Common sterilization and hemostasis dressing for anesthesia and preparation method of common sterilization and hemostasis dressing

Abstract

The invention relates to the field of medical dressings, and discloses a common sterilization and hemostasis dressing for anesthesia and a preparation method thereof. According to the method, PVA is used as a base material, chitosan and a traditional Chinese medicine fermentation extracting solution are introduced into the base material, and the hydrogel dressing is prepared by firstly freezing and thawing and then irradiating; the traditional Chinese medicine fermentation extracting solution is obtained by selecting clove, astragalus membranaceus and white mulberry root-bark, carrying out combined fermentation by adopting lactobacillus acidophilus CICC 6082 and bifidobacterium longum CICC 6191 and extracting. According to the preparation method, the polyvinyl alcohol and the chitosan are subjected to mutual strong crosslinking in the gelling process by combining freeze thawing and irradiation, and meanwhile, the traditional Chinese medicine fermentation extracting solution is uniformly dispersed in a polymer or forms hydrogen bonds or intermolecular acting force with polymer molecules, so that the mechanical property of the hydrogel is improved. Syzygium aromaticum, astragalus membranaceus and white mulberry root-bark are combined with a traditional Chinese medicine fermentation extracting solution obtained by using specific strains of lactobacillus acidophilus CICC 6082 and bifidobacterium longum CICC 6191, the traditional Chinese medicine fermentation extracting solution and chitosan have a synergistic effect, and the effects of remarkably enhancing the antibacterial property, hemostasis and wound healing promotion of the dressing are achieved.

Description

Technical Field

[0001] The invention relates to the field of medical dressings, in particular to a commonly used bactericidal and hemostatic dressing for anesthesia and a preparation method thereof. Background Art

[0002] Anesthesia is an important part of clinical medicine. It can ensure that patients can receive surgical treatment smoothly under painless and safe conditions, eliminate the pain of patients, and thus smoothly perform the operation. A large amount of antibacterial hemostatic excipients are required during wound hemostasis during anesthesia surgery and postoperative wound healing. The functions of the excipients are to stop bleeding, prevent bacterial infection, and promote wound healing.

[0003] Medical antibacterial hemostatic dressings are an important medical auxiliary tool during anesthesia. They can help stop bleeding quickly and effectively prevent wounds from being infected by bacteria and causing inflammation. Traditional hemostatic dressings are mainly gauze dressings made of cotton, linen, and cotton. Such dressings have poor hemostatic effects and poor antibacterial properties during use. During the healing process, patients are affected by bacteria and suffer secondary damage to the wound surface. Therefore, innovation in medical hemostatic gauze is a relatively important research in the current development of medicine.

[0004] The invention uses PVA as a substrate, introduces chitosan CS and a traditional Chinese medicine fermentation extract therein, and adopts a cyclic freeze-thaw method to prepare a hydrogel dressing. The chitosan CS and the traditional Chinese medicine fermentation extract improve the performance of the PVA hydrogel, and the addition of the traditional Chinese medicine fermentation extract gives the hydrogel good antibacterial, hemostatic and wound healing performance. Summary of the invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a commonly used bactericidal hemostatic dressing for anesthesia and a preparation method thereof, so as to solve the problems raised in the above technical background.

[0006] To achieve the above object, the present invention is implemented through the following technical solutions:

[0007] In a first aspect, the present invention provides a method for preparing a commonly used bactericidal hemostatic dressing for anesthesia, comprising the following steps:

[0008] S1. Preparation of bacterial liquid: Lactobacillus acidophilus CICC 6082 was inoculated into MRS agar medium and cultured in a 37°C incubator for 48 h to obtain activated Lactobacillus acidophilus CICC 6082, which was then transferred to a shake flask containing 1000 mL of MRS liquid medium and fermented for 8-12 h to obtain a concentration of 3.6-4.5×10 8cfu / mL of Lactobacillus acidophilus CICC 6082 culture medium; Bifidobacterium longum CICC 6191 was inoculated into RCM agar medium and cultured in a 37°C incubator for 48 h to obtain activated Bifidobacterium longum CICC 6191, which was transferred to a shake flask containing 1000 mL of RCM liquid culture medium and fermented for 8-12 h to obtain a concentration of 1.2-2.0×10 8 cfu / mL Bifidobacterium longum CICC 6191 bacterial solution;

[0009] S2, preparation of fermentation: clove, astragalus, mulberry bark, wash, dry and crush into 80-100 mesh particles, first weigh 18-22 parts of clove, 18-22 parts of astragalus, 3-7 parts of mulberry bark by weight, mix evenly to obtain a mixture, then add 1-2 times the amount of water of the mixture, mix well to obtain a suspension, add Lactobacillus acidophilus CICC 6082 bacterial solution and Bifidobacterium longum CICC 6191 to the suspension at an inoculum amount of 3-5%, respectively, and ferment at 37° C. and anaerobic for 12-24 hours to obtain a fermentation product;

[0010] S3, preparation of Chinese medicine fermentation extract: take the fermented product in step S2, add 6 to 8 times the amount of 70% ethanol for 30-60 min reflux extraction, extract 3 times, discard the residue, combine the filtrate, recover the alcohol, and concentrate until there is no alcohol taste to obtain the Chinese medicine fermentation extract;

[0011] S4, preparing 1% by mass chitosan solution and 10% by mass polyvinyl alcohol solution;

[0012] S5, mixing the Chinese medicine fermentation extract obtained in step S3, the chitosan solution obtained in step S4, and the polyvinyl alcohol solution obtained in step S4 in proportion, and stirring for 1 hour in an oil bath at 85° C. to obtain a dressing solution;

[0013] S6. Defoam the dressing solution (homogenize, sonicate for 2 hours, or let stand at room temperature for 8-12 hours) and transfer it to a mold, freeze-thaw repeatedly, and then irradiate to obtain a hydrogel dressing, i.e., a sterilizing and hemostatic dressing.

[0014] Among them, MRS liquid culture medium: casein peptone 10.0g, beef extract powder 10.0g, yeast extract powder 5.0g, glucose 20.0g, Tween 80 1.0g, sodium acetate 5.0g, ammonium citrate tribasic 2.0g, K2HPO4 2.0g, MgSO4·7H2O 0.2g, MnSO4·H2O 0.05g, agar 15.0g, distilled water 1000.0mL, pH 6.2-6.5; MRS agar culture medium is based on MRS liquid culture medium, with 15.0g agar added.

[0015] RCM liquid medium: beef extract powder 10.0g, peptone 5.0g, yeast extract powder 3.0g, glucose 5.0g, soluble starch 1.0g, NaCl 5.0g, NaAc 3.0g, L-cysteine ​​hydrochloride 0.5g, agar 15.0g, distilled water 1000.0mL, pH 6.8. RCM agar medium is based on RCM liquid medium, with 15.0g agar added.

[0016] Preferably, in step S2, 20 parts of cloves, 20 parts of astragalus, and 5 parts of white mulberry bark are weighed by weight.

[0017] Preferably, in step S4, chitosan powder is dissolved in 1% acetic acid to prepare a 1% by mass chitosan solution.

[0018] Preferably, in step S4, 5 g of polyvinyl alcohol is soaked in deionized water at room temperature for 20 min, and then fully dissolved in a 95° C. oil bath for 1.5-2 h until transparent, to obtain a polyvinyl alcohol solution with a mass fraction of 10%.

[0019] Preferably, in step S5, the amount of chitosan added is 4-8% of the amount of polyvinyl alcohol.

[0020] Preferably, in step S5, the amount of the Chinese medicinal fermentation extract added is 0.05-0.1% of the amount of polyvinyl alcohol.

[0021] Preferably, in step S6, the repeated freezing and thawing method is: freezing at -30 to -20°C for 18 to 24 hours, thawing at 20 to 30°C for 2 to 6 hours, and repeating 3 to 7 times.

[0022] In step S6, the irradiation is carried out by 60 Coγ-ray irradiation has an irradiation dose of 10 to 80 kGy and an irradiation time of 1 to 10 minutes.

[0023] In a second aspect, the present invention provides a commonly used bactericidal hemostatic dressing for anesthesia prepared by the method for preparing the commonly used bactericidal hemostatic dressing for anesthesia.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] 1. In the dressing prepared by the present invention, polyvinyl alcohol and chitosan are combined by freeze-thawing followed by irradiation, and strong cross-linking occurs between them during the gelation process, so that the formed hydrogel has good tensile properties. The Chinese medicine fermentation extract is evenly dispersed in the polymer, or forms hydrogen bonds or intermolecular forces with the polymer molecules, thereby increasing the mechanical properties of the hydrogel.

[0026] 2. The synergistic effect of chitosan and Chinese medicine fermentation extract has significant effects in antibacterial, hemostatic and wound healing.

[0027] 3. The Chinese medicinal fermentation extract obtained by combining clove, astragalus and mulberry bark with specific strains of Lactobacillus acidophilus CICC 6082 and Bifidobacterium longum CICC6191 can significantly enhance the antibacterial, hemostatic and wound healing effects of dressings. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The bar graph shows the effect of different dressings on the inhibition zone diameter of S. aureus and E. coli;

[0029] Figure 2 The figure is a bar graph of the coagulation index of different dressings. DETAILED DESCRIPTION

[0030] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0031] PVA: alcoholysis degree is 98% to 99%, Shanghai Aladdin Biochemical Technology Co., Ltd.

[0032] Chitosan: specification ≥95%, Shanghai McLean Biochemical Technology Co., Ltd.;

[0033] Lactobacillus acidophilus CICC 6082, Lactobacillus acidophilus CICC 6091, Bifidobacterium longum CICC 6191 and Bifidobacterium longum CICC 6187 were purchased from China Industrial Microbiological Culture Collection Center.

[0034] Example 1

[0035] The present invention provides a method for preparing a commonly used bactericidal hemostatic dressing for anesthesia, comprising the following steps:

[0036] S1. Preparation of bacterial solution: The concentration is 4.5×10 8 cfu / mL of Lactobacillus acidophilus CICC 6082 and a concentration of 1.2×10 8 cfu / mL of Bifidobacterium longum CICC 6191 bacterial solution;

[0037] S2, preparation of fermentation: clove, astragalus, mulberry bark, wash, dry and crush into 100 mesh particles, first weigh 18 parts of clove, 22 parts of astragalus, 7 parts of mulberry bark by weight, mix well to obtain a mixture, then add 2 times the amount of water of the mixture, mix well to obtain a suspension, add Lactobacillus acidophilus CICC 6082 bacterial solution and Bifidobacterium longum CICC 6191 to the suspension at an inoculum amount of 3% v / v, respectively, and ferment at 37° C. and anaerobic for 12 h to obtain a fermentation product;

[0038] S3, preparation of Chinese medicine fermentation extract: taking the fermented product in step S2, adding 8 times the amount of 70% ethanol to carry out 60min reflux extraction, extracting 3 times, discarding the residue, combining the filtrate, recovering the alcohol, and concentrating until there is no alcohol taste, to obtain the Chinese medicine fermentation extract;

[0039] S4. Prepare a 1% mass fraction chitosan solution: dissolve chitosan powder in 1% acetic acid to prepare a 1% mass fraction chitosan solution; prepare a 10% mass fraction polyvinyl alcohol solution: soak 5g of polyvinyl alcohol in deionized water at room temperature for 20 minutes, and then fully dissolve it in a 95°C oil bath for 2 hours until it becomes transparent, to obtain a 10% mass fraction polyvinyl alcohol solution;

[0040] S5. The Chinese herbal medicine fermentation extract obtained in step S3, the chitosan solution obtained in step S4 and the polyvinyl alcohol solution obtained in step S4 are mixed in proportion, and stirred in an oil bath at 85° C. for 1 hour to obtain a dressing solution; the amount of chitosan added is 4% of the amount of polyvinyl alcohol, and the amount of the Chinese herbal medicine fermentation extract added is 0.1% of the amount of polyvinyl alcohol.

[0041] S6. After defoaming the dressing solution (standing at room temperature for 12 hours), transfer it to a mold, perform freeze-thaw treatment repeatedly, and then irradiate it to obtain a hydrogel dressing, i.e., a sterilizing and hemostatic dressing.

[0042] The method of repeated freezing and thawing is: freezing at -20℃ for 24h, thawing at 25℃ for 4h, and repeating 6 times. 60 Coγ-ray irradiation, the irradiation dose is 50 kGy, and the irradiation time is 6 minutes.

[0043] Example 2

[0044] The present invention provides a method for preparing a commonly used bactericidal hemostatic dressing for anesthesia, comprising the following steps:

[0045] S1. Preparation of bacterial solution: The concentration is 3.6×10 8 cfu / mL of Lactobacillus acidophilus CICC 6082 and a concentration of 2.0×10 8 cfu / mL of Bifidobacterium longum CICC 6191 bacterial solution;

[0046] S2, preparation of fermentation product: clove, astragalus, mulberry bark, wash, dry and crush into 100 mesh particles, first weigh 22 parts of clove, 18 parts of astragalus, 3 parts of mulberry bark by weight, mix well to obtain a mixture, then add 2 times the amount of water of the mixture, mix well to obtain a suspension, add Lactobacillus acidophilus CICC 6082 bacterial solution and Bifidobacterium longum CICC 6191 to the suspension at an inoculum amount of 5% v / v, respectively, and ferment at 37° C. and anaerobic for 24 hours to obtain a fermentation product;

[0047] S3, preparation of Chinese medicine fermentation extract: taking the fermented product in step S2, adding 8 times the amount of 70% ethanol to carry out 60min reflux extraction, extracting 3 times, discarding the residue, combining the filtrate, recovering the alcohol, and concentrating until there is no alcohol taste, to obtain the Chinese medicine fermentation extract;

[0048] S4. Prepare a 1% mass fraction chitosan solution: dissolve chitosan powder in 1% acetic acid to prepare a 1% mass fraction chitosan solution; prepare a 10% mass fraction polyvinyl alcohol solution: soak 5g of polyvinyl alcohol in deionized water at room temperature for 20 minutes, and then fully dissolve it in a 95°C oil bath for 2 hours until it becomes transparent, to obtain a 10% mass fraction polyvinyl alcohol solution;

[0049] S5. The Chinese herbal medicine fermentation extract obtained in step S3, the chitosan solution obtained in step S4 and the polyvinyl alcohol solution obtained in step S4 are mixed in proportion, and stirred in an oil bath at 85° C. for 1 hour to obtain a dressing solution; the amount of chitosan added is 8% of the amount of polyvinyl alcohol, and the amount of the Chinese herbal medicine fermentation extract added is 0.05% of the amount of polyvinyl alcohol.

[0050] S6. After defoaming the dressing solution (standing at room temperature for 12 hours), transfer it to a mold, perform freeze-thaw treatment repeatedly, and then irradiate it to obtain a hydrogel dressing, i.e., a sterilizing and hemostatic dressing.

[0051] The method of repeated freezing and thawing is: freezing at -20℃ for 24h, thawing at 25℃ for 4h, and repeating 6 times. 60 Coγ-ray irradiation, the irradiation dose is 50 kGy, and the irradiation time is 6 minutes.

[0052] Example 3

[0053] The present invention provides a method for preparing a commonly used bactericidal hemostatic dressing for anesthesia, comprising the following steps:

[0054] S1. Preparation of bacterial solution: The concentration is 4.0×10 8 cfu / mL of Lactobacillus acidophilus CICC 6082 and a concentration of 1.5×10 8 cfu / mL of Bifidobacterium longum CICC 6191 bacterial solution;

[0055] S2, preparation of fermentation product: clove, astragalus, mulberry bark, wash, dry and crush into 100 mesh particles, first weigh 20 parts of clove, 20 parts of astragalus, 5 parts of mulberry bark by weight, mix well to obtain a mixture, then add 2 times the amount of water of the mixture, mix well to obtain a suspension, add Lactobacillus acidophilus CICC 6082 bacterial solution and Bifidobacterium longum CICC 6191 to the suspension at an inoculum amount of 4% v / v, respectively, and ferment at 37° C. and anaerobic for 18 h to obtain a fermentation product;

[0056] S3, preparation of Chinese medicine fermentation extract: taking the fermented product in step S2, adding 8 times the amount of 70% ethanol to carry out 60min reflux extraction, extracting 3 times, discarding the residue, combining the filtrate, recovering the alcohol, and concentrating until there is no alcohol taste, to obtain the Chinese medicine fermentation extract;

[0057] S4. Prepare a 1% mass fraction chitosan solution: dissolve chitosan powder in 1% acetic acid to prepare a 1% mass fraction chitosan solution; prepare a 10% mass fraction polyvinyl alcohol solution: soak 5g of polyvinyl alcohol in deionized water at room temperature for 20 minutes, and then fully dissolve it in a 95°C oil bath for 2 hours until it becomes transparent, to obtain a 10% mass fraction polyvinyl alcohol solution;

[0058] S5. The Chinese herbal medicine fermentation extract obtained in step S3, the chitosan solution obtained in step S4 and the polyvinyl alcohol solution obtained in step S4 are mixed in proportion, and stirred in an oil bath at 85° C. for 1 hour to obtain a dressing solution; the amount of chitosan added is 6% of the amount of polyvinyl alcohol, and the amount of the Chinese herbal medicine fermentation extract added is 0.08% of the amount of polyvinyl alcohol.

[0059] S6. After defoaming the dressing solution (standing at room temperature for 12 hours), transfer it to a mold, perform freeze-thaw treatment repeatedly, and then irradiate it to obtain a hydrogel dressing, i.e., a sterilizing and hemostatic dressing.

[0060] The method of repeated freezing and thawing is: freezing at -20℃ for 24h, thawing at 25℃ for 4h, and repeating 6 times. 60 Coγ-ray irradiation, the irradiation dose is 50 kGy, and the irradiation time is 6 minutes.

[0061] Comparative Example 1

[0062] This embodiment is similar to Embodiment 3, except that the Chinese medicine fermentation extract is removed.

[0063] Comparative Example 2

[0064] This embodiment is similar to embodiment 3, except that chitosan is removed.

[0065] Comparative Example 3

[0066] This embodiment is similar to Embodiment 3, except that the freeze-thaw cycle step is removed in step S6.

[0067] Comparative Example 4

[0068] This embodiment is similar to Embodiment 3, except that the irradiation treatment step is removed in step S6.

[0069] Comparative Example 5

[0070] This embodiment is similar to embodiment 3, except that cloves are removed from the Chinese medicine fermentation extract.

[0071] Comparative Example 6

[0072] This embodiment is similar to embodiment 3, except that astragalus is removed from the traditional Chinese medicine fermentation extract.

[0073] Comparative Example 7

[0074] This embodiment is similar to embodiment 3, except that the mulberry bark is removed from the traditional Chinese medicine fermentation extract.

[0075] Comparative Example 8

[0076] This embodiment is similar to embodiment 3, except that the Chinese medicine fermentation extract is directly fermented by clove, astragalus, and mulberry bark without using a strain.

[0077] Comparative Example 9

[0078] This embodiment is similar to embodiment 3, except that the Chinese medicine fermentation extract only uses Lactobacillus acidophilus CICC6082.

[0079] Comparative Example 10

[0080] This embodiment is similar to embodiment 3, except that the Chinese medicine fermentation extract only uses Bifidobacterium longum CICC6191.

[0081] Comparative Example 11

[0082] This embodiment is similar to the embodiment 3, except that: the traditional Chinese medicine fermentation extract uses Lactobacillus acidophilus CICC 6082 and other Bifidobacterium longum, and the other Bifidobacterium longum is Bifidobacterium longum CICC 6187.

[0083] Comparative Example 12

[0084] This embodiment is similar to the embodiment 3, except that: the Chinese medicine fermentation extract only uses other Lactobacillus acidophilus and Bifidobacterium longum CICC 6191, and other Lactobacillus acidophilus is selected from Lactobacillus acidophilus CICC 6091.

[0085] 1. Mechanical tensile properties test of dressing

[0086] The hydrogels obtained in Examples 1-3 and Comparative Examples 1-4 were respectively made into dumbbell shapes with a total length of 40 mm and a width of 15 mm, with dumbbell parts on both sides and a sample part in the middle. The sample length was 30 mm, the sample width (i.e., the shortest distance in the middle of the dumbbell) was 10 mm, and the sample thickness was 1 mm. At room temperature of 25° C., a stretching rate of 20 mm / min was set, and a C43-104 microcomputer-controlled electronic universal testing machine was used to measure the tension (N) and elongation (mm) of the sample at break. Each sample was repeated 5 times and the average value was taken to calculate the tensile strength and elongation at break of the hydrogel dressing. The results are shown in Table 1.

[0087] The tensile strength of the hydrogel (σ) is the ratio of the maximum tensile stress of the gel to the cross-sectional area of ​​the sample, that is, Where: σ is the tensile strength MPa, F max is the maximum tensile stress N, A is the cross-sectional area of ​​the sample (sample width × sample thickness) mm 2 ;

[0088] The elongation at break (ε) is the ratio of the deformation length of the sample to the initial length at break, that is, Among them, ε (%) is the elongation at break, L0 and L max are the initial and fracture lengths of the sample in mm, respectively.

[0089] Table 1 Tensile strength and elongation at break of hydrogel dressings

[0090]

[0091]

[0092] As shown in Table 1, compared with Comparative Example 1, the tensile strength and elongation at break of Examples 1-3 increased significantly, which may be due to the uniform dispersion of the Chinese medicine fermentation extract in the polymer, or the formation of hydrogen bonds or intermolecular forces between the polymer molecules, which increased the mechanical properties of the hydrogel. Compared with Comparative Examples 2-4, the tensile strength and elongation at break of Examples 1-3 increased significantly, indicating that the removal of chitosan, or the removal of repeated freeze-thaw or irradiation processes, will affect the mechanical properties of the hydrogel. This may be because polyvinyl alcohol and chitosan are combined by freeze-thaw first and then irradiation, and strong cross-linking occurs between each other during the gelation process, so that the formed hydrogel has good mechanical properties.

[0093] 2. Cytotoxicity Test

[0094] The dressings obtained in Examples 1-3 and Comparative Examples 1-4 were placed in sterile DMEM culture medium at an extraction ratio of 0.2 g / mL, incubated at 37°C for 24 hours, and the dressings were taken out to obtain the extract. L929 fibroblasts (purchased from Shanghai Bohu Biotechnology Co., Ltd.) were cultured and configured into a 1×105 / ml cell suspension, which was inoculated into a 96-well plate, 100 μL of cell suspension was added to each well, and the 96-well plate was placed in a cell culture incubator at 37°C and 5% CO2 for 24 hours to allow the cells to adhere to the wall, the culture medium was discarded, and the extracts obtained in Examples 1-3 and Comparative Examples 1-4 were added, 100 μL per well, and an equal amount of DMEM culture medium was added as a control, and the 96-well plate was returned to the cell culture incubator for further culturing for 24 hours. 20 μL of MTT reagent (5 mg / mL) was added and the culture was continued for 4 hours. The culture medium was discarded, 100 μL of DMSO was added, and the crystals were dissolved by shaking for 10 minutes. The absorbance value was read at a wavelength of 490 nm using an ELISA reader. The cell viability was calculated, and the cell viability of the extracts obtained in Examples 1-3 and Comparative Examples 1-4 was calculated based on the absorbance values. The results are shown in Table 2, and the calculation formula is as follows: Cell viability (%) = (absorbance value of the experimental group / absorbance value of the control group) × 100. When the cell viability is ≥ 90%; 70-90%; 50-70%; 30-50%; <30%, the corresponding cytotoxicity ratings are 0 (no cytotoxicity); 1 (no cytotoxicity); 2 (mild cytotoxicity); 3 (moderate cytotoxicity); 4 (severe cytotoxicity).

[0095] Table 2

[0096] Group Cell survival rate (%) Example 1 109.5±2.1 Example 2 101.5±2.1 Example 3 104.5±2.1 Comparative Example 1 93.6±2.3 Comparative Example 2 107.2±3.5 Comparative Example 3 102.4±2.7 Comparative Example 4 101.1±1.8

[0097] As shown in Table 2, the cell survival rates of Examples 1-3 and Comparative Examples 1-4 are all greater than 90%, indicating that the materials are non-cytotoxic. At the same time, the cell survival rates of Examples 1-3 are all greater than 100%, indicating that the dressings can promote cell growth to a certain extent.

[0098] 3. Antibacterial properties of dressings

[0099] During the wound healing process, infection of the wound by bacteria and other microorganisms is a common cause of delayed wound healing, among which S. aureus and E. coli are common pathogens of wound infection.

[0100] 3.1 Escherichia coli (CICC 10899) and Staphylococcus aureus (CICC 21600) were used to test the antibacterial activity of the hydrogel dressing. Staphylococcus aureus (S. aureus, strain number CICC 21600) and Escherichia coli (E. coli, strain number CICC 10899) were purchased from the China Industrial Microbiological Culture Collection Administration Center.

[0101] 3.2 Grouping: 12 groups were set, including 1 blank control group and 11 experimental groups, wherein the experimental groups were the hydrogels obtained in Example 3, Comparative Examples 1-2, and Comparative Examples 5-12;

[0102] 3.3 Test method: The antibacterial properties of the hydrogel against S. aureus and E. coli were determined using the inhibition zone method. The hydrogel was cut into a 10 mm diameter cylinder and sterilized with UV light. 7 The cells were cultured on a solid medium (nutrient broth agar for both S. aureus and E. coli, with the formula: 5.0 g peptone, 3.0 g beef extract powder, 5.0 g NaCl, 15.0 g agar, 1000.0 ml distilled water, pH 7.0) at 37 °C for 24 h in a water-insulated electric constant temperature incubator, and the inhibition zone was observed. The results of the inhibition zone diameters of S. aureus and E. coli by hydrogel dressing are shown in Tables 3 and Figure 1 As shown:

[0103] Table 3

[0104]

[0105] From Table 3 and Figure 1 It can be seen that compared with Comparative Examples 1-2 and Comparative Examples 5-12, Example 3 has a significant inhibitory effect on Escherichia coli and Staphylococcus aureus, indicating that chitosan and the traditional Chinese medicine fermentation extract work synergistically and have a significant antibacterial effect on Escherichia coli and Staphylococcus aureus, which shows that the dressing of the present invention has a significant antibacterial effect.

[0106] IV. In vitro coagulation performance: Proving the coagulation efficiency of hydrogel dressings

[0107] The hydrogel dressing samples obtained in Example 3, Comparative Examples 1-2, and Comparative Examples 5-12 were cut into 1cm×1cm×0.3cm, and the 4 layers of gauze were cut into 1cm×1cm in size, respectively placed in a 24-well plate, placed in a CO2 incubator, and preheated at 37°C for 5 minutes. Take out the well plate, put 50μL of anticoagulated rabbit whole blood and drop it on the sample surface, and then add 10μL of 0.25mol / L CaCl2 solution. Subsequently, the well plate was incubated at 37°C for 3 minutes, and 3mL of deionized water was added to the well with the sample, and incubated in a 37°C constant temperature incubator for 10min. Take 1mL of supernatant from each well and centrifuge at 5000rpm for 5min. Collect the supernatant by centrifugation, take 100μL and use an enzyme marker to measure the absorbance at 540nm. 50μL of anticoagulated whole blood + 10μL of 0.1MCaCl2 solution + 3mL of deionized water was used as the blank control group, and 5 replicates were set for each sample. The blood coagulation index (BCI) of the hydrogel dressing can be calculated by the following formula:

[0108] BCI / %=As / A0×100

[0109] Among them, As is the absorbance of the supernatant of the sample group, and A0 is the absorbance of the supernatant of the blank control group. Obviously, BCI increases with the increase of the absorbance value of the hemoglobin solution. The higher the BCI, the weaker the blood coagulation.

[0110] BCI is an important indicator to measure the hemostatic potential of a material. The blood coagulation index (BCI) method was used to evaluate the in vitro hemostatic performance of the hydrogel dressing, and cotton gauze (CG) was used as the negative control group. The results are shown in Table 4 and Figure 2 A lower BCI corresponds to a smaller percentage of uncoagulated blood, thus indicating that the hemostatic material has a higher hemostatic performance.

[0111] Table 4

[0112]

[0113] From Table 4 and Figure 2 It can be seen that the BCI of Example 3 is significantly lower than that of the other groups of dressings, which confirms that the hydrogel dressing of Example 3 has significant hemostatic performance and has better potential for aggregating red blood cells, indicating that the synergistic effect of the Chinese medicine fermentation extract and chitosan has the effect of enhancing the hemostatic function of the dressing. At the same time, the Chinese medicine fermentation extract obtained by combining cloves, astragalus, and mulberry bark with specific strains of Lactobacillus acidobacillus CICC 6082 and Bifidobacterium longum CICC 6191 has a significant enhancement of the hemostatic function of the dressing.

[0114] 5. Effect of dressing on skin wound repair in mice

[0115] Sixty SPF-grade ICR mice, male, 18-22 g, purchased from Nanjing Junke Biotechnology Co., Ltd. were used. Before the experiment, the mice were fed with clean water and standard feed for one week.

[0116] The mice were randomly divided into 12 groups, with 5 mice in each group, of which 1 group was a model group and 11 groups were experimental groups; both the model group and the experimental group used modeling rats, the model group was not treated, and the 11 experimental groups respectively used the hydrogels obtained in Example 3, Comparative Examples 1-2, and Comparative Examples 5-12.

[0117] Mouse skin wound model: The mice were fasted for 12 hours in advance, and 5% chloral hydrate was injected at a dose of 0.1 mL / 10 g to anesthetize the mice. After the mice entered the anesthetized state, the mice were fixed, the hair on the back of the mice was removed, and the skin was wiped and disinfected with a cotton ball of 0.9% saline solution. A circular skin tissue with a diameter of 1 cm was cut off in the depilated area, and the depth was to the fascia layer to establish a mouse skin wound model. The wounds of the mice in the model group were smeared with 0.9% saline solution every day, and the corresponding dressings were changed every day for the wounds of the mice in the embodiment group and the comparative group. The dressings were changed once a day, and the mice were fed freely for 14 consecutive days.

[0118] During the skin wound treatment process, the appearance of the wound was observed with the naked eye and photographed on the 0th day, 3rd day, 7th day, 10th day and 14th day. The wound healing rates on the 3rd day, 7th day, 10th day and 14th day were calculated respectively, as shown in Table 5. The wound healing effect was expressed by the wound healing rate, which was (initial wound area - wound area on the nth day) / initial wound area × 100%, where n = 3, 7, 10, 14.

[0119] Table 5

[0120]

[0121] As shown in Table 5, compared with the model group, the wound healing rate was significantly increased after using the chitosan dressings of Example 3, Comparative Examples 1-2, and Comparative Examples 5-12; compared with the Comparative Example 1-2 group, the wound healing rate was significantly increased after using the dressing of Example 3, because the addition of chitosan reduces the growth of bacteria on the wound surface, which is conducive to the formation of scabs, and the addition of the Chinese medicine fermentation extract in the hydrogel promotes the formation of granulation, which is more beneficial to the healing of wound wounds. Compared with the Comparative Example 5-12 group, the wound healing rate was significantly increased after using the dressing of Example 3, indicating that the Chinese medicine fermentation extract obtained by combining cloves, astragalus, and mulberry bark with specific strains of Lactobacillus acidobacillus CICC 6082 and Bifidobacterium longum CICC 6191 has a significant enhancement of the wound repair function of the dressing.

[0122] The above-mentioned embodiments only express the specific implementation of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. A method for preparing a commonly used bactericidal hemostatic dressing for anesthesia, characterized in that: The following steps are involved: S1. Preparation of bacterial solution: the concentration is 3.6-4.5×10 8 cfu / mL of Lactobacillus acidophilus solution and a concentration of 1.2-2.0×10 8 cfu / mL of Bifidobacterium longum bacterial solution; S2, preparation of fermented product: clove, astragalus, mulberry bark, wash, dry and crush into 80-100 mesh particles, first weigh 18-22 parts of clove, 18-22 parts of astragalus, 3-7 parts of mulberry bark by weight, mix well to obtain a mixture, then add 1-2 times the amount of water of the mixture, mix well to obtain a suspension, add Lactobacillus acidophilus liquid and Bifidobacterium longum to the suspension at an inoculum amount of 3-5%, respectively, and ferment at 37° C. and anaerobic for 12-24h to obtain a fermented product; S3, preparation of Chinese medicine fermentation extract: take the fermented product in step S2, add 6 to 8 times the amount of 70% ethanol for 30-60 min reflux extraction, extract 3 times, discard the residue, combine the filtrate, recover the alcohol, and concentrate until there is no alcohol taste to obtain the Chinese medicine fermentation extract; S4, preparing 1% by mass chitosan solution and 10% by mass polyvinyl alcohol solution; S5, mixing the Chinese medicine fermentation extract obtained in step S3, the chitosan solution obtained in step S4, and the polyvinyl alcohol solution obtained in step S4 in proportion, and stirring for 1 hour in an oil bath at 85° C. to obtain a dressing solution; S6. Defoam the dressing solution and transfer it to a mold, perform freeze-thaw treatment repeatedly, and then irradiate it to obtain a hydrogel dressing, i.e., a sterilizing and hemostatic dressing.

2. A commonly used bactericidal hemostatic dressing for anesthesia according to claim 1, characterized in that: In step S1, Lactobacillus acidophilus is selected from Lactobacillus acidophilus CICC 6082, and Bifidobacterium longum is selected from Bifidobacterium longum CICC 6191.

3. A commonly used bactericidal hemostatic dressing for anesthesia according to claim 1, characterized in that: In step S2, 20 parts of cloves, 20 parts of astragalus, and 5 parts of white mulberry bark are weighed by weight.

4. The commonly used bactericidal hemostatic dressing for anesthesia according to claim 1, characterized in that: In step S4, chitosan powder is dissolved in 1% acetic acid to prepare a 1% by mass chitosan solution.

5. The commonly used bactericidal hemostatic dressing for anesthesia according to claim 1, characterized in that: In step S4, 5 g of polyvinyl alcohol is soaked in deionized water at room temperature for 20 min, and then fully dissolved in a 95° C. oil bath for 1.5-2 h until transparent, to obtain a polyvinyl alcohol solution with a mass fraction of 10%.

6. The commonly used bactericidal hemostatic dressing for anesthesia according to claim 1, characterized in that: In step S5, the amount of chitosan added is 4-8% of the amount of polyvinyl alcohol.

7. A commonly used bactericidal hemostatic dressing for anesthesia according to claim 6, characterized in that: In step S5, the amount of the Chinese medicinal fermentation extract added is 0.05-0.1% of the amount of polyvinyl alcohol.

8. The commonly used bactericidal hemostatic dressing for anesthesia according to claim 1, characterized in that: In step S6, the repeated freezing and thawing method is: freezing at -30 to -20°C for 18 to 24 hours, thawing at 20 to 30°C for 2 to 6 hours, and repeating 3 to 7 times.

9. The commonly used bactericidal hemostatic dressing for anesthesia according to claim 1, characterized in that: In step S6, the irradiation is carried out by 60 Coγ-ray irradiation has an irradiation dose of 10 to 80 kGy and an irradiation time of 1 to 10 minutes.

10. The bactericidal hemostatic dressing commonly used for anesthesia prepared by the method for preparing the bactericidal hemostatic dressing commonly used for anesthesia according to any one of claims 1 to 9.

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