Wound surface nursing dressing after gynecological operation and preparation method thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a gynecological postoperative wound nursing dressing and a preparation method thereof. The wound surface nursing dressing after the gynecologic operation is prepared from the following components in percentage by mass: 0.002 to 0.004 percent of stem cell factor freeze-dried powder, 0.005 to 0.01 percent of phosphoethanolamine, 0.05 to 0.1 percent of cecropin antibacterial peptide, 0.03 to 0.08 percent of aralia elata acid, 2 to 6 percent of carboxymethyl chitosan, 0.1 to 0.5 percent of polyethylene glycol, 0.5 to 1.5 percent of glycerol and the balance of water. The wound dressing not only can inhibit the growth of harmful bacteria, but also does not influence the growth of beneficial bacteria such as lactobacillus, is beneficial to the stabilization and regulation of genital tract micro-ecology, creates a healthier and more stable environment for wound healing, plays a role in the wound repairing process, accelerates wound healing and shortens the treatment time.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and particularly relates to a wound care dressing for gynecological surgery and a preparation method thereof. Background Art

[0002] Gynecological surgeries mostly involve the female reproductive system, which communicates with the outside world, making the postoperative wounds in this area more vulnerable to the invasion of pathogens such as bacteria and viruses. In particular, those surgeries performed through the vagina or exposed to the vaginal environment are classified as "clean - contaminated" surgeries, and their infection risks are particularly prominent. Therefore, for the wound care after gynecological surgery, not only should the wound healing speed be concerned, but more attention should be paid to the regulation of the wound microenvironment to reduce the infection risk and promote the rapid recovery of patients.

[0003] However, for postoperative gynecological wounds, such as perineal laceration wounds during childbirth, their particularity makes it difficult to apply traditional wound care products such as band - aids or gauzes. At the same time, some existing liquid dressings, although performing well in antibacterial aspects, are likely to cause the pH imbalance of the vaginal environment, over - remove beneficial bacteria, disrupt the microbial ecological balance in the vagina, and even cause more serious infections. In view of this, it is particularly important to develop a wound care dressing for gynecological surgery that can ensure the healing efficiency and effectively regulate the wound microenvironment. Summary of the Invention

[0004] In order to overcome the deficiencies of the prior art, one of the purposes of the present invention is to provide a wound care dressing for gynecological surgery. This dressing can inhibit the growth of harmful bacteria and does not affect the growth of beneficial bacteria, providing a healthy and stable environment for the wound, helping to shorten the healing time and accelerate the wound healing.

[0005] Another purpose of the present invention is to provide a preparation method for the wound care dressing for gynecological surgery, which is simple and efficient and is conducive to production and preparation.

[0006] The purpose one of the present invention is realized by adopting the following technical scheme:

[0007] A wound care dressing for gynecological surgery, calculated by mass percentage, the wound care dressing for gynecological surgery includes: stem cell factor freeze - dried powder 0.002 - 0.004%, phosphoethanolamine 0.005 - 0.01%, cecropin antimicrobial peptide 0.05 - 0.1%, aralia continentalis acid 0.03 - 0.08%, carboxymethyl chitosan 2 - 6%, polyethylene glycol 0.1 - 0.5%, glycerol 0.5 - 1.5%, and the balance is water.

[0008] Furthermore, the wound care dressing after gynecological surgery comprises: 0.003% of stem cell factor lyophilized powder, 0.0075% of phosphoethanolamine, 0.08% of cecropin antimicrobial peptide, 0.05% of aralia continentalis kitag. acid, 4% of carboxymethyl chitosan, 0.3% of polyethylene glycol, 1% of glycerol, and the balance being water.

[0009] Furthermore, the preparation method of the stem cell factor lyophilized powder is as follows:

[0010] S1. Obtaining primary mesenchymal stem cells;

[0011] S2. Subculturing the primary mesenchymal stem cells in step S1, and collecting the subcultured mesenchymal stem cells;

[0012] S3. Culturing the subcultured mesenchymal stem cells under hypoxic conditions until the cell growth density reaches 80-90%, centrifuging, collecting the cell supernatant, filtering the supernatant to collect the filtrate, subjecting the filtrate to dialysis and filtration to obtain a stem cell factor concentrate, adding trehalose, and freeze-drying to obtain the product.

[0013] Furthermore, the mesenchymal stem cells are any one of umbilical cord mesenchymal stem cells, adipose mesenchymal stem cells, and dental pulp mesenchymal stem cells.

[0014] Furthermore, in step S3, the hypoxic conditions are: 5-8% O2, 37°C.

[0015] Furthermore, the specific operation process of the freeze-drying is: first cooling the stem cell factor concentrate at a rate of 5-10°C / h to -25 to -30°C, and then cooling at a rate of 1-5°C / h to -45 to -55°C / h, and holding for 10-14 h.

[0016] The second object of the present invention is achieved by the following technical solution:

[0017] A preparation method of a wound care dressing after gynecological surgery, comprising the following steps:

[0018] (1) Adding carboxymethyl chitosan, polyethylene glycol, and glycerol to one-third of the total amount of water, stirring and mixing evenly to obtain solution A;

[0019] (2) Stirring and mixing evenly the stem cell factor lyophilized powder, phosphoethanolamine, and one-third of the total amount of water to obtain solution B;

[0020] (3) Adding cecropin antimicrobial peptide and aralia continentalis kitag. acid to the remaining water, stirring and mixing evenly to obtain solution C;

[0021] (4) Adding solution B and solution C to solution A in step (1), stirring and mixing evenly, and irradiating and sterilizing to obtain a wound care dressing after gynecological surgery.

[0022] Further, the stirring speed in both step (1) and step (2) is 100 - 200 rpm, and the time is 1 - 2 h for both.

[0023] Further, the stirring speed in both step (3) and step (4) is 50 - 100 rpm, and the time is 1.5 - 3 h for both.

[0024] Further, the dose of the irradiation sterilization is 15 - 20 kGy.

[0025] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0026] 1. The present invention provides a wound care dressing for gynecological surgery. Phosphoethanolamine is added to this dressing. This component can effectively maintain the stability and activity of stem cell growth factor in the dressing, and further promote the role of stem cell growth factor in the wound repair process, accelerating wound healing. At the same time, the dressing of the present invention also adds the combination of araloside A and cecropin antimicrobial peptide. It not only enhances the antibacterial effect of the dressing against harmful bacteria, but also does not affect the growth of beneficial bacteria such as Lactobacillus, contributing to the stability and regulation of the reproductive tract microecology, creating a more healthy and stable environment for wound healing.

[0027] 2. The present invention provides a preparation method of a wound care dressing for gynecological surgery. This preparation method is easy to operate and has high production efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is the morphology diagram of the umbilical cord mesenchymal stem cells of the present invention;

[0029] Figure 2 It is the result diagram of the average healing time when the wound dressing of the present invention is used for wound healing. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0030] Next, in combination with the drawings and the specific embodiments, the present invention will be further described. It should be noted that, on the premise of no conflict, any combination can be formed between the following described embodiments or technical features. The specific conditions not specified in the embodiments are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used, unless otherwise specified, are all conventional products obtained through commercial channels.

[0031] Preparation Example 1

[0032] This preparation example provides a preparation method of a freeze-dried powder of umbilical cord mesenchymal stem cell factor, and the steps are as follows:

[0033] S1. Obtaining umbilical cord mesenchymal stem cells: The umbilical cord tissue was rinsed twice with a solution containing 1% penicillin-streptomycin, and then cut into tissue blocks of 2 mm 3 in size with surgical scissors. It was resuspended in DMEM / F12 medium containing 10% fetal bovine serum, then placed in a sterile culture dish and cultured in an incubator at 37°C and 5% CO2. When the cell confluence reached 90%, the medium was aspirated, gently washed twice with PBS buffer, 0.25% trypsin solution was added, and digestion was carried out in a 37°C incubator for 3 min to collect primary umbilical cord mesenchymal stem cells.

[0034] S2. Subculture of cells: Sterilized PBS buffer was added to the collected primary umbilical cord mesenchymal stem cells to make a cell suspension. The cell suspension was centrifuged at 2000 rpm for 10 min, the supernatant was discarded, and the cells were collected. Then, subculture was carried out at a ratio of 1:2, and P5 generation cells were collected. Their growth morphology was observed under a microscope, and the results are as Figure 1 shown.

[0035] S3. Preparation of freeze-dried stem cell factor: P5 generation mesenchymal stem cells were inoculated into the medium (DMEM / F12, 8% fetal bovine serum, 7 μg / mL L-glutamine, 95 U / mL penicillin, 95 μg / mL streptomycin, 7 μg / mL vitamin C), and then cultured under hypoxic conditions at 37°C and 5% O2. When the cell growth density reached 90%, it was centrifuged at 1500 rpm for 10 min to collect the cell supernatant. The supernatant was filtered through a 0.22 μm filter membrane to collect the filtrate. The filtrate was dialyzed through a 50 KD ultrafiltration membrane to collect the dialysate, and then filtered through a 2 KD ultrafiltration membrane again to obtain the concentrated umbilical cord mesenchymal stem cell factor; Trehalose was added to the concentrate, and the concentration of trehalose in the concentrate was 15 mM. It was cooled to -28°C at a rate of 8°C / h, and then cooled to -50°C at a rate of 3°C / h and incubated for 12 h for freeze-drying to obtain the product.

[0036] Example 1

[0037] A wound care dressing for gynecological surgery, by mass percentage, the wound care dressing for gynecological surgery includes: 0.003% of the freeze-dried umbilical cord mesenchymal stem cell factor prepared in Preparation Example 1, 0.0075% of phosphoethanolamine, 0.08% of cecropin antimicrobial peptide, 0.05% of aralia elata acid, 4% of carboxymethyl chitosan, 0.3% of polyethylene glycol, 1% of glycerol, and the balance is water.

[0038] A preparation method of a wound care dressing for gynecological surgery includes the following steps:

[0039] (1) Add carboxymethyl chitosan, polyethylene glycol, and glycerol to one-third of the total amount of water, stir at a speed of 150 rpm for 1.5 h, and mix evenly to obtain Solution A;

[0040] (2) Add the freeze-dried umbilical cord mesenchymal stem cell factor, phosphoethanolamine, and one-third of the total amount of water, stir at a speed of 100 rpm for 2 h, and mix evenly to obtain Solution B;

[0041] (3) Add cecropin antibacterial peptide and aralia continentalis kitag. acid to the remaining water, stir at a speed of 50 rpm for 3 h, and mix evenly to obtain Solution C;

[0042] (4) Add Solution B and Solution C to Solution A in step (1), stir at a speed of 75 rpm for 2 h, mix evenly, and obtain a wound care dressing for gynecological surgery after cobalt-60 irradiation sterilization (the dose of irradiation sterilization is 18 kGy).

[0043] Example 2

[0044] A wound care dressing for gynecological surgery, calculated by mass percentage, the wound care dressing for gynecological surgery includes: 0.002% of the freeze-dried umbilical cord mesenchymal stem cell factor prepared in Preparation Example 1, 0.005% of phosphoethanolamine, 0.05% of cecropin antibacterial peptide, 0.03% of aralia continentalis kitag. acid, 2% of carboxymethyl chitosan, 0.1% of polyethylene glycol, 0.5% of glycerol, and the balance is water.

[0045] A preparation method of a wound care dressing for gynecological surgery, including the following steps:

[0046] (1) Add carboxymethyl chitosan, polyethylene glycol, and glycerol to one-third of the total amount of water, stir at a speed of 100 rpm for 2 h, and mix evenly to obtain Solution A;

[0047] (2) Add the freeze-dried umbilical cord mesenchymal stem cell factor, phosphoethanolamine, and one-third of the total amount of water, stir at a speed of 200 rpm for 1 h, and mix evenly to obtain Solution B;

[0048] (3) Add cecropin antibacterial peptide and aralia continentalis kitag. acid to the remaining water, stir at a speed of 100 rpm for 1.5 h, and mix evenly to obtain Solution C;

[0049] (4) Add Solution B and Solution C to Solution A in step (1), stir at a speed of 50 rpm for 3 h, mix evenly, and obtain a wound care dressing for gynecological surgery after cobalt-60 irradiation sterilization (the dose of irradiation sterilization is 15 kGy).

[0050] Example 3

[0051] A wound care dressing for gynecological surgery, calculated by mass percentage, the wound care dressing for gynecological surgery includes: freeze-dried umbilical cord mesenchymal stem cell factor prepared in Preparation Example 1, 0.004%, phosphoethanolamine 0.01%, cecropin antimicrobial peptide 0.1%, aralia elata acid 0.08%, carboxymethyl chitosan 6%, polyethylene glycol 0.5%, glycerol 1.5%, and the balance is water.

[0052] A preparation method of a wound care dressing for gynecological surgery, comprising the following steps:

[0053] (1) Add carboxymethyl chitosan, polyethylene glycol, and glycerol to one-third of the total amount of water, stir at a speed of 200 rpm for 1 h, mix evenly to obtain Solution A;

[0054] (2) Stir the freeze-dried umbilical cord mesenchymal stem cell factor, phosphoethanolamine and one-third of the total amount of water at a speed of 150 rpm for 1.5 h, mix evenly to obtain Solution B;

[0055] (3) Add cecropin antimicrobial peptide and aralia elata acid to the remaining water, stir at a speed of 75 rpm for 2 h, mix evenly to obtain Solution C;

[0056] (4) Add Solution B and Solution C to Solution A in step (1), stir at a speed of 100 rpm for 3 h, mix evenly, and obtain a wound care dressing for gynecological surgery after cobalt-60 irradiation sterilization (the dose of irradiation sterilization is 20 kGy).

[0057] Comparative Example 1

[0058] This Comparative Example 1 is basically the same as Example 1, the difference is that phosphoethanolamine is omitted.

[0059] Comparative Example 2

[0060] This Comparative Example 2 is basically the same as Example 1, the difference is that cecropin antimicrobial peptide is omitted, and the dosage of aralia elata acid is adjusted to 0.13%.

[0061] Comparative Example 3

[0062] This Comparative Example 3 is basically the same as Example 1, the difference is that aralia elata acid is omitted, and the dosage of cecropin antimicrobial peptide is adjusted to 0.13%.

[0063] Test Example 1

[0064] Stability test of stem cell factor: Use an ELISA kit to detect the contents of fibroblast growth factor (FGF), epidermal growth factor (EGF), and vascular endothelial growth factor (VEGF) in the wound care dressings prepared in Examples 1 - 3 and Comparative Examples 1 - 3. After storing the dressings in each group at 4°C for 3 months, detect the above factors again. The test results are shown in Table 1.

[0065] Table 1

[0066]

[0067] From the detection results in Table 1, it can be seen that after storing for 3 months, the cytokine contents in the gynecological postoperative wound care dressings of Examples 1 - 3 changed little. However, after omitting phosphoethanolamine in Comparative Example 1, the cytokine contents decreased significantly, indicating that the wound care dressings of the present invention can maintain the stability of cytokines by adding phosphoethanolamine, avoid damage to cytokines during storage, and thus prolong the activity of cytokines.

[0068] Test Example 2

[0069] To test the antibacterial effects of the wound dressings of Examples 1 - 3 and Comparative Examples 1 - 3 against Escherichia coli, Staphylococcus aureus, and Lactobacillus, the antibacterial test was carried out by the antibacterial zone method. The specific test method is as follows:

[0070] Prepare bacterial suspensions of Escherichia coli, Staphylococcus aureus, and Lactobacillus with a concentration of 10 8 CFU / mL respectively. Take 100 μL of the above bacterial suspensions and spread them evenly on petri dishes (nutrient agar medium for Escherichia coli and Staphylococcus aureus, MRS medium for Lactobacillus). Place Oxford cups with an inner diameter of 6 mm, an outer diameter of 8 mm, and a height of 10 mm vertically on the culture media of each group, gently press the Oxford cups, and then add 50 μL of the wound dressings described in Examples 1 - 3 and Comparative Examples 1 - 3 into the Oxford cups. Set 3 parallel experiments for each group. Incubate the petri dishes coated with Escherichia coli and Staphylococcus aureus at 37°C for 24 h, and incubate the petri dishes coated with Lactobacillus at 37°C for 48 h. Remove the Oxford cups, observe and measure the size of the antibacterial zone diameter. If the antibacterial zone diameter is greater than 10 mm, it indicates good antibacterial effect. The results are shown in Table 2.

[0071] Table 2

[0072]

[0073] As can be seen from Table 2, the wound dressings of Examples 1-3 of the present invention have good antibacterial effects against Escherichia coli and Staphylococcus aureus, but basically have no inhibitory effect on Lactobacillus. Compared with Example 1, Comparative Example 2 omitted cecropin antimicrobial peptide and adjusted the dosage of araloside A to the sum of the two; Comparative Example 3 omitted cecropin antimicrobial peptide and adjusted the dosage of araloside A to the sum of the two. The inhibitory effects of Comparative Examples 2 and 3 on Escherichia coli and Staphylococcus aureus became worse, indicating that araloside A and cecropin antimicrobial peptide have a synergistic effect and can inhibit the growth of harmful bacteria (such as Escherichia coli and Staphylococcus aureus), but have a very weak inhibitory ability on beneficial bacteria (Lactobacillus).

[0074] In summary, the combination of araloside A and cecropin antimicrobial peptide added in the dressing of the present invention not only enhances the antibacterial effect of the dressing against harmful bacteria, but also does not affect the growth of beneficial bacteria such as Lactobacillus, which helps to stabilize and regulate the reproductive tract microecology and creates a healthier and more stable environment for wound healing.

[0075] Test Example 3

[0076] Ten-week-old SD rats were selected as experimental animals and randomly divided into 6 groups, with 10 rats in each group, for a total of 60 rats. They were adaptively fed in the laboratory for 7 days with free access to water and food. The rats were anesthetized with an anesthetic (Zoletil, dose 50 mg / kg), the back hair was shaved, and then disinfected with 75% ethanol and iodophor. Then, a circular wound with a diameter of 10 mm was created on the back of the rats, and the wound dressings of Examples 1-3 and Comparative Examples 1-3 were used to cover the wound. The dressings were changed once in the morning and once in the evening every day, and the average healing time (unit: d) of each group was counted. The results are as Figure 2 shown.

[0077] As Figure 2 can be seen, the wound healing time of Examples 1-3 of the present invention is shorter, that of Comparative Examples 2 and 3 is slightly reduced, and that of Comparative Example 1 is the worst. This result is attributed to the phosphoethanolamine added in Examples 1-3, which can effectively maintain the stability and activity of stem cell growth factor in the dressing, and then promote the role of umbilical cord stem cell growth factor in the wound repair process and accelerate wound healing. At the same time, the dressing of the present invention also adds a combination of araloside A and cecropin antimicrobial peptide, which not only enhances the antibacterial effect of the dressing against harmful bacteria, but also does not affect the growth of beneficial bacteria such as Lactobacillus, which helps to stabilize and regulate the reproductive tract microecology and creates a healthier and more stable environment for wound healing.

[0078] The above embodiments are only the preferred embodiments of the present invention and cannot be used to limit the scope of protection of the present invention. Any non-substantive changes and substitutions made by those skilled in the art based on the present invention belong to the scope of protection required by the present invention.

Claims

1. A wound care dressing for gynecological surgery, characterized in that, By mass percentage, the wound care dressing after gynecological surgery comprises: 0.002 - 0.004% of stem cell factor freeze-dried powder, 0.005 - 0.01% of phosphoethanolamine, 0.05 - 0.1% of cecropin antimicrobial peptide, 0.03 - 0.08% of aralia elata acid, 2 - 6% of carboxymethyl chitosan, 0.1 - 0.5% of polyethylene glycol, 0.5 - 1.5% of glycerol, and the balance is water.

2. The wound care dressing for gynecological surgery according to claim 1, wherein By mass percentage, the wound care dressing after gynecological surgery comprises: 0.003% of stem cell factor freeze-dried powder, 0.0075% of phosphoethanolamine, 0.08% of cecropin antimicrobial peptide, 0.05% of aralia elata acid, 4% of carboxymethyl chitosan, 0.3% of polyethylene glycol, 1% of glycerol, and the balance is water.

3. The wound care dressing for gynecological surgery according to claim 1 or 2, characterized in that, The preparation method of the stem cell factor freeze-dried powder is as follows: S1. Obtaining primary mesenchymal stem cells; S2. Subculturing the primary mesenchymal stem cells in step S1, and collecting the subcultured mesenchymal stem cells; S3. Culturing the subcultured mesenchymal stem cells under hypoxic conditions until the cell growth density reaches 80 - 90%, centrifuging, collecting the cell supernatant, filtering the supernatant and collecting the filtrate, subjecting the filtrate to dialysis and filtration to obtain a stem cell factor concentrate, adding trehalose, and freeze-drying to obtain the product.

4. The wound care dressing for gynecological surgery according to claim 3, characterized in that, The mesenchymal stem cells are any one of umbilical cord mesenchymal stem cells, adipose mesenchymal stem cells, and dental pulp mesenchymal stem cells.

5. The wound care dressing for gynecological surgery according to claim 3, characterized in that, In step S3, the hypoxic conditions are: 5 - 8% O2, 37°C.

6. The wound care dressing for gynecological surgery according to claim 3, characterized in that, The specific operation process of the freeze-drying is: first cooling the stem cell factor concentrate at a rate of 5 - 10°C / h to -25 - -30°C, and then cooling at a rate of 1 - 5°C / h to -45 - -55°C / h, and holding for 10 - 14 h.

7. The preparation method of the wound care dressing after gynecological surgery according to claim 1, wherein, It includes the following steps: (1) Adding carboxymethyl chitosan, polyethylene glycol, and glycerol to one-third of the total amount of water, stirring and mixing evenly to obtain solution A; (2) Stirring and mixing evenly the stem cell factor freeze-dried powder, phosphoethanolamine, and one-third of the total amount of water to obtain solution B; (3) Adding cecropin antimicrobial peptide and aralia elata acid to the remaining water, stirring and mixing evenly to obtain solution C; (4) Adding solution B and solution C to solution A in step (1), stirring and mixing evenly, and obtaining the wound care dressing after gynecological surgery after irradiation sterilization.

8. The preparation method of the wound care dressing after gynecological surgery according to claim 7, characterized in that, In steps (1) and (2), the stirring speed is 100 - 200 rpm, and the time is 1 - 2 h.

9. The preparation method of the wound care dressing after gynecological surgery according to claim 7, wherein, In steps (3) and (4), the stirring speed is 50 - 100 rpm, and the time is 1.5 - 3 h.

10. The preparation method of the wound care dressing after gynecological surgery according to claim 7, characterized in that, The dose of the irradiation sterilization is 15 - 20 kGy.

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