CN116370690A - Composite preparation of fetal disc decidua, preparation method thereof, medical dressing containing composite preparation and application of composite preparation - Google Patents

Composite preparation of fetal disc decidua, preparation method thereof, medical dressing containing composite preparation and application of composite preparation Download PDF

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CN116370690A
CN116370690A CN202310228258.6A CN202310228258A CN116370690A CN 116370690 A CN116370690 A CN 116370690A CN 202310228258 A CN202310228258 A CN 202310228258A CN 116370690 A CN116370690 A CN 116370690A
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rich plasma
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CN116370690B (en
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高东
卢瑞珊
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Guangzhou Zhengyuan Biotechnology Co ltd
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L15/00Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/40Bandages, 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
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14Blood; Artificial blood
    • A61K35/16Blood plasma; Blood serum
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48Reproductive organs
    • A61K35/50Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
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    • A61LMETHODS 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
    • A61L15/00Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/42Use of materials characterised by their function or physical properties
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L15/00Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/42Use of materials characterised by their function or physical properties
    • A61L15/44Medicaments
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00Drugs for dermatological disorders
    • A61P17/02Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2300/00Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/30Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2300/00Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412Tissue-regenerating or healing or proliferative agents
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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Abstract

The invention belongs to the technical field of medical biological materials, and in particular relates to a placenta decidua composite preparation, a preparation method thereof, a medical dressing containing the placenta decidua composite preparation and application of the placenta decidua composite preparation. The composite preparation comprises the basal decidua of the fetal disc and platelet-rich plasma. According to the invention, the decidua and the platelet-rich plasma are used in a compound way for the first time, and are applied to the wound surface, and cytokines in the platelet-rich plasma are released, so that the tissue inflammation is inhibited to play a role in field planting and repairing. The decidua basal lamina promotes the adhesion and migration of epithelial cells, induces epithelial differentiation, prevents epithelial apoptosis, improves the regeneration capacity of tissues, promotes the repair of the original tissues, and further plays a synergistic repair role in cooperation with platelet-rich plasma. The invention can be used for repairing wound surfaces caused by various reasons, has obvious repairing effect, no skin irritation, convenient use, no need of intravenous reinjection and only need of being applied to the wound surfaces.

Description

Composite preparation of fetal disc decidua, preparation method thereof, medical dressing containing composite preparation and application of composite preparation
Technical Field
The invention belongs to the technical field of medical biological materials, and in particular relates to a placenta decidua composite preparation, a preparation method thereof, a medical dressing containing the placenta decidua composite preparation and application of the placenta decidua composite preparation.
Background
Wound surface is the damage of normal skin (tissue) caused by external injury factors such as surgery, external force, heat, current, chemical substances, low temperature and internal factors of organism such as inflammation, local blood supply disorder and the like. Often accompanied by a disruption of the integrity of the skin and a loss of a certain amount of normal tissue, while at the same time the normal function of the skin is impaired. For example, acne is a chronic inflammatory skin disease of the pilo-sebaceous unit and is characterized by polymorphic lesions that develop on the face, acne, papules, pustules, nodules, etc. The existing treatment mainly uses western medicines, wherein the oral medicine can be selected from melamycin (minocycline), azithromycin and the like, and the external medicine can be selected from 2.5% -10% benzoyl peroxide gel, 0.1% adapalene gel, clindamycin solution and the like. Although the medicines can play a certain role in repairing wound surfaces, the repairing effect is still not ideal, the repairing speed is low, certain stimulus can be caused to the wound surfaces, and allergic reaction is easy to occur to physique sensitive receptors. These problems are also present in other wound treatment drugs and need to be solved.
Disclosure of Invention
The invention aims at solving the technical problems in the prior art and provides a fetal disc and basal decidua composite preparation with good wound repair effect and high skin use safety, a preparation method thereof, a medical dressing containing the same and application thereof.
The invention adopts the following technical scheme:
the invention provides a placenta decidua composite preparation, which comprises the placenta decidua and platelet-rich plasma. Preferably, the platelet rich plasma contains not less than 1000X 10 9 Platelets at/L concentration. Preferably, in the composite preparation, the volume percentage of the platelet-rich plasma is 20-40%.
The preparation method of the placenta decidua composite preparation comprises the following steps:
(1) Separating decidua from placenta, shearing into tissue blocks, and freeze-drying to obtain freeze-dried decidua; preferably, the size of the tissue mass is 10-30 cm 2 Freeze drying is performed by freezing at-20deg.C24 h, then drying 3 h under the conditions of-50 ℃ and vacuum degree of 1 Pa;
(2) Taking peripheral blood, performing first centrifugation, and taking supernatant to obtain first blood plasma; subjecting the first plasma to a second centrifugation to obtain a platelet pellet and a second plasma supernatant, mixing a portion of the second plasma with the platelet pellet and adjusting the platelet concentration to not less than 1000×10 9 L, obtaining platelet-rich plasma; mixing platelet-rich plasma with activator, and heating in water bath to obtain activated platelet-rich plasma; preferably, the first centrifugation is at 1500 rpm for 10min and the second centrifugation is at 2500rpm for 15 min; preferably, the activator is prepared by: dissolving thrombin freeze-dried powder by 0.1 g/mL calcium gluconate solution, and regulating the final concentration of thrombin to be 100U/mL; the volume ratio of the platelet rich plasma to the activator is 1:10; the temperature of water bath heating is 37 ℃, and the time of water bath heating is 30min;
(3) And soaking and freeze-drying the activated platelet-rich plasma to obtain the fetal disc decidua composite preparation.
The composite preparation provided by the invention, wherein platelet rich plasma (platelet rich plasma, PRP) is originally derived from a transfusion medical concept, and is platelet concentrate separated from blood by a centrifugal method. At present, the traditional Chinese medicine composition is widely applied to various disciplines, such as orthopedics, sports medicine, oral and maxillofacial surgery, plastic and cosmetology, dermatology, wound repair special department, gynaecology and obstetrics, traditional Chinese medicine department and the like. PRP can promote repair of various tissues such as bone, cartilage, muscle, tendon, ligament, fat, blood vessel, nerve fiber, mucous membrane, skin, etc. PRP contains high concentration of platelets and small amounts of white and red blood cells, and the platelets release a large amount of active factors after activation. Platelets are the most important component of PRP and play an important role in the physiological and pathological processes of hemostasis, coagulation, inflammatory reactions, etc. Platelets contain a large number of growth factors and cytokines required for tissue repair, and their abundance of angiopoietin-1 (Ang 1) and other angiogenic factors can stimulate vascular endothelial cell growth and migration.
In addition, placenta has become an important source of biofilm material of interest to many scholars, who have succeeded in separating and extracting amniotic membrane and chorion from placenta and in developing biological patches, but have studied the decidua basal on the maternal side. The decidua basal membrane extracted from placenta has the advantages of abundant stem cells with low rejection reaction after transplantation, no ethical obstacle in clinic and the like. The two major advantages of decidua bottom in skin reconstruction are: promote epithelialization, inhibit inflammatory reaction and fibrosis. Mainly realized by the following mechanism:
(1) The decidua basal can promote the adhesion and migration of epithelial cells, induce epithelial differentiation, and prevent epithelial apoptosis. The patch can also protect new epithelial tissue from instantaneous eyelid scraping while reducing inflammatory cells and tear protein contact with the corneal stroma when used as a mask.
(2) The decidua basal can secrete bFGF, EGF, HGF, KGF and other growth factors to promote the growth of epithelium.
(3) The decidua basal can inhibit the expression of interleukin, regulate the expression of inflammatory chemotactic factors, induce apoptosis of polynuclear leucocytes, and reduce the expression of matrix metalloproteinases l, 2 and 9, thereby reducing inflammatory reaction.
The invention combines the decidua with platelet-rich plasma for use, and is suitable for developing into various medical dressings and other wound repair medicines. Therefore, the invention also relates to a medical dressing prepared by adding auxiliary materials on the basis of the placenta decidua composite preparation and application of the placenta decidua composite preparation in preparing medicines for repairing wound surfaces, including but not limited to the medicines for treating difficult-to-heal wound surfaces such as facial acne, diabetic foot and the like, and has excellent 'positioning' repairing effect.
The beneficial effects of the invention are as follows:
the composite preparation provided by the invention is used for the first time by compounding the decidua bottom and the platelet-rich plasma, and is applied to a wound surface, and cytokines in the platelet-rich plasma are released to inhibit tissue inflammation and play a role in field planting repair. The decidua basal lamina promotes the adhesion and migration of epithelial cells, induces epithelial differentiation, prevents epithelial apoptosis, improves the regeneration capacity of tissues, promotes the repair of the original tissues, and further plays a synergistic repair role in cooperation with platelet-rich plasma. The invention can be used for repairing wound surfaces caused by various reasons, has obvious repairing effect, no skin irritation, convenient use, no need of intravenous reinjection and only need of being applied to the wound surfaces.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, based on the examples herein, which are within the scope of the invention as defined by the claims, will be within the scope of the invention as defined by the claims.
The invention provides a placenta decidua composite preparation, which is prepared by the following steps:
1. separation and extraction of decidua bottom
1.1 The collected placenta and maternal blood are received, photographed by the placenta and then placed into a sample transmission window inlet, and the maternal blood is sent to a quality control part for virus (HIV, HBV, HCV, syphilis) detection.
1.2 Entering the clean zone according to the standard operation rules of personnel entering and exiting the clean zone.
1.3 The ultra-clean bench is sterilized by ultraviolet rays for 30min in advance, and the cell cleaning liquid is placed at room temperature.
1.4 Turning off the ultraviolet lamp, turning on ventilation for 2-5min, and wiping the disinfection super clean bench with sterile gauze from inside to outside in one direction.
1.5 Opening the decidua bottom to prepare an outer package, sterilizing with 75% alcohol, and placing into an ultra clean bench. And (5) sterilizing the cell cleaning liquid sample and related consumable materials by using 75% alcohol, and placing the cell cleaning liquid sample and the related consumable materials into an ultra-clean bench. Opening a collection box filled with placenta, sucking the collection liquid of the placenta immersed in 1 mL by a Pasteur pipette, subpackaging into 2 EP pipes, 0.5 mL/pipe, reserving one part of sample, and sending one part of sample to a quality inspection part for bacteria and fungus detection.
1.6 The placenta is clamped by tissue forceps and hemostat, the placenta is transferred from a collection box to a sterile tray, umbilical cord tissues on the placenta are cut off by direct shearing, the whole decidua is cut off by direct shearing and put into a 10cm culture dish, the blood clots and the parts with serious damage are cut off, the decidua is transferred to three 10cm culture dishes respectively filled with cell cleaning liquid, and the total rinsing is carried out for 3-4 times until the cell cleaning liquid is clarified.
1.7 Cutting the decidua basal using direct shear into 10-30 cm 2 Tissue of a size. Spreading in sterile tray, and absorbing the moisture of decidua with sterile nonwoven paper; sucking the cleaning solution on the surface of the decidua bottom.
1.8 The decidua bottom was frozen at-20℃for 24 h.
1.9 The sample was frozen 24 and h and then freeze-dried under vacuum: starting a refrigerator, taking out the pre-frozen material, placing the material on a drying rack when the temperature of a cold trap is reduced to-50 ℃, placing the material above the cold trap, covering an organic glass cover, closing an inflation valve, opening a vacuum pump and a vacuum gauge, reducing the vacuum value from 50 pa to 1 pa, and vacuum drying 3 h until the decidua is completely dried; packaging aseptically, and storing at-20deg.C for a long time.
2. PRP preparation
2.1 Cleaning the sterile operation table with 75% ethanol, placing 60 mL peripheral blood into the sterile operation table, comparing the blood collection amount of each blood collection tube, performing balance treatment, and sampling to count the whole blood platelets.
2.2 Transferring the balanced blood collection tube to a centrifuge, and centrifuging at 1500 rpm for 10min
2.3 Transferring the centrifuged peripheral blood to a sterile operating table, and transferring the upper plasma layer to a 15mL centrifuge tube by using a Pasteur pipette; the tube containing the plasma was equilibrated and sampled for the first platelet count.
2.4 Based on the first platelet count, the final volume of the formulation was calculated.
2.5 The plasma with the same volume as the PRP final preparation is left in a 15mL centrifuge tube for standby; after the remaining plasma was equilibrated, it was transferred to a centrifuge for centrifugation at 2500rpm for 15 min.
2.6 After centrifugation, sucking plasma above the centrifugation by using a disposable 10mL screw syringe to form a PPP preparation, and attaching a bar code; the refrigerator was kept at 4 ℃.
2.7 Sucking the standby plasma by using a Pasteur pipette, gently blowing the platelet sediment along the wall after centrifugation, and fully and uniformly mixing to obtain the PRP preparation. Subpackaging according to 1-3 mL/tube, and placing in a refrigerator at-20 ℃ for standby.
2.8 Preparing an activating agent: taking 0.1 g/mL of calcium gluconate solution, sucking 2mL of the calcium gluconate solution by a 5mL disposable syringe, injecting into thrombin freeze-dried powder (200 units/bottle), and fully and uniformly mixing to obtain the activator. According to the activator PRP preparation (V: V=1:10), mixing well, covering the cover, and putting into a disposable thin film glove. And (3) water bath activation: inactivating at 37deg.C for 30min; gel was removed with a Pasteur pipette to form the final PRP formulation, which was stored in a refrigerator at 4 ℃.
3. Preparation of the composite preparation: cutting the freeze-dried decidua to a proper size according to the size of a wound, and adding the liquid PRP preparation obtained in the step 2.8 into the freeze-dried decidua according to the volume ratio to obtain the liquid PRP preparation.
TABLE 1
Figure SMS_1
Experimental example 1
90 mice were taken and wound model preparation and grouping were performed: after the mice are routinely raised by 7 d, the mice are anesthetized by intraperitoneal injection of 30 mg/kg of 1% pentobarbital sodium, the back hair is shaved, a 1 cm ×1 cm wound surface is prepared in the middle of the back, and the mice are prepared into a mechanical injury mouse model subcutaneously. The samples are respectively stuck on the wound surface, after the samples are fixed by a medical adhesive tape for 3 days, the residual dressing on the wound surface is taken down, the wound surface is continuously wrapped by sterile gauze, and the healing degree of the wound surface is observed after 7 days.
TABLE 2 results of wound repair test on mice
Figure SMS_2
*P<0.05 represents a significant difference compared to the control group; * P<0.001, indicating a very significant difference compared to the control group; # P<0.05 represents a significant difference compared to the positive control group; ## P<0.001 represents a very significant difference compared to the positive control group; P<0.05 representsThere were significant differences compared to the comparative examples.
As can be seen from the table, the positive control had a wound healing rate of 42.47%, the comparative example had a wound healing rate of 75.46%, the examples 1, 2, and 3 had wound healing rates of 86.93%, 87.42%, 88.71%, respectively, all of which were significantly different from the negative control group having a wound healing rate of 21.25%, and P <0.001; the wound healing effect of the positive control, the comparative example and examples 1-3 is better than that of the negative control group;
comparative example 1 had significant differences, P <0.05, examples 1-3 all had very significant differences, P <0.001, compared to the positive control group. The effect of adding the freeze-dried decidua is better than that of not adding the freeze-dried decidua.
Experimental example 2
The preparation and grouping of wound models were performed on 48 SD rats: after the rats are routinely raised by 7 d, the rats are anesthetized by intraperitoneal injection of 30 mg/kg of 1% pentobarbital sodium, the back hair is shaved, a 1.5 cm ×1.5 cm wound surface is prepared in the middle of the back, and the rats are prepared into a mechanical injury rat model from deep to the skin. The samples are respectively applied to the wound surface, after the samples are fixed by a medical adhesive tape for 3 days, the residual dressing on the wound surface is taken down, the wound surface is continuously wrapped by sterile gauze, the healing degree of the wound surface is observed after 7 days, and the healing rate = the healing area/the original area of the wound surface.
TABLE 3 results of rat wound repair test
Figure SMS_3
* P <0.05, indicating a significant difference compared to the negative control group; * P <0.001, indicating a very significant difference compared to the negative control group.
As shown in the table, the wound healing rates of the negative control group are 23.14%, the wound healing rates of the examples 4 and 5 are 51.28% and 65.50%, respectively, and compared with 23.14% of the blank group, the negative control group has significant differences, and P <0.05, which indicates that the wound healing effects of the examples 4 and 5 are better than those of the blank group;
the wound healing rates of examples 6, 7 and 8 are 85.42%, 87.16% and 88.09%, respectively, and are very significantly different from those of the blank 23.14%, and P <0.001, which indicates that the wound healing effects of examples 6, 7 and 8 are significantly better than those of the blank;
from the experimental results, the wound healing effect of examples 6, 7 and 8 is significantly better than that of examples 4 and 5, indicating that the concentration of PLT is higher than 1000×10 9 The effect is good at/L. In addition, examples 6, 7 and 8 were not different in wound healing, so they were selected to be not less than 1000X 10 9 PLT concentration/L is the preferred embodiment of the formulation.
The foregoing is merely illustrative of the present invention, and the present invention is not limited thereto, and any person skilled in the art will readily recognize that variations or substitutions are within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims (9)

1. The placental decidua composite preparation is characterized by comprising the placental decidua and platelet rich plasma.
2. The placental decidua complex formulation according to claim 1, wherein the platelet rich plasma contains not less than 1000 x 10 9 Platelets at/L concentration.
3. The placental decidua composite formulation of claim 2, wherein the platelet rich plasma is present in the composite formulation in a volume percentage of 20-40%.
4. A method for producing a fetal disc decidua composite preparation as claimed in claim 2 or 3, characterized by comprising the steps of:
(1) Separating decidua from placenta, shearing into tissue blocks, and freeze-drying to obtain freeze-dried decidua;
(2) Taking peripheral blood, performing first centrifugation, and taking supernatant to obtain first blood plasma; subjecting the first plasma to a second centrifugation to obtain a platelet pellet and a second plasma supernatantMixing part of the second plasma and platelet precipitate, and regulating platelet concentration to 1000×10 or more 9 L, obtaining platelet-rich plasma; mixing platelet-rich plasma with activator, and heating in water bath to obtain activated platelet-rich plasma;
(3) And soaking and freeze-drying the activated platelet-rich plasma to obtain the fetal disc decidua composite preparation.
5. The method for preparing a placental decidua complex formulation according to claim 4, wherein in step (1), the size of the tissue mass is 10-30 cm 2 The freeze drying is to freeze 24 h at-20deg.C, and then dry 3 h at-50deg.C under vacuum of 1 Pa.
6. The method for preparing a placental decidua composite formulation according to claim 4, wherein in step (2), the first centrifugation is at 1500 rpm for 10min and the second centrifugation is at 2500rpm for 15 min.
7. The method of preparing a placental decidua complex formulation according to claim 4, wherein in step (2), the activator is prepared by: dissolving thrombin freeze-dried powder by 0.1 g/mL calcium gluconate solution, and regulating the final concentration of thrombin to be 100U/mL; the volume ratio of the platelet rich plasma to the activator is 1:10; the temperature of the water bath heating is 37 ℃, and the time of the water bath heating is 30min.
8. A medical dressing comprising the fetal disc decidua composite formulation of any of claims 1-3.
9. Use of the composite preparation of the placental decidua of one of claims 1-3 for the preparation of a medicament for wound repair.
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US20200239836A1 (en) * 2016-03-14 2020-07-30 Plakous Therapeutics, Inc. Acellular placental preparations
CN111575229A (en) * 2020-04-13 2020-08-25 广东华夏健康生命科学有限公司 Separation method of placenta decidua stem cells
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