure 7 Wound staining with hematoxylin–eosin and Crossmon's trichrome. Histopathological evaluation of the region; re-epithelialization (arrows), 15 Inflammatory cell infiltration (square), angiogenesis (arrowheads), collagen Production and granulation tissue (star) are shown. Crossmon's Trichrome staining (first and second rows), Hematoxylin–Eosin Staining (third and fourth rows). Scale bar: 200 µm. Figure 8 Immunohistochemical representation of TGF-β and VEGF-A expression. 20 TGF-β immunohistochemical staining (first and second rows), VEGF-β immunohistochemical staining (third and fourth rows), TGF-β and VEGF-A positive cells (arrows). Streptavidin-biotin-peroxidase Method; Scale bar: 100 µm Figure 9 AATCC 147 Parallel Line Method of cotton fabric (P0) and composite 25 Evaluation of the antibacterial activity of the wound dressing (P1): (a) against Escherichia coli and (b) Staphylococcus aureus bacteria Inhibition images. Figure 10 Antibacterial properties of cotton fabric (Control) and composite wound dressing (P1). Evaluation of activity: (a) Against Escherichia coli bacteria 30 (b) Antibacterial effect image against Staphylococcus aureus bacteria. (c) Antibacterial effect image and (c) AATCC 100 Quantitative According to the Antibacterial Test Method, logarithmic decrease in bacterial load. graph. Illustrations that will help understand this invention are shown in the attached image. They are numbered and listed below with their names. 5 Detailed Description of the Invention: The terminology used here is intended solely to describe specific applications. and does not limit the scope of the invention. The number 10 used here The term "and / or" refers to any of the items listed as related. It includes one and all combinations thereof. Also, the singular "one" used here, "one" The terms "number" and "specified" are used in their plural forms unless the context explicitly indicates otherwise. It is designed to include singular forms such as those mentioned above. Furthermore, the terms used in this specification... The terms "includes" and / or "contains" refer to the specified features, steps, processes, 15 It indicates the presence of elements and / or components, but one or more other features, steps, processes, elements, components and / or groups thereof It will be understood that this does not exclude its existence or addition. Unless otherwise noted, all terms used herein (including technical and scientific terms) are copyrighted under Article 20 of the Turkish Penal Code. in the sense that a person with general knowledge in the field to which this invention belongs would generally understand it They have the same meaning. Furthermore, as defined in commonly used dictionaries... The terms will have a meaning consistent with the context of the relevant field and this explanation. it should be interpreted as idealized unless otherwise explicitly defined here. or it will be understood that it will not be interpreted in an overly formal sense. 25 The description of the invention will reveal a series of techniques and steps involved. These are: Each of them provides benefits individually, and at the same time, one or more of them can be used together. In some cases, all of the other techniques described may be used in combination. Accordingly, To ensure clarity, each step in the disclosure of the invention should be presented in 30 steps if possible. By doing this, unnecessary repetition of all combinations will be avoided. 6 Together, the specification and claims, such combinations fully constitute an invention and claims. It should be read with the understanding that it falls within its scope. The invention involves combining silver nanoparticles with naturally derived components in a cotton-based carrier. antibacterial, hemostatic, biocompatible and wound-resistant, when combined on the surface. a multifunctional composite wound dressing with properties that support healing It is related to this. The composite wound dressing developed within the scope of the invention is primarily cotton-based. a carrier surface, sodium alginate, Aloe vera, Hypericum perforatum, Achillea silver nanoparticles with millefolium and Equisetum arvense plant extracts. It contains a sodium alginate composite structure with biocompatible and liquid absorbent 10. When forming a structure, Aloe vera and the aforementioned plant extracts promote wound healing. They are included as supporting components. Silver nanoparticles are used in composite wound healing. It provides antibacterial properties to the dressing. The subject of the invention is a composite wound dressing. In an application aimed at realizing the covering, the surface is used as the carrier surface. It has undergone desizing and bleaching processes to remove impurities, 15 100% cotton plain weave fabric without optical brighteners is used. The preparation of plant extracts is as follows; they are used in composite wound dressings. The soil of Hypericum perforatum, Achillea millefolium and Equisetum arvense plants The upper parts are cut into appropriately sized pieces. Each plant specimen is individually divided into 20... In a hydro-alcoholic solvent system with a MeOH₂O ratio of 1:1 at 60 °C It undergoes an extraction process. The product obtained after the extraction process... The extracts are filtered, and the solvent phase is removed, thus concentrating the extracts. The process continues and then the solvent is completely removed. The dried plant... The extracts are re-dissolved in pure water and made ready for use. 25 The prepared extracts are stored at 4°C until use. is being done. The composite wound dressing that is the subject of the invention is prepared as follows: composite wound Before preparing the cover, wash 100% cotton fabric with non-ionic detergent for 30 minutes. It is washed three times using this method, rinsed, and then cooked at 121°C for 45 minutes. It is sterilized in an autoclave for a certain period of time. Coating to be applied to cotton fabric. 7 To prepare the solution, 2 g of sodium alginate was mixed with 100 mL of pure water. It is dissolved. 10 mL of fresh aloe vera gel is added to the resulting sodium alginate solution. The mixture is added and homogenized for 1 hour using a magnetic stirrer. It is processed. Following the homogenization process, Hypericum perforatum, Achillea 5 of each extract prepared from the plants millefolium and Equisetum arvense 100 mg is added to the mixture and the mixing process is continued for another hour. The process continues. Next, 0.003 mol of AgNO₃ is added to the prepared mixture. The system is stirred for 1 hour. The silver is present in the mixture. To enable the reduction of silver ions (Ag⁺) to metallic silver (Ag⁰), 0.05 was added to the system. 1 mL of M NaBH₄ solution is added and the mixture is stirred for another hour. 10 Thus, a coating containing silver nanoparticles and the aforementioned natural components. A solution is obtained. The prepared coating solution is applied to sterilized cotton fabric. It is placed inside and impregnated at room temperature for 2 hours. This At the end of the time, the fabric is removed from the coating solution and placed in the machine under 5 bar pressure. It undergoes an impregnation-drying-curing process underneath. The coating process takes a total of 15 days. This is repeated five times. The coated fabric is placed in a stenter at 80°C for 5 minutes. It is dried in an oven and then cured at 180 °C for 5 minutes. It is subjected to a process. Thanks to these processes, the coating solution is produced. adhesion and fixation of the delivering components to the cotton-based carrier surface is provided. The composite fabric, which is the subject of the invention, is obtained after the curing process. 20 It is washed using pure water and then reheated at 80°C for 5 minutes. It is dried. Thus, a composite wound dressing suitable for application to the wound surface is prepared. is obtained. The structural properties of the composite wound dressing, which is the subject of the invention, were determined using FTIR, XRD, SEM, and EDS 25. It has been evaluated through analyses. The FTIR analysis results for the composite wound dressing made of pure cotton fabric are shown in Figure 1. This is shown. The spectrum of pure cotton fabric is approximately 3332 cm⁻¹. The observed broadband cellulose structure exhibits –OH stretching vibrations, 2896 cm⁻¹ 30 The band at level 1430, 1366 and is associated with aliphatic C–H stretching vibrations. The bands observed around 1314 cm⁻¹ are C–H bending and O–H bending, characteristic of the cellulosic structure. 8 While associated with vibrations, the bands in the range of 1200–1027 cm⁻¹ are C–O–C and C–O strain. It shows the vibrations. The invention concerns a composite wound dressing in which the base bands of cotton fabric are preserved. However, the intensities of these bands and some band locations are 5 Changes appear to have occurred, particularly in the broadband of –OH groups. The change that occurred was due to sodium alginate, aloe vera, and plant extracts found in these substances. This shows that hydroxyl groups are interacting with the cotton surface. Furthermore, changes in the 1600-1400 cm⁻¹ range were observed in sodium alginate and plant-based components. Interaction of carboxylate and phenolic groups with silver nanoparticles found in 10 It supports the fact that polysaccharide bands are located in the range of 1200-1000 cm⁻¹. The preservation means that the cotton-based structure is protected after the coating process. This shows that the FTIR analysis given in Figure 1 shows that sodium alginate, aloe vera, With extracts of Hypericum perforatum, Achillea millefolium and Equisetum arvense This supports the integration of silver nanoparticles into the surface of cotton fabric. 15 The XRD spectrum of the composite wound dressing, which is the subject of this invention, is shown in Figure 2. In the XRD analysis, characteristic peaks were observed at 2θ values ​​of 15.26°, 23.11° and 34.77°. It is compatible with the cellulose I crystalline structure of cotton fabric. Peak value of approximately 15.26°. Crystallographic plane of cellulose (101) and peak (002) crystallographic plane with a value of 23.11° 20 The plane is associated with the crystallographic plane and the peak around 34.77° is associated with the (023) crystallographic plane. The peak observed at approximately 16.86° is associated with the semi-crystalline nature of the cellulosic structure. It consists of sodium alginate, plant extracts, and silver nanoparticles applied to the cotton surface. It is related to structural interactions that occur after immobilization. Approximately The peak observed at a value of 2θ≈21° was determined as the characteristic peak of sodium alginate. 25 In addition, the peaks observed at 32.85°, 38.63° and 65.01° indicate metallic Ag. They correspond to planes (122), (111) and (220) respectively. SEM of surface morphologies of pure cotton fabric and composite wound dressing. The images are shown in Figure 3. The surface of pure cotton fabric seen in Figure 3a is 30 It has a relatively regular, homogeneous, and smooth morphology. In contrast, Figure In the composite wound dressing shown in 3b, the surface of the cotton fibers is rougher and 9 It exhibits a heterogeneous morphology. Bright, even colors are distributed on the fiber surface. The point formations are associated with silver nanoparticles immobilized on the surface. Fiber The coating layer that forms on its surface also contains sodium alginate, aloe vera, and plant extracts. This shows that the extracts adhere to the cotton surface. EDS spectra of pure cotton fabric and composite wound dressing are shown in Figure 4. This is shown. The EDS spectrum of pure cotton fabric is predominantly composed of C and O. While peaks belonging to the elements are observed, C and O peaks are added to the composite wound dressing. Characteristic Ag peaks were identified in the approximately 2.9-3.2 keV region. SEM Point structures detected in the images and Ag 10 obtained in EDS analysis Combined evaluation of signals shows that silver nanoparticles are used on a cotton-based carrier. This indicates that it is immobilized on the surface. To determine the biocompatibility of the composite wound dressing that is the subject of this invention. MTT analysis was performed using the L929 mouse fibroblast cell line. The cells were 15 In DMEM medium containing 10% FBS and 1% penicillin / streptomycin, at 37 °C and 5% The fabrics were cultured in a CO2 atmosphere. Prior to analysis, the fabric samples were subjected to UV light. It was sterilized and applied to cell culture medium. The cells were placed along with the samples. They were incubated for 24, 48, and 72 hours. At the end of the incubation process, MTT The solution was added, and the resulting formazan crystals were placed in DMSO for 20 minutes. The solution was dissolved and absorbance values ​​were measured at a wavelength of 492 nm. Cell viability was calculated using the following equation:  % Viability = (Sample / Control) × 100 25 Cell viability analysis results for pure cotton fabric and composite wound dressing are shown in the figure. This is shown in section 5. The analysis results show a control and composite wound with a diameter of 1 mm. Cytotoxic effect of coating samples on L929 cells at 24, 48 and 72 hours. 30 that did not form and all cell viability values ​​were approximately above 70% It has been determined. In addition, cell viability obtained in the composite wound dressing at 24 hours It was found that the level of increase was significantly higher compared to control DMEM and control fabric samples. The in vitro wound healing potential of composite wound dressing in humans was determined (p<0.05). Scratch analysis performed using dermal fibroblast cells The cells were evaluated. They formed a single layer in 24-well culture plates. cultured in this way and an artificial wound area was created using a sterile pipette tip. It was brought in. Cellular debris was removed using PBS and the control and application 5 Fabric extracts belonging to this group were added to the cell medium. Wound areas Images were taken at 0, 24, and 48 hours, and wound closure was monitored using ImageJ software. The rates have been determined. The aforementioned Scratch analysis images and wound closure The percentages are shown in Figures 6a and 6b. The results obtained are for composite wound care. its covering did not show cytotoxic effects and wound closure occurred through cell proliferation 10 This shows that it supports the process. The composite wound dressing described in this invention has hemostatic properties for in vivo bleeding. It was evaluated using a model. Under general anesthesia, the distal part of the mouse tail... A wound dressing was applied to the bleeding area and 15 minutes after the bleeding started The time it took for it to stop completely was measured. The composite material was evaluated. Bleeding stopped approximately 3.70 seconds after the dressing was applied. It has been determined that it stopped working. This result indicates that the composite wound dressing has rapid hemostatic properties. It has been shown that the composite structure exhibits antibacterial and wound-relieving properties. With its healing properties and ability to provide rapid bleeding control, the wound dressing is 20 It supports a multifunctional structure. Excisional surgery was performed to determine the wound healing effectiveness of composite wound dressings. A wound model was used. The mice used in the study were divided into control and treatment groups. They were divided into two groups. In the control group, any 25 wound areas were injected. The wound areas in the treatment group that did not have wound dressings applied were the subject of the invention. Composite wound dressing was applied. Wound areas were evaluated at points 0, 7, 14, and Standardized digital images obtained on day 21 were used. Wound areas Wound contraction rates were measured using ImageJ software;  Wound contraction (%) = [(A₀ - Aₜ) / A₀] × 100 11 It was calculated using the equality. Here, A₀ is the initial wound area, and Aₜ is the relevant area. It refers to the wound area measured over time. The subject of the control and invention is composite wound Changes in wound area in the groups that received the dressing are shown in Table 1. Day Control Group Invention topic wound dressing p-value 0 2.24 ± 0.28 2.43 ± 0.35 0.328 7 0.55 ± 0.22 1.38 ± 0.20 <0.001 14 0.24 ± 0.07 1.03 ± 0.34 0.006 21 0.14 ± 0.08 0.07 ± 0.11 0.239 Table 1 - Wound dressings in control and subject groups during the experimental period. changes in area (cm²) Data are expressed as mean ± standard deviation. In intergroup comparisons... Welch corrected independent samples t-test was used, and p<0.05 values ​​were considered statistically insignificant. This was considered significant. Wound contraction percentages are shown in Table 2. It is shown: Day Control group Subject of invention wound dressing p-value 7 75.52 ± 8.06 41.51 ± 16.35 0.006 14 88.92 ± 4.17 55.45 ± 19.72 0.018 21 93.56 ± 3.52 97.35 ± 3.75 0.118 Table 2 - Wound dressings in control and experimental groups during the trial period 15 contraction percentage (%) changes Wound in the group that received composite wound dressing at the end of the 21st postoperative day It was determined that the contraction level reached 97.35%. At the end of the experimental period, the results obtained from the control and composite wound dressing groups... Hematological findings are shown in Table 3. 12 10 20 Table 3 - Control and experimental wound dressing groups at the end of the trial period. hematological parameters As a result of the evaluation of the measured hematological parameters, the subject of the invention No significant systemic toxicity was observed in the experimental group where the composite wound dressing was applied. 25 It has been determined that none were observed. The invention aims to determine the effect of a composite wound dressing on wound healing using tissue. The samples were evaluated histopathologically. Tissue samples were placed in 10% neutral buffer. Formalin-fixed, embedded in paraffin blocks, and approximately 5 µm 30 Sections of a certain thickness were prepared. Hematoxylin-Eosin and Modified Acid were added to the sections. Crossmon Trichrome staining was applied. Histopathological examinations were performed. Parameter Control Group Subject of the invention: Wound dressing p-value WBC (×10⁹ / L) 6.97 ± 4.03 13.08 ± 0.79 0.013 LYM# (×10⁹ / L) 4.52 ± 2.36 8.46 ± 0.58 0.008 MON# (×10⁹ / L) 0.18 ± 0.10 0.32 ± 0.08 0.037 GRAN# (×10⁹ / L) 2.30 ± 1.60 4.30 ± 0.56 0.027 LYM (%) 65.42 ± 5.06 64.66 ± 2.74 0.772 MON (%) 2.87 ± 0.51 2.52 ± 0.71 0.369 GRAN (%) 31.87 ± 4.72 32.82 ± 3.21 0.711 RBC (×10¹² / L) ​​5.19 ± 1.59 7.05 ± 0.26 0.034 HGB (g / dL) 10.20 ± 2.97 13.64 ± 0.42 0.036 HCT (%) 28.17 ± 8.45 39.94 ± 1.68 0.018 MCV (fL) 54.38 ± 0.75 56.66 ± 0.69 <0.001 MCH (pg) 19.93 ± 1.30 18.48 ± 0.26 0.037 MCHC (g / dL) 36.65 ± 1.86 33.82 ± 0.50 0.010 RDW (%) 14.10 ± 1.39 15.10 ± 0.27 0.140 PLT (×10⁹ / L) 419.83 ± 575.03 1232.60 ± 21.82 0.018 MPV (fL) 5.28 ± 0.26 5.04 ± 0.11 0.089 PDW 16.95 ± 0.84 15.70 ± 0.16 0.014 PCT (%) 0.17 ± 0.20 0.556 ± 0.026 0.005 13 inflammatory cell infiltration, edema, granulation tissue formation, re-epithelialization and angiogenesis parameters were evaluated. The obtained histopathological scores This is shown in Table 4. Table 4 - Histopathological scores of control and inventory wound dressing groups 5 Histopathological evaluation revealed that the composite wound dressing in question... In the group where it was applied, there was a decrease in inflammatory cell infiltration and edema values. Increased parameters in granulation tissue formation, re-epithelialization, and angiogenesis. It has been determined. Hematoxylin-Eosin and 10 related to the aforementioned histopathological evaluation. Crossmon's trichrome images are shown in Figure 7. Figure 7 shows re- epithelialization, inflammatory cell infiltration, angiogenesis, collagen production, and The findings regarding granulation tissue formation are shown. The subject of the invention is a composite wound dressing with properties related to wound healing and tissue regeneration. TGF-β and VEGF-A to evaluate their effects on biological processes. Their expression was examined immunohistochemically. Immunohistochemical TGF-beta 1 and VEGFA primary antibodies were used in staining, and DAB was used as the chromogen. The procedure was applied and counterstaining was performed with Mayer hematoxylin. Staining intensity; Semi-20 will have 0 no coloring, 1 light coloring, 2 medium coloring, and 3 intense coloring. Inflammatory Cell infiltration Edema Granulation Tissue Formation Re-epithelialization Angiogenesis Day Group n Median p- value Median p- value Median p- value Median p- value Median p- value 7 Control 6 3.00a 0.005 2.00a 0.027 1.00b 0.026 0.00b 0.018 1.00b 0.026 Subject of invention wound dressing 6 2.00b 1.00b 2.00a 1.00a 2.00a 14 Control 6 3.00a 0.014 1.50a 0.043 1.50b 0.019 2.00 0.06 2.00b 0.030 Subject of invention wound dressing 6 2.00b 1.00b 2.50a 3.00 2.50a 21 Control 6 2.00a 0.019 1.00 0.176 2.00 0.269 2.50 0.241 2.00 0.338 Subject of invention wound dressing 6 1.00b 1.00 3.00 3.00 2.00 14 The results were evaluated quantitatively. The obtained VEGF-A and TGF-β scores are shown in Table 5. It is shown. VEGFA TGF-β Day Group n Median p-value Median p-value 7 Control 6 1.00b 0.018 1.00b 0.005 Subject of invention wound dressing 6 2.00a 2.00a 14 Control 6 2.00b 0.019 2.00b 0.018 Subject of invention wound dressing 6 3.00a 3.00a 21 Control 6 2.00 0.269 2.00 0.248 Subject of invention wound dressing 6 3.00 2.50 Table 5 - VEGF-β and TGF-β 5 of the control and subject wound dressing groups. immunohistochemical scores Values ​​with different superscript letters in the same column are grouped at the relevant time point. It indicates that there is a statistically significant difference between them (p<0.05). In the group treated with composite wound dressing, VEGF-A and TGF-β expression levels were 10% higher. It was determined that TGF-β and VEGF-A levels increased according to immunohistochemical evaluation. Expression images are shown in Figure 8. The results obtained are related to composite wound care. the processes of angiogenesis, tissue regeneration and wound healing in its covering This shows that he supports it. The invention concerns the antibacterial activity of composite wound dressing against Gram-positive bacteria. Staphylococcus aureus (ATCC 29213) and Gram-negative Escherichia coli (ATCC 25922) bacteria were used for evaluation. In antibacterial evaluations AATCC 147 Parallel Line Method and AATCC 100 Quantitative Antibacterial Test The method was used. The AATCC 147 Parallel Line Method resulted in pure cotton fabric 20 composite wound dressing against Escherichia coli and Staphylococcus aureus The antibacterial behavior of the composite wound dressing is shown in Figure 9. In samples where the coating was applied, the growth of both bacterial species was lower than in the control group. compared to that, bacterial density was suppressed and decreased in fabric contact areas. It has been determined. Fabric sterilized according to the AATCC 100 Quantitative Antibacterial Test Method. Samples were treated with bacterial suspensions and incubated at 37°C for 24 hours. The bacteria on the fabric surface were incubated into a suitable solution. The mixture was transferred and the suspensions were added to agar medium. The results obtained at the end of incubation... The incoming colonies were counted and CFU / mL values ​​were calculated, and the composite subject to the invention was determined. The wound dressing was compared with pure cotton fabric in the control group. The results of the quantitative antibacterial analysis are shown in Figure 10. Escherichia coli 10 a reduction of approximately 4.88 log₁₀ and over 99.99% antibacterial effect, For Staphylococcus aureus, the reduction was approximately 2.00 log₁₀ and approximately 99.00%. Antibacterial efficacy has been achieved. These results relate to the composite wound healing compound that is the subject of this invention. Its coating is antibacterial against both Gram-negative and Gram-positive bacteria. This reveals that it exhibits this characteristic. 15 The characterization described above is considered together with in vitro and in vivo evaluations. when taken, sodium alginate, Aloe vera, Hypericum perforatum, Achillea millefolium and Equisetum arvense extracts and silver nanoparticles on a cotton-based carrier As a result of being brought together on the surface, it is biocompatible, antibacterial, fast 20 A multifunctional composite with hemostatic effect and promotes wound healing. Wound dressings are produced. 30

Claims

16 REQUESTS 1- The invention relates to a cotton-based carrier surface and the surface built upon it. It is an immobilized composite wound dressing, composed of sodium alginate and aloe vera. Hypericum perforatum extract, Achillea millefolium extract, Equisetum arvense 5 It is characterized by containing extract and silver nanoparticles. 2- The composite wound dressing mentioned in Claim 1 is characterized by its cotton-based composition. It is characterized by its carrier surface being 100% cotton plain weave fabric. It is done. 10 3- The composite wound dressing mentioned in Claim 1 or 2 is characterized by its... sodium alginate, Aloe vera, Hypericum perforatum extract, Achillea millefolium cotton-based extract of Equisetum arvense extract and silver nanoparticles 15 characterized by being present as a coating layer on the carrier surface. It is done. 4- Composite wound dressing according to either of claims 1-3, with the characteristic of being AgNO3. silver ions obtained from the source are converted into metallic silver using NaBH4. 20 characterized by containing silver nanoparticles formed by reduction. It is done. 5- The invention relates to a cotton-based carrier surface and the surface built upon it. It is a method of producing an immobilized composite wound dressing, the characteristic of which is;  Dissolving sodium alginate in pure water, 25  Aloe vera is added to the resulting sodium alginate solution and homogenized. being done,  Add Hypericum perforatum, Achillea millefolium and Equisetum arvense to the mixture. addition of extracts,  Adding AgNO3 to the mixture, 30  Addition of NaBH4 to reduce silver ions to metallic silver, 17  Impregnation of the cotton-based carrier surface with the resulting coating solution to be done  Impregnation-drying-curing process applied to the impregnated carrier surface It includes the steps of the process. 6- The method mentioned in claim 5, its characteristic is; 2 g of sodium alginate in 100 mL of pure water dissolved in the solution and add 10 mL of aloe vera gel to the resulting solution. It is characterized by... 7- The method mentioned in claim 5 or 6, its characteristic being; Hypericum perforatum, Achillea 10 100 mg each of millefolium and Equisetum arvense extracts It is characterized by its addition to the mixture. 8- The method according to either of claims 5-7, and its characteristic is; adding 0.003 mol of AgNO3 to the mixture. 0.05 M NaBH4 15 was added for the purpose of reducing silver ions. It is characterized by the addition of 1 mL of the solution. 9- The method is based on either of the requirements 5-8, and its characteristic is; cotton-based carrier. impregnation of the surface in the coating solution at room temperature for 2 hours. It is characterized by... 20 10- A method according to either of claims 5-9, characterized by its impregnation-drying process. The curing process is carried out in the machine under 5 bar pressure and the coating It is characterized by the process being repeated a total of five times. 11- A method according to either of claims 5-10, characterized by its type; the coated carrier. drying the surface at 80 °C for 5 minutes and then at 180 °C for 5 minutes It is characterized by being subjected to a curing process. 12- A method according to either of claims 5-11, characterized by; curing process 30 Afterwards, the composite structure is washed with pure water and re-treated at 80°C for 5 minutes. It is characterized by its drying process.

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