A composite biological dressing, its preparation method and application

Abstract

This invention provides a composite biological dressing, its preparation method, and its application, belonging to the field of medical dressing technology. By employing a specific preparation method, this invention successfully combines components such as endometrial stem cells, human fibrinogen, human plasma cryoprecipitate, and thrombin solution with a silk scaffold material. The resulting composite biological dressing overcomes the problems of slow repair speed and poor efficacy of existing medical dressings. The composite biological dressing provided by this invention has a significant repair effect, avoiding unnecessary damage to the body. Repair is achieved simply by application, and it also enhances tissue regeneration and promotes the repair of existing tissue, providing comprehensive and rapid wound repair. The composite biological dressing obtained by this invention gradually dissolves after application, releasing endometrial stem cells that differentiate into new tissue, overcoming the drawback of significant cell loss after intravenous injection of existing stem cell injections.

Description

Technical Field

[0001] This invention relates to the field of medical dressing technology, and in particular to a composite biological dressing, its preparation method, and its application. Background Technology

[0002] Dressings are medical materials that temporarily protect wounds, prevent infection, and promote healing. Dressings can be categorized by material into traditional dressings such as absorbent cotton gauze, biological dressings, and synthetic dressings. Biological dressings are a new type of wound repair and protection material developed based on the "moist healing" theory of wound repair proposed by Winter et al. Compared to traditional dressings, biological dressings offer advantages such as reduced infection, improved wound healing quality, reduced patient suffering, and ease of use for medical staff, thus gaining popularity.

[0003] Vaginitis and cervicitis are common diseases in women, clinically occurring after injury to the vagina or cervix (such as childbirth, miscarriage, or surgery), leading to infection by pathogens. Clinically, colposcopy in patients with vaginitis and cervicitis reveals varying degrees of congestion and edema of the vaginal or cervical mucosa, as well as glandular and stromal hyperplasia, resulting in enlargement of the vagina and cervix. Microscopically, inflammatory cell infiltration, including lymphocytes, plasma cells, and monocytes, can be observed in the subepithelial layer of the cervical endometrium; local mucosal degeneration, necrosis, or sloughing may also occur, forming ulcers.

[0004] Current technologies mostly use medical hydrogels such as gelatin for wound repair. While these can provide some wound healing, the repair process is slow. Adding additional medications or preservatives can irritate the wound, and sensitive patients may experience allergic reactions, thus failing to achieve the desired repair effect. Summary of the Invention

[0005] The purpose of this invention is to provide a composite biological dressing, its preparation method, and its application, in order to solve the problems of slow repair speed and poor effect of existing medical dressings.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides a method for preparing a composite biological dressing, comprising the following steps:

[0008] The endometrial stem cell suspension was mixed with composite solution 1 to obtain a cell-fibrin gel suspension;

[0009] A mixture was obtained by soaking a silk scaffold in a cell-fibrin gel suspension;

[0010] The mixture was mixed with composite solution 2 and crosslinked to obtain a composite biological dressing;

[0011] The composite solution 1 contains human fibrinogen and human plasma cryoprecipitate;

[0012] The composite solution 2 contains thrombin solution and calcium salt solution;

[0013] The volume ratio of the endometrial stem cell suspension to the composite solution 1 is 1:2~3.

[0014] Preferably, the soaking time is 5-8 minutes;

[0015] The weight ratio of human fibrinogen to human plasma cryoprecipitate is 1:0.8~1.2;

[0016] The concentration of the composite solution 1 is 20~40g / L.

[0017] Preferably, the concentration of the thrombin solution is 500~800 U / mL;

[0018] The calcium salt solution is calcium gluconate or calcium chloride;

[0019] The concentration of the calcium salt solution is 4~6 mg / mL;

[0020] The volume ratio of the thrombin solution to the calcium salt solution is 1:0.8~1.2.

[0021] Preferably, the volume ratio of the cell-fibrin gel suspension to the composite solution 2 is 8~12:1;

[0022] The crosslinking time is 1~3 min.

[0023] Preferably, the silk scaffold further includes a pretreatment before being soaked in the cell-fibrin gel suspension, the pretreatment comprising the following steps:

[0024] The silk scaffold was prepared by soaking in sodium carbonate aqueous solution and then dried.

[0025] The concentration of the sodium carbonate aqueous solution is 0.01~0.03 mol / L;

[0026] The soaking time is 40-80 minutes, and the soaking temperature is 90-100℃.

[0027] Preferably, the drying temperature is 35~40℃ and the drying time is 20~40min;

[0028] The sodium carbonate aqueous solution is replaced every 15-25 minutes during the soaking process.

[0029] The density of the silk scaffold is 20-22 needles / cm;

[0030] The pore size of the silk scaffold is 0.8~1.2mm. 2 .

[0031] Preferably, the endometrial stem cell suspension is obtained by a preparation method including the following steps:

[0032] Dilute the menstrual blood sample to obtain diluted blood;

[0033] Diluted blood was added to the top layer of the lymphocyte separation medium, and the first centrifugation was performed to obtain the supernatant containing a mononuclear cell layer.

[0034] The supernatant containing the mononuclear cell layer was mixed with physiological saline and centrifuged a second time to obtain the cell pellet.

[0035] The cell pellet obtained by resuspending in complete culture medium was used to obtain the primary cell suspension;

[0036] The primary cell suspension was sequentially seeded, digested, and passaged to obtain progeny endometrial stem cells.

[0037] The progeny endometrial stem cells were mixed with physiological saline containing albumin to obtain an endometrial stem cell suspension.

[0038] Preferably, the rotation speed of the first centrifugation is 2200~2800 rpm, and the time of the first centrifugation is 8~12 min;

[0039] The second centrifugation is performed at a speed of 1500~2000 rpm for a duration of 3~8 min.

[0040] The complete culture medium is Youkang complete culture medium;

[0041] The concentration of the primary cell suspension was 5 × 10⁻⁶. 4 ~5×10 5 cells / mL;

[0042] The progeny endometrial stem cells are 2nd to 5th generation endometrial stem cells;

[0043] The weight percentage of albumin in the saline solution is 3-8%.

[0044] The present invention also provides a composite biological dressing obtained by the above preparation method.

[0045] This invention also provides the application of the above-mentioned composite biological dressing in the preparation of a medicament for treating inflammatory diseases of the female external genital system.

[0046] The technical effects and advantages of this invention are as follows:

[0047] This invention employs a specific preparation method to successfully combine components such as endometrial stem cells, human fibrinogen, human plasma cryoprecipitate, and thrombin solution with silk scaffold materials. The resulting composite biological dressing exhibits significant repair effects, avoids unnecessary damage to the body, and achieves repair effects simply by applying it.

[0048] The composite biological dressing provided by this invention can improve the regenerative capacity of tissues and promote the repair of existing tissues, thus playing a comprehensive and rapid role in wound repair. The product gradually dissolves after application, releasing the endometrial stem cells, which then differentiate into new tissues, overcoming the disadvantage of existing stem cell injections that result in a large loss of cells after intravenous reinjection. Attached Figure Description

[0049] Figure 1 The morphology of second-generation endometrial stem cells;

[0050] Figure 2 This describes the morphology of fifth-generation endometrial stem cells. Detailed Implementation

[0051] This invention provides a method for preparing a composite biological dressing, comprising the following steps: mixing an endometrial stem cell suspension with a composite solution 1 to obtain a cell-fibrin glue suspension; soaking a silk scaffold in the cell-fibrin glue suspension to obtain a mixture; mixing the mixture with a composite solution 2 and crosslinking to obtain a composite biological dressing, wherein the composite solution 1 contains human fibrinogen and human plasma cryoprecipitate; and the composite solution 2 contains thrombin solution and calcium salt solution.

[0052] In this invention, the endometrial stem cell suspension is preferably obtained by a preparation method comprising the following steps: diluting a menstrual blood sample to obtain diluted blood; adding the diluted blood to the supernatant of a lymphocyte separation medium and performing a first centrifugation to obtain a supernatant containing a mononuclear cell layer; mixing the supernatant containing the mononuclear cell layer with physiological saline and performing a second centrifugation to obtain a cell pellet; resuspending the cell pellet in a complete culture medium to obtain a primary cell suspension; sequentially seeding, digesting, and passaged the primary cell suspension to obtain progeny endometrial stem cells; and mixing the progeny endometrial stem cells with physiological saline containing albumin to obtain an endometrial stem cell suspension. The preferred speed for the first centrifugation is 2200-2800 rpm, more preferably 2400-2600 rpm, and the preferred centrifugation time is 8-12 min, more preferably 9-11 min. The preferred speed for the second centrifugation is 1500-2000 rpm, more preferably 1600-1800 rpm, and the preferred centrifugation time is 3-8 min, more preferably 4-6 min. The preferred complete culture medium is Youkang complete culture medium. The preferred concentration of the primary cell suspension in this invention is 5×10⁴-5×10⁵ cells / mL, more preferably 8×10⁵ cells / mL. 4 ~2×10 5 The number of cells / mL, wherein the progeny endometrial stem cells are preferably second to fifth generation endometrial stem cells; the weight percentage of albumin in the physiological saline is preferably 3 to 8%, and more preferably 4 to 6%.

[0053] In this invention, the volume ratio of the endometrial stem cell suspension to the composite solution 1 is 1:2~3, preferably 1:2.3~2.7. The composite solution 1 contains human fibrinogen and human plasma cryoprecipitate. The dosage form of the human fibrinogen and human plasma cryoprecipitate is preferably lyophilized powder. The weight ratio of the human fibrinogen and human plasma cryoprecipitate is preferably 1:0.8~1.2, more preferably 1:0.9~1.1. The solvent of the composite solution 1 is preferably physiological saline. The concentration of the composite solution 1 is preferably 20~40 g / L, more preferably 25~35 g / L. The volume ratio of the cell-fibrin gel suspension to the silk scaffold is preferably 1:0.8~1.2, more preferably 1:0.9~1.1. The soaking time is preferably 5~8 min, more preferably 6~7 min.

[0054] In this invention, the silk scaffold preferably undergoes a pretreatment before being soaked in a cell-fibrin gel suspension. The pretreatment includes the following steps: soaking the silk scaffold in a sodium carbonate aqueous solution, followed by drying; the density of the silk scaffold is preferably 20-22 needles / cm, and the pore size of the silk scaffold is preferably 0.8-1.2 mm²; the concentration of the sodium carbonate aqueous solution is preferably 0.01-0.03 mol / L, more preferably 0.015-0.025 mol / L; the soaking time is preferably 40-80 min, more preferably 50-70 min; the soaking temperature is preferably 90-100℃, more preferably 93-97℃; the drying temperature is preferably 35-40℃, more preferably 37-39℃; the drying time is preferably 20-40 min, more preferably 25-35 min; and the sodium carbonate aqueous solution is preferably replaced every 15-25 min during the soaking process.

[0055] In this invention, the concentration of the thrombin solution is preferably 500-800 U / mL, more preferably 600-700 U / mL; the calcium salt solution is preferably calcium gluconate or calcium chloride, and the concentration of the calcium salt solution is preferably 4-6 mg / mL, more preferably 4.5-5.5 mg / mL; the volume ratio of the thrombin solution to the calcium salt solution is preferably 1:0.8-1.2, more preferably 1:0.9-1.1; the volume ratio of the cell-fibrin glue suspension to the composite solution 2 is preferably 8-12:1, more preferably 9-11:1; and the cross-linking time is preferably 1-3 min, more preferably 1.5-2.5 min.

[0056] The present invention also provides a composite biological dressing obtained by the above preparation method, wherein the composite biological dressing is preferably stored under low temperature conditions, wherein the low temperature is preferably 0~10℃.

[0057] The present invention also provides the application of the above-mentioned composite biological dressing in the preparation of a drug for treating inflammatory diseases of the female external genital system, wherein the composite biological dressing is a topical drug, preferably applied to the wound by means of a cauterization.

[0058] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0059] Reagent source:

[0060] Fibrinogen: Purchased from Jiangxi Boya Biopharmaceutical Co., Ltd., National Drug Approval Number: S20013006;

[0061] Freeze-dried human plasma cryoprecipitate: Fresh frozen plasma was provided by the Guangdong Provincial Blood Bank; the white flocculent precipitate formed after the frozen plasma thaws at low temperature (2°C) mainly contains coagulation factor VIII, von Willebrand factor (vWF), fibrinogen, factor XIII, and fibronectin. The obtained plasma cryoprecipitate is freeze-dried to form freeze-dried powder.

[0062] Example 1

[0063] Clean the sterile work surface with 75% ethanol. Place the diluted menstrual blood into the sterile work surface and into two EP tubes. Take a 15mL centrifuge tube, add 5mL of Ficoll meglumine lymphocyte separation medium (density 1.007g / L), add 1mL of diluted blood to the surface of the lymphocyte separation medium, and centrifuge at 2500rpm for 10min.

[0064] After centrifugation, the supernatant containing the mononuclear cell layer was aspirated and transferred to another 50 mL centrifuge tube. Physiological saline was added to a final volume of 40 mL, and the cells were resuspended. The tube was then centrifuged at 1800 rpm for 8 minutes. The supernatant was discarded, and the cells were resuspended in 10 mL of Youkang complete culture medium to adjust the cell concentration to 7.5 × 10⁻⁶ cells / mL. 4 / mL, inoculated into 25cm 2 The cells were cultured in culture flasks using standard methods (37℃, 5% CO2, 95% humidity). The medium was changed for the first time after 48 hours, and then changed every 3 days. Once the cell confluence rate reached 80%, the cells were passaged using trypsin digestion and cultured until the third generation of daughter cells was obtained.

[0065] The silk support frame (purchased from Zhejiang Cathaya International Co., Ltd., warp knitting pattern; specifications: 12 knitting machine yarn (1 yarn / filament), density 21 stitches / cm, pore size 1mm) was used. 2 Immerse it in a 0.02 mol / L sodium carbonate aqueous solution and soak it at 95℃ for 60 min, changing the solution every 20 min during the soaking period. After soaking, remove it and dry it at 37℃ for 30 min for later use.

[0066] Prepare compound reagent 1 and compound reagent 2 at 37°C:

[0067] Compound reagent 1: Weigh refined fibrinogen and lyophilized human plasma cryoprecipitate in a weight ratio of 1:1, add them to physiological saline, and the total concentration is 30 g / L.

[0068] Compound reagent 2: Take 1 mL each of 600 U / mL thrombin solution and 5 mg / mL calcium gluconate aqueous solution, and mix them to obtain compound reagent 2.

[0069] The endometrial stem cell suspension and compound reagent 1 were mixed at a volume ratio of 1:2 to form a cell-fibrin gel suspension.

[0070] The silk scaffold and cell-fibrin gel suspension were mixed at a volume ratio of 1:1 and soaked for 6 minutes.

[0071] Add compound reagent 2 to the system at a concentration of 10% of the cell-fibrin glue suspension to promote cross-linking of the silk scaffold and fibrin, so that endometrial stem cells are evenly dispersed in the cross-linked network. After standing for 2 minutes, the composite biological dressing is obtained.

[0072] Example 2

[0073] Clean the sterile work surface with 75% ethanol. Place the diluted menstrual blood into the sterile work surface and into two EP tubes. Take a 15mL centrifuge tube, add 5mL of Ficoll meglumine lymphocyte separation medium (density 1.007g / L), add 1mL of diluted blood to the surface of the lymphocyte separation medium, and centrifuge at 2500rpm for 10min.

[0074] After centrifugation, the supernatant containing the mononuclear cell layer was aspirated and transferred to another 50 mL centrifuge tube. Physiological saline was added to a final volume of 40 mL, and the cells were resuspended. The tube was then centrifuged at 1600 rpm for 10 min. The supernatant was discarded, and the cells were resuspended in 10 mL of Youkang complete culture medium to adjust the cell concentration to 9 × 10⁶ cells / mL. 4 / mL, inoculated into 25cm 2 Cultured in culture flasks using standard methods (37℃, 5% CO2, 95% humidity), with the first medium change after 48 hours, followed by medium changes every 3 days. Once cell confluence reaches 80%, cells are passaged using trypsin digestion until the second generation of daughter cells is obtained. Figure 1 As shown, by Figure 1 It can be seen that endometrial stem cells in P2 cells have a spindle-shaped cell morphology and maintain vigorous proliferative capacity.

[0075] The silk support frame (purchased from Zhejiang Cathaya International Co., Ltd., warp knitting pattern; specifications: 12 knitting machine yarn (1 yarn / filament), density 21 stitches / cm, pore size 1mm) was used. 2 Immerse it in a 0.02 mol / L sodium carbonate aqueous solution and soak it at 90℃ for 60 min, changing the solution every 20 min during the soaking period. After soaking, remove it and dry it at 37℃ for 30 min for later use.

[0076] Prepare compound reagent 1 and compound reagent 2 at 37°C:

[0077] Compound reagent 1: Weigh refined fibrinogen and lyophilized human plasma cryoprecipitate in a weight ratio of 1:1, add them to physiological saline, and the total concentration is 20 g / L.

[0078] Compound reagent 2: Take 1 mL each of 500 U / mL thrombin solution and 4 mg / mL calcium gluconate aqueous solution, and mix them to obtain compound reagent 2.

[0079] The endometrial stem cell suspension and compound reagent 1 were mixed at a volume ratio of 1:2 to form a cell-fibrin gel suspension.

[0080] The silk scaffold and cell-fibrin gel suspension were mixed at a volume ratio of 1:1.2 and soaked for 6 minutes.

[0081] Add compound reagent 2 to the system at a concentration of 10% of the cell-fibrin glue suspension to promote cross-linking of the silk scaffold and fibrin, so that endometrial stem cells are evenly dispersed in the cross-linked network. After standing for 2 minutes, the composite biological dressing is obtained.

[0082] Example 3

[0083] Clean the sterile work surface with 75% ethanol. Place the diluted menstrual blood into the sterile work surface and into two EP tubes. Take a 15mL centrifuge tube, add 5mL of Ficoll meglumine lymphocyte separation medium (density 1.007g / L), add 1mL of diluted blood to the surface of the lymphocyte separation medium, and centrifuge at 2500rpm for 10min.

[0084] After centrifugation, the supernatant containing the mononuclear cell layer was aspirated and transferred to another 50 mL centrifuge tube. Physiological saline was added to a final volume of 40 mL, and the cells were resuspended. The tube was then centrifuged at 1800 rpm for 8 minutes. The supernatant was discarded, and the cells were resuspended in 10 mL of Youkang complete culture medium to adjust the cell concentration to 5 × 10⁻⁶ cells / mL. 5 / mL, inoculated into 25cm 2 Cultured in culture flasks using standard methods (37℃, 5% CO2, 95% humidity), with the medium changed for the first time after 48 hours, and then every 3 days thereafter. Once the cell confluence reaches 80%, the cells are passaged using trypsin digestion. This process is repeated until the 5th generation of daughter cells is obtained. Figure 2 As shown in the figure, the cells are spindle-shaped, small and uniform in size; they have strong refractive properties, abundant cytoplasm, and visible nuclei, indicating that the P5 generation endometrial cells still maintain vigorous proliferative capacity and good cell condition.

[0085] The silk support frame (purchased from Zhejiang Cathaya International Co., Ltd., warp knitting pattern; specifications: 12 knitting machine yarn (1 yarn / filament), density 21 stitches / cm, pore size 1mm) was used. 2 Immerse it in a 0.02 mol / L sodium carbonate aqueous solution and soak it at 100℃ for 60 min, changing the solution every 20 min during the soaking. After soaking, remove it and dry it at 37℃ for 30 min for later use.

[0086] Prepare compound reagent 1 and compound reagent 2 at 37°C:

[0087] Compound reagent 1: Weigh refined fibrinogen and lyophilized human plasma cryoprecipitate in a weight ratio of 1:1, add them to physiological saline, and the total concentration is 40 g / L.

[0088] Compound reagent 2: Take 1 mL each of 800 U / mL thrombin solution and 6 mg / mL calcium gluconate aqueous solution, and mix them to obtain compound reagent 2.

[0089] The endometrial stem cell suspension and compound reagent 1 were mixed at a volume ratio of 1:3 to form a cell-fibrin gel suspension.

[0090] The silk scaffold and cell-fibrin gel suspension were mixed at a volume ratio of 1:0.8 and soaked for 6 minutes.

[0091] Add compound reagent 2 to the system at a concentration of 12% of the cell-fibrin gel suspension to promote cross-linking of the silk scaffold and fibrin, so that endometrial stem cells are evenly dispersed in the cross-linked network. After standing for 2 minutes, the composite biological dressing is obtained.

[0092] Example 4

[0093] The only difference from Example 1 is that the calcium gluconate aqueous solution is replaced with a calcium chloride aqueous solution of the same concentration.

[0094] Experimental Example 1

[0095] Establishment of a cervical erosion model: Thirty healthy female rats (purchased from the Animal Center of Southern Medical University) were randomly divided into a blank group, a model group, and an experimental group. Phenol slurry was prepared by mixing 30 mL of phenol, 40 g of gum arabic, and 50 mL of distilled water. The phenol slurry (0.2 mL / time / 3 days) was injected into the rat vagina through a conical venous incision needle and injected for a total of 4 times. Thickening and keratinization of the vaginal epithelium, inflammatory cell infiltration, and vasodilation and congestion were observed in the rats, indicating that the model was successfully established.

[0096] The control group and model group were given physiological saline, while the experimental group was given the preparation obtained in Example 1 of this invention (administered via a conical vein incision needle into the rat vagina at a dose of 0.3 mL). Treatment was administered continuously for 7 days. On the 8th day, the animals were sacrificed, and their vaginas and uteruses were examined macroscopically and histopathologically. The results are shown in Table 1 below:

[0097] Table 1. Pathological results of rats with uterine erosion model (rats)

[0098]

[0099] Table 1 shows that the animal models of cervical erosion used in both the model group and the experimental group were successfully established. Compared to the model group, which had 7 rats with epithelial thickening and keratinization, the experimental group had 2 rats with 9 rats with inflammatory cell infiltration. The experimental group had 10 rats with vasodilation and congestion, while the experimental group had only 2. This indicates that the composite biological dressing obtained in Example 1 of this invention has a good therapeutic effect on cervical erosion.

[0100] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method of preparing a composite biological dressing, characterized by, Includes the following steps: The endometrial stem cell suspension was mixed with composite solution 1 to obtain a cell-fibrin gel suspension; A mixture was obtained by soaking a silk scaffold in a cell-fibrin gel suspension; The mixture was mixed with composite solution 2 and crosslinked to obtain a composite biological dressing; The composite solution 1 contains human fibrinogen and human plasma cryoprecipitate; The composite solution 2 contains thrombin solution and calcium salt solution; The volume ratio of the endometrial stem cell suspension to the composite solution 1 is 1:2~3; The silk scaffold further includes a pretreatment before being immersed in the cell-fibrin gel suspension, and the pretreatment includes the following steps: The silk scaffold was impregnated with a sodium carbonate aqueous solution and then dried. The concentration of the sodium carbonate aqueous solution is 0.01~0.03 mol / L; The immersion time is 40-80 minutes, and the immersion temperature is 90-100°C. The drying temperature is 35~40℃, and the drying time is 20~40min; The sodium carbonate aqueous solution is replaced every 15-25 minutes during the impregnation process. The density of the silk scaffold is 20-22 needles / cm; The silk scaffold has a pore size of 0.8-1.2 mm 2 ; The endometrial stem cell suspension was obtained by a preparation method including the following steps: Dilute the menstrual blood sample to obtain diluted blood; Diluted blood was added to the top layer of the lymphocyte separation medium, and the first centrifugation was performed to obtain the supernatant containing a mononuclear cell layer. The supernatant containing the mononuclear cell layer was mixed with physiological saline and centrifuged a second time to obtain the cell pellet. The cell pellet obtained by resuspending in complete culture medium was used to obtain the primary cell suspension; The primary cell suspension was sequentially seeded, digested, and passaged to obtain progeny endometrial stem cells. The progeny endometrial stem cells were mixed with physiological saline containing albumin to obtain an endometrial stem cell suspension. The soaking time is 5-8 minutes; The weight ratio of human fibrinogen to human plasma cryoprecipitate is 1:0.8~1.2; The concentration of the composite solution 1 is 20~40 g / L; The concentration of the thrombin solution is 500~800 U / mL; The calcium salt solution is calcium gluconate or calcium chloride; The concentration of the calcium salt solution is 4~6 mg / mL; The volume ratio of the thrombin solution to the calcium salt solution is 1:0.8~1.2; The volume ratio of the cell-fibrin gel suspension to the composite solution 2 is 8~12:1; The crosslinking time is 1~3 min; The first centrifugation speed is 2200~2800 rpm, and the first centrifugation time is 8~12 min; The second centrifugation is performed at a speed of 1500~2000 rpm for a duration of 3~8 min. The complete culture medium is Youkang complete culture medium; The concentration of the primary cell suspension is 5 x 10 4 5 x 10 5 cells / mL; The progeny endometrial stem cells are 2nd to 5th generation endometrial stem cells; The weight percentage of albumin in the saline solution is 3-8%.

2. The composite biological dressing obtained by the preparation method according to claim 1.

3. The use of the composite biological dressing according to claim 2 in the preparation of a medicament for treating inflammatory diseases of the female external genital system.

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