WO2020208651A1

WO2020208651A1 - A dressing material for wound healing

Info

Publication number: WO2020208651A1
Application number: PCT/IN2020/050340
Authority: WO
Inventors: R Mala; A S Ruby CELSIA; N Hari PRASATH
Original assignee: R Mala
Priority date: 2019-04-10
Filing date: 2020-04-10
Publication date: 2020-10-15

Abstract

The present invention relates to dressing material for wound hearing purpose, comprising one ormore hydrogel layers, wherein a layer made up of soft silicon mesh forms the first layer, ahydrogel layer containing a hemostat and natural immune booster prepared from the plant C.hirsutus forms the second layer, a hydrogel layer containing hydroxyapatite nanorods, antibiotics and antibiotic adjuvant prepared from the plant T.purpureaI forms the third layer, ahydrogel layer containing a natural anti-inflammatory agents and selective matrix metalloproteinase 9 inhibitors prepared from the plant T.purpurea forms the fourth layer, ahydrogel layer containing zinc oxide nanomaterial, and an anti- inflammatory agent prepared from T.purpurea forms the fifth layer, a layer made up of non-woven silk cotton forms the sixthlayer, a layer made up of soft silicon forms the seventh layer.

Description

A DRESSING MATERIAL FOR WOUND HEALING

TECHNICAL FIELD OF INVENTION:

The present invention relates to a dressing material for wound healing purposes. More particularly it relates to a dressing material that is helpful in rapid healing of chronic wounds such as diabetic wounds and burn wounds.

BACKGROUND OF THE INVENTION

Wound is a silent epidemic that affects millions of people globally. Wound healing is a complex cascade of overlapping physiological events such as hemostasis, inflammation, proliferation and migration and remodeling stage. Different materials have been used as wound dressings since ancient times. Wound dressings used before many decades were passive dressings. They protect the wound from external environments. With technological advancement, a vast array of wound care products are available. Many existing wound care products carry medicaments that play a vital role in enhancing wound healing. Most commonly used medicaments are antimicrobial agents. The antimicrobial agent containing wound care products are formulated as gel, ointment, cream, lotion, foam, hydrocolloids, hydrogels etc. Hydrogels are an ideal component of wound dressings as they provide a cool and moist environment necessary for wound healing, and at the same time they can carry and release the antimicrobial or active components to the wound for healing. However, the existing polymers being used to prepare the hydrogel are synthetic polymers causing several side effects like central nervous system toxicity, hyperosmolarity, hemolysis, lactic acidosis etc. Therefore, there is a need in the existing art, for a dressing material, having a hydrogel prepared from a different type of polymer, which shall be devoid of these side effects, as well acting as an effective agent for quick healing of the wounds.

Most of the dressings have to be applied multiple times a day/week based on the severity of the wound. Repeated dressing of the wound causes trauma, pain and discomfort to the patient. In order to overcome these difficulties, it's desirable that dressings that can be retained at the wound site for a longer period of time. Moreover, the dressing material that uses insitu gel formation for the hydrogels, should be preferred, so that it facilitates gel formation only after being applied to the wound. Therefore, there is a need in the art for a dressing material that contains both the ideal hydrogel devoid of side effects and at the same time it should be suitable for in situ gel formation.

Most of the wound care products currently available contain antibiotics or antimicrobial agents for their topical delivery. They don’t function in the inflammatory, proliferation and remodeling phase of the wound healing.

Under normal physiological conditions, there is a well orchestrated balance between the generation of reactive oxygen species and their scavenging by antioxidant enzymes and biomolecules. In contrast, in case of chronic wounds, the oxidative stress is exacerbated and the inflammation and cellular damage are beyond the normal regulation. In chronic wounds like burn wound and diabetic wound, many genes are upregulated and down regulated in haphazard manner. Among the proteins, matrix metalloproteinase balance is deranged with increased expression of matrix metalloproteinase 9. This blocks the reconstruction of the extracellular matrix and the proliferation and migration of fibroblasts. With the above said complications, wound healing does not follow the normal healing cascade and each phase is delayed leading to delayed healing. Therefore, there is also need in the art for dressing material that contain medicaments that address the above complications in chronic wound healing.

During the healing process, the wound has to be prepared for debridement to remove the dead tissues and to facilitate reepithilialization. Wound debridement removes senescent cells and biofilm. Generally wound debridement is mediated by mechanical or enzymatic means. For this, a separate wound dressings is generally used. Therefore, there is also need in the art for dressing material which addresses this issue.

In the present invention, a novel dressing material has been developed that addresses all the above requirements of the existing art as described in the preceding paragraphs, that minimizes pain, provides a desirable moist healing environment, provides a multi layered in situ gelling requirement, releases the active components required for enhancing proliferation of fibroblast, accelerates healing, absorb wound exudates, remodels the wound without scar.

OBJECT OF THE INVENTION: One of the objects of the invention is to provide a dressing material, having a hydrogel prepared from a polymer, that is devoid of side effects associated with existing polymers being used in preparation of the hydrogel.

Another object of the invention is to provide a dressing material that uses a natural polymer suitable for hydrogel formation.

Another object of the invention is to provide a dressing material that can be retained at the wound site for a longer period of time and suitable for in situ gel formation, particularly multilayered in situ gel formation.

Another object of the invention is to provide a dressing material that provides a moist and cool environment suitable for wound healing, and minimizes pain.

Another object of the invention is to provide a dressing material that releases active components required for accelerated wound healing.

Another object of the invention is to provide a dressing material that helps in wound healing without scar left at the place of wound. SUMMARY OF THE INVENTION:

The present invention relates to a dressing material for wound healing purpose, wherein the dressing material comprises of one or more layers of hydrogel, wherein the hydrogel is prepared using powder of gum extracted from the plant T.catappa, and one or more compound helpful in wound healing, wherein the particle size of the powder of gum extracted from the plant T.catappa, ranges between 60-80 nm. Among the layers of hydrogel used in the present invention, one of the layers of hydrogel contains a hemostat and natural immune booster prepared from the plant C.hirsutus, one of the layers of hydrogel contains a. hydroxyapatite nanorods, antibiotics and antibiotic adjuvant prepared from the plant T.purpurea, one of the layers of hydrogel contains natural anti-inflammatory agents and selective matrix metalloproteinase 9 inhibitors prepared from the plant T.purpurea, one of the layers of hydrogel contains zinc oxide nano material, and an anti- inflammatory agent prepared from T.purpurea and an immune booster and natural fibroblast proliferation enhancer prepared from the leaves of C.hirsutus. In the present invention the hydrogel is prepared on the surface of Poly Vinyl Alcohol (PVA). In the present invention the layer wise arrangement of layers of hydrogel can vary and may not be restricted to one arrangement/combination of the layers. However more particularly and preferably the invention comprises of following layers of hydrogel in the following arrangements: a first layer (5) made up of soft silicon mesh which remain in contact with the wound; a second layer (6) covering above the first layer, made up of hydrogel containing a hemostat and natural immune booster prepared from the plant C.hirsutus, a third layer (7) covering above the second layer, made up of hydrogel containing hydroxyapatite nanorods, antibiotics and antibiotic adjuvant prepared from the plant T.purpurea; a fourth layer (8) covering above the third layer, made up of hydrogel containing a natural anti-inflammatory agents and selective matrix metalloproteinase 9 inhibitors prepared from the plant T.purpurea; a fifth layer (9) covering above the fourth layer made up of hydrogel, containing zinc oxide nanomaterial, and an anti- inflammatory agent prepared from T.purpurea and an immune booster and natural fibroblast proliferation enhancer prepared from the leaves of C.hirsutus; a sixth layer (10) covering above the fifth layer, made up of now woven silk cotton; a seventh layer (11) covering above the sixth layer, made up soft silicon.

BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS

Fig. 1 represents the top view of the dressing material as disclosed in the present invention.

Fig. 2 represents the inner view of the dressing material as disclosed in the present invention. Fig. 3 represents different layers associated with the dressing material.

Fig. 4 represents the first layer (5) of the dressing material that is made up of soft silicon, that remains in contact with the wound.

Fig. 5 represents the second layer (6) of the dressing material that remains above the first layer, which provides insitu gel formation that releases hemostat and natural immune booster. Fig. 6 represents the third layer (7) of the dressing material that remains above the second layer which provides insitu gel formation that releases hydroxyapatite nanorods, antibiotics and antibiotic adjuvant. Fig. 7 represents the fourth layer (8) of the dressing material remains above the third layer which provides insitu gel formation that releases natural anti-inflammatory agents and selective matrix metalloproteinase 9 inhibitors.

Fig. 8 represents the fifth layer (9) of the dressing material that remains above the fourth layer which provides insitu gel formation that releases natural fibroblast proliferation enhancer and angiogenesis accelerator.

Fig. 9 represents the sixth layer (10) of the dressing material that remains above the fifth layer which is made up of non-woven silk cotton.

Fig. 10 represents the seventh layer (11) that remains above the sixth layer which is made up of soft silicon mesh.

DETAILED DESCRIPTION OF A PREFERRED EMBODIMENT OF THE INVENTION

The present invention relates to a novel dressing material for wound healing purpose, wherein the dressing material comprises multiple insitu gel forming layers one above the other, wrapped within two soft silicon layers. The present invention including the outer silicon layers comprises seven layers which have been explained below, where their numbering from the first, second and subsequent, is in order of bottom to top order, where the bottom is lowest most layer in contact with the wound, and the top is layer farthest from the wound.

The first layer (5): This layer is made up of soft silicon mesh of thickness of 0.2 mm, and of pore size 1 mm. The other layers (second to seventh) are formed on the first layer, one after the other, in a layerwise manner, involving the process of in situ hydrogel formation.

The second layer (6): This layer is a insitu hydrogel forming layer releasing hemostat and natural immune booster prepared from the plant Cocculus hirsutus prepared.

The third layer (7): This layer is a insitu hydrogel forming layer releasing hydroxyapatite nanorods, antibiotics and antibiotic adjuvant. The antibiotic adjuvant is prepared from the plant Tephrosea purpurea. The fourth layer (8): This layer is a insitu hydrogel forming layer releasing anti-inflammatory agents and selective matrix metalloproteinase 9 inhibitors prepared from the plant Tephrosea purpurea.

The fifth layer (9): This layer is a insitu hydrogel forming layer releasing zinc oxide nanomaterial, natural fibroblast proliferation enhancer and angiogenesis accelerator prepared from the plant Tephrosea purpurea.

The sixth layer (10): This layer is made up of non-woven silk cotton.

The seventh layer (11): This layer is made up of soft silicon mesh of thickness of 0.2 mm, of pore size 1 mm. The method for preparation of each layer is provided as below:

Preparation of gum powder:

The dried mass of gum from the plant Teminalia catappa was prepared as a fine powder in a blender. The coarse powder was sieved in stainless steel mesh of size 1250 (10 micron). The powder was ball milled in a Planetary Ball milling instrument with zirconium balls. In short, 25g of fine powder of gum was mixed with 900g of zirconium balls and loaded onto a 1.5L tank of ball milling apparatus. The fine powder was milled at 450 rpm for 6 hrs. The milled powder was collected and cooled to room temperature. The size of the powder was analyzed in a particle size analyzer. The size of the powder ranged from 60-80 nm. This powder was used as the gel base in all layers of the present wound dressing. Preparation of 3D printed polyvinyl alcohol (PVA) mesh:

PVA mesh (4”x4”) was prepared by 3D printing using a fused filament fabrication desktop printer (Ultimaker 3, Netherlands). PVA filament (AquaSolve, Formfutura, Nijmegen, The Netherlands, 1.75 mm diameter) was printed as a 3D mesh at 200°C with a pore size of 1 mm. The mesh was prepared at 0.15 mm thickness. Preparation of hemostat and immune booster: Fresh leaves of Cocculus hirsutus were washed with sterile water and surface sterilized with 75% ethanol for 5 min. The leaves were shade dried under aseptic conditions till the moisture content becomes zero. The dried leaves were powdered in a blender. The coarse powder was sieved in stainless steel mesh of size 1250 (10 micron). 500 mg of fine powder was mixed with 10 mL of nanopure water stirred in a magnetic stirrer for 5 min. The extract was filtered through muslin cloth and lyophilized to powder.

Preparation of the first layer:

The first layer made up of soft silicon mesh of thickness of 0.2 mm, and pore size of 1 mm, is prepared using an iron mold.

Preparation of Second Layer (in situ gel formation that releases hemostat and natural immune booster):

10 mg of gum powder was mixed with 2 mg of the lyophilized hemostat and immune booster. To the mixture, 1 ml of sterile nano pure water was added to form a gel. The gel is spread over the 3D printed PVA mesh of size 4”x4”.

Preparation of hydroxyapatite nano rods:

Hydroxyapatite nanorods are prepared using calcium chloride, orthophosphoric acid and aqueous extract of T. purpurea. For the preparation of aqueous extract, the roots of T. purpurea were cut into 1 cm size and finely powdered in a blender. The powder is sieved in 1250 mesh (10 micron). The fine powder was boiled with water (cone. lOmg/mL) at 60°C for 30 min. The extract was centrifuged at 10,000 rpm for 10 min. The supernatant was used as the aqueous extract. Hydroxyapatite nanorods was prepared by mixing 100 mL of 500 mM calcium chloride with 40 mL of 1.2 mol orthophosphoric acid and lOmL of aqueous extract prepared from the roots of T.purpurea under vigorous stirring at 500rpm. 100 mL of 0.1 N sodium hydroxide was added drop wise under the same stirring conditions, till the hydroxyapatite was precipitated. The precipitated hydroxyapatite was dried in a hot air oven and calcined at 800°C for 16 hrs. Size of nano hydroxyapatite particles: 10 nm.

Preparation of b lactam antibiotic adjuvant: The adjuvant for improving activity of b lactam antibiotic was prepared from the roots of T. purpurea. The dried root of T. purpurea was prepared as powder in a blender and sieved in 1250 mesh (10 micron). The sieved material was soaked in 75% methanol for 2 hr and extracted in soxhlet apparatus at 50°C for 4hr. The extract was concentrated and dried in a rotary evaporator and used as adjuvant for improving activity of b lactam antibiotic.

Preparation of third layer (in situ gel formation that releases hydroxyapatite nanorods, antibiotics and antibiotic adjuvant:

10 mg of the gum powder was mixed with 10 pg of nano hydroxyapatite, 25 mg of cefdinir and 75 mg of adjuvant prepared from the roots of T. purpurea and mixed with 2 ml of nanopure water and coated onto 3D printed PVA mesh prepared as described earlier.

Preparation of anti inflammatory agent and matrix metalloproteinase 9 inhibitor from the roots of T. purpurea):

The finely sieved root powder of T. purpurea was prepared as described in the preceding paragraph in relation to“preparation of b lactam antibiotic adjuvant”. The fine powder was extracted with 50% ethanol in soxhlet apparatus for 3 hr at 50°C and concentrated and dried in a rotary evaporator. The powder was used as the source of anti-inflammatory agent and matrix metalloproteinase 9 inhibitors.

Preparation of fourth layer (in situ gel formation that releases natural anti-inflammatory agents and matrix metalloproteinase 9 inhibitors. 10 mg of gum powder prepared from T.catappa gum as mentioned earlier was mixed with 2 mg of anti-inflammatory agent and matrix metalloproteinase 9 inhibitors prepared from the roots of T. purpurea as described in the preceding paragraph. To the mixture, 2 ml of sterile nano pure water was added and the same was coated on 3D printed PVA mesh prepared as described earlier. Preparation of Zinc oxide nanomaterial:

The Zinc oxide nano material was prepared by adding 10 mL of T. purpurea extract powder (lOmg/mL) prepared as described in the preceding paragraph relating to“preparation of anti inflammatory agent and matrix metalloproteinase 9 inhibitor” to 90 mL of 0.1 M Zinc acetate solution. Under vigorous stirring, 0.1 N sodium hydroxide was added drop wise till the precipitate was obtained. The precipitate was washed with distilled water five times and dried in a hot air oven at 100°C for 16 hrs. Preparation of fifth layer (in situ gel formation that releases natural fibroblast proliferation enhancer and angiogenesis accelerator): lOmg of gum powder, 3 mg of zinc oxide nano material, 2 mg of the antiinflammatory agent prepared from the roots of T. purpurea plant and 2mg of immune booster prepared from the leaf of C.hirsutus each one as described in the preceding paragraphs, were mixed well. To this mixture, 5 mL of nano pure water was added and the same was coated on to 3D printed PVA mesh as described earlier.

Preparation of sixth layer (non-woven silk cotton)

Preparation of seventh layer (soft silicon mesh)

Activity of the constituents, the hydrogel and the dressing material has been discussed in the following examples:

Example 1: Hemostat activity of lyophilized powder of C.hirsutus used in preparation of the second layer (6)

Hemostat activity of the lyophilized powder prepared from C.hirsutus was assessed by Light Transmission platelet aggregometry. The procedure for assay consisted of collecting blood sample from a healthy volunteer and preparing the platelet rich plasma (PRP). The ability of the lyophilized powder to activate platelet activation and aggregation was assessed by its light transmission capacity in a spectrophotometer. Increased transmission of light indicates increased hemostatic potential of the test sample.

(a) Preparation of Platelet Rich Plasma (PRP) and Platelet Poor Plasma (PPP) 5mL of venous blood was withdrawn from a healthy volunteer in a 3.2% trisodium citrate anticoagulant tube and centrifuged at 180g for 10 min. The plasma was aspirated and centrifuged again at 1550g for 10 min. lmL of plasma and the platelets precipitated at the bottom was used as PRP. The rest of the sample was again centrifuged at 10,000g for 10 min to obtain PPP. The supernatant was used as PPP.

(b) Platelet aggregation Test The test uses PRP and PPP references to set 0% aggregation and 100% aggregation respectively. To 270pL of PRP and PPP in separate tubes 30 pL of phosphate buffered saline was added and stirred in a magnetic stirrer for 15 min. The optical density (OD) was recorded at 595 nm.

For assessing the hemostat potential of lyophilized powder prepared from C.hirsutus, the following procedure was adopted. To 270pL of PRP, 30 pL of ADP (0.1M) was added as a reference agonist of platelet aggregation and stirred in a magnetic stirrer for 15 min and the percentage of optical transmission (%T) was recorded. In another tube 270pL of PRP, 100 pg of lyophilized powder and 30 pL of the ADP (0.1M) were added and mixed in a magnetic stirrer for 15 min. In another tube 270pL of PRP, 100 pg of gelatin powder and 30 pL of the ADP (0.1M) were added and mixed in a magnetic stirrer for 15 min. Platelet aggregation was calculated from the OD. Platelet aggregation = (OD of PRP - OD of hemostat) / (OD of PRP - OD of PPP)*100

Table 1. Hemostat Activity of C.hirsutus

Example 2. Immune booster Activity of C.hirsutus : (a) Leukocyte Migration Assay: The influence of lyophilized powder prepared from C.hirsutus on the immune modulatory properties was assessed through leukocyte migration assay in agarose gel.

Preparation of Peripheral Blood Mononuclear Cells (PBMC)

5mL of blood was collected in a tube containing 3.2% trisodium citrate and 5mL of phosphate buffered saline (PBS) was added to blood in the tube. The sample was centrifuged at 400g for 10 min. The plasma layer was aspirated and removed. The cells were suspended in 15 mL of phosphate buffered saline. The cell suspension was carefully layered onto lOmL of Ficoll hypaque without mixing. The sample was centrifuged at 850g for 45 min. The top most layer containing platelets and plasma was removed. The huffy coat layer containing PBMC was carefully collected. The collected cells were washed with PBS to remove the medium. The number of cells was counted in a hemocytometer. lg of agarose was dissolved in 50 mL of phosphate buffered saline. The suspension were heated upto 70C to solubilize the agarose. After cooling to room temperature, to the agarose solution, 50mL of Dulbeccos Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum, 60U of penicillin and 60pg/mL of streptomycin were added. The agarose solution was poured onto a 60mm glass petri dish. Three wells of 3 mm diameter were punched onto the agarose gel with stainless steel gel puncher. To one well 100 x 10 PBMC were seeded in DMEM medium supplemented with 10% fetal bovine serum. In the second well, 5 pL of DMEM medium was added without PBMC. In the third well, 5 pL of immune modulator (lmg/mL) was added. The plate was incubated in inverted condition in the C0 chamber. The experiment was replicated thrice and the mean of the value was calculated as the final result. The Migration Index of leukocyte was calculated by dividing the distance moved by the cells towards the test sample to the DMEM.

Table 2. Influence of Hemostat prepared from C.hirsutus on Leucocyte migration

Example 3: Assessment of adjuvant activity b lactam antibiotic adjuvant used in preparation of third layer (7)

The adjuvant activity of b lactam antibiotic adjuvant used in preparation of third layer (7) prepared from T. purpurea extract was determined by agar well diffusion assay. Staphylococcus aureus and Pseudomonas aeruginosa were isolated from bum wounds. The bacteria were grown as cultures overnight in nutrient broth. The bacterial cell density was adjusted to lxlOCFU/mL. Mueller and Hinton agar medium was poured onto two different plates. 100 pL of Staphylococcus aureus and Pseudomonas aeruginosa were swabbed on to different plates. Three wells of 8mm were punctured onto medium and loaded with cefdinir (30pg), 120 pg of adjuvant prepared from T. purpurea and combination of cefdinir and adjuvant in separate wells. The plates were incubated at 37°C in the incubator for 24 hr. The inhibition zone was recorded in mm. The experiment was replicated thrice and the result is the mean of three values. The sample is considered to have adjuvant activity if the inhibition zone exhibited by the combination with antibiotic exhibits an inhibition zone greater than 6 mm than when the antibiotic and adjuvant are used alone.

Table 3. Adjuvant activity of b lactam antibiotic adjuvant used in preparation third layer (7)

Example 4. Assessment of anti-inflammatory activity of T. purpurea extract used in fourth layer

(8)

(a) Assessment of anti-inflammatory activity by red blood cells (RBC) membrane stabilization test Fresh whole human blood (5 mL) was collected and transferred into an eppendorf containing 3.2% trisodium citrate. The tube was centrifuged at 3000 rpm for 10 min and washed thrice with 5 mL of phosphate buffered saline. The reaction mixture for RBC stabilization assay consists of 0.5 mL of RBC suspension and 0.5 mL of anti inflammatory agent (lmg/mL). A control was maintained by replacing the anti-inflammatory agent by PBS. Naproxen (lmg/mL) was used as a positive control drug. The tubes were heated at 56°C for 30 min and cooled to room temperature. The reaction mixture was centrifuged at 2500 rpm for 5 min and the supernatant was collected. That absorbance of the supernatant was measured at 560 nm. RBC membrane stabilization was calculated by using formula mentioned below:

RBC Membrane stabilization (%) = [(Control OD - Test OD)/Control OD] * 100 Table 4. Anti-inflammatory activity of T.purpurea extract used in preparation of the fourth layer.

Example 5 Assessment of MetalloProteinase inhibition activity in T.purpurea extract used in preparation of the fourth layer.

The flavonoids present in T.purpurea extract were quantified (45mg/g). The name of the flavonoids were obtained from Phyto Chemical Interaction DataBase (PCIDB) and docked against Matrix MetalloProteinase 9 using AutodockVina and the results are presented in Table

7.

Table 5. Inhibition of MMP9 by extract of T.purpurea

Example 6: Assessment of fibroblast enhancing activity of components used in preparation of the fifth layer (9)

Fibroblast proliferation and migration assay: Human fibroblast cells were grown in DMEM media supplemented with 10% fetal bovine serum. The cells were grown to confluency and disaggregated with trypsin. The cells (2xl0 ) were seeded in plate and incubated at 37°C to form a monolayer. The monolayer was scratched horizontally with a sterile pipette tip. The cells were treated with (lmg/mL) of the constituents used in preparation of the fifth layer. Control plate was maintained without any test sample. A positive control plate was maintained with (lmg/mL) of a widely available hydrogel in the market (“hydroheal” is the trade name of the product that contains Colloidal silver 32 PPM).

Migration rate was calculated from the distance moved by the cells along with the test sample, in comparison to that of the control plate. Table 6. Migration of Fibroblast cells

The efficacy of the dressing material of the present invention has been evaluated in vivo, on the chronic wounds, as indicated below.

Example 7 Invivo healing of bum wounds Healing of bum wounds

The efficiency of the dressing material of the present invention in healing burn wounds was assessed in Wistar rats as per Institutional Animal Ethical Committee guidelines (IAEC/MSEC/BT/P2/Bum Wound,2018). Hair was removed from the rats and anesthetized with ketamine and xylazine. Third degree burn wounds were created in rats on the dorsal surface using a 1cm diameter hot metal rod. The rats were segregated into three groups. One group was applied with neat paraffin without any medicament (negative control), the next group was treated with hydroheal (positive control) and the third group was treated with the dressing material of the present invention. The healing rate was evaluated with reference to the decrease in initial wound size from day 0.

Table 7. Invivo healing of burn wound

Example 8 Invivo healing of diabetic wound

Healing of diabetic wound

The efficiency of the dressing material of the present invention in healing diabetic wounds was assessed as per Wistar rats Institutional Animal Ethical Committee guidelines (IAEC/MSEC/BT/P 1/Diabetic Wound,2018). Diabetic condition was induced by streptozotocin. After induction of diabetes, the hair in the dorsal surface was removed from the rats and anesthetized with ketamine and xylazine. An excision wound of 1cm was created using a biopsy punch. The rats were segregated into three groups. One group was applied with neat paraffin without any medicament (negative control), the next group was treated with hydroheal (positive control) and the third group was treated with the dressing material of the present invention. The healing rate was evaluated with reference to the decrease in initial wound size from day 0.

Table 8. 10 Invivo Healing of diabetic wound

Claims

Claims:

1. A dressing material for wound healing purpose, wherein the dressing material comprises of one or more layers of hydrogel, wherein the hydrogel is prepared using powder of gum extracted from the plant T.catappa, and one or more compound helpful in wound healing, wherein the particle size of the powder of gum extracted from the plant T.catappa, ranges between 60-80 nm.

2. The dressing material as claimed in claim 1, wherein at least one of the layers of hydrogel contains a hemostat and natural immune booster prepared from the plant C.hirsutus.

3. The dressing material as claimed in claim 1, wherein at least one of the layers of hydrogel contains a. hydroxyapatite nanorods, antibiotics and antibiotic adjuvant prepared from the plant T. purpurea.

4. The dressing material as claimed in claim 1, wherein at least one of the layers of hydrogel contains natural anti-inflammatory agents and selective matrix metalloproteinase 9 inhibitors prepared from the plant T. purpurea.

5. The dressing material as claimed in claim 1, wherein at least one of the layers of hydrogel contains zinc oxide nanomaterial, and an anti- inflammatory agent prepared from T. purpurea and an immune booster and natural fibroblast proliferation enhancer prepared from the leaves of C.hirsutus..

6. The dressing material as claimed in claim 1, wherein the hydrogel is prepared on the surface of Poly Vinyl Alcohol (PVA).

7. The dressing material as claimed in claims 1, wherein the layer wise arrangement of layers of hydrogel one above the other is not restricted to any particular arrangement or combination of the layers.

8. The dressing material as claimed in claim 1, wherein the dressing material comprises of a first layer (5) made up of soft silicon mesh;

a second layer (6) covering above the first layer, made up of hydrogel containing a hemostat and natural immune booster prepared from the plant C.hirsutus,

a third layer (7) covering above the second layer, made up of hydrogel containing hydroxyapatite nanorods, antibiotics and antibiotic adjuvant prepared from the plant T. purpurea; a fourth layer (8) covering above the third layer, made up of hydrogel containing a natural anti-inflammatory agents and selective matrix metalloproteinase 9 inhibitors prepared from the plant T. purpurea;

a fifth layer (9) covering above the fourth layer made up of hydrogel, containing zinc oxide nanomaterial, and an anti- inflammatory agent prepared from T. purpurea and an immune booster and natural fibroblast proliferation enhancer prepared from the leaves of C.hirsutus;

a sixth layer (10) covering above the fifth layer, made up of now woven silk cotton;

a seventh layer (11) covering above the sixth layer, made up soft silicon;

wherein, the the hydrogel is prepared from the powder of gum extracted from the plant

T.catappa, wherein the particle size of the powder of gum is between 60-80nm;

wherein, the hydrogel is prepared on the surface of poly vinyl alcohol (PVA);

wherein, the soft silicon mesh is having pore size of 1 mm and thickness of 0.2 mm.