CN114939188A

CN114939188A - Postoperative skull base repairing membrane for transnasal butterfly approach and preparation method thereof

Info

Publication number: CN114939188A
Application number: CN202210471278.1A
Authority: CN
Inventors: 崔云; 宋天喜; 何志敏; 胡艳丽; 朱金亮; 仇志烨; 吴晶晶; 胡刚
Original assignee: Shandong Aojing Biotechnology Co ltd; Aojing Medical Technology Co ltd
Current assignee: Shandong Aojing Biotechnology Co ltd; Aojing Medical Technology Co ltd
Priority date: 2022-04-28
Filing date: 2022-04-28
Publication date: 2022-08-26
Anticipated expiration: 2042-04-28
Also published as: CN114939188B

Abstract

The invention relates to the technical field of repair membranes, in particular to a cranium base repair membrane used after transnasal butterfly approach and a preparation method thereof. A preparation method of a cranium base repair membrane after transnasal sphenoidal approach operation comprises the following steps: preparing a compact layer by adopting a macromolecule or biological protein solution; uniformly mixing the polymer or biological protein solution with the bone meal to obtain a mixture; preparing an osteogenic layer on the surface of the dense layer using the mixture; and preparing a sealing layer on one surface of the osteogenic layer far away from the compact layer by adopting the polymer or biological protein solution to obtain the skull base repairing membrane. The embodiment of the invention provides a skull base repairing film after a transnasal butterfly approach operation and a preparation method thereof, which can effectively repair a skull base after the transnasal butterfly approach operation.

Description

Postoperative skull base repairing membrane for transnasal butterfly approach and preparation method thereof

Technical Field

The invention relates to the technical field of repair membranes, in particular to a cranium base repair membrane used after transnasal butterfly approach and a preparation method thereof.

Background

With the development of microscopic and endoscopic techniques, transnasal butterfly approach can be used for the treatment of trauma of anterior basis cranii, tumor, saddle area occupation and pathological changes of three ventricles of brain and the middle and upper slopes. Neurosurgeons often choose to open the sphenoid sinus, or even a portion of the ethmoid sinus, and abrade bone from the bottom of the saddle to expose the diseased tissue that needs to be excised. The saddle bone acts as an important load-bearing structure, and once opened, the brain contents and cerebrospinal fluid (CSF) are easily leaked into the nasal cavity due to gravity. At the moment, the cranial cavity is not a closed space, but is communicated with the external environment, and needs to be repaired in time in the operation; otherwise, CSF leakage, intracranial pneumatosis, intracranial infection, cerebral expansion and other complications are easily caused, great risks are brought to the operation, and even death is caused in serious cases.

In the related art, the closure method of the skull base defect comprises autologous fat transplantation, dural suture, fascia transplantation and nasal septum vascular flap combined repair by using a rigid material support. However, adipose tissue, as a repair material, may affect the reliability of repair due to liquefaction of fat; the dura mater is used as a repair material, and the repair capability is poor, so that CSF (CSF) is easy to leak again after operation; the fascia is used as a repairing material, and needs a patient to lie in bed for a long time after operation, so long-term complications caused by lying in bed are easy to cause; the nasal septum vascular flap uses a rigid material support in combination with the repair material, which may cause infection or interfere with the formation of epithelium, and if the tumor recurs again, it becomes very difficult for the rigid material support to re-enter. That is, none of the above related arts can effectively repair the skull base after the transnasal sphenoidal approach.

Therefore, in order to overcome the defects, a skull base repairing film after a transnasal butterfly approach and a preparation method thereof are urgently needed.

Disclosure of Invention

The embodiment of the invention provides a skull base repairing film after a transnasal butterfly approach operation and a preparation method thereof, which can effectively repair a skull base after the transnasal butterfly approach operation.

In a first aspect, an embodiment of the present invention provides a method for preparing a cranium base repair membrane after a transnasal sphenoidal approach, including:

preparing a compact layer by adopting a macromolecule or biological protein solution;

uniformly mixing the polymer or biological protein solution with the bone meal to obtain a mixture;

preparing an osteogenic layer on the surface of the dense layer using the mixture;

and preparing a sealing layer on the surface of the osteogenesis layer far away from the compact layer by adopting the polymer or biological protein solution to obtain the skull base repairing film.

In one possible design, the preparing the dense layer using a polymer or bioprotein solution includes:

and drying the polymer or biological protein solution to obtain the compact layer.

In one possible design, the preparing an osteogenic layer at the surface of the dense layer using the mixture includes:

coating the mixture on the surface of the dense layer;

and performing freeze-drying treatment on the mixture coated on the surface of the compact layer to obtain an osteogenic layer.

In a possible design, after the preparing an osteogenic layer on the surface of the dense layer with the mixture and before the preparing a sealing layer on the side of the osteogenic layer away from the dense layer with the polymer or bioprotein solution, the method further comprises:

and carrying out cross-linking treatment on the compact layer and the osteogenesis layer.

In one possible design, the preparing a sealing layer on the side of the osteogenic layer far from the dense layer with the polymer or biological protein solution includes:

coating the high-molecular or biological protein solution on one side of the osteogenesis layer far away from the compact layer;

and carrying out freeze-drying treatment on the polymer or biological protein solution coated on the osteogenic layer to obtain the sealing layer.

In one possible design, the mass ratio of the solute in the polymer or biological protein solution to the bone meal in the mixture is (0.5-1): 99.

In one possible design, the thickness of the compact layer is 0.1-0.2 mm;

and/or the presence of a gas in the gas,

the thickness of the osteogenic layer is 0.2-0.3 mm;

and/or the presence of a gas in the gas,

the thickness of the sealing layer is 0.2-5 mm.

In one possible design, the polymer or bio-protein solution comprises at least one of collagen gel, silk fibroin, polycaprolactone, polyurethane, silicone rubber, polyester fiber, polyvinylpyrrolidone, polyetheretherketone, polymethylmethacrylate, polyvinyl alcohol, polylactic acid, polyethylene, and gelatin.

In one possible design, the bone meal includes at least one of mineralized collagen particles, sintered hydroxyapatite, and β -tricalcium phosphate.

In a second aspect, an embodiment of the present invention provides a cranium fundus repair membrane after a transnasal sphenoid approach, which is prepared by the preparation method described in any one of the first aspect, wherein the compact layer is the side facing the outside of the brain, the sealing layer is the side facing the inside of the brain, and the cranium fundus repair membrane is used for repairing cranium fundus defects after the transnasal sphenoid approach.

Compared with the prior art, the invention at least has the following beneficial effects:

in some embodiments of the invention, a dense layer is prepared by using a polymer or biological protein solution, and the dense layer has high strength and a protection function; the polymer or biological protein solution and the bone powder are uniformly mixed, the bone powder is bonded together by the polymer or biological protein solution to obtain a mixture with certain plasticity, the mixture is coated on the surface of the compact layer to obtain an osteogenic layer with certain plasticity, and the osteogenic layer with plasticity can be subjected to micro-deformation under external force to adapt to the external force, so that the osteogenic layer can be matched with irregular skull base damage; the sealing layer is prepared on the surface, far away from the compact layer, of the osteogenic layer by adopting a macromolecule or biological protein solution, the sealing layer has the property of water absorption and swelling, once cerebrospinal fluid seeps out, the sealing layer swells after absorbing the cerebrospinal fluid, and the swollen sealing layer has pure water pressure, so that the skull base damage can be sealed, and the cerebrospinal fluid leakage is prevented. Therefore, the skull base repairing film consisting of the compact layer, the osteogenesis layer and the sealing layer has the functions of protecting, supporting, promoting the growth of bone tissues and sealing cerebrospinal fluid, and can effectively repair skull base defects after the transnasal butterfly approach.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.

FIG. 1 is a flow chart of a method for preparing a cranium base repair membrane after a transnasal sphenoidal approach according to an embodiment of the present invention;

FIG. 2 is a flow chart of another method for preparing a cranium base repair membrane after a transnasal butterfly approach according to an embodiment of the present invention;

FIG. 3 is a sectional structure diagram of a skull base repairing membrane after a transnasal butterfly approach according to an embodiment of the present invention;

in the figure:

1-a dense layer;

2-an osteogenic layer;

3-sealing layer;

10-a mould.

Detailed Description

To make the objects, technical solutions and advantages of the embodiments of the present invention clearer and more complete, the technical solutions in the embodiments of the present invention will be described below with reference to the drawings in the embodiments of the present invention, it is obvious that the described embodiments are some, but not all embodiments of the present invention, and based on the embodiments of the present invention, all other embodiments obtained by a person of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

In the description of the embodiments of the present invention, unless explicitly specified or limited otherwise, the terms "first", "second", and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance; the term "plurality" means two or more unless specified or indicated otherwise; the terms "connected," "fixed," and the like are to be construed broadly and may, for example, be fixedly connected, detachably connected, integrally connected, or electrically connected; may be directly connected or indirectly connected through an intermediate. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.

In the description of the present invention, it should be understood that the terms "upper" and "lower" as used in the description of the embodiments of the present invention are used in the angle shown in the drawings, and should not be construed as limiting the embodiments of the present invention. In addition, in this context, it will also be understood that when an element is referred to as being "on" or "under" another element, it can be directly on "or" under "the other element or be indirectly on" or "under" the other element via an intermediate element.

As shown in fig. 1, an embodiment of the present invention provides a method for preparing a cranium base repair membrane after a transnasal sphenoidal approach, including:

step 100, preparing a compact layer by adopting a macromolecule or biological protein solution;

step 102, uniformly mixing a polymer or biological protein solution and bone meal to obtain a mixture;

104, preparing an osteogenic layer on the surface of the compact layer by adopting the mixture;

and step 106, preparing a sealing layer on the surface, far away from the compact layer, of the osteogenic layer by adopting a high molecular or biological protein solution to obtain the skull base repairing film.

In some embodiments of the invention, a high molecular or biological protein solution is used to prepare the dense layer, which has high strength and a protective function; the method comprises the following steps of uniformly mixing a polymer or biological protein solution with bone meal, bonding the bone meal together by the polymer or biological protein solution to obtain a mixture with certain plasticity, coating the mixture on the surface of a compact layer to obtain an osteogenic layer with certain plasticity, wherein the osteogenic layer with plasticity can be subjected to micro-deformation under the action of external force so as to adapt to the external force, so that the osteogenic layer can be matched with irregular skull base damage; the sealing layer is prepared on the surface, far away from the dense layer, of the osteogenic layer by adopting a polymer or biological protein solution, the sealing layer has the property of water absorption and swelling, once cerebrospinal fluid seeps out, the sealing layer swells after absorbing the cerebrospinal fluid, and the swollen sealing layer has pure water pressure, can seal skull base damage and prevent leakage of the cerebrospinal fluid. Therefore, the skull base repairing film consisting of the compact layer, the osteogenesis layer and the sealing layer has the functions of protecting, supporting, promoting the growth of bone tissues and sealing cerebrospinal fluid, and can effectively repair skull base defects after the transnasal butterfly approach.

The solute mass fraction of the polymer or bioprotein solution is 1 to 1.5%.

It should be noted that the sealing layer has the effect similar to fat, but compared with fat, the sealing layer does not slip, does not liquefy, and is more stable and durable.

In some embodiments of the present invention, the dense layer, the osteogenic layer and the sealing layer may be prepared using a mold, for example, a polymer or bioprotein solution may be coated on a bottom layer of the mold, an upper layer of the mold is covered and dried to obtain a dense layer uniformly spread, the dense layer is placed on the bottom layer of the mold after obtaining the dense layer, the mixture is coated on the dense layer, the upper layer of the mold is covered and dried to obtain an osteogenic layer uniformly bonded to the dense layer and spread, the dense layer and the osteogenic layer are bonded to each other and placed on the bottom layer of the mold, wherein the dense layer is attached to the bottom layer of the mold, the polymer or bioprotein solution is coated on the osteogenic layer, and the upper layer of the mold is covered and dried to obtain a sealing layer uniformly bonded to the osteogenic layer and spread.

The drying treatment may be a drying treatment or a freeze-drying treatment.

In some embodiments of the present invention, the distance between the bottom mold layer and the top mold layer is adjustable, and the dense layer, the osteogenic layer and the sealing layer are prepared in different thicknesses by controlling the distance between the bottom mold layer and the top mold layer.

In order to better fit the skull base damage after the transnasal butterfly approach, a mould with a curved surface inside can be selected to prepare a compact layer, an osteogenic layer and a sealing layer with the curved surface.

In some embodiments of the invention, the material from which the mold is made comprises polytetrafluoroethylene and a metal.

In some embodiments of the invention, when the skull base repairing film provided by the invention is used for repairing skull base damage, biological glue can be coated on the edge of the skull base repairing film so as to enable the skull base repairing film and the skull base to be tightly bonded.

According to some preferred embodiments, the dense layer is prepared using a solution of macromolecules or bioproteins, comprising:

In some embodiments of the invention, the polymer or biological protein solution is dried, so that the prepared compact layer is more compact and has stronger mechanical strength, wherein the drying temperature is 40 ℃, and the drying time is 12-24 h.

According to some preferred embodiments, the osteogenic layer is prepared at the surface of the dense layer using a mixture comprising:

coating the mixture on the surface of the compact layer;

and performing freeze-drying treatment on the mixture coated on the surface of the compact layer to obtain the osteogenic layer.

In some embodiments of the present invention, the freeze-drying process can fix the shape of the osteogenic layer, increase the hardness, increase the pore size of the microporous structure in the osteogenic layer, increase the support strength of the osteogenic layer, and, in addition, bond the dense layer and the osteogenic layer together because both layers contain a polymer or a bioprotein.

In some embodiments of the present invention, the lyophilization process comprises a prefreezing stage, a first sublimation stage, a second sublimation stage, and a cooling stage, wherein the process conditions of each stage are as follows:

a pre-freezing stage: the target temperature is-12 to-8 ℃, the speed is 3 to 4.0 ℃/min, and the constant temperature duration is 280 to 320 min;

a first sublimation stage: vacuumizing, wherein the air is mixed at 90-110 Pa, the target temperature is-4 to-2 ℃, the speed is 0.6-0.8 ℃/min, and the constant temperature duration is 1300-1340 min;

and in the second sublimation stage, vacuumizing is performed, and 90-110 Pa of air is aerated, and the method comprises five temperature rising steps which are respectively as follows:

the temperature is between-1 and 1 ℃, the speed is 0.2 to 0.3 ℃/min, and the constant temperature duration is 110 to 130 min;

the temperature is 8-12 ℃, the speed is 1.0-1.2 ℃/min, and the constant temperature duration is 110-130 min;

the temperature is 18-22 ℃, the speed is 1.0-1.2 ℃/min, and the constant temperature duration is 110-130 min;

the temperature is 28-32 ℃, the speed is 1.0-1.2 ℃/min, and the constant temperature duration is 110-130 min;

38-42 ℃, the speed is 1.0-1.2 ℃/min, the constant temperature duration is as follows: judging the end point every 10 minutes until the end point is qualified; the end point is judged to be less than or equal to 0.9Pa/10 min;

and (3) cooling: cooling to room temperature at a rate of 1.4-1.6 deg.C/min.

According to some preferred embodiments, after preparing the osteogenic layer on the surface of the dense layer using the mixture and before preparing the sealant layer on the side of the osteogenic layer away from the dense layer using the polymer or bioprotein solution, the method further comprises:

and carrying out crosslinking treatment on the compact layer and the osteogenesis layer.

In some embodiments of the invention, the crosslinking treatment can prevent the compact layer and the osteogenic layer from swelling when meeting water, prevent the dense layer and the osteogenic layer from delaminating due to swelling, increase the stability of the skull base repair membrane and increase the degradation period of the skull base repair membrane.

In some embodiments of the invention, the process of cross-linking treatment comprises:

a. preparing a cross-linking agent solution: measuring edible alcohol with a certain volume by using a measuring cylinder, transferring the edible alcohol into a reaction kettle, transferring a glutaraldehyde solution with a certain volume by using a liquid transfer gun (the mass fraction of glutaraldehyde is 50%), and mixing the glutaraldehyde solution with the liquid transfer gun according to the volume ratio of the edible alcohol to the glutaraldehyde solution of 100: preparing alcohol solution of glutaral at a ratio of 0.05;

b. soaking the materials in the crosslinking solution, wherein the stirring speed of the reaction kettle is 20-30r/min, and the uppermost layer of the materials can rotate with stirring, and crosslinking is carried out for 48 h. (note: after the material adsorbed the crosslinking agent and sunk during crosslinking, if the material was below the paddle, the paddle did not contact the material, the stirring speed was reduced to 10r/min.

c. Water washing step

(1) Taking out the cross-linked material from the cross-linking agent solution;

(2) the centrifuge was loaded with filter bags and centrifuged once (5 seconds); the rotating speed of the centrifugal machine is 3000r/min by default;

(3) soaking in 3% hydrogen peroxide solution at a ratio of 1:80 for 42 + -1 h, i.e. 3% hydrogen peroxide solution volume (ml) × 80 times the product mass (g), completely soaking the product in hydrogen peroxide solution.

(4) Carrying out ultrasonic treatment on purified water for 30min, and centrifuging once (5s) after ultrasonic treatment; repeating for 2 times;

(5) ultrasonic treating with edible alcohol for 5min, with the volume of edible alcohol based on the principle of immersing the product. After the ultrasonic treatment is finished, the centrifugal operation is carried out once (5 seconds);

d. vacuum drying

And (3) putting the centrifuged material into a vacuum drying oven for vacuum drying, wherein the vacuum degree is not higher than-0.08 Mpa, the temperature is set to be 50 ℃, and the drying is carried out for at least 24 hours.

According to some preferred embodiments, the sealant layer is prepared from a polymer or bioprotein solution on a side of the osteogenic layer remote from the dense layer, comprising:

coating a macromolecule or biological protein solution on one surface of the osteogenic layer far away from the compact layer;

and (3) carrying out freeze-drying treatment on the polymer or biological protein solution coated on the osteogenesis layer to obtain the sealing layer.

In some embodiments of the invention, the lyophilization process may also bond the dense layer, the osteogenic layer, and the sealing layer together. The process of the freeze-drying treatment is the same as above, and is not described herein.

The sealing layer is prepared by drying the dense layer, the obtained dense layer is in a dense sheet shape, and the sealing layer is prepared by freeze-drying, and the obtained sealing layer is in a loose sponge shape.

According to some preferred embodiments, the mass ratio of solute to bone meal in the polymer or bioprotein solution in the mixture is (0.5-1): 99 (e.g., may be 0.5:99, 0.6:99, 0.7:99, 0.8:99, 0.9:99, or 1: 99).

In some embodiments of the invention, the mass ratio of the solute of the polymer or biological protein solution to the bone meal is (0.5-1): 99, wherein the polymer or biological protein solution has a binding effect and can bind the bone meal together to form a mixture with plasticity and viscosity, and if the mass ratio of the solute of the polymer or biological protein solution to the bone meal is less than 0.5:99, the bone meal cannot be completely bound together; if the mass ratio of the solute of the polymer or biological protein solution to the bone meal is more than 1:99, the pore diameter and porosity of the mixture after freeze-drying are higher, the spongy feeling is stronger, and the supporting effect is weakened.

According to some preferred embodiments, the dense layer has a thickness of 0.1 to 0.2mm (e.g., may be 0.1mm, 0.11 mm, 0.12 mm, 0.13 mm, 0.14mm, 0.15mm, 0.16mm, 0.17mm, 0.18mm, 0.19mm, or 0.2 mm);

and/or the presence of a gas in the gas,

the thickness of the osteogenic layer is 0.2-0.3 mm (for example, 0.2mm, 0.21mm, 0.22mm, 0.23mm, 0.24mm, 0.25mm, 0.26mm, 0.27mm, 0.28mm, 0.29mm or 3mm can be used);

and/or the presence of a gas in the gas,

the sealing layer has a thickness of 0.2 to 5mm (for example, 0.2mm, 0.5mm, 1mm, 1.5mm, 2mm, 2.5mm, 3mm, 3.5mm, 4mm, 4.5mm, or 5 mm).

In some embodiments of the invention, the dense layer, the osteogenic layer, and the sealant layer are all relatively thin in thickness to meet the need for repairing skull base damage, and the sealant layer is relatively thick relative to the other layers, with the thicker sealant layer further increasing the sealing effect of the swollen sealant layer.

According to some preferred embodiments, the polymer or bioprotein solution includes at least one of collagen gel, silk fibroin, polycaprolactone, polyurethane, silicone rubber, polyester fiber, polyvinylpyrrolidone, polyetheretherketone, polymethylmethacrylate, polyvinylalcohol, polylactic acid, polyethylene, and gelatin.

In some embodiments of the invention, collagen gel, silk fibroin, polycaprolactone, polyurethane, silicone rubber, polyester fiber, polyvinylpyrrolidone, polyetheretherketone, polymethylmethacrylate, polyvinyl alcohol, polylactic acid, polyethylene, and gelatin are well-established in application technology and have good biocompatibility. In some embodiments of the present invention, a method of preparing a collagen sol comprises:

(1) removing redundant fascia, fat, muscle and the like on the bovine achilles tendon, washing with tap water, neatly arranging in a freezing box, freezing at-20 ℃ for at least 12 h;

(2) cutting frozen ox Achilles tendon into slices of about 1mm, and washing with a filter screen until the liquid is clear;

(3) enzymolysis: carrying out enzymolysis on the cleaned bovine achilles tendon slices, and fully stirring for not less than 72 h; wherein the mass ratio of the enzymolysis liquid to the bovine achilles tendon is 130: 1, the volume ratio of purified water to acetic acid in the enzymolysis liquid is 25: 1, the mass ratio of the purified water to the pepsin is 15: 1.

(4) salting out: and centrifuging the solution after enzymolysis, taking supernatant, adding the supernatant into a sodium chloride solution to separate out white flocculent collagen, filtering and washing, and draining water.

(5) And (3) dialysis: filling the salted-out material into a dialysis bag, wherein the volume of the salted-out material is about 1/3 of the dialysis bag; placing the dialysis bag in a dialysate of 0.057mol/L acetic acid solution for 6 days, surreptitiously heating to 10-20 deg.C, and replacing the dialysate every 3 days; then placing the dialysis bag in 0.00057mol/L acetic acid solution for dialysis for 5 days, wherein the dialysis temperature is 10-20 ℃, and the dialysate is replaced every 1 day; dialyzing in 0.0000057mol/L acetic acid solution at pH of 5.0-5.5 from day 12 at 10-20 deg.C, and changing the dialysate once a day as required.

(6) The collagen gel was removed and tested for solids content.

In some embodiments of the invention, a method of preparing silk fibroin comprises:

adding mulberry silk into 0.02mol/L Na at 100 deg.C ₂ CO ₃ Boiling the solution twice, each time for 30min,washing with tap water and distilled water for several times to completely remove sericin, and air drying for later use. Dissolving degummed silk in 9.3mol/L LiBr solution, dissolving for 4h at 60 ℃, dialyzing for 3d (the molecular weight cutoff is 12000-14000) to obtain a fibroin solution with the concentration of about 8%, centrifuging for 15min at 12000rpm, concentrating a low-concentration PEG solution for a certain time to obtain a high-concentration fibroin solution, centrifuging for 15min at 12000rpm, centrifuging and measuring the concentration by a dry constant weight method. Storing in a refrigerator at 4 deg.C.

According to some preferred embodiments, the bone meal comprises at least one of mineralized collagen particles, sintered hydroxyapatite, and β -tricalcium phosphate.

In some embodiments of the present invention, mineralized collagen particles, sintered hydroxyapatite, and β -tricalcium phosphate are well-established in the art, are biocompatible, have a microporous structure, and are capable of promoting bone tissue growth.

In some embodiments of the invention, the method of preparing mineralized collagen particles comprises:

step 1, dissolving collagen in any one of hydrochloric acid, nitric acid or acetic acid to prepare an acid solution of the collagen, wherein the concentration of the collagen is 0.01-0.2 g/ml;

step 2, dropwise adding a calcium salt solution into the acid solution of the collagen, wherein the addition amount of calcium ions is 0.1-2 mol per gram of the collagen;

step 3, dropwise adding a phosphoric acid solution into the solution obtained in the step 2, wherein the molar ratio of the addition amount of phosphate ions to the addition amount of calcium ions in the step 2 is 1/1-2/1;

step 4, dropwise adding a NaOH solution into the solution obtained in the step 3 to form a mixed solution, and adjusting the pH value to 6-8;

step 5, standing the mixed solution obtained in the step 4 for 4-12 hours, centrifuging at the speed of 3000-6000 r/min to obtain a precipitate, and performing air blast drying at the temperature of 50-70 ℃ for 24-72 hours to obtain mineralized collagen particles;

step 6, a crushing procedure, namely putting the mineralized collagen particles into a small crusher, and crushing the mineralized collagen particles into fine particles for 0.5 s;

step 7, a screening process, namely screening the crushed materials by using a vibrating screen or a stainless steel screen, wherein the mesh number of an upper layer screen is about 18 meshes (the particle size of the corresponding material is less than 1000 mu m), and the mesh number of a lower layer screen is about 60 meshes for interception (the particle size of the corresponding material is more than 250 mu m); the particles retained by the 60 mesh screen were collected and the 18 mesh retained material was subjected to the comminution procedure, which was repeated.

As shown in fig. 2, an embodiment of the present invention further provides another method for preparing a skull base repair membrane after a transnasal butterfly approach, including:

step 200, drying the polymer or biological protein solution to obtain a compact layer;

202, uniformly mixing a polymer or biological protein solution and bone meal to obtain a mixture;

step 204, coating the mixture on the surface of the compact layer;

step 206, performing freeze-drying treatment on the mixture coated on the surface of the compact layer to obtain an osteogenic layer;

step 208, performing cross-linking treatment on the compact layer and the osteogenesis layer;

step 210, coating a macromolecule or biological protein solution on one surface of the osteogenic layer, which is far away from the compact layer;

and step 212, carrying out freeze-drying treatment on the polymer or biological protein solution coated on the osteogenic layer to obtain a sealing layer.

As shown in fig. 3, an embodiment of the present invention further provides a cranium fundus repair membrane after transnasal sphenoid approach, which is prepared by any one of the above preparation methods, wherein the compact layer 1 is the side facing the outside of the brain, the sealing layer 3 is the side facing the inside of the brain, and the cranium fundus repair membrane is used for repairing cranium fundus defects after transnasal sphenoid approach.

The basis is that the skull base repairing membrane after the nasal butterfly approach is prepared by the preparation method of the skull base repairing membrane after the nasal butterfly approach, and the two are based on the same invention concept, so the same technical effect can be obtained.

In order to more clearly illustrate the technical scheme and advantages of the invention, a craniofacial repair film after transnasal sphenoidal approach and a preparation method thereof are described in detail through a plurality of embodiments.

Example 1

Drying the collagen gel to obtain a compact layer;

uniformly mixing the collagen gel and the sintered hydroxyapatite to obtain a mixture, wherein the mass ratio of solute in the collagen gel to the sintered hydroxyapatite is 0.5: 99;

coating the mixture on the surface of the compact layer;

freeze-drying the mixture coated on the surface of the compact layer to obtain an osteogenic layer;

performing cross-linking treatment on the compact layer and the osteogenic layer;

coating collagen gel on one side of the osteogenic layer far away from the compact layer;

freeze-drying the collagen gel coated on the osteogenesis layer to obtain a sealing layer;

wherein, the thickness of the compact layer is 0.1mm, the thickness of the osteogenesis layer is 0.2mm, and the thickness of the sealing layer is 0.2 mm.

Example 2

Drying the silk fibroin solution to obtain a compact layer;

uniformly mixing silk fibroin solution and mineralized collagen particles to obtain a mixture, wherein the mass ratio of solute in the silk fibroin solution to the mineralized collagen particles is 1: 99;

coating the mixture on the surface of the compact layer;

coating the silk fibroin solution on one surface of the osteogenesis layer far away from the compact layer;

freeze-drying the silk fibroin solution coated on the osteogenesis layer to obtain a sealing layer;

wherein, the thickness of the compact layer is 0.2mm, the thickness of the osteogenesis layer is 0.3mm, and the thickness of the sealing layer is 5 mm.

Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.

Claims

1. A preparation method of a craniofacial repair film after transnasal butterfly approach is characterized by comprising the following steps:

2. The method of claim 1, wherein the step of preparing the dense layer from the solution of the polymer or the bioprotein comprises:

3. The method of claim 1, wherein the preparing an osteogenic layer on the surface of the dense layer using the mixture comprises:

coating the mixture on the surface of the dense layer;

4. The method for preparing the bone-forming layer of the dental implant according to claim 1, wherein after preparing the bone-forming layer on the surface of the compact layer by using the mixture and before preparing the sealing layer on the side of the bone-forming layer away from the compact layer by using the polymer or bioprotein solution, the method further comprises the following steps:

5. The method for preparing the bone-forming layer of the bone-forming device of the invention according to claim 1, wherein the step of preparing the sealing layer on the side of the bone-forming layer away from the dense layer by using the polymer or bioprotein solution comprises the following steps:

6. The method according to claim 1, wherein the mass ratio of the solute in the polymer or bioprotein solution to the bone meal in the mixture is (0.5-1): 99.

7. The production method according to claim 1,

the thickness of the compact layer is 0.1-0.2 mm;

and/or the presence of a gas in the gas,

the thickness of the osteogenic layer is 0.2-0.3 mm;

and/or the presence of a gas in the gas,

the thickness of the sealing layer is 0.2-5 mm.

8. The method of claim 1, wherein the polymer or bio-protein solution comprises at least one of collagen gel, silk fibroin, polycaprolactone, polyurethane, silicone rubber, polyester fiber, polyvinylpyrrolidone, polyetheretherketone, polymethylmethacrylate, polyvinyl alcohol, polylactic acid, polyethylene, and gelatin.

9. The method of claim 1, wherein the bone meal comprises at least one of mineralized collagen particles, sintered hydroxyapatite, and β -tricalcium phosphate.

10. A cranium base repair membrane after a transnasal sphenoid approach operation, which is prepared by the preparation method of any one of claims 1 to 9, wherein the compact layer is the side facing the outside of the brain, the sealing layer is the side facing the inside of the brain, and the cranium base repair membrane is used for repairing cranium base defects after the transnasal sphenoid approach operation.