CN112121223A - Sericin medical dressing and production method thereof - Google Patents
Sericin medical dressing and production method thereof Download PDFInfo
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- CN112121223A CN112121223A CN202011108152.5A CN202011108152A CN112121223A CN 112121223 A CN112121223 A CN 112121223A CN 202011108152 A CN202011108152 A CN 202011108152A CN 112121223 A CN112121223 A CN 112121223A
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- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/28—Polysaccharides or their derivatives
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
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- A61L15/40—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing ingredients of undetermined constitution or reaction products thereof, e.g. plant or animal extracts
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Abstract
The invention discloses a medical dressing of sericin and a production method thereof, wherein the scheme comprises the following steps: 1) dissolving silk fibroin powder into ionized water, and 2) modified soybean protein isolate for later use; 3) mixing the paste for later use; 4) mixing the solution; 5) dissolving the modified soybean protein isolate in the step 2) into a phosphate buffer solution with the pH value of 7.4, adding the sericin solution into the solution, adding the mixed paste while stirring, adding a cross-linking agent into the mixed solution, putting the mixed solution into a refrigerator, reducing the temperature to 2-5 ℃, adding growth factor freeze-dried powder, mixing medical alginate fibers into the mixture, heating the mixture to 40-55 ℃, stirring the mixture, adding algal polysaccharide into the mixture after reducing the temperature, repeatedly freezing the mixture for multiple times and then thawing the mixture to obtain the dressing.
Description
Technical Field
The invention relates to the technical field of dry preparations, in particular to a production method of a medical dressing of sericin.
Background
With the increasing standard of living, wound treatment and care are receiving attention. For some wounds which are difficult to heal, the common bandaging treatment is often difficult to heal, and special means are needed. The wounds difficult to heal generally comprise chronic difficult wounds such as pressure sores, diabetic feet, arteriovenous ulcers, malignant tumors and the like, surgical wounds such as traffic wounds, wound acute wounds, burn and plastic wounds and the like, and surgical wounds such as cardiac stent operations, fat liquefaction, abscess incision and drainage and the like. The wound is difficult to heal, and some systemic diseases cause the damage of immune cells of a human body; the old and the infirm, malnutrition and the like cause insufficient cell regeneration capacity; wound infection, inflammatory reaction, etc. The root causes of this are poor collagen synthesis and impaired fibroblast metabolism. Some clinicians are also dedicated to the treatment of wounds which are difficult to heal, and the usage amount of wound dressings in some hospitals can reach more than 70 ten thousand times per year, so that the demand is extremely high.
The wound dressings which are clinically used at present mainly comprise traditional dressings and novel wet dressings. The traditional dressings, namely gauze, iodoform gauze and the like, are used for dressing change treatment by taking the traditional principle of keeping the wound surface dry as a guide principle, and have no healing promotion effect. The novel wet dressing includes a covering type outer dressing represented by a transparent film type dressing; inner layer-packed dressings represented by alginate dressings and wound gels; the antibacterial dressing represented by silver ion dressing and the like can create a moist environment favorable for cell growth by taking moist healing as a principle, and the dressing closed or semi-closed wound can keep the wound at a constant temperature, prevent bacteria and external granular foreign matters from invading, reduce the infection chance of the wound, can not generate mechanical damage when the dressing is replaced, and promote the wound to heal. However, the characteristics and permeability of the drug for absorbing the exudate in a humid environment are poor, and finally, many infection situations appear, which becomes a medical problem.
In recent years, third generation dressings, known as the future technology type, have become a hot spot for development. The method is characterized in that: the intelligent automatic wound moistening degree adjusting device has the performance of intelligently and automatically adjusting the wound moistening degree, so that tissues and cells are in the optimal growth environment; the dressing has the components capable of providing nutrition for cell and tissue growth and inhibiting bacteria reproduction. The wound dressing with the vacuum reservoir, which is invented by D, Fenke and the like, which is most representative, can drain wound exudate in time, keep the wound from being infected, and the wound surface material has a nutritional function, is quick in wound healing, but is extremely high in price, and is difficult for patients to bear.
Disclosure of Invention
The invention provides a medical dressing of sericin and a production method thereof.
The scheme of the invention is as follows:
a production method of a medical dressing of sericin comprises the following steps:
1) dissolving silk fibroin powder into ionized water to prepare silk fibroin solution for later use;
2) dissolving soybean protein isolate in phosphate solution with pH of 7.4 at 40-80 ℃, adding vinyltrimethoxysilane and N-hydroxysuccinimide for mixing, adjusting the pH value of a reaction system to 4-7 after stirring, adding dopamine and fluoxetine after stirring and reacting for 30-60 min at 20-50 ℃, maintaining the pH value of the reaction system to 4-7, continuing stirring and reacting for 10-36 h at 20-50 ℃, and finally dialyzing and freeze-drying to obtain modified soybean protein isolate for later use;
3) adding Concha Ostreae powder into Aloe fresh gel, stirring, and sequentially adding docosahexaenoic acid, ascorbic acid, titanium dioxide, oleum Camelliae Japonicae and linoleic acid during stirring to obtain mixed paste;
4) mixing chitosan, xanthan gum, dencichine, quercetin, D-arbutin, carbomer, urea capsule and the medicinal extract, adding appropriate amount of water into the obtained mixture, and performing ultrasonic vibration to obtain mixed solution;
5) dissolving the modified soybean protein isolate in the step 2) in a phosphate buffer solution with the pH value of 7.4, adding the sericin solution, adding the mixed paste while stirring, adding a cross-linking agent, adding the mixed solution, putting the mixed solution into a refrigerator, reducing the temperature to 2-5 ℃, adding the growth factor freeze-dried powder, mixing the medical alginate fibers, heating to 40-55 ℃, stirring, reducing the temperature, adding the algal polysaccharide, repeatedly freezing and thawing to obtain the dressing.
As a preferable technical scheme, sericin solution with the mass volume percentage concentration of 2.0-5.0% is prepared in the step 1).
As a preferred technical scheme, the feeding proportion of the soy protein isolate, the vinyltrimethoxysilane, the N-hydroxysuccinimide, the dopamine and the fluoxetine in the step 2) is 10: 2-3: 1-2: 4-6: 2 to 4.
According to a preferable technical scheme, the feeding proportion of the oyster shell powder, the aloe vera fresh gel, the docosahexaenoic acid, the ascorbic acid, the titanium dioxide, the camellia oil and the linoleic acid in the step 3) is 12-15: 30-40: 20-30: 0.5-1.2: 2-5: 2-3.
As a preferred technical scheme, the feeding proportion of the chitosan, the xanthan gum, the dencichine, the quercetin, the D-arbutin, the carbomer, the urea capsule and the medicine extract in the step 4) is 2-5: 30-43: 1-2: 1-1.5: 0.3-0.8: 3-4: 1-2: 15-20.
According to a preferable technical scheme, the mixed solution with the mass volume percentage concentration of 1.5-3.0% is prepared in the step 4).
The invention also provides a medical dressing of sericin, which comprises the following raw materials in parts by weight:
the preferable technical scheme comprises the following raw materials in parts by weight:
as a preferred technical scheme, the mixed paste comprises oyster shell powder, aloe fresh gel, docosahexaenoic acid, ascorbic acid, titanium dioxide, camellia oil and linoleic acid; the cross-linking agent is one or more of glutaraldehyde, malondialdehyde and genipin; the mixed solution comprises chitosan, xanthan gum, dencichine, quercetin, D-arbutin, carbomer, urea capsule and medicinal extract.
As a preferred technical scheme, the medicine extract comprises pearl powder, a mulberry leaf extract, a common perilla extract, an angelica dahurica extract and a raw liquorice extract.
Due to the adoption of the technical scheme, the medical dressing of sericin comprises the following steps: 1) dissolving silk fibroin powder into ionized water to prepare silk fibroin solution for later use; 2) dissolving soybean protein isolate in phosphate solution with pH of 7.4 at 40-80 ℃, adding vinyltrimethoxysilane and N-hydroxysuccinimide for mixing, adjusting the pH value of a reaction system to 4-7 after stirring, adding dopamine and fluoxetine after stirring and reacting for 30-60 min at 20-50 ℃, maintaining the pH value of the reaction system to 4-7, continuing stirring and reacting for 10-36 h at 20-50 ℃, and finally dialyzing and freeze-drying to obtain modified soybean protein isolate for later use; 3) adding Concha Ostreae powder into Aloe fresh gel, stirring, and sequentially adding docosahexaenoic acid, ascorbic acid, titanium dioxide, oleum Camelliae Japonicae and linoleic acid during stirring to obtain mixed paste; 4) mixing chitosan, xanthan gum, dencichine, quercetin, D-arbutin, carbomer, urea capsule and the medicinal extract, adding appropriate amount of water into the obtained mixture, and performing ultrasonic vibration to obtain mixed solution; 5) dissolving the modified soybean protein isolate in the step 2) in a phosphate buffer solution with the pH value of 7.4, adding the sericin solution, adding the mixed paste while stirring, adding a cross-linking agent, adding the mixed solution, putting the mixed solution into a refrigerator, reducing the temperature to 2-5 ℃, adding the growth factor freeze-dried powder, mixing the medical alginate fibers, heating to 40-55 ℃, stirring, reducing the temperature, adding the algal polysaccharide, repeatedly freezing and thawing to obtain the dressing.
The invention has the advantages that:
the antibacterial whitening cream has the advantages of strong biodegradability and antibacterial property, whitening and scar repairing, no stimulation to skin, no anaphylactic reaction, good affinity with skin, effective absorption by skin, complementary absorption by skin, and long-term storage under the normal-temperature aseptic condition;
the dressing is not easy to adhere to the wound, and the degradation rate, porosity, mechanical property and the like of the dressing are excellent; the medicine has the advantages of quick healing, itching relieving, painless dressing change, shortened nurse working time, bacteriostasis, high cost performance, no pain and less dressing change times, and can be used for tissue repair, ulcer, bedsore and chronic wound;
the sericin protein has the natural structure and activity of sericin, keeps the original ecology characteristic of sericin, and has popularization; the porous structure is good in air permeability; has good mechanical strength and high biocompatibility; the sericin wound dressing has good biocompatibility and cell adhesion, and can support the survival, proliferation and migration of cells for a long time; can be used for supporting cell growth and promoting nutrient exchange by extracellular matrix, and can be used for repairing various tissue injuries and treating diseases.
Detailed Description
In order to make up for the above deficiencies, the present invention provides a sericin medical dressing and a production method thereof to solve the problems in the background art.
A production method of a medical dressing of sericin comprises the following steps:
1) dissolving silk fibroin powder into ionized water to prepare silk fibroin solution for later use;
2) dissolving soybean protein isolate in phosphate solution with pH of 7.4 at 40-80 ℃, adding vinyltrimethoxysilane and N-hydroxysuccinimide for mixing, adjusting the pH value of a reaction system to 4-7 after stirring, adding dopamine and fluoxetine after stirring and reacting for 30-60 min at 20-50 ℃, maintaining the pH value of the reaction system to 4-7, continuing stirring and reacting for 10-36 h at 20-50 ℃, and finally dialyzing and freeze-drying to obtain modified soybean protein isolate for later use;
3) adding Concha Ostreae powder into Aloe fresh gel, stirring, and sequentially adding docosahexaenoic acid, ascorbic acid, titanium dioxide, oleum Camelliae Japonicae and linoleic acid during stirring to obtain mixed paste;
4) mixing chitosan, xanthan gum, dencichine, quercetin, D-arbutin, carbomer, urea capsule and the medicinal extract, adding appropriate amount of water into the obtained mixture, and performing ultrasonic vibration to obtain mixed solution;
5) dissolving the modified soybean protein isolate in the step 2) in a phosphate buffer solution with the pH value of 7.4, adding the sericin solution, adding the mixed paste while stirring, adding a cross-linking agent, adding the mixed solution, putting the mixed solution into a refrigerator, reducing the temperature to 2-5 ℃, adding the growth factor freeze-dried powder, mixing the medical alginate fibers, heating to 40-55 ℃, stirring, reducing the temperature, adding the algal polysaccharide, repeatedly freezing and thawing to obtain the dressing.
The sericin solution with the mass volume percentage concentration of 2.0-5.0% is prepared in the step 1).
The feeding proportion of the soy protein isolate, the vinyl trimethoxy silane, the N-hydroxysuccinimide, the dopamine and the fluoxetine in the step 2) is 10: 2-3: 1-2: 4-6: 2 to 4.
The feeding proportion of the oyster shell powder, the aloe vera fresh gel, the docosahexaenoic acid, the ascorbic acid, the titanium dioxide, the camellia oil and the linoleic acid in the step 3) is 12-15: 30-40: 20-30: 0.5-1.2: 2-5: 2-3.
In the step 4), the feeding proportion of the chitosan, the xanthan gum, the dencichine, the quercetin, the D-arbutin, the carbomer, the urea capsule and the medicine extract is 2-5: 30-43: 1-2: 1-1.5: 0.3-0.8: 3-4: 1-2: 15-20.
And (4) preparing a mixed solution with the mass volume percentage concentration of 1.5-3.0%.
The invention also provides a medical dressing of sericin, which comprises the following raw materials in parts by weight:
comprises the following raw materials in parts by weight:
the mixed paste comprises oyster shell powder, aloe vera fresh gel, docosahexaenoic acid, ascorbic acid, titanium dioxide, camellia oil and linoleic acid; the cross-linking agent is one or more of glutaraldehyde, malondialdehyde and genipin; the mixed solution comprises chitosan, xanthan gum, dencichine, quercetin, D-arbutin, carbomer, urea capsule and medicinal extract.
The medicinal extract comprises pearl powder, mulberry leaf extract, common perilla extract, angelica dahurica extract and raw licorice extract.
In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further described with the specific embodiments.
Example 1:
1) dissolving silk fibroin powder into ionized water to prepare silk fibroin solution for later use;
2) dissolving soybean protein isolate in phosphate solution with pH of 7.4 at 40-80 ℃, adding vinyltrimethoxysilane and N-hydroxysuccinimide for mixing, adjusting the pH value of a reaction system to 4-7 after stirring, adding dopamine and fluoxetine after stirring and reacting for 30-60 min at 20-50 ℃, maintaining the pH value of the reaction system to 4-7, continuing stirring and reacting for 10-36 h at 20-50 ℃, and finally dialyzing and freeze-drying to obtain modified soybean protein isolate for later use;
3) adding Concha Ostreae powder into Aloe fresh gel, stirring, and sequentially adding docosahexaenoic acid, ascorbic acid, titanium dioxide, oleum Camelliae Japonicae and linoleic acid during stirring to obtain mixed paste;
4) mixing chitosan, xanthan gum, dencichine, quercetin, D-arbutin, carbomer, urea capsule and the medicinal extract, adding appropriate amount of water into the obtained mixture, and performing ultrasonic vibration to obtain mixed solution;
5) dissolving 30% by weight of modified soybean protein isolate obtained in the step 2) in a phosphate buffer solution with the pH value of 7.4, adding 10% by weight of sericin solution, adding 8% by weight of mixed paste while stirring, adding 5% by weight of cross-linking agent, adding 25% by weight of mixed solution, refrigerating to reduce the temperature to 2-5 ℃, adding 2% by weight of growth factor freeze-dried powder, mixing with 1% by weight of medical alginate fibers, heating to 40-55 ℃, stirring, reducing the temperature, adding 4% by weight of algal polysaccharide, repeatedly freezing for many times, and thawing to obtain the dressing.
In the step 1), sericin solution with the mass volume percentage concentration of 2.0% is prepared.
The feeding proportion of the soy protein isolate, the vinyl trimethoxy silane, the N-hydroxysuccinimide, the dopamine and the fluoxetine in the step 2) is 10: 2: 1: 4: 2.
the feeding proportion of the oyster shell powder, the aloe vera fresh gel, the docosahexaenoic acid, the ascorbic acid, the titanium dioxide, the camellia oil and the linoleic acid in the step 3) is 12:30:20:0.5:2:2: 2.
In the step 4), the feeding proportion of the chitosan, the xanthan gum, the dencichine, the quercetin, the D-arbutin, the carbomer, the urea capsule and the medicine extract is 5: 43: 2:1.5:0.8: 4:2:20.
And step 4) preparing a mixed solution with the mass volume percentage concentration of 1.5%.
Example 2:
1) dissolving silk fibroin powder into ionized water to prepare silk fibroin solution for later use;
2) dissolving soybean protein isolate in phosphate solution with pH of 7.4 at 40-80 ℃, adding vinyltrimethoxysilane and N-hydroxysuccinimide for mixing, adjusting the pH value of a reaction system to 4-7 after stirring, adding dopamine and fluoxetine after stirring and reacting for 30-60 min at 20-50 ℃, maintaining the pH value of the reaction system to 4-7, continuing stirring and reacting for 10-36 h at 20-50 ℃, and finally dialyzing and freeze-drying to obtain modified soybean protein isolate for later use;
3) adding Concha Ostreae powder into Aloe fresh gel, stirring, and sequentially adding docosahexaenoic acid, ascorbic acid, titanium dioxide, oleum Camelliae Japonicae and linoleic acid during stirring to obtain mixed paste;
4) mixing chitosan, xanthan gum, dencichine, quercetin, D-arbutin, carbomer, urea capsule and the medicinal extract, adding appropriate amount of water into the obtained mixture, and performing ultrasonic vibration to obtain mixed solution;
5) dissolving 40% of modified soybean protein isolate in the step 2) in phosphate buffer solution with the pH value of 7.4, adding 20% of sericin solution by weight, adding 14% of mixed paste by weight while stirring, adding 8% of cross-linking agent by weight, adding 35% of mixed solution by weight, putting into a refrigerator to reduce the temperature to 2-5 ℃, adding 6% of growth factor freeze-dried powder by weight, mixing with 2% of medical alginate fiber by weight, heating to 40-55 ℃, stirring, adding 10% of algal polysaccharide by weight after reducing the temperature, repeatedly freezing and thawing for multiple times to obtain the dressing.
In the step 1), a sericin solution with the mass volume percentage concentration of 5.0% is prepared.
The feeding proportion of the soy protein isolate, the vinyl trimethoxy silane, the N-hydroxysuccinimide, the dopamine and the fluoxetine in the step 2) is 10: 3:2: 6: 4.
the feeding proportion of the oyster shell powder, the aloe vera fresh gel, the docosahexaenoic acid, the ascorbic acid, the titanium dioxide, the camellia oil and the linoleic acid in the step 3) is 15:40:30:1.2:5:3: 3.
In the step 4), the feeding proportion of the chitosan, the xanthan gum, the dencichine, the quercetin, the D-arbutin, the carbomer, the urea capsule and the medicine extract is 5: 43: 2:1.5:0.8: 4:2:20.
And step 4) preparing a mixed solution with the mass volume percentage concentration of 3.0%.
Example 3:
1) dissolving silk fibroin powder into ionized water to prepare silk fibroin solution for later use;
2) dissolving soybean protein isolate in phosphate solution with pH of 7.4 at 40-80 ℃, adding vinyltrimethoxysilane and N-hydroxysuccinimide for mixing, adjusting the pH value of a reaction system to 4-7 after stirring, adding dopamine and fluoxetine after stirring and reacting for 30-60 min at 20-50 ℃, maintaining the pH value of the reaction system to 4-7, continuing stirring and reacting for 10-36 h at 20-50 ℃, and finally dialyzing and freeze-drying to obtain modified soybean protein isolate for later use;
3) adding Concha Ostreae powder into Aloe fresh gel, stirring, and sequentially adding docosahexaenoic acid, ascorbic acid, titanium dioxide, oleum Camelliae Japonicae and linoleic acid during stirring to obtain mixed paste;
4) mixing chitosan, xanthan gum, dencichine, quercetin, D-arbutin, carbomer, urea capsule and the medicinal extract, adding appropriate amount of water into the obtained mixture, and performing ultrasonic vibration to obtain mixed solution;
5) dissolving 35% by weight of modified isolated soy protein obtained in the step 2) in a phosphate buffer solution with the pH value of 7.4, adding 12% by weight of sericin solution, adding 10% by weight of mixed paste while stirring, adding 6% by weight of cross-linking agent, adding 27% by weight of mixed solution, refrigerating to reduce the temperature to 2-5 ℃, adding 3% by weight of growth factor freeze-dried powder, adding 1% by weight of medical alginate fibers, heating to 40-55 ℃, stirring, reducing the temperature, adding 6% by weight of algal polysaccharide, repeatedly freezing for many times, and thawing to obtain the dressing.
In the step 1), a sericin solution with the mass volume percentage concentration of 3.2% is prepared.
The feeding proportion of the soy protein isolate, the vinyl trimethoxy silane, the N-hydroxysuccinimide, the dopamine and the fluoxetine in the step 2) is 10: 2: 1: 5: 3.
the feeding proportion of the oyster shell powder, the aloe vera fresh gel, the docosahexaenoic acid, the ascorbic acid, the titanium dioxide, the camellia oil and the linoleic acid in the step 3) is 12:35:25:0.8:3:2: 2.
In the step 4), the feeding proportion of the chitosan, the xanthan gum, the dencichine, the quercetin, the D-arbutin, the carbomer, the urea capsule and the medicine extract is 3: 32: 1:1:0.4: 3:1:16.
And step 4) preparing a mixed solution with the mass volume percentage concentration of 2.2%.
Example 4
The dressing prepared in example 3 was given to participate in the following experiment:
30 patients with scars for more than one year are selected, the dressing is respectively applied twice a day for thirty days, after 30 days, the scars of 12 patients recover and slightly color-difference with the skin around the scars, the scars of 16 patients appear pink, the color of the scars is obviously lightened, and the color of the scars of 2 patients is slightly lightened.
In addition, 15 of 30 patients had dark skin, and the skin was improved in all 15 patients after use.
Example 5
The dressing prepared in example 3 was given to participate in the following experiment:
(1) cell viability assay for mouse dermodermablastic fibroblasts (NIH3T3) and human skin keratinocytes (Hacat) sericin wound dressing
The sericin wound dressing is firstly cut into slices, then washed with sterilized PBS for 3 times, soaked with 75% ethanol for 1 hour, finally washed with sterilized PBS for 1 time, and placed at room temperature for later use.
Suspending and blowing off cells collected from a cell culture bottle, planting the cells in a 96-hole cell culture plate, transferring the pretreated sericin wound dressing into the cell-containing culture plate after the cells adhere to the wall, and placing one piece of sericin wound dressing in each hole.
Cell culture wells without sericin wound dressing placed served as controls.
The culture medium used for cell culture is DMEM high-sugar medium, and the cells are cultured in a cell culture box (37 ℃, the concentration of CO2 is 5%, and the humidity is 100%).
The activity of the cells was examined 2 and 3 days after planting by the CCK8 method. The results show that: compared with the human skin keratinocyte (Hacat) sericin wound dressing group and the control group, the mouse skin fibroblast (NIH3T3) sericin wound dressing group has no significant difference in cell viability, and the result shows that the sericin wound dressing has good cell compatibility.
(2) Cell migration detection of sericin wound dressing
The cell line used was mouse skin fibroblast (NIH3T 3).
The sericin wound dressing is firstly cut into a circular sheet shape, then washed with sterilized PBS for 3 times, soaked with 75% ethanol for 1 hour, finally washed with sterilized PBS for 1 time, and placed in a 24-hole cell culture plate.
And (3) suspending and blowing off the cells collected from the cell culture bottle, planting the cells in the cell culture plate containing the sericin wound dressing in the step (1), and counting the number of the initially planted cells. After the cells were attached to the wall, the sericin wound dressing was transferred to a new 24-well cell culture plate. And counting the number of the cells remained in each hole of the original culture plate to obtain the number of the cells initially planted on the sericin wound dressing.
Thereafter, the sericin wound dressing was transferred to a new cell culture plate once every other day, and the number of cells remaining in the corresponding well in the original culture plate was counted and recorded. And finally, carrying out statistical analysis. The culture medium used for cell culture was DMEM high-sugar medium, and the cells were cultured in a cell culture chamber (37 ℃, 5% CO2 concentration, 100% humidity).
Cells that migrated from the sericin wound dressing exhibited a phenomenon of continued growth within 24 days on a statistical basis. The sericin wound dressing has good cell compatibility and can support the survival, proliferation and migration of cells for a long time.
(3) 3D cell culture of sericin wound dressing
The cell line used was the GFP-stabilized human neuroblastoma cell line SHSY 5Y.
The sericin wound dressing is firstly cut into a circular sheet shape, then washed with sterilized PBS for 3 times, soaked with 75% ethanol for 1 hour, finally washed with sterilized PBS for 1 time, and placed in a 24-hole cell culture plate. Then, the cells collected from the cell culture flask are suspended and blown off, and then planted in a cell culture plate containing a sericin wound dressing. After 2 days the samples containing the sericin wound dressing were taken out for observation and photographing with a laser confocal lens. The cell growth state in the sericin wound dressing is good. The result shows that the sericin wound dressing has good cell compatibility and can be used as a carrier of cells.
Example 6
The sericin wound dressing of example 3 is used for repairing the skin injury of mice
(1) The sericin wound dressing is firstly cut into a circular sheet with the diameter of 1cm, then washed with sterilized PBS for 3 times, soaked with 75% ethanol for 1 hour, finally washed with sterilized PBS for 1 time, dried and placed at room temperature for later use.
(2) A mouse scald model is prepared, and the selected animal is a Kunming mouse with the age of 6 weeks. Firstly, anesthetizing and unhairing the mice; the mice were then hot-ironed on their backs for 5 seconds using a metal rod (cylinder 1cm in diameter) at 100 ℃. Then, the scalded skin is cut off, the sericin wound dressing pretreated in the step (1) is arranged on the wound surface, and a mouse which is not treated by the wound dressing is used as a control. The mouse skin scab using the sericin wound dressing dropped out of granulation after 10 days, the control scab was firm, and the mouse skin using the sericin wound dressing recovered after 20 days with slight color difference from the peripheral skin, with the control exposed granulation. Therefore, the invention has strong repairing effect on skin injury.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (10)
1. A production method of a medical dressing of sericin is characterized by comprising the following steps:
1) dissolving silk fibroin powder into ionized water to prepare silk fibroin solution for later use;
2) dissolving soybean protein isolate in phosphate solution with pH of 7.4 at 40-80 ℃, adding vinyltrimethoxysilane and N-hydroxysuccinimide for mixing, adjusting the pH value of a reaction system to 4-7 after stirring, adding dopamine and fluoxetine after stirring and reacting for 30-60 min at 20-50 ℃, maintaining the pH value of the reaction system to 4-7, continuing stirring and reacting for 10-36 h at 20-50 ℃, and finally dialyzing and freeze-drying to obtain modified soybean protein isolate for later use;
3) adding Concha Ostreae powder into Aloe fresh gel, stirring, and sequentially adding docosahexaenoic acid, ascorbic acid, titanium dioxide, oleum Camelliae Japonicae and linoleic acid during stirring to obtain mixed paste;
4) mixing chitosan, xanthan gum, dencichine, quercetin, D-arbutin, carbomer, urea capsule and the medicinal extract, adding appropriate amount of water into the obtained mixture, and performing ultrasonic vibration to obtain mixed solution;
5) dissolving the modified soybean protein isolate in the step 2) in a phosphate buffer solution with the pH value of 7.4, adding the sericin solution, adding the mixed paste while stirring, adding a cross-linking agent, adding the mixed solution, putting the mixed solution into a refrigerator, reducing the temperature to 2-5 ℃, adding the growth factor freeze-dried powder, mixing the medical alginate fibers, heating to 40-55 ℃, stirring, reducing the temperature, adding the algal polysaccharide, repeatedly freezing and thawing to obtain the dressing.
2. The method for producing a sericin medical dressing according to claim 1, wherein: the sericin solution with the mass volume percentage concentration of 2.0-5.0% is prepared in the step 1).
3. The method for producing a sericin medical dressing according to claim 1, wherein: the feeding proportion of the soy protein isolate, the vinyl trimethoxy silane, the N-hydroxysuccinimide, the dopamine and the fluoxetine in the step 2) is 10: 2-3: 1-2: 4-6: 2 to 4.
4. The method for producing a sericin medical dressing according to claim 1, wherein: the feeding proportion of the oyster shell powder, the aloe vera fresh gel, the docosahexaenoic acid, the ascorbic acid, the titanium dioxide, the camellia oil and the linoleic acid in the step 3) is 12-15: 30-40: 20-30: 0.5-1.2: 2-5: 2-3.
5. The method for producing a sericin medical dressing according to claim 1, wherein: in the step 4), the feeding proportion of the chitosan, the xanthan gum, the dencichine, the quercetin, the D-arbutin, the carbomer, the urea capsule and the medicine extract is 2-5: 30-43: 1-2: 1-1.5: 0.3-0.8: 3-4: 1-2: 15-20.
6. The method for producing a sericin medical dressing according to claim 1, wherein: and (4) preparing a mixed solution with the mass volume percentage concentration of 1.5-3.0%.
7. The medical dressing of sericin according to claim 1, which comprises the following raw materials in parts by weight:
8. the medical dressing of sericin according to claim 7, which comprises the following raw materials in parts by weight:
9. a sericin medical dressing as claimed in claim 7 or 8, wherein: the mixed paste comprises oyster shell powder, aloe vera fresh gel, docosahexaenoic acid, ascorbic acid, titanium dioxide, camellia oil and linoleic acid; the cross-linking agent is one or more of glutaraldehyde, malondialdehyde and genipin; the mixed solution comprises chitosan, xanthan gum, dencichine, quercetin, D-arbutin, carbomer, urea capsule and medicinal extract.
10. The medical dressing of sericin according to claim 9, wherein: the medicinal extract comprises pearl powder, mulberry leaf extract, common perilla extract, angelica dahurica extract and raw licorice extract.
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