CN112915256B

CN112915256B - Method for preparing human dura mater repair material with imitated ECM (extracellular matrix) components and structure by long fiber network framework preforming method

Info

Publication number: CN112915256B
Application number: CN202110028425.3A
Authority: CN
Inventors: 牟永告; 赵光磊
Original assignee: South China University of Technology SCUT; Sun Yat Sen University Cancer Center
Current assignee: South China University of Technology SCUT; Sun Yat Sen University Cancer Center
Priority date: 2021-01-11
Filing date: 2021-01-11
Publication date: 2022-09-16
Anticipated expiration: 2041-01-11
Also published as: CN112915256A

Abstract

The invention discloses a method for preparing an ECM (extracellular matrix) -simulated human dura mater repair material with a structure by a long fiber network skeleton preforming method. The method comprises the following steps: firstly, adopting swelling liquid to pretreat chitosan fiber, adopting Na ₂ CO ₃ The solution carries out collagen removal treatment on the silk fiber; then cleaning chitosan fiber and silk fiber, and pulping (grinding); diluting the beaten (ground) fiber slurry into fiber suspension, and respectively preparing thin-layer long fiber interwoven framework materials and short fiber interwoven base materials from the fiber slurry with different lengths; compounding the two materials into a chitosan-silk paper base material; finally, the human dura mater repair material can be obtained after non-chemical cross-linking physical dipping and coating treatment. The material prepared by the method has good toughness and sewability, is easy to industrialize and has good application prospect.

Description

Method for preparing human dura mater repair material with imitated ECM (extracellular matrix) components and structure by long fiber network framework preforming method

Technical Field

The invention relates to the field of human tissue engineering repair materials, in particular to a method for preparing an ECM (extracellular matrix) -imitated human dura mater repair material with a structure by a long fiber network framework preforming method.

Background

The dura mater of a human body is a thick and tough double-layer membrane, the outer layer of the dura mater is loosely attached to a skull cap, and the thick and tough inner layer of the dura mater is connected with the dura mater at the position of a large hole of an occipital bone, so that the dura mater is an important barrier for protecting brain tissues, and has important significance for maintaining the structure and the function of a brain nervous system. Craniocerebral trauma and operations can cause dural membrane defects, and dural membrane reconstruction has reached a consensus in cranial nerve surgery, and becomes an important and basic operation step in brain surgery. The defected dura mater is reconstructed by cranial nerve surgery to seal the subdural cavity, and the integrity and compatibility of the reconstructed dura mater are closely related to the postoperative recovery of a wound (patient). The healing of the wound can be accelerated after the dura mater is repaired by the operation, and the cerebrospinal fluid leakage and the intracranial infection are obviously reduced or prevented; reducing the diffusion of seepage from the scalp or muscle layer into the subarachnoid space to stimulate brain tissue to cause inflammation or other meningeal stimulation symptoms; reduce the complication of epilepsy caused by the adhesion of scalp and brain tissue. The traditional dura mater reconstruction material is mainly taken from autologous tissues, has limited material sources and aggravates the pain of patients, and the artificial dura mater repair material has been widely accepted in recent years. The tissue repair materials commonly used at present mainly include the following types: allogenic meninges repairing material, xenogenic meninges repairing material, synthetic meninges repairing material and natural meninges repairing material. The allogenic meninges repairing material is taken from human skin, so that the allogenic meninges repairing material is not easy to take and is high in price, and has some ethical problems. The xenogenic dura mater repair material is widely applied clinically in recent years, the material mainly adopts bovine pericardium, ovine pericardium, porcine peritoneal membrane, porcine small intestine mucosa and the like, and the current commercialized products are expensive and have the risks of causing infection and spreading viruses. The biocompatibility of the synthetic material is relatively poor due to lack of an ECM three-dimensional structure, and the reaction of local tissues can cause stimulation to brain tissues, form scar or wrapping reaction, meningitis symptoms or bleeding and other serious complications. The existing meninges repairing material taking collagen as a main raw material still has the problems of quick degradation, poor mechanical property and the like.

Extracellular matrix (ECM) plays an important role in connecting tissue structures, regulating tissue generation and physiological activities of cells. The intercellular space is large in connective tissue and is filled with much ECM, so it determines the properties of connective tissue. ECM is generally considered to be composed of various macromolecular Glycosaminoglycans (Glycosaminoglycans), proteoglycans (proteoglycans), structural proteins (e.g., collagen and elastin), and adhesion proteins (also called fibronectin), which form a complex having a complex three-dimensional lattice structure with collagen fibers and proteoglycans as basic skeletons, and are linked to or attached to a cell surface receptor through adhesion proteins or the like. With the intensive research on the ECM, the ECM can combine various growth factors and hormones, can affect the cell morphology, movement and differentiation, and further regulate the functions of tissues and organs, and is an essential part for completing the physiological functions of cells. Thus, ECM is considered to be the "soil" upon which cells and tissues live, and if ECM structural tissue is defective in the human body, repair and regeneration of body tissues is severely affected. The biomedical material with a three-dimensional (3D) microstructure similar to human tissue Extracellular matrix (ECM) is provided, when the biomedical material is implanted into a human body as a tissue reconstruction and regeneration medical instrument, the three-dimensional tissue engineering scaffold structure provided by the biomedical material can better simulate a cell microenvironment, is beneficial to cell adhesion, differentiation, migration and Extracellular matrix deposition, and is gradually degraded along with time due to the degradability of the material, and meanwhile, new vessels and nerves grow along the deposition matrix to gradually remodel or regenerate host tissue defect parts.

Chitosan (Chitosan) is also called soluble chitin, Chitosan and the like, has a chemical name of 2-amino-beta-1, 4-glucan, is a natural cationic polysaccharide obtained after deacetylation of chitin, and has excellent characteristics of good biocompatibility, degradability, hygroscopicity, no toxicity, antibacterial property, fiber forming property, film forming property, certain antibacterial property, tumor resistance and the like. The chitosan is widely distributed in the nature, is the second largest biological resource next to plant fibers on the earth, wherein the generation amount of marine organisms is more than 100 hundred million t, and can be regarded as an inexhaustible biological resource. Has the name of omnipotent polysaccharide, and is a cheap and easily available raw material. At present, chitosan is widely applied in the fields of food, cosmetics, wastewater treatment, heavy metal recovery, biology, medicine, textile, printing and dyeing, paper making and the like. In recent years, as people fully utilize the biological functionality, biocompatibility, low toxicity, biodegradability and edibility of chitosan, the markets of chitosan in the aspects of biology, medicine and medicine are continuously expanded, and the application of chitosan is greatly developed. The biological applications are mainly as carriers of immobilized enzymes, microcarriers for cell culture, plant physiological agents, biological membranes for producing substances with physiological activity and the like. Medical applications are mainly the promotion of blood coagulation and wound healing, as slow release matrices for drugs, as artificial tissues and organs, etc. In industrial application, chitosan has good film forming property, selectivity and permeability, and can be used for preparing reverse osmosis membranes, ultrafiltration membranes or microporous filtration membranes, food preservative films and the like. The silk is an animal protein long fiber existing in nature and is composed of sericin at the outer layer and fibroin at the inner layer, wherein the fibroin accounts for 70-80% of the total mass of the silk. The silk fiber is used as a medical silk suture line because of the excellent performance of the silk fiber, and is very early applied to the medical industry. In recent years, silk fibroin is widely applied to industries such as textile, cosmetics and the like as protein fiber, and is also applied to the fields of biological tissue scaffold materials, drug slow release, biosensing and the like as functional fiber.

Chinese patent CN107354800A discloses a method for preparing a human tissue substitute material with ECM structure based on paper making method. The method comprises the steps of treating a fiber material pretreated by water, dilute acid or dilute alkali by adopting a pulping or beating method, diluting the obtained fiber slurry into a suspension, adding a dispersing agent and a reinforcing agent, fully mixing and dispersing, dewatering on a papermaking former, drying, and coating by adopting a chemical crosslinking coating liquid to prepare the human tissue substitute material with the ECM structure. The method has mature production process and equipment, and the prepared human substitute material has good biocompatibility and mechanical strength, but the material has relatively low suturability and toughness due to the adoption of the fiber raw material with the length of common fiber raw material (about 3mm) in the preparation process of the method, and can not fully meet the requirements of surgical operations such as suturing in the process of dura mater repair surgery. In addition, the biocompatibility of the material can be affected by the subsequent chemical crosslinking coating process, such as reactive crosslinking coating.

The Chinese invention patent CN108815578A discloses an artificial biological dura mater which is prepared by an electrostatic spinning method based on bacterial cellulose and collagen as raw materials and has good biocompatibility and good suture performance under the condition of not adding any cross-linking agent. The artificial biological dura mater related to the patent application is characterized by having good suture performance, low endotoxin and clinical use convenience, thus not only mechanically protecting tissues of brain parenchyma parts from invasion interference of other peripheral tissues, but also effectively preventing cerebrospinal fluid from leaking due to no pore structure, thereby meeting the requirement of surgical suture in the clinical use process. However, the artificial dura mater prepared by the method of electrostatic spinning based on the bacterial cellulose has certain limitation in neurosurgery because no enzyme capable of degrading the bacterial cellulose exists in a human body, the yield of the artificial dura mater prepared by the method of electrostatic spinning is low, and the spinning solution relates to a harmful organic solvent, and is not easy to industrialize.

The Chinese invention patent CN111420123A discloses a preparation method of an anti-adhesion double-layer dura mater patch, which comprises a compact layer (chitosan-bacterial cellulose) and a porous layer (o-carboxymethyl chitin and bacterial cellulose); the compact layer faces the side of the brain tissue when the patch is used, is used for preventing cerebrospinal fluid leakage and brain tissue adhesion, and can be sutured; the porous layer faces the skull when the patch is used, is used for supporting the growth, migration, proliferation and differentiation of autologous dura mater cells, promoting the regeneration of autologous dura mater, and generates a tissue adhesion effect after water absorption and gelation so as to improve the sealing compactness. The dura mater patch related to the patent application is characterized by good flexibility, good mechanical property and histocompatibility, capability of remarkably promoting the regeneration of damaged meningeal tissues, anti-adhesion, easy storage, disinfection and preoperative preparation, and full play of the advantages of biological polysaccharide materials in the aspect of tissue repair. However, the preparation of the porous layer of the dura mater patch needs to use glutaraldehyde which is a cross-linking agent and has certain toxicity, and the glutaraldehyde has a very adverse effect on the use in a human body if not cleaned; and the bacterial cellulose is adopted as the reinforcing material of the chitosan-based meninges, so that the defect that no enzyme capable of degrading the bacterial cellulose exists in a human body is overcome, and the method is equivalent to the method that foreign matters need to be removed by a secondary operation.

Disclosure of Invention

In order to overcome the defects in the prior art, the invention aims to provide a method for preparing an ECM-like component and structural human dura mater repair material by a long fiber network skeleton preforming method.

The method provided by the invention is a method for preparing the human dura mater repair material with good toughness and suturable imitative ECM component and structure based on a long fiber network skeleton preforming method.

The invention provides a method for preparing a human dura mater repair material with good toughness and suturable imitative ECM components and structure by a long fiber network skeleton preforming method. The method takes chitosan fiber and silk as main raw materials, adopts the basic process of papermaking production, has the advantages of simple operation, continuous production, rich raw materials, low cost and the like, and overcomes the defects that the existing products are complex to operate and can not be produced in large quantity continuously; meanwhile, the chitosan fiber paper-based material produced based on the method has good physical strength, a fibrous network structure with rich layers and non-chemical crosslinking physical coating, so that the material has good toughness and sewability and good biocompatibility, the papermaking process technology and equipment are mature, most of the participating media can be recycled, the cost is reduced, and the industrial production is realized.

The method provided by the invention uses chitosan fiber and silk as basic raw materials, and comprises the steps of firstly carrying out pretreatment on the raw materials by using clear water for moistening, alkali or acid solution, and then carrying out respective or mixed grinding (pulping) treatment on the treated fiber raw materials. The processed raw materials can be used for preparing a chitosan-silk fiber paper base material by adopting continuous or intermittent multilayer forming, drying and squeezing equipment in the production of the paper industry according to the production scale, and the mechanical and physiological properties are further improved by physical dipping coating and other operations to prepare the human dura mater repair material with high toughness and sewability. The method is relatively simple to operate, is expected to realize continuous production, can greatly reduce the production cost of the tissue material, and has industrial application value and social benefit.

The purpose of the invention is realized by at least one of the following technical solutions.

The method for preparing the human dura mater repair material with the imitated ECM components and the structure by the long fiber network skeleton preforming method comprises the following steps:

(1) respectively soaking a chitosan long fiber material (10mm-50mm) and a chitosan short fiber material (3mm-5mm) in a swelling solution, and performing pretreatment (soaking pretreatment) to swell the fibers so as to facilitate the subsequent treatment process, thereby obtaining a pretreated chitosan long fiber material and a pretreated chitosan short fiber material; soaking silk long fiber (10mm-50mm) and silk short fiber (3mm-5mm) in Na respectively ₂ CO ₃ In the solution, performing collagen removal treatment to obtain silk long fibers and silk short fibers after the collagen removal;

(2) mixing the pretreated chitosan long fibers and the silk long fibers after the collagen is removed in the step (1), washing the mixture to be neutral by using deionized water or clear water, and then performing pulping treatment or pulping treatment by using a groove type pulping machine or a PFI mill to obtain chitosan/silk long fiber mixed pulp; diluting the chitosan/silk long fiber mixed slurry with water, and performing fluffing treatment to obtain suspension (1-10 g/m) ² Quantitative) is put on a paper-making former for dehydration treatment to obtain a thin-layer long fiber interwoven framework material (the dryness is 10-15 percent of wet)Paper base material in the state);

(3) cleaning the pretreated chitosan short fibers and the silk short fibers subjected to collagen removal in the step (1) to be neutral, then carrying out pulp grinding treatment or pulping treatment to obtain chitosan/silk short fiber mixed pulp, diluting the chitosan/silk short fiber mixed pulp with water, and carrying out defibering treatment to obtain suspension (1-10 g/m) ² Quantitative), dewatering in a former to obtain short fiber interwoven base material (paper base material with dryness of 10-15% in wet state);

(4) compounding the thin-layer long fiber interwoven skeleton material obtained in the step (2) with the short fiber interwoven base material obtained in the step (3), performing squeezing and primary drying treatment to obtain a chitosan-silk paper base material, then performing dip coating treatment (non-chemical cross-linking physical dip coating treatment) on an acidic collagen solution on the chitosan-silk paper base material, and performing secondary drying treatment to obtain the human dura mater repair material (the human dura mater repair material with the imitated ECM component and structure) with the good toughness and sewability.

Further, the swelling solution in the step (1) is water (clear water or deionized water), a dilute acid solution or a dilute alkali solution, and the concentration of the dilute acid solution or the dilute alkali solution is 0wt% -2 wt%; the pretreatment time is 1-8 h.

Preferably, the concentration of the dilute acid solution or the dilute alkali solution in the step (1) is 0.05 wt% to 2 wt%.

Further preferably, the dilute alkali solution in step (1) is a 1wt% sodium hydroxide solution.

Preferably, the chitosan long fiber material and the chitosan short fiber material in the step (1) are respectively soaked in the swelling solution for 2-8 hours.

Further, said Na of step (1) ₂ CO ₃ The concentration of the solution is 0.5-2 wt%, the temperature of the collagen removing treatment is 80-100 ℃, and the time of the collagen removing treatment is 1-2 h; the length of the chitosan long fiber material is 10mm-50mm, the length of the chitosan short fiber material is 3mm-5mm, the length of the silk long fiber material is 10mm-50mm, and the silkThe length of the short fiber is 3mm-5 mm.

Preferably, the length of the chitosan long fiber material and the length of the silk long fiber in the step (1) are both 20-40 mm. The fiber length is far beyond what is normally required for papermaking.

Further preferably, the length of the chitosan long fiber material in the step (1) is 30 mm; the length of the silk long fiber is 30 mm.

Preferably, the width of the chitosan long fiber material and the chitosan short fiber material in the step (1) is 10-20 μm.

Further, the mass ratio of the chitosan long fibers after pretreatment in the step (2) to the silk long fibers after collagen removal is 10: 90 to 90: 10; the revolution of the grinding (adopting a PFI mill) is 10000-50000 revolutions, the pressure of the grinding (adopting the PFI mill) is 1-5N/mm, and the time of the grinding (adopting the PFI mill) is 10-50 min; the rotation speed of the pulping treatment (adopting a groove type pulping machine) is 10r/s-30r/s, the time of the pulping treatment (adopting the groove type pulping machine) is 10min-50min, and the pressure load of the pulping treatment (adopting the groove type pulping machine) is 1kg/cm ² -3kg/cm ² (ii) a When the pulping treatment (adopting a PFI mill) is selected in the step (2), the concentration of the obtained chitosan/silk long fiber mixed pulp is 10-20%; when the pulping treatment (groove type pulping machine) is adopted in the step (2), the concentration of the obtained chitosan/silk long fiber mixed pulp is 1-10%.

Preferably, when the beating treatment is selected in the step (2), the concentration of the obtained chitosan/silk long fiber mixed pulp is 5wt% (concentration of absolute dry pulp).

Preferably, the concentration of the refining treatment in the step (2) is 12-18%, the pressure is 3-4N/mm, the concentration of the pulping treatment is 3-7%, and the pulping pressure is 1kg/cm ² -2kg/cm ² 。

Preferably, in the step (2), the pretreated chitosan long fiber material and the collagen-removed silk long fiber material are respectively subjected to pulping treatment and then are uniformly mixed, or the two materials are mixed and then subjected to pulping treatment.

Preferably, the mass ratio of the pretreated chitosan long fiber material to the silk long fiber after the collagen removal in the step (2) is 50:50, 60:40, 70:30, 80:20 or 90: 10.

Further preferably, the mass ratio of the pretreated chitosan long fiber material to the silk long fiber after the collagen removal in the step (2) is 80: 20.

Further, the concentration of the suspension in the step (2) is 0.01wt% to 0.1 wt%; the dryness of the thin-layer long fiber interwoven skeleton material is 10% -15%.

Further, the mass ratio of the pretreated chitosan short fibers to the silk short fibers after the collagen removal in the step (3) is 10: 90 to 90: 10; the rotation number of the grinding is 10000-50000 revolutions, the pressure of the grinding is 1-5N/mm, and the time of the grinding is 10-50 min; the rotation speed of the pulping treatment is 10r/s-30r/s, the pulping treatment time is 10min-50min, and the pulping treatment pressure is 1kg/cm ² -3kg/cm ² (ii) a When the pulping treatment is selected in the step (3), the concentration of the obtained chitosan/silk short fiber mixed pulp is 10-20%; when pulping is selected in the step (3), the concentration of the obtained chitosan/silk short fiber mixed pulp is 1-10%.

Preferably, the mass ratio of the pretreated chitosan short fiber material in the step (3) to the degummed silk short fiber is 50:50, 60:40, 70:30, 80:20 or 90: 10.

Further preferably, the mass ratio of the pretreated chitosan short fiber material to the degummed silk short fiber in the step (3) is 80: 20.

Preferably, when the beating treatment is selected in the step (3), the concentration of the obtained chitosan/silk long fiber mixed pulp is 5wt% (concentration of absolute dry pulp).

Preferably, the concentration of the refining treatment in the step (3) is 12-18%, the pressure is 2-4N/mm, the concentration of the pulping treatment is 3-7%, and the pulping pressure is 1.5kg/cm ² -2.5kg/cm ² 。

Preferably, in the step (3), the pretreated chitosan short fiber material and the degummed silk short fiber material can be respectively subjected to pulping treatment and then uniformly mixed, or the two materials can be mixed and then subjected to pulping treatment.

Further, the concentration of the suspension in the step (3) is 0.01wt% to 0.1 wt%; the dryness of the thin-layer long fiber interwoven skeleton material is 10% -15%.

Preferably, in the step (2) and the step (3), the beating (grinding) pulp adopts continuous beating equipment in the papermaking process, such as a disc grinder, a conical refiner or a cylindrical refiner; batch beating equipment such as PFI mills or trough beaters and the like may also be employed. And (3) according to the requirements of finished products, the beating (grinding) pulp is subjected to cutting, fibrillation and other treatments on the fibers by regulating and controlling the beating revolution and beating pressure to obtain fiber pulp, and the beating degree of the mixed long fiber pulp is 10-30 DEG SR, and the beating degree of the short fiber pulp is 40-60 DEG SR.

Preferably, in the step (2) and the step (3), the beating (grinding) pulp adopts a beating (grinding) pulp processing method in the papermaking process. The pulping process is completed by a groove type pulping machine, the pulping degree of the long fiber pulp is 15-degree SR, the pulping concentration is 1.5%, the rotating speed is 20r/s, the pulping time is 25min, and the pulping pressure load is 1.5kg/cm ² (ii) a The beating degree of the short fiber pulp is 50-SR, the beating concentration is 5.0%, the rotating speed is 20r/s, the beating time is 25min, and the beating pressure load is 2kg/cm ² (ii) a Or the refining process is completed by a PFI mill, wherein the beating concentration of the long fiber is 15 percent, the revolution is 30000r, and the pressure is 3.33N/mm; the beating concentration of short fiber is 10%, the revolution is 15000 and the pressure is 3.33N/mm.

Preferably, in the steps (2) and (3), the paper former is an intermittent paper sheet machine capable of performing conventional dewatering or vacuum-assisted dewatering, and a continuous paper sheet machine such as a cylinder machine, a fourdrinier machine, or a slant wire machine. Preferably using a batch sheet former; the forming concentration of long fibers of the paper sheet material is 0.01-0.1 wt%, preferably 0.05 wt%, and the forming concentration of short fibers is 0.1-1.0 wt%, preferably 0.3 wt%; during compounding, the dryness of the two formed materials is 10-15%, and the preferred dryness is 12%.

Further, the oven-dry mass ratio of the thin-layer long fiber interwoven skeleton material to the short fiber interwoven base material in the step (4) is 1:19-1: 4; the squeezing mode is roller type continuous squeezing or intermittent squeezing, and the squeezing pressure is 1.0kg/cm ² -3.0kg/cm ² Pressing to increase bonding between the materials and further remove moisture; the first drying treatment and the second drying treatment are drying by a drying cylinder, drying by a hot air box, auxiliary infrared drying or freeze drying adopted in the paper industry; when the drying mode adopts drying cylinder drying, hot air box drying or auxiliary infrared drying adopted in the paper industry, the drying temperature is 40-120 ℃; when the drying method adopts freeze drying, the drying temperature is-30 ℃ to-70 ℃.

Preferably, the oven dry mass ratio of the thin-layer long fiber interwoven skeleton material to the short fiber interwoven base material in the step (4) is 1: 9;

preferably, the pressing pressure in step (4) is 1.5kg/cm ² ；

Preferably, the temperature of the first drying treatment in step (4) is 90 ℃ or 80 ℃.

Preferably, the temperature of the second drying treatment in the step (4) is 90 ℃ or 80 ℃.

Further, the acidic collagen solution obtained in the step (4) is a solution obtained by uniformly mixing collagen and an acidic solution, wherein the acidic solution is sulfuric acid, hydrochloric acid or acetic acid, the concentration of the collagen is 0-20wt%, and the pH value of the acidic collagen solution is 1.0-4.0; the time of the dip coating treatment is 1s-90 s.

Preferably, the time of the dip coating treatment in the step (4) is 1 to 30 s. The dipping coating treatment is continuously carried out in the papermaking process or intermittently carried out after the paper making of the base paper is finished.

Preferably, the coater used for coating is used for carrying out dip coating treatment on the sample in the step (4) according to different requirements of the base material, and the components of the dip coating liquid can be single components or composite components.

Preferably, the dipping solution in the step (4) can be a collagen solution with 10 wt% -20 wt%.

Further preferably, in the step (4), the sample is subjected to dip coating treatment, and the treatment may be carried out by using a gelatin dipping solution prepared by an acid method with a concentration of 10 wt% alone.

The invention provides a human dura mater repair material with imitated ECM components and structure, which is prepared by the preparation method.

The method comprises the steps of firstly selecting chitosan regenerated fibers and silk fibers as basic raw materials to respectively simulate glycosaminoglycan and protein components in human ECM, and then establishing and simulating a three-dimensional nano network structure of the ECM through fiber raw material grinding (pulping) and wet papermaking. The chitosan-silk fiber human dura mater repair material prepared based on the long fiber network skeleton preforming method can well simulate the components and the structure of a human dura mater (dura mater repair and the like in the process of neurosurgery operation), and the prepared material has good toughness, good suturability and good biocompatibility and is suitable for clinical operation. The whole preparation process basically adopts the common equipment and process in the pulping and papermaking process, so that good feasibility is provided for the large-scale production of the materials. Until now, no report related to the preparation of human dura mater repair materials with imitated ECM components and structures by using chitosan fibers and silk as basic raw materials based on a paper making method is found.

Compared with the prior art, the invention has the following advantages and beneficial effects:

(1) according to the preparation method provided by the invention, physical interweaving and toughness connection are increased mainly by a long fiber preforming method and introduction of a skeleton of a long fiber network, and subsequent physical dipping and coating treatment further improves the toughness and the suturability of the hard film repair material, ensures the original good biocompatibility of the hard film repair material, and can adjust the micro/nano proportion of chitosan and silk fibers or improve the microstructure of the hard film repair material by adjusting parameters such as pulping revolution, pressure and quantification;

(2) according to the chitosan-silk fiber paper-based dura mater repair material (human dura mater repair material with imitated ECM (extracellular matrix) component and structure), because chitosan, silk, collagen and other widely-used raw materials with excellent biocompatibility in the field of biomedical materials are adopted as basic raw materials in the forming process, and the raw materials are not modified by any chemical crosslinking method in the preparation process, the safety and biocompatibility of the chitosan-silk fiber paper-based dura mater material are well guaranteed;

(3) the invention prepares chitosan-silk imitated ECM component and structural dura mater repair material (human dura mater repair material of imitated ECM component and structure) by long fiber network skeleton preforming method, compared with the material obtained by other methods, after the treatment of the method, chitosan fiber and silk fiber are obviously divided, part of fiber is nano-sized, has similar structure with collagen fiber in ECM, high tensile strength and large tensile strain under wet state, the introduction of the long fiber network skeleton provides good support for suture thread, and the material shows good elasticity and toughness without chemical crosslinking treatment, is easy to cut and sew in the operation process, can bear higher external pressure (dura mater and brain pressure), has good strength and fibrous porous structure with rich layers, can induce and promote the formation of human body self tissue, in addition, the in vivo degradation time can be adjusted by adjusting the components and the proportion of the raw materials, and the material is suitable for being used as a repair material for human dura mater surgery;

(4) the preparation method provided by the invention adopts mature process methods and equipment in the paper industry, is easy to realize the control of thickness, structure and function, is expected to realize large-scale industrial production, reduces the product cost and relieves the medical and family burden of patients.

Drawings

FIG. 1 is an SEM image of the fiber skeleton and fiber morphology in the examples;

FIG. 2 is an SEM image of the fiber interlacing morphology in the human dura mater repair material of the imitated ECM composition and structure of the example;

FIG. 3 is an SEM image of cross-sectional fiber interlacing morphology in the human dura mater repair material of the imitated ECM composition and structure in the example;

fig. 4 is an SEM image of surface morphology in the human dura mater repair material of the mock ECM composition and structure after dip coating in the examples.

Detailed Description

The following examples are presented to further illustrate the practice of the invention, but the practice and protection of the invention is not limited thereto. It is noted that the processes described below, if not specifically described in detail, are all realizable or understandable by those skilled in the art with reference to the prior art. The reagents or apparatus used are not indicated to the manufacturer, and are considered to be conventional products available by commercial purchase.

Example 1

A preparation method of a human dura mater repair material imitating ECM components and structures comprises the following steps:

(1) respectively soaking a chitosan long fiber material (with the length of 2cm) and a chitosan short fiber material (with the length of 3mm) in swelling solution (0.5 wt% of sodium hydroxide solution), performing pretreatment (water absorption swelling for 8 hours), and washing the slurry with deionized water until the pH value of slurry washing liquor is neutral to obtain a pretreated chitosan long fiber material and a pretreated chitosan short fiber material; soaking silk long fiber (length of 2cm) and silk short fiber (length of 3mm) in Na respectively ₂ CO ₃ Solution (mass percentage concentration is 1wt%, silk fiber and Na) ₂ CO ₃ The solution material-liquid ratio is 1:5g/mL), the collagen removing treatment is carried out under the stirring state (the water bath state is carried out, the temperature is 100 ℃, the time is 1h), a large amount of deionized water with the temperature of 80 ℃ is used for washing the fibers after the water bath, then the collagen removing treatment is repeated again (the time of the collagen removing treatment is 0.5h), and a large amount of deionized water with the temperature of 80 ℃ is used for washing the fibers again after the water bath, so that the silk long fibers after the collagen removing and the silk short fibers after the collagen removing are obtained.

(2) Mixing the pretreated chitosan long fibers obtained in the step (1) with the silk long fibers after the collagen removal, wherein the mass ratio of the pretreated chitosan long fibers to the silk long fibers after the collagen removal is 4:1, cleaning the mixture until the mixture is neutral (the dryness of the mixed pulp after cleaning is 3wt percent), and then adopting a groove typePulping with a pulping machine (rotation speed of 20r/s, pulping time of 10min, pressure load of 1.5 kg/cm) ² Beating degree of about 17 ° SR), obtaining chitosan/silk long fiber mixed slurry; diluting the chitosan/silk long fiber mixed slurry into a suspension by using water, carrying out defibering treatment (the suspension is defibered into the suspension with the concentration of 0.05 wt%), and placing the suspension on a papermaking former for dehydration treatment to obtain a thin-layer long fiber interwoven framework material (a long fiber network framework material with the dryness of 10%);

(3) mixing the pretreated chitosan short fibers and the collagen-removed silk short fibers in the step (1) according to the mass ratio of 4:1, cleaning to be neutral (the dryness of the mixed pulp after cleaning is 3 wt%), and pulping by using a trough type pulping machine (the rotating speed is 20r/s, the pulping time is 15min, and the pressure load is 2 kg/cm) ² Beating degree of about 48-SR) to obtain chitosan/silk short fiber mixed slurry, carrying out defibering treatment (defibering to form suspension with concentration of 0.3 wt%), diluting the chitosan/silk short fiber mixed slurry to form suspension with water, placing the suspension on a manual paper sheet making machine for dewatering treatment, and carrying out paper making and forming to obtain a short fiber interwoven base material (short fiber network framework material with dryness of 11%);

(4) compounding the thin-layer long fiber interwoven framework material in the step (2) with the short fiber interwoven base material in the step (3), wherein the oven dry mass ratio of the thin-layer long fiber interwoven framework material to the short fiber interwoven base material is 1:9, and performing rolling treatment, wherein the pressing pressure of the rolling treatment is 1.5kg/cm ² Transferring to a dryer at 80 ℃ for drying treatment (time is 20min) to obtain a chitosan-silk paper base material, then soaking the chitosan-silk paper base material in an acidic collagen solution (a solution obtained by uniformly mixing collagen and a hydrochloric acid solution, wherein the pH is 2.0, and the concentration of collagen is 15 wt%) for dip coating treatment (the dipping time is 45s), and drying at 80 ℃ for 20min to obtain the human dura mater repair material imitating the ECM component and structure. The human dura mater repair material with the imitated ECM components and structure has the quantitative rate of 110g/m ² The thickness is 3 mm. The fiber interweaving form and structure are shown in FIG. 1, FIG. 2 and FIG. 3 (same below), and the inner part of the membraneThe skeleton fiber with larger thickness (figure 1) and the micro/nano-distributed fine fiber after treatment can be observed obviously in the partial structure; the surface morphology of the membrane is shown in figure 4 (the same below), one side of the membrane surface after dip coating treatment is smooth and has no obvious micropores, and the prepared membrane material can be ensured to have good tightness.

The human dura mater repair material with the imitated ECM components and the structure is placed in a constant temperature and humidity chamber to balance moisture for 24h, and an INSTRON 5565 tensile compression material testing machine is used for testing the tensile stress and the tensile strain of the human dura mater repair material, wherein the tensile stress is about 15Mpa and the tensile strain is 40% under the condition that the moisture content is 20%. The 3/0 atraumatic silk thread is adopted, the circular needle 4-gauge thread is installed at the distance of 4mm, the suture condition is observed after the conventional force suture of the dura mater operation, the material is found to be sutured, the needle hole is tightly jointed with the silk thread, and no obvious tearing trace exists at the joint.

Example 2

A preparation method of a human dura mater repair material with imitated ECM components and structure comprises the following steps:

(1) respectively soaking a chitosan long fiber material (the length is 3cm) and a chitosan short fiber material (the length is 4mm) in swelling solution (1 wt% of sodium hydroxide solution), performing pretreatment (water absorption swelling for 8 hours), and washing the slurry with deionized water until the pH value of slurry washing liquor is neutral to obtain a pretreated chitosan long fiber material and a pretreated chitosan short fiber material; soaking silk long fiber (length of 3cm) and silk short fiber (length of 4mm) in Na respectively ₂ CO ₃ Solution (mass percentage concentration is 1wt%, silk fiber and Na) ₂ CO ₃ The material-liquid ratio of the solution is 1:5), performing collagen removal treatment under a stirring state (performed under a water bath state, the temperature is 100 ℃, and the time is 1h), washing the fibers with a large amount of 80 ℃ deionized water after the water bath, then repeating the collagen removal treatment again (the time of the collagen removal treatment is 0.5h), and washing the fibers with a large amount of 80 ℃ deionized water again after the water bath to obtain the silk long fibers after the collagen removal and the silk short fibers after the collagen removal.

(2) The pretreated chitosan long fiber in the step (1) and the silk after the collagen removalMixing long fibers, wherein the mass ratio of the pretreated chitosan long fibers to the silk long fibers subjected to collagen removal is 3:1, cleaning to neutrality (the dryness of the mixed pulp after cleaning is 5 wt%), and then pulping by using a groove type pulping machine (the rotating speed is 20r/s, the pulping time is 15min, and the pressure load is 1 kg/cm) ² Beating degree of about 13 ° SR), obtaining chitosan/silk long fiber mixed slurry; diluting the chitosan/silk long fiber mixed slurry into a suspension with water, carrying out untwining treatment (untwining into a suspension with the concentration of 0.05 wt%), and placing on a papermaking forming machine for dehydration treatment to obtain a thin-layer long fiber interwoven skeleton material (a long fiber network skeleton material with the dryness of 12%);

(3) mixing the pretreated chitosan short fibers and the silk short fibers after the collagen removal in the step (1), wherein the mass ratio of the pretreated chitosan short fibers to the silk short fibers after the collagen removal is 4:1, cleaning the mixture until the mixture is neutral (the dryness of the mixed pulp after cleaning is 5 wt%), and then performing pulping treatment by adopting a groove type pulping machine (the rotating speed is 20r/s, the pulping time is 15min, and the pressure load is 1.5 kg/cm) ² Beating degree of about 43-SR) to obtain chitosan/silk short fiber mixed slurry, carrying out defibering treatment (defibering to form suspension with concentration of 0.5 wt%), diluting the chitosan/silk short fiber mixed slurry to form suspension with water, placing the suspension on a manual paper sheet making machine for dewatering treatment, and carrying out paper making and forming to obtain a short fiber interwoven base material (the dryness is 12% short fiber network framework material);

(4) compounding the thin-layer long fiber interwoven framework material in the step (2) with the short fiber interwoven base material in the step (3), wherein the oven dry mass ratio of the thin-layer long fiber interwoven framework material to the short fiber interwoven base material is 1:8, and performing rolling treatment, wherein the pressing pressure of the rolling treatment is 1kg/cm ² Transferring to a drier at 80 deg.C for drying for 20min to obtain chitosan-silk paper base material, soaking in acidic collagen solution (pH of 3.0 and collagen concentration of 10 wt%) for soaking for 30s, and drying at 90 deg.C for 20min to obtain the human dura mater repairing material with ECM-like component and structure. The human dura mater repair material with the imitated ECM components and structure has the quantitative rate of 140g/m ² The thickness is 3.5 mm.

The material is placed in a constant temperature and humidity chamber to balance moisture for 24h, and an INSTRON 5565 tensile compression material testing machine is used for testing the tensile stress and the tensile strain of the material, wherein under the condition that the moisture content is 30%, the tensile stress is about 12Mpa, and the tensile strain is 35%. The 3/0 atraumatic silk thread is adopted, the circular needle with the number 4 thread gauge is installed on the dura mater, the suture condition is observed after the conventional force suture, the material is sutured, the needle hole is tightly jointed with the silk thread, and no obvious tearing trace exists at the joint.

Example 3

(1) respectively soaking a chitosan long fiber material (with the length of 4cm) and a chitosan short fiber material (with the length of 3mm) in a swelling solution (1.5 wt% of sodium hydroxide solution), performing pretreatment (water absorption swelling for 8 hours), and washing the slurry with deionized water until the pH value of a slurry washing solution is neutral to obtain a pretreated chitosan long fiber material and a pretreated chitosan short fiber material; soaking silk long fiber (length of 4cm) and silk short fiber (length of 3mm) in Na respectively ₂ CO ₃ Solution (mass percentage concentration is 1wt%, silk fiber and Na) ₂ CO ₃ The material-liquid ratio of the solution is 1:5), performing the collagen removal treatment under a stirring state (performed under a water bath state, the temperature is 100 ℃, and the time is 1h), washing the fibers with a large amount of 80 ℃ deionized water after the water bath, then repeating the collagen removal treatment (the time of the collagen removal treatment is 0.5h), and washing the fibers with a large amount of 80 ℃ deionized water again after the water bath to obtain the silk long fibers after the collagen removal and the silk short fibers after the collagen removal.

(2) Mixing the pretreated chitosan long fibers and the silk long fibers after the collagen removal in the step (1), wherein the mass ratio of the pretreated chitosan long fibers to the silk long fibers after the collagen removal is 2:1, cleaning the mixture until the mixture is neutral (the dryness of the mixed pulp after cleaning is 6wt percent), and then pulping the mixture by using a groove type pulping machine (the rotating speed is 20r/s and the pulping speed is 6wt percent)The time is 10min, and the pressure load is 1.5kg/cm ² Beating degree of about 15 ° SR), obtaining chitosan/silk long fiber mixed slurry; diluting the chitosan/silk long fiber mixed slurry into a suspension by using water, carrying out defibering treatment (the suspension is defibered into the suspension with the concentration of 0.08 wt%), and placing the suspension on a papermaking former for dehydration treatment to obtain a thin-layer long fiber interwoven framework material (a long fiber network framework material with the dryness of 11%);

(3) mixing the pretreated chitosan short fibers and the silk short fibers after the collagen removal in the step (1), wherein the mass ratio of the pretreated chitosan short fibers to the silk short fibers after the collagen removal is 2:1, cleaning the mixture until the mixture is neutral (the dryness of the mixed pulp after cleaning is 6 wt%), and then pulping the mixture by using a groove type pulping machine (the rotating speed is 20r/s, the pulping time is 15min, and the pressure load is 2 kg/cm) ² Beating degree of about 51-SR) to obtain chitosan/silk short fiber mixed slurry, carrying out defibering treatment (defibering to form suspension with concentration of 0.5 wt%), diluting the chitosan/silk short fiber mixed slurry to form suspension with water, placing the suspension on a manual paper sheet making machine for dewatering treatment, and carrying out paper making and forming to obtain a short fiber interwoven base material (short fiber network framework material with dryness of 13%);

(4) compounding the thin-layer long fiber interwoven framework material in the step (2) with the short fiber interwoven base material in the step (3), wherein the oven dry mass ratio of the thin-layer long fiber interwoven framework material to the short fiber interwoven base material is 1:10, and performing rolling treatment, wherein the pressing pressure of the rolling treatment is 1.5kg/cm ² Transferring to a dryer at 80 ℃ for drying treatment (time is 20min) to obtain a chitosan-silk paper base material, then soaking the chitosan-silk paper base material in an acidic collagen solution (a solution obtained by uniformly mixing collagen and the acidic solution, wherein the pH is 4.0, and the concentration of collagen is 15 wt%) for dip coating treatment (the dipping time is 45s), and drying at 100 ℃ for 20min to obtain the human dura mater repair material imitating the ECM component and structure. The human dura mater repair material with the imitated ECM component and structure has the quantitative rate of 115g/m ² The thickness is 3 mm.

The material is placed in a constant temperature and humidity chamber to balance moisture for 24h, and an INSTRON 5565 tensile compression material tester is used for testing the tensile stress and the tensile strain of the material, wherein under the condition that the moisture content is 20%, the tensile stress is about 15Mpa, and the tensile strain is 40%. The 3/0 atraumatic silk thread is adopted, the circular needle with the number 4 thread gauge is installed on the dura mater, the suture condition is observed after the conventional force suture, the material is sutured, the needle hole is tightly jointed with the silk thread, and no obvious tearing trace exists at the joint.

Example 4

(1) respectively soaking a chitosan long fiber material (the length is 3cm) and a chitosan short fiber material (the length is 4mm) in swelling solution (2 wt% of sodium hydroxide solution), performing pretreatment (water absorption swelling for 8 hours), and washing the slurry with deionized water until the pH value of slurry washing liquor is neutral to obtain a pretreated chitosan long fiber material and a pretreated chitosan short fiber material; soaking silk long fiber (length of 3cm) and silk short fiber (length of 4mm) in Na respectively ₂ CO ₃ Solution (mass percentage concentration is 1wt%, silk fiber and Na) ₂ CO ₃ The material-liquid ratio of the solution is 1:5), performing collagen removal treatment under a stirring state (performed under a water bath state, the temperature is 100 ℃, and the time is 1h), washing the fibers with a large amount of 80 ℃ deionized water after the water bath, then repeating the collagen removal treatment again (the time of the collagen removal treatment is 0.5h), and washing the fibers with a large amount of 80 ℃ deionized water again after the water bath to obtain the silk long fibers after the collagen removal and the silk short fibers after the collagen removal.

(2) Mixing the pretreated chitosan long fibers and the silk long fibers after the collagen removal in the step (1), wherein the mass ratio of the pretreated chitosan long fibers to the silk long fibers after the collagen removal is 1:1, cleaning the mixture until the mixture is neutral (the dryness of the mixed pulp after cleaning is 3.0 wt%), and then pulping the mixture by using a trough type pulping machine (the rotating speed is 20r/s, the pulping time is 10min, and the pressure load is 1.0 kg/cm) ² Beating degree of about 16 ° SR), obtaining chitosan/silk long fiber mixed slurry; diluting the chitosan/silk long fiber mixed slurry into suspension with water, and performing defibering treatment (the concentration of the defibered solution is 0.03 w)t% suspension liquid), placing on a papermaking former for dehydration treatment to obtain a thin-layer long fiber interwoven framework material (the dryness of the long fiber network framework material is 11%);

(3) mixing the pretreated chitosan short fibers and the silk short fibers after the collagen removal in the step (1), wherein the mass ratio of the pretreated chitosan short fibers to the silk short fibers after the collagen removal is 1:1, cleaning the mixture until the mixture is neutral (the dryness of the mixed pulp after cleaning is 3.0 wt%), and then pulping the mixture by using a groove type pulping machine (the rotating speed is 20r/s, the pulping time is 15min, and the pressure load is 2.0 kg/cm) ² Beating degree of about 50 ° SR) to obtain chitosan/silk short fiber mixed slurry, performing defibering treatment (defibering to form suspension with concentration of 0.5 wt%), diluting the chitosan/silk short fiber mixed slurry with water to form suspension, placing the suspension on a manual paper sheet making machine for dewatering treatment, and performing paper making and forming to obtain a short fiber interwoven substrate (dryness is 12% short fiber network framework material);

(4) compounding the thin-layer long fiber interwoven framework material in the step (2) with the short fiber interwoven base material in the step (3), wherein the oven dry mass ratio of the thin-layer long fiber interwoven framework material to the short fiber interwoven base material is 3:25, and performing rolling treatment, wherein the pressing pressure of the rolling treatment is 2.0kg/cm ² Transferring to a dryer at 90 ℃ for drying treatment (time is 20min) to obtain a chitosan-silk paper base material, then soaking the chitosan-silk paper base material in an acidic collagen solution (a solution obtained by uniformly mixing collagen and the acidic solution, wherein the pH is 3.0, and the concentration of collagen is 15 wt%) for dip coating treatment (the dipping time is 25s), and drying at 100 ℃ for 20min to obtain the human dura mater repair material imitating the ECM component and structure. The human dura mater repair material with the imitated ECM components and structure has the quantitative of 110g/m ² The thickness is 3 mm.

The material is placed in a constant temperature and humidity chamber to balance moisture for 24h, and an INSTRON 5565 tensile compression material tester is used for testing the tensile stress and the tensile strain of the material, wherein under the condition that the moisture content is 30%, the tensile stress is about 6Mpa, and the tensile strain is 47%. The 3/0 atraumatic silk thread is adopted, the circular needle 4-gauge thread is installed at the distance of 4mm, the suture condition is observed after the conventional force suture of the dura mater operation, the material is found to be sutured, the needle hole is tightly jointed with the silk thread, and no obvious tearing trace exists at the joint.

Example 5

(1) respectively soaking a chitosan long fiber material (with the length of 2cm) and a chitosan short fiber material (with the length of 3mm) in swelling solution (1 wt% of sodium hydroxide solution), performing pretreatment (water absorption swelling for 8 hours), and washing the slurry with deionized water until the pH value of slurry washing liquor is neutral to obtain a pretreated chitosan long fiber material and a pretreated chitosan short fiber material; soaking silk long fiber (length of 2cm) and silk short fiber (length of 3mm) in Na respectively ₂ CO ₃ Solution (mass percentage concentration is 1wt%, silk fiber and Na) ₂ CO ₃ The material-liquid ratio of the solution is 1:5), performing collagen removal treatment under a stirring state (performed under a water bath state, the temperature is 100 ℃, and the time is 1h), washing the fibers with a large amount of 80 ℃ deionized water after the water bath, then repeating the collagen removal treatment again (the time of the collagen removal treatment is 0.5h), and washing the fibers with a large amount of 80 ℃ deionized water again after the water bath to obtain the silk long fibers after the collagen removal and the silk short fibers after the collagen removal.

(2) Mixing the pretreated chitosan long fibers and the silk long fibers after the collagen removal in the step (1), wherein the mass ratio of the pretreated chitosan long fibers to the silk long fibers after the collagen removal is 1:1, washing the mixture to be neutral (the dryness of the mixed pulp after washing is 15 wt%), and then performing pulp grinding treatment by adopting a PFI mill (the number of revolutions is 10000 revolutions, the pressure is 3.33N/mm, and the beating degree is about 20 DEG SR) to obtain chitosan/silk long fiber mixed pulp; diluting the chitosan/silk long fiber mixed slurry into a suspension by using water, carrying out defibering treatment (the suspension is defibered into the suspension with the concentration of 0.03 wt%), and placing the suspension on a papermaking former for dehydration treatment to obtain a thin-layer long fiber interwoven framework material (a long fiber network framework material with the dryness of 11%);

(3) mixing the pretreated chitosan short fibers and the silk short fibers after the collagen removal in the step (1), wherein the mass ratio of the pretreated chitosan short fibers to the silk short fibers after the collagen removal is 1:1, washing to be neutral (the dryness of the mixed pulp after washing is 15 wt%), then performing pulp grinding treatment by adopting a PFI (the number of revolutions is 30000r, the pressure is 3.33N/mm, and the beating degree is about 60 DEG SR) to obtain chitosan/silk short fiber mixed pulp, performing defibering treatment (the mixed pulp is defibered into suspension with the concentration of 0.5 wt%), diluting the chitosan/silk short fiber mixed pulp into suspension by using water, placing the suspension on a manual paper sheet making machine for dehydration treatment, and performing paper making and forming to obtain a short fiber interwoven substrate (a short fiber network framework material with the dryness of 10%);

(4) compounding the thin-layer long fiber interwoven framework material in the step (2) with the short fiber interwoven base material in the step (3), wherein the oven dry mass ratio of the thin-layer long fiber interwoven framework material to the short fiber interwoven base material is 3:25, and performing rolling treatment, wherein the pressing pressure of the rolling treatment is 1.5kg/cm ² Transferring to a dryer at 90 ℃ for drying treatment (time is 20min) to obtain a chitosan-silk paper base material, then soaking the chitosan-silk paper base material in an acidic collagen solution (a solution obtained by uniformly mixing collagen and the acidic solution, wherein the pH is 4.0, and the concentration of collagen is 15 wt%) for dip coating treatment (the dipping time is 45s), and drying at-55 ℃ for 6h to obtain the human dura mater repair material imitating the ECM component and structure. The human dura mater repair material with the imitated ECM components and structure has the quantitative rate of 110g/m ² The thickness is 8 mm.

The material is placed in a constant temperature and humidity chamber to balance moisture for 24h, and an INSTRON 5565 tensile compression material tester is used for testing the tensile stress and the tensile strain of the material, wherein under the condition that the moisture content is 35 percent, the tensile stress is about 5.5Mpa, and the tensile strain is 43 percent. The 3/0 atraumatic silk thread is adopted, the circular needle with the number 4 thread gauge is installed on the dura mater, the suture condition is observed after the conventional force suture, the material is sutured, the needle hole is tightly jointed with the silk thread, and no obvious tearing trace exists at the joint.

The above examples are only preferred embodiments of the present invention, which are intended to be illustrative and not limiting, and those skilled in the art should understand that they can make various changes, substitutions and alterations without departing from the spirit and scope of the invention.

Claims

1. A method for preparing an ECM (extracellular matrix) -imitated and structural human dura mater repair material by a long fiber network skeleton preforming method is characterized by comprising the following steps of:

(1) respectively soaking the chitosan long fiber material and the chitosan short fiber material in swelling liquid, and performing pretreatment to obtain a pretreated chitosan long fiber material and a pretreated chitosan short fiber material; soaking long silk fiber and short silk fiber in Na respectively ₂ CO ₃ In the solution, performing collagen removal treatment to obtain silk long fibers and silk short fibers after the collagen removal; the swelling solution is water, dilute acid solution or dilute alkali solution, and the concentration of the dilute acid solution or the dilute alkali solution is 0-2 wt%; the pretreatment time is 1-8 h; the Na is ₂ CO ₃ The concentration of the solution is 0.5-2 wt%, the temperature of the collagen removing treatment is 80-100 ℃, and the time of the collagen removing treatment is 1-2 h; the length of the chitosan long fiber material is 10mm-50mm, the length of the chitosan short fiber material is 3mm-5mm, the length of the silk long fiber material is 10mm-50mm, and the length of the silk short fiber material is 3mm-5 mm;

(2) mixing the pretreated chitosan long fibers in the step (1) with the silkworm silk long fibers after the collagen is removed, cleaning to be neutral, and then carrying out jordaning treatment or pulping treatment to obtain chitosan/silk long fiber mixed slurry; diluting the chitosan/silk long fiber mixed slurry with water, carrying out defibering treatment to obtain a suspension, and carrying out dehydration treatment to obtain a thin-layer long fiber interwoven framework material;

(3) cleaning the pretreated chitosan short fibers and the silk short fibers after collagen removal in the step (1) to be neutral, then carrying out jordaning treatment or pulping treatment to obtain chitosan/silk short fiber mixed slurry, diluting the chitosan/silk short fiber mixed slurry with water, carrying out defibering treatment to obtain suspension, and carrying out dehydration treatment to obtain a short fiber interweaving base material;

(4) compounding the thin-layer long fiber interwoven skeleton material in the step (2) with the short fiber interwoven base material in the step (3), performing squeezing and primary drying treatment to obtain a chitosan-silk paper base material, then performing dipping coating treatment on an acidic collagen solution on the chitosan-silk paper base material, and performing secondary drying treatment to obtain the human dura mater repair material imitating the ECM component and structure.

2. The method for preparing the ECM-like component and the structural human dura mater repair material by the long fiber network skeleton preforming method according to claim 1, wherein the mass ratio of the pretreated chitosan long fibers to the silk long fibers after the collagen removal in the step (2) is 10: 90 to 90: 10; the pulping treatment is PFI pulping treatment; the rotation number of the grinding is 10000-50000 revolutions, and the pressure of the grinding is 1-5N/mm; the rotation speed of the pulping treatment is 10r/s-30r/s, the pulping treatment time is 10min-50min, and the pulping treatment pressure is 1kg/cm ² -3kg/cm ² (ii) a When PFI pulping treatment is selected in the step (2), the concentration of the obtained chitosan/silk long fiber mixed pulp is 10-20%; when pulping is selected in the step (2), the concentration of the obtained chitosan/silk long fiber mixed slurry is 1-5%.

3. The method for preparing the ECM-imitated material and the structural human dura mater repair material by the long fiber network skeleton preforming method according to claim 1, wherein the concentration of the suspension in the step (2) is 0.01-0.1 wt%; the dryness of the thin-layer long fiber interwoven skeleton material is 10-15%.

4. The method for preparing the ECM-like component and the structural human dura mater repair material by the long fiber network skeleton preforming method according to claim 1, wherein the mass ratio of the pretreated chitosan short fibers to the silk short fibers after the collagen removal in the step (3) is 10: 90 to 90: 10; the pulping treatment is PFI pulping treatment; the rotation number of the grinding is 10000-50000 revolutions, the pressure of the grinding is 1-5N/mm, and the time of the grinding is 10-50 min;the rotation speed of the pulping treatment is 10r/s-30r/s, the pulping treatment time is 10min-50min, and the pressure load of the pulping treatment is 1kg/cm ² -3kg/cm ² (ii) a When the pulping treatment is selected in the step (3), the concentration of the obtained chitosan/silk short fiber mixed pulp is 10-20%; when pulping is selected in the step (3), the concentration of the obtained chitosan/silk short fiber mixed pulp is 1-5%.

5. The method for preparing the ECM-imitated material and the structural human dura mater repair material by the long fiber network skeleton preforming method according to claim 1, wherein the concentration of the suspension in the step (3) is 0.01-0.1 wt%; the dryness of the short fiber interwoven base material is 10-15%.

6. The method for preparing the ECM-imitated component and the structural human dura mater repair material by the long fiber network skeleton preforming method according to claim 1, wherein the oven dry mass ratio of the thin-layer long fiber interwoven skeleton material to the short fiber interwoven base material in the step (4) is 1:19-1: 4; the squeezing mode is roller type continuous squeezing or intermittent squeezing, and the squeezing pressure is 1.0kg/cm ² -5.0 kg/cm ² (ii) a The first drying treatment and the second drying treatment are drying by a drying cylinder, drying by a hot air box, auxiliary infrared drying or freeze drying adopted in the paper industry; when the drying mode adopts drying cylinder drying, hot air box drying or auxiliary infrared drying adopted in the paper industry, the drying temperature is 40-120 ℃; when freeze drying is adopted as the drying mode, the drying temperature is-30 ℃ to-70 ℃.

7. The method for preparing ECM-like composition and structural human dura mater repair material by the long fiber network skeleton preforming method according to claim 1, wherein the acidic collagen solution of step (4) is a solution obtained by uniformly mixing collagen with an acidic solution, the acidic solution is sulfuric acid, hydrochloric acid or acetic acid, the concentration of the collagen is 0 to 20wt%, and the pH of the acidic collagen solution is 1.0 to 4.0; the time of the dip coating treatment is 1s-30 s.

8. A human dura mater repair material of simulated ECM composition and structure prepared by the method of any of claims 1-7.