CN115154658A - Preparation method and application of water-absorbing self-adhesive dura mater patch - Google Patents
Preparation method and application of water-absorbing self-adhesive dura mater patch Download PDFInfo
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- CN115154658A CN115154658A CN202210426474.7A CN202210426474A CN115154658A CN 115154658 A CN115154658 A CN 115154658A CN 202210426474 A CN202210426474 A CN 202210426474A CN 115154658 A CN115154658 A CN 115154658A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/16—Macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
- D04H1/72—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged
- D04H1/728—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged by electro-spinning
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/32—Materials or treatment for tissue regeneration for nerve reconstruction
Abstract
The invention belongs to the technical field of biomedical materials, and particularly relates to a preparation method and application of a water-absorbing self-adhesive dura mater patch. The preparation method comprises the steps of firstly preparing a polylactide-co-caprolactone spinning film by an electrostatic spinning technology, taking the polylactide-co-caprolactone spinning film as a substrate layer, taking particle powder obtained by mixing polyethyleneimine with polyacrylic acid solution, freeze-drying and grinding the mixture as an adhesive layer, and then combining the particle powder of the adhesive layer with the substrate layer spinning film to prepare the double-layer dura mater patch. The obtained double-layer dura mater patch can be firmly combined with tissues through hydrogen bonds and electrostatic action after contacting wet tissues, and can quickly exert self-adhesive sealing effect without surgical suture and long-time pressing. In addition, the dura mater patch has good tensile property and excellent swelling resistance, can bear the pressure of cerebrospinal fluid when being applied to dura mater repair, can not cause the risk of pressing nerves due to overhigh swelling rate, and has great practical value in the aspect of clinical dura mater repair.
Description
Technical Field
The invention belongs to the technical field of biomedical materials, and particularly relates to a preparation method and application of a water-absorbing self-adhesive dura mater patch.
Background
Dura mater is a dense connective tissue mainly composed of collagen fibers, and the injuries of dura mater are very common in clinic and mainly divided into traumatic injuries and iatrogenic injuries, and the incidence rate of dura mater is different according to the operation position and the operation mode. Dural trauma, if handled improperly, can cause a series of complications, such as cerebrospinal fluid leakage, low intracranial pressure syndrome, poor wound healing, sinus tract formation, dural pseudocyst, and infectious meningitis, which can be serious and even life threatening. Therefore, how to treat dural damage efficiently and rapidly to reduce the incidence of complications becomes a clinically urgent problem to be solved.
Different treatments are usually clinically used depending on the degree of dural damage. For general linear lesions without significant defect of dura mater, the treatment is usually performed by direct suturing or by combined use of sealing adhesives (e.g. fibrin glue, bio-protein glue, polyethylene glycol hydrogel, etc.). For severe injuries with blocky defects, autograft, allogeneic, xenogeneic and artificial dura mater patches are usually used for repair. Among them, the dura mater autograft has advantages of no rejection immunity, no disease transmission and no inflammatory reaction, but increases additional trauma at the donor site while prolonging the operation time. Allogeneic strains are at risk of transmitting diseases such as Creutzfeldt-Jakob and are abandoned basically at present. The common xenogenic materials (such as bovine heel aponeurosis, porcine pericardium, porcine small intestine submucosa and the like) have the problems of rejection reaction, immunogenicity and the like, and limit the application of the xenogenic materials in dura mater repair. In recent years, the biomimetic patch prepared based on high polymer materials (polyurethane, polycaprolactone, polylactic acid, silk fibroin and the like) has the advantages of wide source, simplicity in synthesis, good biocompatibility, controllable mechanical properties and the like, and has become a research hotspot of the artificial dura mater spinalis patch. However, the traditional high molecular dura mater patch has no adhesiveness, and still needs to be sutured in clinical use. Due to the restriction of narrow spinal canal space, the suture difficulty is increased when the polymer patch is adopted to repair the dura mater spinalis, and the infection risk of the operation part is easily increased by prolonging the operation time. With the continuous research and development of bionic wet adhesive hydrogel, more and more wet adhesives based on high polymer materials are applied to the biomedical fields of wound sealing, hemostasis, tissue repair and the like, such as polyethylene glycol hydrogel, chitosan hydrogel, cyanoacrylate adhesives and the like, but the defects of high swelling rate, insufficient adhesive sealing property, high biotoxicity, weak biomechanical strength and the like still exist, so that the clinical treatment requirements on quick sealing and long-term use of dura mater spinalis injury cannot be met. Therefore, the development of a novel dura mater patch with low swelling degree, rapid self-adhesion, good mechanical properties and biocompatibility is of great significance for clinical dura mater repair application.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide the preparation method of the water-absorbing self-adhesive dura mater patch, the patch prepared by the method can quickly generate a strong self-adhesive sealing effect after absorbing water, and meanwhile, the patch has the advantages of good biocompatibility, low expansibility and tensile resistance, and has higher clinical practical value in the aspect of dura mater repair.
In order to realize the purpose, the invention is realized by the following technical scheme:
the invention provides a preparation method of a water-absorbing self-adhesive dura mater patch, which comprises the following steps:
s1, dissolving polylactide-co-caprolactone (namely polylactide caprolactone) in an organic solvent to prepare spinning solution, and performing electrostatic spinning on the spinning solution to prepare a spinning membrane;
s2, uniformly mixing the polyacrylic acid solution and the polyethyleneimine solution, and then freeze-drying and grinding to obtain adhesion layer powder;
and S3, spreading the powder of the adhesion layer of the S2 on the surface, soaked in the wet spraying manner, of the spinning film in the step S1 in advance, and compacting and freeze-drying to obtain the water-absorbing self-adhesive double-layer hard ridge film patch.
Preferably, the concentration of the polylactide-co-caprolactone in the organic solvent is from 10 to 15wt%.
Preferably, the intrinsic viscosity of the polylactide-co-caprolactone is 1.5dl/g.
Preferably, during electrostatic spinning, the spinning voltage is 12-20kV, the liquid supply rate is 1.0-2.0mL/h, the spinning time is 1-6h, the roller rotation speed is 1000-3000rpm, the temperature is 22-25 ℃, and the humidity is 25-60%.
Preferably, the weight average molecular weight (M) of the polyethyleneimine w ) 40000-70000, weight average molecular weight (M) of polyacrylic acid w ) Is 240000-450000。
Preferably, the concentration of the polyacrylic acid solution and the polyethyleneimine solution is 8-15wt%, and the volume ratio of the polyacrylic acid solution to the polyethyleneimine solution is (1-3) to (1-3). Further preferably, the volume ratio of the polyacrylic acid solution to the polyethyleneimine solution is 1.
Further preferably, the solvents of the polyacrylic acid solution and the polyethyleneimine solution are both water.
Preferably, the organic solvent includes, but is not limited to, 6-fluoroisopropanol.
Preferably, the compacting is repeated 3-5 times by using a roller.
The invention also provides the water-absorbing self-adhesive dura mater patch prepared by the preparation method.
The water-absorbing self-adhesive dura mater patch prepared by the method can generate a strong adhesive sealing effect with wet dura mater tissues; has low swelling performance and no risk of swelling to press nerves; the external bursting pressure is 79.56 +/-6.56 mmHg and can bear the maximum value of the cerebrospinal fluid pressure; the tensile strength can reach 1.13 +/-0.05 MPa; has good adhesive capacity, and the shear strength can reach 18.43 +/-2.73 kPa.
The invention also provides application of the water-absorbing self-adhesive dura mater patch in preparation of a dura mater repair medical material.
The water-absorbing self-adhesive dura mater patch prepared by the method can generate a quick and strong adhesive sealing effect after absorbing water, has excellent swelling resistance and can be applied to dura mater repair.
Compared with the prior art, the invention has the beneficial effects that:
the invention discloses a preparation method of a novel water-absorbing self-adhesive double-layer dura mater patch, which comprises the steps of preparing a polylactide-co-caprolactone spinning film by an electrostatic spinning technology, taking the polylactide-co-caprolactone spinning film as a substrate layer, taking particle powder which is formed by mixing polyethyleneimine with a polyacrylic acid solution and then freeze-drying and grinding the mixture as an adhesive layer, and then combining the particle powder of the adhesive layer with a substrate layer spinning film to prepare the double-layer dura mater patch. After the double-layer dura mater patch prepared by the invention is contacted with wet tissues, the double-layer dura mater patch can be firmly combined with the tissues through hydrogen bonds and electrostatic action, and the self-adhesion sealing effect can be quickly exerted without surgical suture and long-time pressing. In addition, the film basal layer has good tensile property and excellent swelling resistance, can bear the pressure of cerebrospinal fluid when being applied to the repair of the dura mater, can not cause the risk of pressing nerves because of too high swelling ratio, and has great practical value in the aspect of repairing the clinical dura mater.
Drawings
FIG. 1 is a schematic appearance of a double-layer dura mater patch (A before freeze-drying and B after freeze-drying);
FIG. 2 shows the in vitro burst pressure test results for the bilayer dura patch of examples 1-3 (p < 0.01 after statistical analysis and p < 0.0001 after statistical analysis);
FIG. 3 is a graph of (A) swelling ratio and (B) thickness change ratio of a two-layer dura patch at different soaking times;
FIG. 4 is a schematic representation of a two-layer dura mater patch adhered to bovine dura mater (A before patch adhesion and B after patch adhesion);
fig. 5 is an SEM image of a bilayer dura patch.
Detailed Description
The following further describes embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
The experimental procedures in the following examples were carried out by conventional methods unless otherwise specified, and the test materials used in the following examples were commercially available by conventional methods unless otherwise specified.
Example 1 preparation method of water-absorbing self-adhesive double-layer dura mater patch
(1) Dissolving 1.0g of polylactide-co-caprolactone (namely polylactide caprolactone with the intrinsic viscosity of 1.5 dl/g) in 6-fluoroisopropanol solvent, stirring for 12h at room temperature to prepare a uniform spinning solution with the concentration of 10wt%, then adding the spinning solution into a 10mL syringe for electrostatic spinning, and collecting the spinning film by using a roller with the rotating speed of 2500 rpm. Wherein the spinning voltage is 16kV, the liquid supply rate is 1.5mL/h, the spinning time is 6h, the spinning temperature and the spinning humidity are respectively 25 ℃ and 45%, and finally, the film is dried in a vacuum oven at 60 ℃ for 24h to prepare the spinning film.
(2) A 25% strength by weight aqueous solution (M) of polyacrylic acid W = 240000) was added to deionized water to dilute to a 15wt% aqueous solution of polyacrylic acid, followed by mixing the aqueous solution of polyacrylic acid with a 15wt% polyethyleneimine (M) w = 70000) aqueous solution was uniformly mixed at a volume ratio of 1.
(3) And (3) spreading the powder of the adhesion layer on the surface of the spinning film which is soaked in advance and sprayed with water, repeatedly compacting by a roller, spraying water again until the spinning film is completely wet, then placing the spinning film loaded with the powder between glass flat plates, and freeze-drying for 24h to obtain the water-absorbing self-adhesive double-layer dura mater patch, wherein the appearance of the water-absorbing self-adhesive double-layer dura mater patch is shown in figure 1.
Example 2 preparation method of water-absorbing self-adhesive double-layer dura mater patch
The preparation method is the same as that of example 1, except that in the step (2), polyacrylic acid aqueous solution with the concentration of 15wt% and polyethylene imine aqueous solution with the concentration of 15wt% are uniformly mixed according to the volume ratio of 1.
Example 3 preparation method of water-absorbing self-adhesive double-layer dura mater patch
The preparation method is the same as example 1, except that in the step (2), polyacrylic acid aqueous solution with the concentration of 15wt% and polyethyleneimine aqueous solution with the concentration of 15wt% are uniformly mixed according to the volume ratio of 3.
Example 4 preparation method of water-absorbing self-adhesive double-layer dura mater patch
The preparation method is the same as example 1, except that in step (2), polyacrylic acid aqueous solution with the concentration of 15wt% is freeze-dried to prepare adhesive layer powder. The result shows that after the powder prepared from the polyacrylic acid aqueous solution is spread on the surface of the electrospun film and water is sprayed, the particles are dissolved to become liquid hydrogel, and the preparation requirement of the double-layer dura mater patch particle layer is not met.
Example 5 preparation method of water-absorbing self-adhesive double-layer dura mater patch
The preparation method was the same as example 1, except that in the step (2), the powder of the adhesive layer was prepared by freeze-drying a 15wt% aqueous solution of polyethyleneimine. The results show that freeze-drying of the polyethyleneimine aqueous solution results in a viscous colloidal liquid, and the desired granular powder cannot be obtained for preparing the double-layer dura mater patch.
Experimental example 1 Performance testing or appearance characterization
(1) And (3) testing the external explosion pressure:
the bovine dura mater is taken as a test tissue, a circular bovine dura mater tissue with the diameter of 30mm is taken, and a round hole is made in the central area of the bovine dura mater tissue by using a puncher with the diameter of 3 mm. The dural patch described in examples 1-3 was trimmed to a circular shape with a diameter of 15mm and adhered to the surface-wetted bovine dural defect area, and then the sample was fixed to the test board. An artificial cerebrospinal fluid solution (containing 124.0mM NaCl,26mM NaHCO) was loaded by a micro syringe pump 3 ,2.5mM KCl,2.0mM CaCl 2 ,1.0mM MgCl 2 ,1.25mM NaH 2 PO 4 10.0mM anhydrous glucose) and operated at a constant rate of 2mL/min, the maximum value of the pressure display was observed and recorded. The results are shown in FIG. 2.
Fig. 2 shows that the mean value of the burst pressure test of the dura mater patch prepared in example 2 is 79.56 ± 6.56mmHg, the maximum value can reach 86mmHg, and the maximum value is greater than the upper limit of 20mmHg of the cerebrospinal fluid pressure, so that the patch can meet the repair requirement of dura mater.
(2) And (3) testing swelling resistance:
the double-layer dura mater patch prepared in example 2 was trimmed to a square shape and initial data (e.g., length, width and thickness) of the patch was recorded, which was then soaked in PBS buffer and the thickness and volume changes of the patch were measured and recorded every 12 h. Swelling data for the dural patch at various times are shown in fig. 3.
Fig. 3 shows that the thickness increase of the dura mater patch after soaking for 48 hours is 7.4%, and the volume swelling ratio is 33.6%, which shows that the dura mater patch has excellent swelling resistance and does not have the risk of pressing nerves when applied to dura mater repair.
(3) Dural damage leakage adhesion test:
a section of bovine dura mater is taken and the bottom end is tightened to ensure that no water seeps out, and a sharp knife is used to make a longitudinal incision with the length of about 1cm at the bottom of the dura mater. PBS buffer was injected through the upper end of bovine dura mater and flowed out of the bottom incision, and the flow rate of the water injection tube was slowly adjusted. The double-layered dura patch prepared in example 2 was lightly attached to the incised dura surface and observed for adhesiveness.
As shown in fig. 4, when the dura patch is attached to the liquid-permeable dura, a quick self-adhesive sealing effect can be achieved, and no liquid flows out from the incision.
(4) And (3) testing by a scanning electron microscope:
the micro-topography of the two-layer dura mater patch prepared in example 2 was observed using a cold field emission scanning electron microscope (HITACHI S-4800). As shown in fig. 5, the back surface of the dura mater patch exhibited a uniform fiber diameter morphology (a), and the front surface of the film to which the granular powder was adhered exhibited a rough asperity morphology (B).
(5) And (3) testing tensile property:
the double-layer dura mater patch prepared in example 2 was cut into a dumbbell shape, parameters such as width (W), thickness (d), length (L) were measured, the patch was soaked in deionized water and fixed to both ends of an electronic universal tester (WD-5A), a 50N sensor was used for the test, the tensile rate of the universal tester was set to 20mm/min, and the tensile strength and elongation at break of the sample were recorded, and the tensile strength = F/(W × d). The result shows that the mean value of the tensile strength of the dura mater patch is 1.13 +/-0.05 MPa, the elongation at break is 133 +/-0.94 percent, and the elastic modulus is 2.51 +/-0.74 MPa, which indicates that the dura mater patch has enough tensile resistance.
(6) And (3) testing the shear strength:
the double-layer dura mater patch prepared in example 2 was manufactured to a size of 20mm × 50mm, and a strip-shaped pigskin having a size of 25mm × 50mm was prepared, first, the subcutaneous adipose tissues of the pigskin were scraped clean, then, the patch adhesive layer was adhered to the wet pigskin, and the sheet was left to stand for 30min, and the adhesion length (L) and width (W) were measured, and then, one ends of the patch and the pigskin were respectively fixed to both ends of an electronic universal tester (WD-5A), a 50N sensor was used for the measurement, the tensile rate of the universal tester was set to 20mm/min, and the shear strength was recorded, and the shear strength = F/(W × L). The results showed that the shear strength of the dura mater patch was 18.43. + -. 2.73kPa.
The embodiments of the present invention have been described in detail, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, and the scope of protection is still within the scope of the invention.
Claims (9)
1. A preparation method of a water-absorbing self-adhesive dura mater patch is characterized by comprising the following steps:
s1, dissolving polylactide-co-caprolactone in an organic solvent to prepare a spinning solution, and performing electrostatic spinning on the spinning solution to prepare a spinning membrane;
s2, uniformly mixing the polyacrylic acid solution and the polyethyleneimine solution, and then freeze-drying and grinding to obtain adhesion layer powder;
and S3, spreading the powder of the adhesion layer of the S2 on the surface, soaked in the wet spraying manner, of the spinning film in the step S1 in advance, and compacting and freeze-drying to obtain the water-absorbing self-adhesive double-layer hard ridge film patch.
2. The method for preparing a water-absorbing self-adhesive dura mater patch according to claim 1, wherein the concentration of the polylactide-co-caprolactone in the organic solvent is 10-15wt%.
3. The preparation method of the water-absorbing self-adhesive dura mater patch according to claim 1, wherein in the electrostatic spinning, the spinning voltage is 12-20kV, the liquid supply rate is 1.0-2.0mL/h, the spinning time is 1-6h, the drum rotation speed is 1000-3000rpm, and the humidity is 25-60%.
4. The preparation method of the water-absorbing self-adhesive dura mater patch according to claim 1, wherein the weight-average molecular weight of the polyethyleneimine is 40000 to 70000, and the weight-average molecular weight of the polyacrylic acid is 240000 to 450000.
5. The method for preparing the water-absorbing self-adhesive dura mater patch according to claim 1, wherein the concentration of the polyacrylic acid solution and the polyethyleneimine solution is 8-15wt%, and the volume ratio of the polyacrylic acid solution to the polyethyleneimine solution is (1-3) to (1-3).
6. The method for preparing the water-absorbing self-adhesive dura mater patch according to claim 1, wherein the organic solvent comprises 6-fluoroisopropanol.
7. The preparation method of the water-absorbing self-adhesive dura mater patch according to claim 1, wherein the compacting is repeated 3-5 times by using a roller.
8. The water-absorbing self-adhesive dura mater patch prepared by the preparation method of any one of claims 1 to 7.
9. Use of the water-absorbing self-adhesive dura mater patch according to claim 8 for preparing a dura mater repair medical material.
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| CN116370687B (en) * | 2023-06-01 | 2023-08-04 | 四川大学 | First-aid hemostatic tissue patch based on acrylic acid copolymer and preparation method thereof |
| CN116942908A (en) * | 2023-07-31 | 2023-10-27 | 鹏拓生物科技(杭州)有限公司 | Absorbable biological isolation composite membrane material and preparation method thereof |
| CN116942908B (en) * | 2023-07-31 | 2024-02-23 | 鹏拓生物科技(杭州)有限公司 | Absorbable biological isolation composite membrane material and preparation method thereof |
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