CN114479205A - Temperature-sensitive hydrogel for treating wounds and preparation method thereof - Google Patents
Temperature-sensitive hydrogel for treating wounds and preparation method thereof Download PDFInfo
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Abstract
A temperature-sensitive hydrogel for treating wounds and a preparation method thereof relate to the field of hydrogel dressings, in particular to a temperature-sensitive hydrogel and a preparation method thereof. The preparation method aims to solve the problems that existing hydrogel wound dressings prepared from chitosan are poor in solubility and limited by pH. The hydrogel is prepared from N-2-hydroxypropyl trimethyl ammonium chloride chitosan, lactic acid and sodium glycerophosphate solution. The method comprises the following steps: firstly, preparing N-2-hydroxypropyl trimethyl ammonium chloride chitosan; and secondly, dropwise adding a sodium glycerophosphate solution into the N-2-HACC lactic acid solution under ice-bath stirring, and reacting for 30-40 min to obtain the N-2-HACC hydrogel solution. The invention is used for preparing the temperature-sensitive hydrogel for treating the wound.
Description
Technical Field
The invention relates to the field of hydrogel dressings, in particular to a temperature-sensitive hydrogel and a preparation method thereof.
Background
With the development of times, the occurrence of wounds everywhere is seen, and the problem of wound healing is more and more important in clinic. The different degrees of injury of the wound caused by different conditions are different, and the wound is also easy to infect and destroy the stable environment in the body. The complex environment in which the skin is exposed makes it difficult or even impossible for conventional wound dressings to meet the need for high quality wound repair and management. Therefore, there is an urgent need for new biomedical dressings that can accelerate wound healing.
The hydrogel serving as a wound auxiliary material can play a barrier function, prevent exogenous foreign matters from invading, provide a moist and acidic environment for the wound, accelerate the wound healing and reduce the dressing change times and the use amount; the hydrogel also has good water absorption, biocompatibility and air permeability, promotes the dissolution of necrotic tissues and dead skin, and effectively plays a role in autolysis debridement; meanwhile, the hydrogel can be tightly attached to an uneven wound, but cannot be adhered, and can be easily removed from the wound without causing secondary damage; in addition, various medicines and growth factors can be introduced into the three-dimensional network structure of the hydrogel, so that the healing of the wound is further accelerated; in addition, the hydrogel dressing is convenient to use, can be directly smeared on wounds, and has the advantages of easiness in cleaning and no residue. Therefore, the hydrogel wound dressing or the hydrogel wound dressing serving as a drug carrier has a very wide application prospect in the aspect of promoting wound healing.
Chitosan has multiple performances of biocompatibility, biodegradability, nontoxicity, broad-spectrum antibacterial property, hemostasis, anti-inflammation, promotion of wound healing and the like, is a common material of the existing hydrogel wound dressing, but is limited by pH because of poor solubility, can only be dissolved under the condition that the pH is lower than 6, and greatly limits the wide application of the chitosan.
Disclosure of Invention
The invention aims to solve the problems of poor solubility and pH limitation of the existing hydrogel wound dressing prepared by chitosan, and provides a temperature-sensitive hydrogel for treating wounds and a preparation method thereof.
The temperature-sensitive hydrogel for treating the wound is prepared from N-2-hydroxypropyl trimethyl ammonium chloride chitosan, lactic acid and sodium glycerophosphate solution. The sodium glycerophosphate solution contains alpha-sodium glycerophosphate and beta-sodium glycerophosphate.
The invention also provides a preparation method of the temperature-sensitive hydrogel for treating the wound, which comprises the following steps:
dissolving chitosan in an acetic acid solution, stirring, dropwise adding a NaOH solution, adjusting the pH to 9, soaking for 0.5-1 h, performing suction filtration, washing and precipitating with deionized water to be neutral, and performing vacuum freeze drying until the mass is m to obtain the treated chitosan;
dispersing the treated chitosan into isopropanol, uniformly stirring, heating to 80-85 ℃, dropwise adding an isopropanol solution of 2, 3-epoxypropyltrimethylammonium chloride, stirring at a constant temperature of 80-85 ℃ for 9-10 hours at a stirring speed of 500-550 r/min after dropwise adding is completed within 30min, cooling to room temperature, standing for 1-2 hours, adding 4 ℃ absolute ethyl alcohol, soaking for 0.5-1 hour, performing suction filtration, and performing vacuum freeze drying to constant weight to obtain the N-2-hydroxypropyltrimethylammonium chloride chitosan (N-2-HACC).
And secondly, dropwise adding a sodium glycerophosphate solution into the N-2-HACC lactic acid solution under ice-bath stirring, and reacting for 30-40 min to obtain the N-2-HACC hydrogel solution.
Furthermore, the volume ratio of the acetic acid to the deionized water in the acetic acid solution in the first step is (1-15): 20-40.
Furthermore, in the first step, the volume ratio of the mass of the chitosan to the volume of the acetic acid in the acetic acid solution is 1g (1-15) mL.
Furthermore, the mass m ratio of the chitosan to the mass after freeze drying in the step one is 1 (4-10).
Further, the ratio of the mass of the chitosan treated in the step one to the volume of the isopropanol is 1g (5-15) mL.
Further, the volume ratio of the mass of the chitosan treated in the step one to the volume of the epoxypropyltrimethylammonium chloride isopropyl alcohol solution is 1g (5-15) mL; the concentration of the epoxypropyltrimethylammonium chloride isopropanol solution was 0.18 g/mL.
Further, in the first step, the volume ratio of the mass of the chitosan to the volume of the absolute ethyl alcohol is 1g (10-30) mL.
Further, the mass ratio of the sodium glycerophosphate to the ultrapure water in the sodium glycerophosphate solution in the second step is 1.5 (1.5-7.5), wherein the sodium glycerophosphate is composed of alpha-sodium glycerophosphate and beta-sodium glycerophosphate according to the mass ratio of 1: 2.
Furthermore, the volume ratio of the mass of the N-2-HACC in the lactic acid solution of the N-2-HACC in the step II to the volume of the lactic acid is 0.4g (5-10) mL, and the concentration of the lactic acid is 0.1 mol/L.
Furthermore, the volume ratio of the N-2-HACC lactic acid solution to the sodium glycerophosphate solution in the second step is (1-2) to 1.
The invention has the beneficial effects that:
the invention adopts the chitosan derivative, namely the N-2-hydroxypropyl trimethyl ammonium chloride chitosan, and compared with the chitosan, the chitosan derivative has the advantages of better solubility under neutral and weakly acidic conditions, is more suitable for preparing gel and can reduce the irritation to the skin.
The structure of the N-2-HACC hydrogel is a porous three-dimensional reticular structure, and the structure is favorable for the exchange of water, gas and some small molecular substances, so that the N-2-HACC hydrogel is guaranteed to be used as a wound dressing or a wound repair drug; at the same time, the three-dimensional network structure is suitable for being used as a carrier for delivering drugs. The method introduces quaternary ammonium salt with positive charges, further introduces a three-dimensional self-assembly function formed by the attraction of the positive charges and the negative charges on the basis of forming gel by chitosan ion crosslinking, so that the formed gel has better porosity, and the porous three-dimensional network structure has large specific surface area and strong adsorption force, thereby being more suitable for drug loading.
The N-2-HACC hydrogel can store a large amount of water and has good water storage performance, which indicates that the N-2-HACC hydrogel can keep a wound moist, provides a good environmental foundation for cell proliferation, tissue hyperplasia and the like at the wound and is beneficial to wound healing; meanwhile, the characteristic also provides feasible support for carrying soluble drugs or vaccine antigens by the N-2-HACC hydrogel. The N-2-HACC hydrogel can store a large amount of water and has better water storage performance, because compared with chitosan, the N-2-HACC hydrogel has better hydration compatibility, and meanwhile, a porous three-dimensional structure formed by self-assembly and ionic crosslinking has high specific surface area, so that the gel obtained by the invention can store a large amount of water.
N-2-HACC, alpha-sodium glycerophosphate and beta-sodium glycerophosphate which are adopted by the invention are all non-toxic and side-effect-free materials, cytotoxicity tests show that hydrogel freeze-dried powder does not show obvious toxicity to PK-15 cells, and the cell activity can still reach (88.72% +/-3.39)%, when the maximum concentration is 1000 mu g/mL, which shows that the N-2-HACC hydrogel has good safety.
The N-2-HACC hydrogel synthesized by the invention has excellent wound promoting effect.
Drawings
FIG. 1 is the morphology of the N-2-HACC hydrogel in example 1 under a scanning electron microscope;
FIG. 2 is a graph showing the measurement of PK-15 cytotoxicity of N-2-HACC hydrogel dry powder at various concentrations in example 1;
FIG. 3 is a graph showing the determination of PK-15 cytotoxicity of various concentrations of N-2-HACC hydrogel solutions;
FIG. 4 shows the wound healing of SD rats after administration in example 1;
FIG. 5 is a graph showing the wound healing rate of SD rats after administration in example 1;
FIG. 6 is a pathological observation image of the skin tissue wound of the rat in example 1.
Detailed Description
The technical solution of the present invention is not limited to the following specific embodiments, but includes any combination of the specific embodiments.
The first embodiment is as follows: the temperature-sensitive hydrogel for treating the wound is prepared from N-2-hydroxypropyl trimethyl ammonium chloride chitosan, lactic acid and sodium glycerophosphate solution. The sodium glycerophosphate solution contains alpha-sodium glycerophosphate and beta-sodium glycerophosphate.
The invention synthesizes hydrophilic quaternized chitosan N-2-HACC by chemical modification, and prepares temperature-sensitive quaternized chitosan Hydrogel (N-2-HACC Hydrogel) by taking the N-2-HACC as a raw material and alpha-sodium glycerophosphate and beta-sodium glycerophosphate as cross-linking agents.
The second embodiment is as follows: the preparation method of the temperature-sensitive hydrogel for treating the wound comprises the following steps:
firstly, preparing N-2-HACC with controllable degree of substitution
1. Soaking treatment of chitosan
Dissolving chitosan in an acetic acid solution, dropwise adding a NaOH solution after stirring, adjusting the pH to 9, soaking for 0.5-1 h, carrying out suction filtration, washing and precipitating with deionized water to be neutral, and carrying out vacuum freeze drying until the mass is m to obtain the treated chitosan; the concentration of the NaOH solution is 15 mol/L;
2. ring opening reaction of chitosan and epoxypropyl trimethyl ammonium chloride
Dispersing the treated chitosan into isopropanol, uniformly stirring, heating to 80-85 ℃, dropwise adding an isopropanol solution of 2, 3-epoxypropyltrimethylammonium chloride, stirring at a constant temperature of 80-85 ℃ for 9-10 hours at a stirring speed of 500-550 r/min after dropwise adding is completed within 30min, cooling to room temperature, standing for 1-2 hours, adding 4 ℃ absolute ethyl alcohol, soaking for 0.5-1 hour, performing suction filtration, and performing vacuum freeze drying to constant weight to obtain the N-2-hydroxypropyltrimethylammonium chloride chitosan (N-2-HACC).
Secondly, preparing N-2-HACC hydrogel
And (3) dropwise adding a sodium glycerophosphate solution into the N-2-HACC lactic acid solution under ice-bath stirring, carrying out an ionic crosslinking reaction, and reacting for 30-40 min to obtain the N-2-HACC hydrogel solution.
The method introduces quaternary ammonium salt with positive charges, further introduces three-dimensional self-assembly action formed by attraction of the positive charges and the negative charges on the basis of forming gel by chitosan ion crosslinking, so that the formed gel has better porosity, and the porous three-dimensional network structure has large specific surface area and strong adsorption force, thereby being more suitable for drug loading.
The N-2-HACC hydrogel can store a large amount of water and has better water storage performance, because compared with chitosan, the N-2-HACC hydrogel has better hydration compatibility, and meanwhile, a porous three-dimensional structure formed by self-assembly and ionic crosslinking has high specific surface area, so that the gel obtained by the invention can store a large amount of water.
The third concrete implementation mode: the second embodiment is different from the first embodiment in that: the volume ratio of acetic acid to deionized water in the acetic acid solution in the first step is (1-15): 20-40. The rest is the same as the second embodiment.
The fourth concrete implementation mode: the second embodiment is different from the first embodiment in that: in the first step, the volume ratio of the mass of the chitosan to the volume of the acetic acid in the acetic acid solution is 1g (1-15) mL. The rest is the same as the second embodiment.
The fifth concrete implementation mode: the second embodiment is different from the first embodiment in that: in the first step, the mass m ratio of the chitosan to the mass m after freeze drying is 1 (4-10). The rest is the same as the second embodiment.
Because the chitosan added is completely dried, the freeze drying in the first step is stopped to a certain extent, and the chitosan is not completely dried, certain moisture is reserved so that the porous structure is reserved, and the reserved moisture can be used as a solvent of the product in the next reaction. Therefore, the chitosan after freeze drying treatment has larger mass than the original chitosan.
The sixth specific implementation mode: the second embodiment is different from the first embodiment in that: the volume ratio of the chitosan treated in the step one to the isopropanol is 1g (5-15) mL. The rest is the same as the second embodiment.
The seventh embodiment: the second embodiment is different from the first embodiment in that: the volume ratio of the mass of the chitosan treated in the step one to the volume of the epoxypropyltrimethylammonium chloride isopropyl alcohol solution is 1g (5-15) mL; the concentration of the epoxypropyltrimethylammonium chloride isopropanol solution was 0.18 g/mL. The rest is the same as the second embodiment.
The specific implementation mode is eight: the second embodiment is different from the first embodiment in that: in the first step, the volume ratio of the chitosan to the absolute ethyl alcohol is 1g (10-30) mL. The rest is the same as the second embodiment.
The specific implementation method nine: the second embodiment is different from the first embodiment in that: and secondly, the mass ratio of the sodium glycerophosphate to the ultrapure water in the sodium glycerophosphate solution is 1.5 (1.5-7.5), wherein the sodium glycerophosphate is composed of alpha-sodium glycerophosphate and beta-sodium glycerophosphate according to the mass ratio of 1: 2. The rest is the same as the second embodiment.
The detailed implementation mode is ten: the second embodiment is different from the first embodiment in that: and in the second step, the volume ratio of the mass of the N-2-HACC in the N-2-HACC lactic acid solution to the lactic acid is 0.4g (5-10) mL, and the concentration of the lactic acid is 0.1 mol/L. The rest is the same as the second embodiment.
The concrete implementation mode eleven: the second embodiment is different from the first embodiment in that: in the second step, the volume ratio of the N-2-HACC lactic acid solution to the sodium glycerophosphate solution is (1-2) to 1. The rest is the same as the second embodiment.
The following examples are given to illustrate the present invention, and the following examples are carried out on the premise of the technical solution of the present invention, and give detailed embodiments and specific procedures, but the scope of the present invention is not limited to the following examples.
Example 1:
the preparation method of the temperature-sensitive hydrogel for treating the wound comprises the following steps:
firstly, preparing N-2-HACC;
soaking chitosan
Dissolving 6g of chitosan in an acetic acid solution, stirring for 1 hour by using a constant-speed stirrer, then dropwise adding 15mol/L NaOH solution, adjusting the pH value to 9, soaking for 0.5 hour, performing suction filtration, washing and precipitating with deionized water to be neutral, and performing vacuum freeze drying to 25g to obtain the soaked chitosan; the acetic acid solution was made from 66.6mL acetic acid and 173.4mL deionized water;
② the ring-opening reaction of chitosan and epoxypropyl trimethyl ammonium chloride
Dispersing the chitosan obtained after the soaking treatment in the step I into 50mL of isopropanol solution, uniformly stirring, carrying out water bath at 80 ℃, dropwise adding the isopropanol solution of 2, 3-epoxypropyltrimethylammonium chloride, finishing dropwise adding within 30min, then carrying out stirring reaction at 80 ℃ for 9h at a constant temperature, wherein the stirring speed is 500r/min, cooling to room temperature, standing for 1h, adding 150mL of 4 ℃ absolute ethyl alcohol, soaking for 0.5h, carrying out suction filtration by using the absolute ethyl alcohol, and carrying out vacuum freeze drying to constant weight to obtain the N-2-hydroxypropyl trimethyl ammonium chloride chitosan. The isopropanol solution of 2, 3-epoxypropyltrimethylammonium chloride was prepared from 9g of 2, 3-epoxypropyltrimethylammonium chloride and 50mL of isopropanol.
Secondly, preparing an N-2-HACC lactic acid solution;
weighing 0.4g N-2-HACC, dissolving in 6mL of 0.1mol/L lactic acid, fully dissolving, and placing in a refrigerator to keep the temperature at 4 ℃;
thirdly, preparing a sodium glycerophosphate solution;
weighing 0.5g of alpha-sodium glycerophosphate and 1g of beta-sodium glycerophosphate, dissolving in 4mL of ultrapure water, fully dissolving, and placing in a refrigerator to keep the temperature at 4 ℃;
fourthly, preparing N-2-HACC hydrogel;
and dropwise adding a sodium glycerophosphate solution into the N-2-HACC lactic acid solution under ice-bath stirring for carrying out an ionic crosslinking reaction, and reacting for 30min to obtain the N-2-HACC hydrogel solution.
FIG. 1 shows the morphology of the N-2-HACC hydrogel in this example under a scanning electron microscope. The structure of the N-2-HACC hydrogel is a porous three-dimensional reticular structure, the structure is favorable for the exchange of water, gas and some small molecular substances, and the guarantee is provided for the N-2-HACC hydrogel as a wound dressing or a wound repair drug; at the same time, the three-dimensional network structure is suitable for being used as a carrier for delivering drugs.
The water content determination result of the N-2-HACC hydrogel in the embodiment shows that the water content of the N-2-HACC hydrogel is (87.27 +/-0.11)%, which indicates that the N-2-HACC hydrogel can store a large amount of water and has better water storage performance, the N-2-HACC hydrogel can keep a wound moist, provides a good environmental basis for cell proliferation, tissue proliferation and the like at the wound and is beneficial to wound healing; meanwhile, the characteristic also provides feasible support for carrying soluble drugs or vaccine antigens by the N-2-HACC hydrogel.
FIG. 2 is a graph showing the measurement of PK-15 cytotoxicity of N-2-HACC hydrogel dry powder at different concentrations in this example. It can be seen that with the increase of the concentration of the N-2-HACC hydrogel freeze-dried powder, although the influence of the N-2-HACC hydrogel freeze-dried powder on the PK-15 cell activity is changed, the influence difference of the N-2-HACC hydrogel freeze-dried powder with different concentrations on the PK-15 cell activity is not significant (p is greater than 0.05), and the cell activity can reach more than 85% under each concentration. Compared with a control group, the N-2-HACC hydrogel freeze-dried powder does not show obvious toxicity to PK-15 cells, and the cell activity can still reach (88.72% +/-3.39)% at the maximum concentration of 1000 mu g/mL, which indicates that the N-2-HACC hydrogel freeze-dried powder does not have obvious cytotoxicity to the PK-15 cells.
FIG. 3 is a graph showing the determination of PK-15 cytotoxicity of various concentrations of N-2-HACC hydrogel solutions. With the increase of the concentration of the N-2-HACC hydrogel solution, the influence of the N-2-HACC hydrogel solution on the activity of PK-15 cells is increased, but the influence on the activity of the cells is not great, and the activity of the PK-15 cells treated by the N-2-HACC hydrogel with the rest concentration can reach more than 80 percent except at the highest concentration. Compared with a control group, the N-2-HACC hydrogel solution does not show obvious toxicity to PK-15 cells, when the maximum concentration of the N-2-HACC hydrogel solution is 12.50%, the cell viability can still reach (74.13 +/-1.51)%, which indicates that the N-2-HACC hydrogel solution has little influence on the growth of the PK-15 cells, and indicates that the N-2-HACC hydrogel has extremely low cytotoxicity.
Randomly dividing the SD rats after punching into four groups, namely a negative Control group (Control), a chitosan hydrogel group (CS group), a carboxynin liquid wound dressing group (SN group) and an N-2-HACC hydrogel group (N-2-HACC group), wherein each group is 14; the negative control group is free from medicine application to the wound, the CS group is chitosan hydrogel, the SN group is a liquid wound dressing containing hydroxynin, and the N-2-HACC group is N-2-HACC hydrogel. FIG. 4 shows the healing of the wound of the SD rat after administration in this example. As can be seen from FIG. 4, the wounds of rats in each treatment group were improved to different degrees after administration. At 3d after administration, the wound surface on the back of rats in each treatment group was dark red, and the wound began to scab; the wound of the rat in the 5d, N-2-HACC hydrogel group changed from red to dark red after the drug application, and the scab began to dry and harden; at 10d, the wounds of the rats with the N-2-HACC hydrogel group scab and fall off, the healing of the wounds is accelerated, at 12d, the skin wounds of the rats with the N-2-HACC hydrogel group are basically and completely healed, and the skin wounds of the rats with the commodity medicine group and the chitosan hydrogel group are not healed. The wound surface of the rat in the negative control group becomes dark and hard at the 7 th scab, the scab falls off at the 10 th day and the wound shrinks at the 15 th day; the chitosan hydrogel group and the commercial medicament of the phentermine group have the advantages that scab becomes hard at the 5 th day after administration, the scab becomes dark red at the 7 th day, the scab becomes dry scab at the same time, the scab becomes small at the 10 th day, the wound surface is further healed, the scab disappears at the 15 th day, scars are left, and the wound is basically healed. In conclusion, at the same time after administration, the wounds of the N-2-HACC hydrogel group were smaller than those of the other groups, and at 12d, the N-2-HACC hydrogel group appeared in rats with completely healed wounds, while none of the other groups appeared; at 15d, 1 rat with completely healed wound appeared in both the chitosan hydrogel group and the commercial drug carboxynine group, while the control group did not have completely healed wound at 15 d.
FIG. 5 shows the wound healing rate of SD rats after administration in this example,
a negative control group was indicated, and,
it represents a group of chitosan hydrogel,
showing a group of the liquid wound dressings of carboxynine,
represents the N-2-HACC hydrogel group. There was no significant difference in the wound healing rate of rats in each treatment group at 1d post-dose. After administration, the average healing rate of the wounds of the rats in the 3d, N-2-HACC hydrogel group is 24.79%, the average healing rate of the wounds of the negative control group is 14.94%, the healing rates of the wounds of the rats in the chitosan hydrogel group and the commercial drug carboxymethyl group are 21.09% and 21.91%, respectively, and the healing rate of the wounds of the rats in the N-2-HACC hydrogel group is obviously higher than that of the negative control group (p is less than 0.001); the healing rate of the N-2-HACC hydrogel group rat wound is not obviously different from that of the chitosan hydrogel group and the carboxymethyl hydrogel group (p)>0.05), which shows that the three groups of drugs have the same effect on healing the skin wound of the rat when continuously administered for 3 d.
The average healing rate of the wounds of rats in the 5d, N-2-HACC hydrogel group was 38.36%, the negative control group was 35.41%, and the chitosan hydrogel group and the commercial drug carboxynine group were 38.67% and 38.71%, respectively, after administration. The wound healing rate of the N-2-HACC hydrogel group rat is obviously higher than that of a negative control group (p is less than 0.001); the healing rate of the wounds of the rats in the N-2-HACC hydrogel group is not obviously different from that of the other two groups (p is greater than 0.05), which indicates that the three groups of the drugs have the same healing effect on the wounds of the skins of the rats when the drugs are continuously administered for 5 days.
The average healing rate of the wound of the rat in the 7d, N-2-HACC hydrogel group after administration was 55.76%, that of the negative control group was 42.47%, and that of the chitosan hydrogel group and the commercial drug carboxynine group was 44.67% and 45.20%, respectively. The wound healing rate of the N-2-HACC hydrogel group rat is obviously higher than that of a negative control group (p is less than 0.001); meanwhile, the wound healing rate of the N-2-HACC hydrogel group of the rat is obviously higher than that of the other two groups (p is less than 0.001), which shows that the wound healing promoting effect of the N-2-HACC hydrogel group is better than that of the chitosan hydrogel group and the commercial medicament carboxynine group when the N-2-HACC hydrogel group is continuously administrated for 7 days.
The average wound healing rate of rats in the 10d, N-2-HACC hydrogel group was 77.69%, 71.88% in the negative control group, and 72.67% and 71.83% in the chitosan hydrogel group and the commercial drug, carboxymethyl group, respectively, after administration. The healing rate of the rat wound of the N-2-HACC hydrogel group is obviously higher than that of the negative control group (p is less than 0.001); meanwhile, the wound healing rate of the N-2-HACC hydrogel group of the rat is obviously higher than that of the other two groups (p is less than 0.05), which shows that the wound healing promoting effect of the N-2-HACC hydrogel group is better than that of the chitosan hydrogel group and the commercial medicament carboxynine group when the N-2-HACC hydrogel group is continuously administrated for 10 days.
The average healing rate of the wounds of the rats in the 15d, N-2-HACC hydrogel group after administration was 100%, 85.92% in the negative control group, 93.89% in the chitosan hydrogel group and 92.28% in the commercial drug, carboxymethyl group, respectively.
FIG. 6 is a view showing the pathological observation of the skin tissue wound of the rat in this example; as can be seen from the figure, the skin tissue around the wound of the rats in the N-2-HACC hydrogel group showed an earlier inflammatory cell infiltration time, and new fibroblasts and new blood vessels began to appear at 5d after administration, and the epidermis was formed at an earlier time, compared with the pathological section of the skin tissue around the wound of the rats in the negative control group. Compared with the skin tissues around the wounds of rats in the N-2-HACC hydrogel group, the chitosan hydrogel group and the carboxynine group, the skin tissues around the wounds of the three groups of rats have inflammatory cell infiltration time at the 3 rd after administration, inflammatory cell infiltration is increased at the 5 th after administration, and new blood vessels and fibroblasts appear in dermal tissues, which shows that the three groups of rats have similar effects in the inflammation stage and the aspect of promoting the formation of the new blood vessels. However, from 7d after administration, the N-2-HACC hydrogel group exhibited a significant predominance in promoting epithelization, granulation tissue proliferation, neovascularization, and fibroblast, compared to the skin tissue changes around the wound in rats in the chitosan hydrogel group and the carboxynine group, indicating that the N-2-HACC hydrogel has a stronger chemotactic effect on fibroblast and an effect of promoting epithelization, granulation tissue proliferation, neovascularization. These results are consistent with the trend of the results from external observations of wound healing and the rate of wound healing.
Claims (10)
1. A temperature-sensitive hydrogel for treating wounds is characterized in that the hydrogel is prepared from N-2-hydroxypropyl trimethyl ammonium chloride chitosan, lactic acid and sodium glycerophosphate solution; the sodium glycerophosphate solution contains alpha-sodium glycerophosphate and beta-sodium glycerophosphate.
2. The method for preparing a temperature-sensitive hydrogel for treating wounds according to claim 1, comprising the steps of:
dissolving chitosan in an acetic acid solution, stirring, dropwise adding a NaOH solution, adjusting the pH to 9, soaking for 0.5-1 h, performing suction filtration, washing and precipitating with deionized water to be neutral, and performing vacuum freeze drying until the mass is m to obtain the treated chitosan;
dispersing the treated chitosan into isopropanol, uniformly stirring, heating to 80-85 ℃, dropwise adding an isopropanol solution of 2, 3-epoxypropyltrimethylammonium chloride, stirring at a constant temperature of 80-85 ℃ for 9-10 hours at a stirring speed of 500-550 r/min after dropwise adding is completed within 30min, cooling to room temperature, standing for 1-2 hours, adding 4 ℃ absolute ethyl alcohol, soaking for 0.5-1 hour, carrying out suction filtration, and carrying out vacuum freeze drying to constant weight to obtain the N-2-hydroxypropyl trimethylammonium chloride chitosan;
and secondly, dropwise adding a sodium glycerophosphate solution into the N-2-HACC lactic acid solution under ice-bath stirring, and reacting for 30-40 min to obtain the N-2-HACC hydrogel solution.
3. The preparation method of the temperature-sensitive hydrogel for treating wounds according to claim 2, wherein the volume ratio of the mass of chitosan to acetic acid in the acetic acid solution in the first step is 1g (1-15) mL.
4. The preparation method of the temperature-sensitive hydrogel for treating wounds according to claim 2 or 3, wherein the mass m ratio of the chitosan to the mass m after freeze drying in the first step is 1 (4-10).
5. The preparation method of the temperature-sensitive hydrogel for treating wounds according to claim 4, wherein the volume ratio of the chitosan to the isopropanol after treatment in the step one is 1g (5-15) mL.
6. The preparation method of the temperature-sensitive hydrogel for treating the wounds according to claim 2, 4 or 5, wherein the volume ratio of the mass of the chitosan treated in the step one to the volume of the epoxypropyltrimethylammonium chloride isopropyl alcohol solution is 1g (5-15) mL; the concentration of the epoxypropyltrimethylammonium chloride isopropanol solution was 0.18 g/mL.
7. The preparation method of the temperature-sensitive hydrogel for treating wounds according to claim 6, wherein the volume ratio of the mass of chitosan to the volume of absolute ethyl alcohol in the first step is 1g (10-30) mL.
8. The preparation method of the temperature-sensitive hydrogel for treating wounds according to claim 7, wherein in the second step, the mass ratio of sodium glycerophosphate to ultrapure water in the sodium glycerophosphate solution is 1.5 (1.5-7.5), wherein the sodium glycerophosphate is composed of alpha-sodium glycerophosphate and beta-sodium glycerophosphate according to the mass ratio of 1: 2.
9. The method for preparing a temperature-sensitive hydrogel for treating wounds according to claim 2 or 3, wherein the volume ratio of the mass of N-2-HACC to the volume of lactic acid in the N-2-HACC lactic acid solution in the step two is 0.4g (5-10) mL, and the concentration of lactic acid is 0.1 mol/L.
10. The preparation method of the temperature-sensitive hydrogel for treating wounds according to claim 9, wherein the volume ratio of the N-2-HACC lactic acid solution to the sodium glycerophosphate solution in the second step is (1-2): 1.
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| CN115089510A (en) * | 2022-06-20 | 2022-09-23 | 江南大学 | Intelligent temperature-sensitive permanent wave agent capable of reducing thioglycollic acid substances from entering human body and preparation method thereof |
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| CN115089510A (en) * | 2022-06-20 | 2022-09-23 | 江南大学 | Intelligent temperature-sensitive permanent wave agent capable of reducing thioglycollic acid substances from entering human body and preparation method thereof |
| CN115089510B (en) * | 2022-06-20 | 2023-03-28 | 江南大学 | Intelligent temperature-sensitive permanent wave agent capable of reducing thioglycollic acid substances from entering human body and preparation method thereof |
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