Multilayer wound dressing material that accelerates wound healing

Abstract

The present disclosure relates to spunbond nonwoven fabric surfaces coated with multiple multilayer film layers comprising groups containing hyaluronic acid, chitosan and Theranekron, and to the related production method, to impart functional efficacy to wound dressing materials that are widely used in the healthcare sector.

Description

[0001] MULTILAYER WOUND DRESSING MATERIAL THAT ACCELERATES WOUND HEALING

[0002] Technical Field

[0003] The present disclosure relates to spunbond nonwoven fabric surfaces coated with multiple multilayer film layers comprising groups containing hyaluronic acid, chitosan, and Theranekron, and to the related production method, to impart functional efficacy to wound dressing materials that are widely used in the healthcare sector.

[0004] Prior Art

[0005] Wound healing consists of a dynamic process containing different stages and influenced by numerous factors. In this process, wound healing is generally divided into three main phases. These phases are inflammation, proliferation, and remodelling. This process exhibits healing at different durations depending on disease types and the nature of the wound. Possible disruptions that may occur in the events ensuring the progression of the process and its successful completion may lead to failure of healing or result in an unfavourable outcome. Scientists strive to develop the ideal method by experimenting with active substances or techniques that may influence various steps of these healing phases in their studies.

[0006] In the known art, in the production of cream-type wound dressing materials that are frequently used in wound healing, substances providing antibacterial efficacy and suppressing the severe inflammation picture caused by bacteria such as Staphylococcus aureus, which dominate the skin flora, are used in preferred combinations of active substances. On the other hand, in the proliferation phase, which is an important intermediate step in wound repair, active substances or methods that can increase cellular mitotic activity are tested. Despite the efficacy of these dressing materials, in which a large number of active substances are incorporated into combinations, an almost ideal therapeutic combination has not yet been achieved. Just as repair cannot be achieved with insufficient procoagulant effect, antibacterial activityand proliferative effect, in the presence of increased activities, wound healing that ends with intense scar formation may also be observed.

[0007] As is known, wound healing creates physical, psychological and financial adverse effects on patients and healthcare systems. Problems in the wound care process constitute a challenging situation for both the patient and the hospital. For this reason, wound care must be carried out carefully and systematically, optimised by considering the condition of patients and hospital resources, and alternative wound healing systems must be developed.

[0008] Among the wound treatment methods used today are alginate dressings, polyurethane films, hydrogel dressings, hydrocolloid dressings, foams and bioactive dressings containing hyaluronic acid, collagen, elastin, alginates and chitosan. The high costs of these treatment materials create an economic burden for hospitals and the relatives of patients. Therefore, patients generally have to resort to more traditional treatment methods, and the wound healing process is prolonged. Besides the diversity of wound dressing materials, there are also some applications involving devices that use negative pressure, and these applications help cells that contribute to repair migrate to the lesion area. Due to the high cost of the consumables used during such applications, the economic burden on the public is also quite high, and this situation creates pressure on the healthcare system. From the hospital perspective, infection control measures, inadequacy of healthcare personnel, lack of training and insufficiencies in materials also make wound healing more difficult.

[0009] Consequently, due to the disadvantages and shortcomings mentioned above, a need has arisen for an innovation in the related technical field.

[0010] The Aim of the Invention

[0011] The aim of the present invention relates to a multilayer wound dressing material that accelerates wound healing, meets the requirements mentioned above, eliminates all disadvantages and provides additional advantages.

[0012] The aim of the invention is to set forth a method for coating spunbond nonwoven fabric surfaces with multiple multilayer film layers comprising groups containing hyaluronicacid, chitosan, and Theranekron, to impart functional efficacy to wound dressing materials that are widely used in the healthcare sector.

[0013] The invention aims to provide a wound dressing material that will enable influencing different steps of the phases of wound healing and accelerate the repair process. The invention aims to obtain a wound healing process that may conclude with minimal scar formation.

[0014] The invention aims to provide a wound dressing material that is easy to use and economical.

[0015] To achieve the aims described above, the present disclosure is a wound dressing material used in wound healing in the healthcare sector and made of spunbond fabric, wherein it comprises multilayer films consisting of hyaluronic acid, chitosan and Theranekron from the inside (from the wound surface) to the outside on the said spunbond fabric surface, to enable influencing different steps of the inflammation, proliferation and remodelling phases of wound healing and thereby accelerate the repair process.

[0016] To achieve the aims described above, the present disclosure is a production method of a wound dressing material used in wound healing in the healthcare sector and made of spunbond fabric, wherein, to influence different steps of the inflammation, proliferation and remodelling phases of wound healing and thereby accelerate the repair process, the method comprises the process steps of:

[0017] • rendering the cotton spunbond nonwoven fabric surface that is cleaned with alcohol cationic, keeping the fabric for 10 minutes at room temperature in a 5% polyethyleneimine solution, and then drying at SO'C for 5 minutes and fixing at S'C for 5 minutes,

[0018] • preparing 3% hyaluronic acid (HA) and 0.5% chitosan (C) solutions in order to obtain multilayer coating on mentioned fabric surface, immersion-holding the fabric respectively in the HA solution, distilled water, the K solution and distilled water, drying the multilayer films formed on the fabric at OO'C and fixing them at ISO'C for 3 minutes, and• preparing the Theranekron (T) solution and impregnating the multilayer film- coated fabric for 10 minutes, and then drying the fabric at OO'C and fixing it at ISO'C for 3 minutes.

[0019] The structural and characteristic features of the invention and all advantages will be understood more clearly through the detailed description provided below, and therefore, the assessment should also be made by considering this detailed description.

[0020] Detailed Description of the Invention

[0021] In this detailed description, the multilayer wound dressing material that accelerates wound healing is explained solely for a better understanding of the subject and in a manner that creates no limiting effect.

[0022] The present disclosure relates to a method for coating spunbond nonwoven fabric surfaces with multiple multilayer film layers comprising groups containing hyaluronic acid, chitosan, and Theranekron, to impart functional efficacy to wound dressing materials that are widely used in the healthcare sector. The feature of the present disclosure is to obtain a wound dressing material that enables influencing different steps of the inflammation, proliferation and remodelling phases of wound healing, and thereby accelerates the repair process.

[0023] The wound dressing material introduced within the scope of the present disclosure comprises Theranekron® (T), hyaluronic acid (HA), and chitosan (C) as active ingredients.

[0024] It is known that venoms obtained from poisonous animals have been used as a therapeutic method throughout human history for many years. Theranekron® (T), similarly, is the alcoholic extract of the toxin obtained by processing the venom of the poisonous spider T. cubensis with a 60% alcohol solution, and it is used in veterinary medicine as a medicinal product for wound repair and for the regression of certain vesicular lesions. It is known that the venom encountered following the spider’s bite of humans may also lead to death. In this case, the basis for using medicines for therapeutic purposes is the dosage of the medicine. The same drug may exhibit a particular activity at low doses, while showing the opposite activity at higher doses.Analyses carried out during the determination of the therapeutic doses of this venom have revealed the potential beneficial effects of the drug. In addition to its antiinflammatory, antioxidant and anti-apoptotic effects at the cellular level, it has been proven that it also contributes significantly to wound healing. In cell culture studies, it has been observed that low doses may enhance wound healing by increasing cellular proliferation, and that Theranekron® applied onto open wounds also accelerates healing significantly.

[0025] Hyaluronic acid (HA) is considered a crucial component of the extracellular matrix, and it also plays significant roles in cellular migration, signal transmission, and wound repair. HA is found at high concentrations in the skin and soft connective tissues. Therefore, it is a suitable choice for a matrix that supports dermal regeneration, namely the renewal and strengthening of the skin. It has been observed that shortly after a wound occurs, the presence of HA increases at the wound site. With the increase in HA, cellular migration in the injury area rises, connective tissue formation accelerates, and it has been demonstrated that cross-linking between collagens increases, thereby enhancing the strength of the wound tissue.

[0026] Chitosan (C), on the other hand, is a polymer available abundantly in nature and possesses properties such as being biodegradable, biocompatible, non-toxic and adsorptive. C is a polysaccharide that supports many living organisms and is renewable. Chitin is the skeletal material of invertebrates such as crabs, lobsters and shrimps, and it is obtained from their shells. It is known to be a renewable resource with no waste concerns in sectors such as aquaculture, textiles, food, and ecology. The polycationic nature of C allows its use in many areas. One of these is that it and its derivatives have been proven to have proliferation-enhancing, haemostasis-inducing, antibacterial and antifungal activities. Owing to its haemostatic property, it may be effective in the early phases of wound healing called primary haemostasis. Through its ability to increase the production of growth factors, it allows cellular proliferation to be enhanced. On the other hand, its inhibition of the growth and spread of populations of a wide range of bacteria and yeast species, and its ability to suppress bacterial activity at a high level compared with other disinfectants, may create an appropriate environment for the healthy initiation and continuation of wound repair.In studies conducted within the scope of the present disclosure, it has been proven that T, particularly at doses of 10-50 mcg (low doses), increases the synthesis of growth factors required for proliferation in wound tissue. In samples taken from the wound tissues of rats with experimentally created wound models on day 7, it was demonstrated in the animal study that it increased the expressions of vascular endothelial growth factor, fibroblast growth factor, epidermal growth factor, platelet-derived growth factor, and transforming growth factor beta at doses of 10-50 mcg. The findings of this study are as follows:

[0027] When the histopathological analysis results were examined, after the evaluation of haematoxylin-eosin staining, slow healing was observed in the control group that was left to heal without any intervention, whereas more pronounced healing was observed in the HA-C group. Healing was observed to become more evident in a dosedependent manner in the T10, T20, and T50 groups, whereas in the T100, T200, and T500 groups the amount of defect closure, neovascularisation and connective tissue development decreased. The best healing was observed in the T10 and T20 groups. Healing in the T50 group was similar to that in the T20 group.

[0028] When the blue-coloured connective tissue development and amount were evaluated in Masson trichrome staining, it was observed that the mild connective tissue amount and development in the control group increased markedly and displayed a regular appearance in the HA-C group. The T 10 group constituted the group in which the most pronounced and homogeneous connective tissue development was formed. In the T20 and T50 groups, a moderate level of regional connective tissue development concentrated at the base was observed. In the T100 group, a non-homogeneous connective tissue development was encountered, becoming evident especially in some areas at the base of the defect. In the T200 and T500 groups, connective tissue development was decreased, and in some regions completely immature connective tissue formation was noted. In the T 100 and higher groups, collagen development was observed to be irregular and heterogeneous in distribution.

[0029] In Piero Sirius Red staining performed to evaluate connective tissue formation and maturation, the assessments of red-coloured connective tissue development in the defect regions were carried out. With a polarised filter, the amounts of green (immature) and red (mature) collagen were evaluated. Under the polarised filter, regions giving a green reflection and consisting of fine Type III collagen were observed. Depending onthe groups, thicker Type I collagen formations giving a red-orange reflection were found to increase at varying levels. Type I collagen was observed to increase most prominently in the HA-C group. In the control group, collagen was found to be moderately matured. Connective tissue formation and maturation, which became pronounced in the T 10, T20, and T50 groups, decreased in the T 100, T200, and T500 groups. Additionally, it was shown that HA-C was effective in healing of the defect region, and it was noted that the T10, T20 and T50 groups were significantly effective in healing. In the T100, T200 and T500 groups, inflammation and necrosis were observed to begin to become evident.

[0030] When the genetic analysis results were examined by PCR, the maximum expression level of epidermal growth factor (EGF) gene expression was detected in the T20 group compared with the other groups. The levels of the T20 group showed statistical significance compared with all groups except the T10 group (p<0.001 for all). In the T10 group, a statistically significant increase was detected compared with the expressions observed in all groups except T20 and T50 (p<0.001 for all). In addition, in the T100, T200, and T500 groups, a significant decrease in expression levels was detected compared with the control and HA+C groups (p<0.001 for all).

[0031] In the fibroblast growth factor (FGF) gene expression levels, the maximum expression level was detected in the T20 group compared with the other groups. The levels of the T20 group showed statistical significance compared with all groups (p=0.032 for TW, p=0.015 for T50, and p<0.001 for the others). In the TW group, a statistically significant increase was detected compared with the expressions observed in the control, HA+K, TWO, and higher-dose T groups (p<0.001 for all). In addition, in the TWO, T200, and T500 groups, a significant decrease in expression levels was detected compared with the control and HA+C groups (p<0.001 for all). On the other hand, a significant decrease was also detected in the T500 group compared with the TWO group (p=0.046).

[0032] In the platelet-derived growth factor (PDGF) gene expression levels, the maximum expression level was observed in the T20 group, followed by the TW levels. The levels of the T20 group showed statistical significance compared with all groups except the TW group (p=0.027 for T50, p<0.001 for TWO and the others). In the TW group, although no significance was detected between the TW and T50 groups, a statistically significant difference was found between the TW group and the control and HA+Cgroups and the groups in which T 100 and higher doses were applied (p=0.003 for the control, p<0.001 for the others). In addition, the levels of the T50 group were significantly increased compared with the levels observed in the T 100, T200, and T500 groups (p=0.003, p<0.001 and p<0.001 , respectively).

[0033] In the transforming growth factor-p (TGF-[3) gene expression levels, the maximum expression level was detected in the T20 group and showed statistical significance compared with all other groups (p<0.001 for all). The T10 and T50 values were also detected as statistically significantly higher compared with all groups except T20 (p<0.001 for all). The T500, T200 and T 100 groups showed a decrease compared with the control and HA+K groups, whereas statistical significance was detected between T200 (p=0.022 and p=0.017, respectively) and the control, and between T500 (p=0.001 and p<0.001 , respectively) and the control.

[0034] The second active substance, K, appears to provide important advantages for wound healing owing to its haemostasis-inducing, proliferation-enhancing, and connective tissue formation-enhancing effects, in addition to its antibacterial activity. No incompatibility has been detected in the incorporation of this polymeric structure, which is added to many wound dressing materials, with other active substances. This substance, which can also be used in surface coatings, is easy and inexpensive to obtain.

[0035] The third substance, HA, can be used in many fields such as dermocosmetics, orthopaedics and wound healing, and has found routine use. Its use in all situations where structural integrity is impaired will provide beneficial effects, as it promotes connective tissue formation that will provide the necessary structural support for the wound and create a basis for the approximation of the two wound edges during epithelialisation.

[0036] In addition to the advantages provided by HA and C in many wound dressing materials, the property of T to increase growth factors particularly at doses of 10-50 mcg makes a significant contribution to the effect of these main polymers in repair.

[0037] With the method introduced by the present disclosure, coating the surface of the spunbond nonwoven fabric with the active ingredients Theranekron, hyaluronic acid and chitosan has increased the effectiveness of the wound dressing material. In order to ensure the standardisation of this coating, in the multilayer film coating to be carriedout on the spunbond fabric by the dip-withdraw method within the scope of the present disclosure, the amount of active ingredient embedded in the polymer and the number of layers may be adjusted according to the required amount of active ingredient release per unit time.

[0038] For surface preparation, the cotton spunbond nonwoven fabric surface is first cleaned, and a chemical modification process is carried out to obtain ionic charges. To obtain a cationic structure, polyethyleneimine (PEI, 5 g / L, pH: 10) is impregnated into the cotton nonwoven fabric. The fabric is dried in a laboratory-type oven at SO'C for 5 minutes and subsequently subjected to fixation at 105* for 5 minutes.

[0039] In the continuation of the process, homogeneous solutions of hyaluronic acid (HA) and chitosan (C) are prepared for multilayer coating. HA is prepared at 3 g / L, at pH 6 for use in anionic layers, and chitosan dispersions are prepared at 0.5 g / L, at pH 5, for use in cationic layers. HCI and NaOH are used to adjust the pH values of the solutions used in the coating process. Using the prepared solutions, in order to form two film layers on the fabric, immersion-holding steps are carried out successively in the HA solution, distilled water, the K solution, and distilled water. While the fabric is held for 5 minutes in the HA and K solutions, it is held for 2 minutes in distilled water for rinsing. These process steps are repeated until a multilayer, preferably 10-layer, film is obtained. Each HA and K solution is replaced / refreshed before a new operation. The multilayer films formed on the fabric are dried at 60<C and subsequently subjected to fixation at 130<C for 3 minutes.

[0040] In the final processing step for application of Theranekron, six different doses are taken using micropipettes 10, 20, 50, 100, 200, and 500 mcg added into beakers containing 100 mL distilled water, and each resulting solution is impregnated into the multilayer film-coated fabrics for 10 minutes. The fabrics coated with multilayer films of HA-C-T are dried at GO'C and subsequently subjected to fixation at ISO'C for 3 minutes.

[0041] It has been determined that the low doses of Theranekron, namely T 10, T20 and T50, exhibit beneficial effects on the wound, and the maximum effect was detected in the T20 group in which 20 mcg of active ingredient was used.For the wound dressing material obtained within the scope of the present disclosure, first a 10-layer HA / chitosan structure is prepared and then T is impregnated. Here, the outermost layer of the dressing material, which is the closest part to the wound surface, is specifically selected to be chitosan. It is known that chitosan applied onto the wound surface has haemostasis-inducing properties required in the early phases of wound healing and exerts a bactericidal effect against foreign bacterial agents, primarily Staphylococcus aureus, which are major constituents of the skin flora. Following haemostasis, Theranekron is applied to the intermediate layer to increase the synthesis of growth factors whose release is desired for repair. Hyaluronic acid is also added to support the extracellular matrix formed through the proliferation occurring at the cellular level under the influence of these growth factors, and this is the layer closest to the fabric, meaning the first layer to be coated and the layer farthest from the wound. That is, the order from the spunbond material towards the wound is hyaluronic acid, chitosan, and Theranekron, respectively. In this sequence, the pH of the skin was considered by examining the zeta potential curves of hyaluronic acid and chitosan.

Claims

CLAIMS1. A wound dressing material made of a spunbond fabric and used in wound healing in the healthcare sector characterized by comprising; multilayer films of hyaluronic acid, chitosan, and Theranekron arranged from the inside to the outside on the spunbond fabric surface, enabling influence over different steps of the inflammation, proliferation and remodelling phases of wound healing and thereby accelerating the repair process.

2. A production method of a wound dressing material made of a spunbond fabric and used in wound healing in the healthcare sector, wherein, to enable influencing different steps of the inflammation, proliferation, and remodelling phases of wound healing and thereby accelerate the repair process, the method comprises the process steps of:• rendering the cotton spunbond nonwoven fabric surface that is cleaned with alcohol cationic by keeping the fabric for 10 minutes at room temperature in a 5% polyethyleneimine solution and then drying the fabric at 80C for 5 minutes and fixing it at 105C for 5 minutes,• preparing 3% hyaluronic acid (HA) and 0.5% chitosan (C) solutions to obtain multilayer coating on mentioned fabric surface, subjecting the fabric respectively to immersion-holding in the HA solution, distilled water, the K solution and distilled water, drying the multilayer films formed on the fabric at 60C and fixing them at 130C for 3 minutes, and• preparing the Theranekron (T) solution and impregnating the multilayer film- coated fabric for 10 minutes and then drying the fabric at 60C and fixing it at ISO'C for 3 minutes.

3. The production method according to Claim 2, wherein the solution mentioned in step c is prepared by dissolving 20 mcg of Theranekron in 100 mL of distilled water.

4. The production method according to Claim 2 or Claim 3, wherein the spunbond fabric surface forming the resulting wound dressing material comprises multilayer films of hyaluronic acid, chitosan, and Theranekron arranged from the inside (facing the wound surface) to the outside.