TWI655005B - Thermal wound dressing - Google Patents

Abstract

The thermal sensation wound dressing disclosed in the present invention is prepared by crosslinking a polysaccharide molecule, a hyaluronic acid and a graphene. The thermal wound dressing disclosed by the present invention covers the wound or the injured tissue, which can effectively reduce the wound area and improve the cell growth efficiency, and thus can be used as an effective component of the pharmaceutical composition for promoting wound healing.

Description

Thermal wound dressing

The present invention relates to a medical material, and in particular to a thermal wound dressing.

According to the current clinical solution to deep skin injuries, the following methods are as follows: First, autologous transplantation, which uses surgical methods to transplant skin tissue from other parts to the injured site. Although it has good curative effect, the part that donates the skin will stay. The scar is scarred and has a limited range of use; second, allogeneic transplantation, which transplants the skin of others to the injured site, but an immune reaction occurs, resulting in poor healing; third, xenograft, non-human skin tissue transplanted to The injured part not only triggers an immune reaction, but also has the risk of disease transmission. Fourth, cell culture, using its own cells to culture epidermal tissue in vitro, and then transplanting, although it can avoid immune response, but the culture time is too long, can not To cope with emergencies, and in vitro culture can not cultivate dermal tissue; Fifth, water-based dressings, which are prepared through biological materials, are convenient to use and can be used in mass production. Therefore, it is currently the most clinically Often used in the treatment of wounds.

However, the current commercial water-based dressings still have the risk of subcutaneous empyema, and the removal of the dressing will destroy the granulation tissue, causing second damage to the wound and delaying the recovery rate. However, in order to solve the problem of secondary damage of the wound, there are manufacturers who will be allogeneic. As a component of water-based dressings, cells can achieve the effect of no need to change dressings, but the use of allogeneic cells poses a high risk of viral infection and immune response.

The main object of the present invention is to provide a thermal sensation wound dressing which can effectively promote cell growth efficiency and convert light energy into heat energy to achieve the effect of accelerating wound healing.

Another object of the present invention is to provide a thermal wound dressing which has good biocompatibility and can maintain flatness for a long time and covers a wet wound to increase clinical acceptance and application. Sex.

Therefore, in order to achieve the above object, the thermal wound dressing disclosed in the present invention is prepared by crosslinking a polysaccharide molecule, a hyaluronic acid and graphene oxide.

Wherein, the polysaccharide molecule is xanthan gum.

Furthermore, in one embodiment of the invention, the thermal wound dressing is prepared by the following steps:

Step a: preparing an aqueous solution of xanthan gum and an aqueous solution of hyaluronic acid.

Step b: mixing the aqueous solution of xanthan gum and the aqueous solution of hyaluronic acid to form a xanthan gum/hyaluronic acid aqueous solution.

Step c: adding a predetermined amount of graphene oxide to the xanthan gum/hyaluronic acid aqueous solution to obtain a xanthan gum/hyaluronic acid/graphene oxide aqueous solution.

Step d: reacting the xanthan gum/hyaluronic acid/graphene oxide aqueous solution with a crosslinking agent, thinning and drying to obtain a thermal wound dressing.

In order to improve the mechanical strength and sensibility of the thermal wound dressing of the present invention, the weight percentage of the graphene oxide is at least 6% in the raw material of the thermal wound dressing, and if the graphene oxide is When the weight percentage is less than 1%, the efficacy of the present invention cannot be attained.

Specifically, the weight ratio of the graphene oxide, the xanthan gum and the hyaluronic acid is about 1:11:2 to 1:11:3.

Generally, the crosslinking agent is 1-ethyl-3-(3-dimethylaminopropyl)carbonium diimide.

The thermal wound dressing disclosed by the present invention covers the wound or the injured tissue, which can effectively reduce the wound area and improve the cell growth efficiency, and thus can be used as an effective component of the pharmaceutical composition for promoting wound healing.

The first figure is the result of analyzing graphene oxide by an infrared spectrometer.

The second graph is the result of particle size analysis of graphene oxide.

The third figure is the result of the analysis of the interface potential of graphene oxide.

The fourth figure is the appearance of the crosslinked Xn/HA film, Xn/HA/GO (0.003) film and Xn/HA/GO (0.03) film.

The fifth figure is an infrared spectrum of the xanthan gum before and after the crosslinking reaction.

The sixth figure is the infrared spectrum of the hyaluronic acid before and after the cross-linking reaction.

The seventh figure is an infrared spectrum of the hyaluronic acid before and after the crosslinking reaction.

Fig. 8A shows the results of observing the Xn/HA film at a magnification of 5000X with an electron microscope.

Figure 8B shows the results of observing Xn/HA/GO (0.003) film at a magnification of 5000X with an electron microscope.

Figure 8C shows the results of observing the Xn/HA/GO (0.03) film at a magnification of 5000X with an electron microscope.

The ninth panel A shows the appearance of the wound after treatment with Xn/HA film as a dressing for seven days.

Figure IX B shows the appearance of the wound after treatment with Xn/HA/GO (0.03) film as a dressing for seven days.

The ninth panel C is based on a Xn/HA/GO (0.03) film as a dressing and provides infrared light for the appearance of the wound after seven days of treatment.

The tenth figure is a wound dressing with Xn/HA/GO (0.03) film as a dressing and infrared radiation treatment after seven days of treatment.

Fig. 11A shows the Xn/HA film as a dressing, and the wound tissue section after seven days of treatment was subjected to HE staining, and the magnification was 40X.

Figure 11B shows the Xn/HA film as a dressing, and the wound tissue section after seven days of treatment was subjected to HE staining, and the magnification was 100X.

Fig. 12A shows the Xn/HA/GO (0.03) film as a dressing, and the wound tissue section after seven days of treatment was subjected to HE staining, and the magnification was 40X.

Fig. 12B shows the Xn/HA/GO (0.03) film as a dressing, and the wound tissue section after seven days of treatment was subjected to HE staining, and the magnification was 100X.

The thirteenth image A is a Xn/HA/GO (0.03) film as a dressing, and provides infrared irradiation, and the wound tissue section after seven days of treatment is subjected to HE staining, and the magnification is 40X.

Figure 13B shows the Xn/HA/GO (0.03) film as a dressing and provides infrared irradiation. After seven days of treatment, the wound tissue sections were stained with HE and the magnification was 100X.

Fig. 14A shows the Xn/HA film as a dressing, and the wound tissue section after seven days of treatment was subjected to three colors of Meisheng, and the magnification was 40X.

Figure 14B shows the Xn/HA film as a dressing, and the wound tissue section after seven days of treatment was subjected to three colors of Meisheng, and the magnification was 100X.

The fifteenth figure A is a Xn/HA/GO (0.03) film as a dressing, and the wound tissue section after seven days of treatment is subjected to the three colors of Meisheng, and the magnification is 40X.

The fifteenth figure B is the Xn/HA/GO (0.03) film as a dressing, and the wound tissue section after seven days of treatment is the result of the three colors of the plum, the magnification is 100X.

Fig. 16A shows the Xn/HA/GO (0.03) film as a dressing, and provides infrared irradiation. After seven days of treatment, the wound tissue sections were subjected to three colors of Meisheng, and the magnification was 40X.

Figure 16B shows the Xn/HA/GO (0.03) film as a dressing, and provides infrared radiation. After seven days of treatment, the wound tissue sections were subjected to three colors of Meisheng, and the magnification was 100X.

The thermal wound dressing disclosed by the invention is a novel wet dressing which forms a film through the cross-linking reaction to form a film of xanthan gum, hyaluronic acid, graphene oxide and the like, and can directly cover the wound or the affected part. It does not deform or cause unacceptable wounds under prolonged use, and has good biocompatibility. Therefore, it can continuously protect the wound and prevent the wound from being damaged twice. Furthermore, the thermal wound dressing disclosed in the present invention can achieve the effects of reducing the wound area and promoting wound healing under the condition of not containing cells, and the manufacturing cost is very cheap compared with the conventional cell dressing. It can be provided to the public and clinical use.

The xanthan gum disclosed in the present invention is a microbial glycan which is a branched polysaccharide obtained by fermentation of Xanthomonas campestris. Compared with other polysaccharides, the xanthan gum system can make the thermal wound dressing of the present invention have better softness and foldability, thereby enabling the thermal wound dressing of the present invention to be applied to various kinds. Partial injury.

In the following, in order to confirm the efficacy of the thermal wound dressing of the present invention, several examples will be given and further illustrated in detail with the drawings.

Example 1: Preparation of graphene oxide

4 g of graphite powder and 2 g of sodium nitrate were dissolved in 92 ml of concentrated sulfuric acid, stirred on ice for 5 minutes, returned to room temperature and then added with 12 g of potassium permanganate, heated to 35 ° C, stirring was continued for another 30 minutes, and 184 ml was slowly added. After the second water, the reaction temperature was raised to 165-175 ° C for 15 minutes, and then 160 ml of 3% hydrogen peroxide was added and allowed to stand for 1 hour. The liquid was removed by centrifugation, the precipitate was taken out, and the precipitate was suspended in a small amount of secondary water, subjected to ultrasonic vibration, and placed in an oven to obtain a graphene oxide powder. The functional group of graphene oxide was analyzed by an infrared spectrometer, and the results are shown in the first figure. The particle size distribution and interface potential of GO were measured by a dynamic light scattering particle size analyzer and an interface potential analyzer. The results are shown in the second and third figures.

From the results of the first to third figures, the graphene oxide functional signal prepared by the method of the present example is a ketone group (C=O) at 1680 cm-1, and 1640 cm-1 is a carbon-carbon double bond on the benzene ring. (C=C), 1230 cm-1 is an ether group (COC). 3500 cm-1 is a carbon-hydrogen bond (C-H), and its average particle diameter is 1284 nm, and the interface potential is -25.4 mV.

Example 2: Preparation of a film

0.33 g of xanthan gum powder was uniformly dissolved in 33 ml of hot water in water. Another 0.07 g of hyaluronic acid was dissolved in 7 ml of secondary water. The xanthan gum aqueous solution and the hyaluronic acid aqueous solution were uniformly mixed, the bubbles were removed by centrifugation, the mixed solution was poured into a glass dish, and placed in an oven at 45 ° C for about one day to obtain a xanthan gum/hyaluronic acid film (hereinafter referred to as Xn/HA film).

Example 3: Preparation of a thermal wound dressing

0.33 g of Xanthan gum powder was uniformly dissolved in 33 ml of hot water in water. Another 0.07 g of hyaluronic acid was dissolved in 7 ml of secondary water. The xanthan gum aqueous solution is uniformly mixed with the hyaluronic acid aqueous solution, and then 0.003 g and 0.03 g of the graphene oxide powder are separately added and uniformly mixed to obtain a xanthogen. Glue/hyaluronic acid/graphene oxide mixed solution. Pour the xanthan gum/hyaluronic acid/graphene oxide mixed solution into a glass dish and place it in an oven at 45 ° C for about one day to obtain a xanthan gum/hyaluronic acid/graphene oxide film (0.003 g) and obtain xanthan gum/hyaluronic acid/oxidation. A graphene film (0.03 g), hereinafter referred to as Xn/HA/GO (0.003) film and Xn/HA/GO (0.03) film, respectively.

Example 4: Crosslinking reaction

A powder of 1-ethyl-3-(3-dimethylaminopropyl)carbenium diimine was taken and dissolved in 80% alcohol to form an EDC cross-linking solution having a concentration of about 15 mM/L.

The Xn/HA film, Xn/HA/GO (0.003) film and Xn/HA/GO (0.03) film were respectively immersed in the crosslinking solution, and after about 24 hours of reaction, the uncrosslinked polymer on the film was removed by alcohol and the mixture was dispensed. The mixture was placed in an oven at 45 ° C to obtain Xn/HA film, Xn/HA/GO (0.003) film and Xn/HA/GO (0.03) film crosslinked with EDC, respectively, as shown in the fourth figure.

Xanthan gum and hyaluronic acid and EDC cross-linking solution were analyzed by infrared spectrometer before and after cross-linking reaction. As shown in the fifth and sixth figures, the film showing xanthan gum or hyaluronic acid can be cross-linked by EDC. Further, the change of the Xn/HA/GO (0.03) film before and after the crosslinking reaction was analyzed by an infrared spectrometer, and the results are shown in the seventh chart. It can be seen from the seventh figure that after cross-linking, the absorption peaks of 1520 and 1720 cm-1 are still quite obvious, indicating that graphene oxide does not affect the cross-linking of xanthan gum and hyaluronic acid.

Example 5: Film surface analysis

Xn/HA membrane, Xn/HA/GO (0.003) membrane and Xn/HA/GO (0.03) membrane crosslinked with EDC were respectively immersed in phosphate buffer solution to simulate the state of contact with wound tissue, and reused. The surface morphology of each film was observed by an electron microscope, and the results are shown in Figs. 8 to A.

It can be seen from the eighth figure A that after the Xn/HA film is immersed, the surface thereof is in a state of unevenness, indicating that the mechanical strength is not good. Referring to FIG. 8B and FIG. 8C again, the surface of the film having graphene oxide is relatively flat compared to the Xn/HA film, and a sheet-like structure can be found on the surface thereof. Graphene oxide (as indicated by the arrow) in which a higher amount of graphene oxide is contained in the film based on Xn/HA/GO (0.03), and thus the sheet structure density on the surface thereof is also high. It can be seen that the Xn/HA/GO film disclosed in the present invention can maintain the structural flatness after contacting the wound or the injured part, can continue to conform to the tissue, and has better mechanical properties.

Example 6: Water content test

Xn/HA film, Xn/HA/GO (0.003) film and Xn/HA/GO (0.03) film crosslinked with EDC were cut into 2 square centimeters, respectively, and dried and weighed (dry weight). Then, the physiological saline solution is separately placed for about 24 hours, and then the water on the surface of the membrane is separately sucked by the filter paper, and then weighed (wet weight) to calculate the water content of each membrane. The calculation formula is as follows: water content (%)= (wet weight - dry weight) / wet weight X 100%

After calculation, the water content of the Xn/HA film was 87.2%, the water content of the Xn/HA/GO (0.003) film was 85.5%, and the water content of the Xn/HA/GO (0.03) film was 80.0%, indicating oxidation. The graphene film has a lower water content, meaning that it can maintain its structural flatness in a humid environment.

Example 7: Mechanical properties test

The Xn/HA film, the Xn/HA/GO (0.003) film, and the Xn/HA/GO (0.03) film before and after the cross-linking reaction with EDC were cut into a size of 1×5 cm 2 , respectively, and used as a desktop type. The material testing machine clamps 1 cm above and below each film to make the measured area 1×3 cm ^ 2; the tensile tester is used to conduct the tensile test at a tensile rate of 5 mm/min, and the deformation is 2%. Values, the results are shown in Table 1 below.

From the results of Table 1, it is understood that the crosslinking reaction with EDC and the graphene oxide-based system can improve the mechanical strength of each film. Furthermore, the maximum load and maximum stress of the Xn/HA film were 1.30 Kgf and 4.35 Kgf/mm2, respectively, and the maximum load and maximum of Xn/HA/GO (0.003) film were crosslinked with EDC. The stresses were 1.44 Kgf and 4.81 Kgf/mm2, respectively; the maximum load and maximum stress of Xn/HA/GO (0.03) film crosslinked with EDC were 1.61 Kgf and 5.39, respectively. Kgf/mm2. Statistical analysis of the above values revealed that the Xn/HA/GO (0.003) film was crosslinked with EDC and the Xn/HA film system was not significantly different from the EDC, and XN/HA/GO was crosslinked with EDC ( 0.03) The film has a significant difference from the Xn/HA film system in the cross-linking reaction with EDC.

From the above results, it is shown that although graphene oxide can enhance the mechanical strength of the diaphragm, its content must be a predetermined amount. In other words, the thermal wound dressing of the present invention has at least about 0.03 g of graphene oxide, or a weight ratio of graphene oxide, xanthan gum and hyaluronic acid of about 1:11:2 to 1:11:3. That is, the weight percentage of graphene oxide should be higher than 6%.

Example 8: Animal Testing

Nine 6-week-old, male SD rats (Lesco Biotech Co., Ltd.) were randomly divided into three groups. After anesthetizing the rats in each group, a 1 cm2 wound was made in the center of the back (day 0), and the wounds of each group were treated differently, and wound dressing was carried out for seven days. One group covered the wound with Xn/HA membrane; the second group covered the wound with Xn/HA/GO (0.03) membrane; the third group covered the wound with Xn/HA/GO (0.03) membrane and irradiated with wavelength of 805nm per day. The infrared rays were kept for 20 minutes to maintain the wound temperature at 45 °C.

After seven days of the above treatment, the appearance and area of the wounds of each group of rats were observed, and the results are shown in the ninth and tenth figures.

Please refer to Figure 9A. Because Xn/HA film has the characteristics of high water absorbing and moisturizing, hyaluronic acid has rapid degradability, so that the Xn/HA film system is completely disintegrated to form a colloid; see Figure IX and B. Compared with the first group of rats, the appearance of Xn/HA/GO (0.03) film was still intact and no fragmentation after covering for seven days, and there was obvious scabbing around the wound of the third group of rats. The wound is relatively dry and the wound area is significantly reduced.

Furthermore, from the area analysis before and after wound treatment in each group of rats, it was found that the wound area of the first group of rats did not change significantly from the 0th day to the 7th day of the experiment, indicating that the wound healing degree was not good; Although the wound area did not change significantly, the wounds showed a dry state and gradually recovered. The wounds of the third group of rats were significantly reduced, as shown in the tenth figure, and the reduction ratio of each sample in the tenth figure was The following are 33%, 33%, and 43%, respectively, indicating that the thermal-sensing wound dressing provided by the present invention cooperates with the provision of thermal energy to increase the rate of wound healing by at least 1.3 times.

The result shows that the thermal wound dressing of the present invention can protect the wound for a long time without causing disintegration or displacement, and the thermal wound dressing of the present invention can receive external heat source, and Heat is transferred to the wound site to accelerate wound healing.

Each group of rats was sacrificed, and the wound tissue was taken for paraffin-embedded sections, and HE and Meisheng were stained separately to observe the wound tissue. The results are shown in Fig. 11 to Fig. 16.

From the results of the eleventh to thirteenth graphs, no inflammatory cell aggregation was found in the wound tissue sections of the second and third groups of rats, and the fibroblast density of the second and third groups of rats was observed. Compared with the first group, the third group of rats has the highest fiber cell density, and thus it can be seen that the thermal wound dressing of the present invention has good biocompatibility and can convert light energy into heat energy. The heat is then transmitted to the wound to enhance the healing of the wound.

From the results of the fourteenth to sixteenth images, the density of collagen in the wounds of the second and third groups of rats was higher than that of the first group of rats after seven days of treatment, indicating that it contained graphene oxide. The dressing system can increase the strength of the membrane and release the ingredients into the wound, which can protect the wound for a long time and increase the healing efficiency of the wound.

Claims (7)

A thermal wound dressing prepared by the following steps: Step a: preparing a xanthan gum aqueous solution and a hyaluronic acid aqueous solution; Step b: mixing the xanthan gum aqueous solution and the hyaluronic acid aqueous solution to form a xanthan gum/hyaluronic acid An aqueous solution; step c: adding a predetermined amount of graphene oxide to the xanthan gum/hyaluronic acid aqueous solution to obtain a xanthan gum/hyaluronic acid/graphene oxide aqueous solution; step d: the xanthan gum/hyaluronic acid/graphene oxide aqueous solution After reacting with a crosslinking agent, it is thinned and dried to obtain a heat-sensing wound dressing; wherein, the weight ratio of the graphene oxide, the xanthan gum and the hyaluronic acid is about 1:11: 2~1:11:3. The thermal wound dressing according to claim 1, wherein the crosslinking agent is 1-ethyl-3-(3-dimethylaminopropyl)carbonium diimide. A use of a thermal wound dressing according to claim 1 of the patent application for the preparation of a medicament for accelerating the improvement of wound healing efficiency. A thermal wound dressing prepared by the following steps: Step a: preparing a xanthan gum aqueous solution and a hyaluronic acid aqueous solution; Step b: mixing the xanthan gum aqueous solution and the hyaluronic acid aqueous solution to form a xanthan gum/hyaluronic acid An aqueous solution; step c: adding a predetermined amount of graphene oxide to the xanthan gum / hyaluronic acid aqueous solution to obtain a xanthan gum / hyaluronic acid / graphene oxide aqueous solution; Step d: reacting the xanthan gum/hyaluronic acid/graphene oxide aqueous solution with a crosslinking agent, thinning and drying to obtain a thermal sensation wound dressing, wherein the crosslinking agent is 1-ethyl -3-(3-dimethylaminopropyl)carbonium diimine. The thermal wound dressing according to claim 4, wherein the weight ratio of the graphene oxide, the xanthan gum and the hyaluronic acid is about 1:11:2 to 1:11:3. The thermal wound dressing of claim 4, wherein the graphene oxide has a weight percentage of at least 6%. A use of a thermal wound dressing according to claim 4 of the patent application for the preparation of a medicament for accelerating the improvement of wound healing efficiency.