CN110193098B - Multilayer gradient biological membrane and preparation method thereof - Google Patents
Multilayer gradient biological membrane and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a multilayer gradient biomembrane and a preparation method thereof. The composition and structure of the nanofiber biomembrane are in gradient change, the biomembrane with the gradient structure is constructed through the change of the proportion of the gelatin and the polycaprolactone, the regulation and control of mechanical property and degradation behavior are realized, and the upper layer and the lower layer of the biomembrane have the function of synchronously guiding soft and hard tissues to regenerate. The gradient biological membrane has controllable mechanical property, degradation behavior and good biocompatibility, can simultaneously induce the regeneration of soft and hard tissues, and has application prospect in the field of guided tissue regeneration.
Description
Technical Field
The invention belongs to the technical field of nanofiber membrane preparation, and particularly relates to a multilayer gradient biological membrane and a preparation method thereof.
Background
Modeling the structure and composition of human tissue through the use of appropriate materials and techniques is one of the important goals of biomimetics in the fields of biomaterial engineering and tissue engineering. The electrostatic spinning method can simply and effectively prepare the micro-nano fiber support, and the support has a unique microstructure, appropriate mechanical properties and a structure similar to a natural extracellular matrix, so that the bionic structural characteristics can be achieved, and the electrostatic spinning becomes one of the most potential technologies in the aspect of tissue engineering application. In addition, the nanofiber biofilm prepared by the electrospinning method can not only simulate the structure and environment of extracellular matrix, but also adjust the composition, fiber diameter, porosity, and the like. Therefore, the nanofiber membrane material prepared by the electrostatic spinning method has wide application prospect in the field of biomedical materials, and the reported biomedical materials prepared by the electrostatic spinning method comprise biological membranes, wound dressing materials, hemostatic materials, artificial blood vessels, drug and gene delivery, tissue engineering scaffolds and the like.
At present, there are hundreds of polymers capable of preparing superfine nanofibers by an electrostatic spinning method, including natural polymers such as gelatin, collagen, silk fibroin, chitosan, hyaluronic acid, fibrin and the like and artificially synthesized polymers such as polycaprolactone, polylactic acid, polyvinyl alcohol, polyurethane, polyamide, polylactic acid-glycolic acid copolymer and the like.
The natural polymers such as gelatin, collagen and the like are common spinning materials, and have the advantages of good biocompatibility, low antigenicity, biodegradability, no toxic or side effect of in-vivo degradation products and the like. However, the nanofiber membranes prepared from natural polymers such as gelatin and collagen have the defects of low mechanical strength, high degradation speed and the like, and the application of the nanofiber membranes is limited. Therefore, it is necessary to combine synthetic polymer with better mechanical properties to strengthen and improve the properties of nanofiber membranes such as gelatin and collagen, so that the nanofiber membranes can be applied to the field of medical biomaterials. Polycaprolactone has good biocompatibility, good mechanical properties and the like, has been approved by the U.S. food and drug administration, and is widely applied to the field of biomedicine. Therefore, the polycaprolactone, the gelatin, the collagen and the like are blended and electrostatically spun, so that good mechanical properties can be provided for the nanofiber membrane, and the nanofiber membrane has good biocompatibility, so that the performance of the nanofiber membrane is improved, and the nanofiber membrane is more favorable for being applied to the field of biomedicine.
In addition, the biofilms currently used to guide tissue regeneration can be divided into two broad categories: absorbable biological membranes mainly comprise collagen membranes, chitosan membranes, polylactic acid membranes, polyglycolic acid membranes, polylactic acid and polyglycolic acid copolymer membranes and the like, and the absorbable biological membranes have the defects of poor mechanical property, excessive rapid degradation, no isolation effect and the like; the non-absorbable biological membrane mainly comprises a polytetrafluoroethylene membrane, an expanded polytetrafluoroethylene and tetrafluoroethylene hexafluoropropylene copolymer membrane, a titanium membrane and the like, and the membranes have stable physical and chemical properties and better isolation effect, but have the defects of easy tissue flap cracking, membrane exposure, secondary operation and the like. In the prior art, the biomedical material which can be used for tissue induction has better biodegradability, good biocompatibility and low cytotoxicity. However, most of these nanofiber membranes applied to guided tissue regeneration are single membranes for guiding hard tissue or soft tissue regeneration, and it is only reported that the biofilms simultaneously induce the regeneration of both soft and hard tissues.
Because the single material and structure are difficult to meet the requirement of ideal tissue regeneration guidance, the preparation of the biological membrane with different components and structures by combining several materials has great research prospect and development value.
Disclosure of Invention
The invention aims to: the invention provides a multilayer gradient biomembrane and its preparation method, on the one hand through carrying on the blending with other synthetic macromolecules natural macromolecules such as gelatin, collagen, etc. and adopt the appropriate cross-linking agent to carry on the controllable cross-linking to prepare the nanometer fibrous membrane with certain mechanical strength, so as to solve the deficiency that the mechanical property of single natural macromolecule is insufficient and the degradation rate is too fast; on the other hand, the induced tissue regeneration performance of the nanofiber membrane is adjusted by adding calcium phosphate salt, and the calcium phosphate salt is combined with the nanofiber membrane without the calcium phosphate salt to form a multilayer nanofiber membrane, so that the better mechanical property is provided, and the soft and hard tissue regeneration can be synchronously induced in a bidirectional mode.
The technical scheme adopted by the invention is as follows:
a multilayer gradient biomembrane comprises an upper layer biomembrane, a middle layer biomembrane and a lower layer biomembrane, wherein the thickness of the upper layer biomembrane is 0.1-2mm, the upper layer biomembrane is gelatin/polycaprolactone nanofiber biomembrane, the content of gelatin is 40-90wt%, the middle layer biomembrane is gelatin/polycaprolactone nanofiber biomembrane, the content of gelatin is 1-50wt%, the lower layer biomembrane is gelatin/polycaprolactone/apatite biomembrane, the content of gelatin is 10-90wt%, and the content of apatite is 1-40 wt%.
The preparation method of the multilayer gradient biological membrane comprises the following steps:
s1, adding apatite into an organic solvent for ultrasonic dispersion to obtain an organic solvent containing apatite, wherein the mass of the apatite accounts for 5-35% of the total mass of the organic solvent containing apatite; wherein the mass of the apatite accounts for 5-15% of the total mass of the organic solvent containing apatite;
s2, mixing gelatin and polycaprolactone according to the gelatin content of 10-90wt%, and dissolving the gelatin/polycaprolactone mixture in the apatite-containing organic solvent obtained in the step S1 under the action of continuous stirring, wherein the mass-volume ratio of the gelatin/polycaprolactone to the apatite-containing organic solvent is 6-20 g: 100mL, preferably 10-14 g: 100 mL;
s3, dropwise adding 1-3vt% acetic acid solution into the mixed solution obtained in the step S2, and fully stirring to obtain a spinning solution A;
s4, preparing a lower layer film from the spinning solution A obtained in the step S3 by an electrostatic spinning method;
s5, mixing gelatin and polycaprolactone according to the gelatin content of 1-50wt%, and dissolving the gelatin/polycaprolactone mixture into an organic solvent at 25-45 ℃ under the action of continuous stirring, wherein the mass-volume ratio of the gelatin/polycaprolactone to the organic solvent is 6-20 g: 100mL, preferably 10-14 g: 100 mL;
s6, dropwise adding 1-3vt% acetic acid solution into the mixed solution obtained in the step S5, and fully stirring to obtain a spinning solution B;
s7, preparing a middle layer film by an electrostatic spinning method on the basis of the spinning solution B obtained in the step S6;
s8, mixing gelatin and polycaprolactone according to the gelatin content of 40-90wt%, and dissolving the gelatin/polycaprolactone mixture into an organic solvent at 25-45 ℃ under the action of continuous stirring, wherein the mass-volume ratio of the gelatin/polycaprolactone to the organic solvent is 6-20 g: 100mL, preferably 10-14 g: 100 mL;
s9, dropwise adding 1-3vt% acetic acid solution into the mixed solution obtained in the step S8, and fully stirring to obtain a spinning solution C;
s10, preparing an upper layer membrane from the spinning solution C obtained in the step S9 on the basis of the middle layer membrane by an electrostatic spinning method to obtain a three-layer biological membrane;
s11, drying the three-layer biomembrane obtained in the step S10, placing the dried three-layer biomembrane into a crosslinking solution for crosslinking for 0.5-72h, and cleaning and drying the dried three-layer biomembrane to obtain the biological membrane.
The multilayer gradient biomembrane is of a three-layer membrane structure, each layer of biomembrane contains gelatin and polycaprolactone with different proportions, and the multilayer gradient biomembrane has the characteristics of adjustable and controllable mechanical properties, porosity, degradation behavior and the like, and can be used for preparing an asymmetric membrane structure material with a certain gradient. The gradient biomembranes with different degradation speeds and mechanical properties can be designed and prepared according to the change of the content of the gelatin and the polycaprolactone, the higher the content of the gelatin is, the faster the degradation speed is, the mechanical properties are reduced, the higher the content of the polycaprolactone is, the slower the degradation is, and the better the mechanical properties are. The gelatin content of the upper layer is highest, the compatibility of cells and tissues is good, the degradation is fast, and the soft tissue regeneration can be effectively guided; the polycaprolactone content of the middle layer is highest, the degradation is slower, the tensile strength is highest, the mechanical support and barrier effects are realized, the integrity of the membrane structure in the process of guiding tissue regeneration can be maintained, and the polycaprolactone can be used as a transition layer to connect an upper membrane and a lower membrane; the lower layer contains calcium and phosphorus components, which endows the membrane with better biological activity and promotes the regeneration of bone tissues.
Furthermore, the gelatin is one of pig skin gelatin, fish skin gelatin, type I collagen, type II collagen, type III collagen, type IV collagen, type VI collagen, type X collagen and type XI collagen.
Further, the apatite is one of hydroxyapatite, tricalcium phosphate, calcium hydrogen phosphate, calcium pyrophosphate, and octacalcium phosphate, and the particle size of the apatite is 20 to 200nm, preferably 30 to 100 nm.
Further, the organic solvent is trifluoroethanol or hexafluoroisopropanol.
Further, the polycaprolactone may be replaced with one of polylactic acid, polylactic-co-glycolic acid, and polyurethane.
Furthermore, the voltage of electrostatic spinning is 4-12kV, the distance of a receiving plate is 10-20cm, and the injection speed is 0.1-1 mL/h; preferably, the voltage of electrostatic spinning is 6-10kV, the distance of a receiving plate is 13-17cm, and the injection speed is 0.3-0.7 mL/h.
Further, the ultrasonic power in the step S1 is 200-400W, and the ultrasonic time is 30-120 min; preferably, the ultrasonic power is 250-350W, and the ultrasonic time is 30-60 min.
Further, in the step S11, the crosslinking temperature is 0-5 ℃, and the crosslinking solution is an ethanol mixed solution with the crosslinking agent concentration of 5-100 mM; the crosslinking solution can also adopt ethanol water mixed solution with the concentration of the crosslinking agent of 5-100mM, wherein the water content is 5vt percent.
Further, the cross-linking agent is at least one of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide, genipin, glutaraldehyde and vanillin.
In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:
1. the invention prepares three layers of gradient biological membranes by electrostatic spinning technology, the thickness of each layer is 0.1-2mm, the fiber diameter is distributed between 300-500nm, the composite membrane has good biocompatibility, biodegradability and capability of guiding the growth of soft and hard tissues;
2. the upper, middle and lower three-layer films of the invention contain gelatin and polycaprolactone in different proportions, and have adjustable and controllable mechanical properties, porosity, degradation behavior and the like, so that an asymmetric film structure material with a certain gradient can be prepared;
3. the asymmetric structures of the upper layer and the lower layer of the biological membrane can respectively induce the growth of soft tissues and hard tissues, and the middle layer plays a role of a barrier on one hand and prevents soft tissues and connective tissues from growing into a bone defect area; on the other hand, the membrane plays a role in transitional connection and is tightly connected with the upper and lower layers of membranes, so that the upper and lower layers of membranes are prevented from being separated, and simultaneously, better mechanical properties can be provided for the biological membrane;
4. the upper layer gelatin of the biological membrane has the highest content, the degradation is fast, and the regeneration of soft tissues can be effectively induced; the polycaprolactone content of the middle layer is highest, the degradation is slower, the tensile strength is highest, the mechanical support and barrier effects are realized, the integrity of the membrane structure in the process of guiding tissue regeneration can be maintained, and the polycaprolactone can be used as a transition layer to connect an upper membrane and a lower membrane; the lower layer contains calcium and phosphorus components, so that the biological membrane has better biological activity and can promote the regeneration of bone tissues;
5. the biological membrane overcomes the defects of insufficient mechanical property and over-high degradation speed of a single natural polymer, not only provides better mechanical property, but also can synchronously and bidirectionally induce the regeneration of soft and hard tissue tissues, and can be better and more widely applied to the field of guided tissue regeneration.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a scanning electron microscope image of the surface of a multilayer gradient biomembrane prepared in example 1 of the present invention;
FIG. 2 is a scanning electron microscope image of a cross section of a multilayer gradient biomembrane prepared in example 1 of the present invention;
FIG. 3 is a stress-strain curve of polycaprolactone/gelatin biofilms of different gelatin content;
FIG. 4 is a diagram showing the appearance of polycaprolactone/gelatin biofilm after 1 day of in vitro degradation;
FIG. 5 is a diagram showing the appearance of polycaprolactone/gelatin biofilm after 7 days of in vitro degradation;
FIG. 6 is a topographical view of polycaprolactone/gelatin biofilm after 84 days of in vitro degradation;
FIG. 7 is a scanning electron micrograph of mesenchymal stem cells cultured on a three-layer gradient biofilm for 7 days;
FIG. 8 is a scanning electron micrograph of mesenchymal stem cells after cultured on polycaprolactone/gelatin/hydroxyapatite for 7 days;
FIG. 9 is a scanning electron micrograph of mesenchymal stem cells after culturing for 7 days on polycaprolactone/gelatin;
FIG. 10 is a scanning electron micrograph of mesenchymal stem cells cultured on polycaprolactone fibrous membrane for 7 days;
FIG. 11 is a scanning electron micrograph of gingival fibroblasts after being cultured on a three-layered gradient biofilm for 7 days;
FIG. 12 is a scanning electron micrograph of gingival fibroblasts after being cultured on polycaprolactone/gelatin/hydroxyapatite for 7 days;
FIG. 13 is a scanning electron micrograph of gingival fibroblasts after 7 days of culture on polycaprolactone/gelatin;
FIG. 14 is a scanning electron microscope image of gingival fibroblasts cultured on polycaprolactone fiber membrane for 7 days.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the detailed description and specific examples, while indicating the preferred embodiment of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without making any creative effort, shall fall within the protection scope of the present invention.
It is noted that relational terms such as "first" and "second," and the like, may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
The preparation method of the multilayer gradient biomembrane provided by the preferred embodiment of the invention comprises the following specific steps:
dissolving 0.06g of polycaprolactone and 0.54g of gelatin in 5mL of trifluoroethanol at 25 ℃ under the stirring action, adding 0.015mL of acetic acid after the polycaprolactone and the gelatin are completely dissolved, and fully stirring to prepare a polycaprolactone/gelatin spinning solution A; dissolving 0.30g of polycaprolactone and 0.30g of gelatin in 5mL of trifluoroethanol at 25 ℃ under the stirring action, adding 0.015mL of acetic acid after the polycaprolactone and the gelatin are completely dissolved, and fully stirring to prepare a polycaprolactone/gelatin spinning solution B; at 25 ℃, 0.0325g of nano-hydroxyapatite is added into 5mL of trifluoroethanol solution, ultrasonic oscillation is carried out for 60min at 350W, and then 0.455g of polycaprolactone and 0.195g of gelatin are dissolved into the trifluoroethanol solution of the hydroxyapatite under the stirring action, so as to prepare the spinning solution C of the nano-hydroxyapatite/polycaprolactone/gelatin. Under the spinning parameters of the acceptance distance of 15cm, the spinning voltage of 8kV positive electrode, negative electrode-2 kV negative electrode, the injection speed of 0.5mL/h and the like, taking a stainless steel plate pasted with tinfoil paper as a receiving device, firstly spinning the solution C to generate a lower layer biomembrane; spinning the solution B on the lower layer film by taking the acceptance distance of 15cm, the spinning voltage of 6kV positive electrode, the negative electrode of-2 kV negative electrode and the injection speed of 0.5mL/h as spinning parameters to generate an intermediate layer biomembrane; and finally, spinning the solution A on the intermediate layer film by using the acceptance distance of 15cm, the spinning voltage of 8kV positive electrode, the negative electrode of-2 kV negative electrode and the injection speed of 0.5mL/h as spinning parameters to form an upper layer biomembrane. Marking the upper layer and the lower layer of the obtained three-layer biomembrane, drying for 24h at 25 ℃ in vacuum, then crosslinking for 24h in ethanol solution containing 50mM EDC/50mM NHS, washing for a plurality of times by using absolute ethyl alcohol after the crosslinking is finished, drying, packaging, sterilizing and the like to obtain the multilayer gradient biomembrane.
The thickness of the prepared biomembrane is 0.2-0.3mm by measuring with a vernier caliper, and the prepared biomembrane is composed of fibers with the diameter of 300-500nm and forms a porous structure by gold plating and S-450 type scanning electron microscope observation.
Example 2
The preparation method of the multilayer gradient biomembrane provided by the preferred embodiment of the invention comprises the following specific steps:
dissolving 0.18g of polycaprolactone and 0.42g of gelatin in 5mL of trifluoroethanol at 25 ℃ under the stirring action, adding 0.015mL of acetic acid after the polycaprolactone and the gelatin are completely dissolved, and fully stirring to prepare a polycaprolactone/gelatin spinning solution A; dissolving 0.30g of polycaprolactone and 0.30g of gelatin in 5mL of trifluoroethanol at 25 ℃ under the stirring action, adding 0.015mL of acetic acid after the polycaprolactone and the gelatin are completely dissolved, and fully stirring to prepare a polycaprolactone/gelatin spinning solution B; at 25 ℃, 0.065g of nano-hydroxyapatite is added into 5mL of trifluoroethanol solution, ultrasonic oscillation is carried out for 60min at 350W, and then 0.455g of polycaprolactone and 0.195g of gelatin are dissolved into the trifluoroethanol solution of the hydroxyapatite under the stirring action, so as to prepare the spinning solution C of the nano-hydroxyapatite/polycaprolactone/gelatin. Under the spinning parameters of the acceptance distance of 15cm, the spinning voltage of 8kV positive electrode, negative electrode-2 kV negative electrode, the injection speed of 0.5mL/h and the like, taking a stainless steel plate pasted with tinfoil paper as a receiving device, firstly spinning the solution C to generate a lower layer biomembrane; spinning the solution B on the lower layer film by taking the acceptance distance of 15cm, the spinning voltage of 6kV positive electrode, the negative electrode of-2 kV negative electrode and the injection speed of 0.5mL/h as spinning parameters to generate an intermediate layer biomembrane; and finally, spinning the solution A on the intermediate layer film by using the acceptance distance of 15cm, the spinning voltage of 8kV positive electrode, the negative electrode of-2 kV negative electrode and the injection speed of 0.5mL/h as spinning parameters to form an upper layer biomembrane. Marking the upper layer and the lower layer of the obtained three-layer biomembrane, drying for 24h at 25 ℃ in vacuum, then crosslinking for 24h in ethanol solution containing 50mM EDC/50mM NHS, washing for a plurality of times by using absolute ethyl alcohol after the crosslinking is finished, drying, packaging, sterilizing and the like to obtain the multilayer gradient biomembrane.
The thickness of the prepared biomembrane is 0.2-0.3mm by measuring with a vernier caliper, and the prepared biomembrane is composed of fibers with the diameter of 300-500nm and forms a porous structure by gold plating and S-450 type scanning electron microscope observation.
Example 3
The preparation method of the multilayer gradient biomembrane provided by the preferred embodiment of the invention comprises the following specific steps:
dissolving 0.30g of polycaprolactone and 0.30g of gelatin in 5mL of trifluoroethanol at 25 ℃ under the stirring action, adding 0.015mL of acetic acid after the polycaprolactone and the gelatin are completely dissolved, and fully stirring to prepare a polycaprolactone/gelatin spinning solution A; dissolving 0.30g of polycaprolactone and 0.30g of gelatin in 5mL of trifluoroethanol at 25 ℃ under the stirring action, adding 0.015mL of acetic acid after the polycaprolactone and the gelatin are completely dissolved, and fully stirring to prepare a polycaprolactone/gelatin spinning solution B; 0.0975g of nano-hydroxyapatite is added into 5mL of trifluoroethanol solution at 25 ℃, ultrasonic oscillation is carried out for 60min at 350W, and then 0.455g of polycaprolactone and 0.195g of gelatin are dissolved into the trifluoroethanol solution of the hydroxyapatite under the stirring action, so as to prepare the spinning solution C of the nano-hydroxyapatite/polycaprolactone/gelatin. Under the spinning parameters of the acceptance distance of 15cm, the spinning voltage of 8kV positive electrode, negative electrode-2 kV negative electrode, the injection speed of 0.5mL/h and the like, taking a stainless steel plate pasted with tinfoil paper as a receiving device, firstly spinning the solution C to generate a lower layer biomembrane; spinning the solution B on the lower layer film by taking the acceptance distance of 15cm, the spinning voltage of 6kV positive electrode, the negative electrode of-2 kV negative electrode and the injection speed of 0.5mL/h as spinning parameters to generate an intermediate layer biomembrane; and finally, spinning the solution A on the intermediate layer film by using the acceptance distance of 15cm, the spinning voltage of 8kV positive electrode, the negative electrode of-2 kV negative electrode and the injection speed of 0.5mL/h as spinning parameters to form an upper layer biomembrane. Marking the upper layer and the lower layer of the obtained three-layer biomembrane, drying for 24h at 25 ℃ in vacuum, then crosslinking for 24h in ethanol solution containing 50mM EDC/50mM NHS, washing for a plurality of times by using absolute ethyl alcohol after the crosslinking is finished, drying, packaging, sterilizing and the like to obtain the multilayer gradient biomembrane.
The thickness of the prepared biomembrane is 0.2-0.3mm by measuring with a vernier caliper, and the prepared biomembrane is composed of fibers with the diameter of 300-500nm and forms a porous structure by gold plating and S-450 type scanning electron microscope observation.
Example 4
The preparation method of the multilayer gradient biomembrane provided by the preferred embodiment of the invention comprises the following specific steps:
dissolving 0.42g of polycaprolactone and 0.18g of gelatin in 5mL of trifluoroethanol at 25 ℃ under the stirring action, adding 0.015mL of acetic acid after the polycaprolactone and the gelatin are completely dissolved, and fully stirring to prepare a polycaprolactone/gelatin spinning solution A; dissolving 0.30g of polycaprolactone and 0.30g of gelatin in 5mL of trifluoroethanol at 25 ℃ under the stirring action, adding 0.015mL of acetic acid after the polycaprolactone and the gelatin are completely dissolved, and fully stirring to prepare a polycaprolactone/gelatin spinning solution B; at 25 ℃, 0.0325g of nano-hydroxyapatite is added into 5mL of trifluoroethanol solution, ultrasonic oscillation is carried out for 60min at 350W, and then 0.455g of polycaprolactone and 0.195g of gelatin are dissolved into the trifluoroethanol solution of the hydroxyapatite under the stirring action, so as to prepare the spinning solution C of the nano-hydroxyapatite/polycaprolactone/gelatin. Under the spinning parameters of the acceptance distance of 15cm, the spinning voltage of 8kV positive electrode, negative electrode-2 kV negative electrode, the injection speed of 0.5mL/h and the like, taking a stainless steel plate pasted with tinfoil paper as a receiving device, firstly spinning the solution C to generate a lower layer biomembrane; spinning the solution B on the lower layer film by taking the acceptance distance of 15cm, the spinning voltage of 6kV positive electrode, the negative electrode of-2 kV negative electrode and the injection speed of 0.5mL/h as spinning parameters to generate an intermediate layer biomembrane; and finally, spinning the solution A on the intermediate layer film by using the acceptance distance of 15cm, the spinning voltage of 8kV positive electrode, the negative electrode of-2 kV negative electrode and the injection speed of 0.5mL/h as spinning parameters to form an upper layer biomembrane. Marking the upper layer and the lower layer of the obtained three-layer biomembrane, drying for 24h at 25 ℃ in vacuum, then crosslinking for 24h in ethanol solution containing 50mM EDC/50mM NHS, washing for a plurality of times by using absolute ethyl alcohol after the crosslinking is finished, drying, packaging, sterilizing and the like to obtain the multilayer gradient biomembrane.
The thickness of the prepared biomembrane is 0.2-0.3mm by measuring with a vernier caliper, and the prepared biomembrane is composed of fibers with the diameter of 300-500nm and forms a porous structure by gold plating and S-450 type scanning electron microscope observation.
Example 5
The preparation method of the multilayer gradient biomembrane provided by the preferred embodiment of the invention comprises the following specific steps:
dissolving 0.54g of polycaprolactone and 0.06g of gelatin in 5mL of trifluoroethanol at 25 ℃ under the stirring action, adding 0.015mL of acetic acid after the polycaprolactone and the gelatin are completely dissolved, and fully stirring to prepare a polycaprolactone/gelatin spinning solution A; dissolving 0.30g of polycaprolactone and 0.30g of gelatin in 5mL of trifluoroethanol at 25 ℃ under the stirring action, adding 0.015mL of acetic acid after the polycaprolactone and the gelatin are completely dissolved, and fully stirring to prepare a polycaprolactone/gelatin spinning solution B; at 25 ℃, 0.065g of nano-hydroxyapatite is added into 5mL of trifluoroethanol solution, ultrasonic oscillation is carried out for 60min at 350W, and then 0.455g of polycaprolactone and 0.195g of gelatin are dissolved into the trifluoroethanol solution of the hydroxyapatite under the stirring action, so as to prepare the spinning solution C of the nano-hydroxyapatite/polycaprolactone/gelatin. Under the spinning parameters of the acceptance distance of 15cm, the spinning voltage of 8kV positive electrode, negative electrode-2 kV negative electrode, the injection speed of 0.5mL/h and the like, taking a stainless steel plate pasted with tinfoil paper as a receiving device, firstly spinning the solution C to generate a lower layer biomembrane; spinning the solution B on the lower layer film by taking the acceptance distance of 15cm, the spinning voltage of 6kV positive electrode, the negative electrode of-2 kV negative electrode and the injection speed of 0.5mL/h as spinning parameters to generate an intermediate layer biomembrane; and finally, spinning the solution A on the intermediate layer film by using the acceptance distance of 15cm, the spinning voltage of 8kV positive electrode, the negative electrode of-2 kV negative electrode and the injection speed of 0.5mL/h as spinning parameters to form an upper layer biomembrane. Marking the upper layer and the lower layer of the obtained three-layer biomembrane, drying for 24h at 25 ℃ in vacuum, then crosslinking for 24h in ethanol solution containing 50mM EDC/50mM NHS, washing for a plurality of times by using absolute ethyl alcohol after the crosslinking is finished, drying, packaging, sterilizing and the like to obtain the multilayer gradient biomembrane.
The thickness of the prepared biomembrane is 0.2-0.3mm by measuring with a vernier caliper, and the prepared biomembrane is composed of fibers with the diameter of 300-500nm and forms a porous structure by gold plating and S-450 type scanning electron microscope observation.
Experimental example 1
Scanning electron microscope observation is respectively carried out on the surface and the section of the multilayer gradient biomembrane prepared in the embodiment 1 of the invention, and the result is shown in figure 1 and figure 2, and the visible fiber has good appearance and is in a typical multilayer structure.
Experimental example 2
The polycaprolactone/gelatin biomembranes with different gelatin contents prepared by the preparation method of the invention are subjected to stress-strain curve measurement, and the result is shown in figure 3, which shows that the tensile strength of the fibrous membrane is the maximum when the polycaprolactone content is 90% and the gelatin content is 10%.
Experimental example 3
In vitro degradation tests were performed on the polycaprolactone/gelatin biofilm prepared by the preparation method of the present invention, and the morphology was observed in 1 day, 7 days, and 84 days, respectively, and the results are shown in fig. 4-6, which indicates that the fiber pores became large and the morphology remained good after a long time.
Experimental example 4
And (3) culturing the bone marrow mesenchymal stem cells on the three-layer gradient biomembrane (A), the polycaprolactone/gelatin/hydroxyapatite (B), the polycaprolactone/gelatin (C) and the polycaprolactone fibrous membrane (D) respectively, and observing by using a scanning electron microscope after culturing for 7 days, wherein the results are shown in figures 7-10, and the bone marrow mesenchymal stem cells grow well on the gradient biomembrane.
Experimental example 5
The gingival fibroblasts are cultured on the three layers of the gradient biomembrane (A), the polycaprolactone/gelatin/hydroxyapatite (B), the polycaprolactone/gelatin (C) and the polycaprolactone fiber membrane (D) respectively, and the scanning electron microscope observation is carried out after the culture for 7 days, so that the results are shown in figures 11-14, and the gingival fibroblasts can be known to grow well on the gradient biomembrane.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (7)
1. A multilayer gradient biofilm, comprising: the biological film comprises biological films with the thicknesses of an upper layer, a middle layer and a lower layer of 0.1-2mm, wherein the upper layer is a gelatin/polycaprolactone nanofiber biological film, the gelatin content is 40-90wt%, the middle layer is a gelatin/polycaprolactone nanofiber biological film, the gelatin content is 1-50wt%, the lower layer is a gelatin/polycaprolactone/apatite biological film, the gelatin content is 10-90wt%, and the apatite content is 1-40 wt%;
the biological membrane consists of fibers with the diameter of 300-500 nm;
the preparation method comprises the following steps:
s1, adding apatite into an organic solvent for ultrasonic dispersion to obtain an organic solvent containing apatite, wherein the mass of the apatite accounts for 5-35% of the total mass of the organic solvent containing apatite;
s2, mixing gelatin and polycaprolactone according to the gelatin content of 10-90wt%, and dissolving the gelatin/polycaprolactone mixture in the apatite-containing organic solvent obtained in the step S1 under the action of continuous stirring, wherein the mass-volume ratio of the gelatin/polycaprolactone to the apatite-containing organic solvent is 6-20 g: 100 mL;
s3, dropwise adding 1-3vt% acetic acid solution into the mixed solution obtained in the step S2, and fully stirring to obtain a spinning solution A;
s4, preparing a lower layer film from the spinning solution A obtained in the step S3 by an electrostatic spinning method;
s5, mixing gelatin and polycaprolactone according to the gelatin content of 1-50wt%, and dissolving the gelatin/polycaprolactone mixture into an organic solvent at 25-45 ℃ under the action of continuous stirring, wherein the mass-volume ratio of the gelatin/polycaprolactone to the organic solvent is 6-20 g: 100 mL;
s6, dropwise adding 1-3vt% acetic acid solution into the mixed solution obtained in the step S5, and fully stirring to obtain a spinning solution B;
s7, preparing a middle layer film by an electrostatic spinning method on the basis of the spinning solution B obtained in the step S6;
s8, mixing gelatin and polycaprolactone according to the gelatin content of 40-90wt%, and dissolving the gelatin/polycaprolactone mixture into an organic solvent at 25-45 ℃ under the action of continuous stirring, wherein the mass-volume ratio of the gelatin/polycaprolactone to the organic solvent is 6-20 g: 100 mL;
s9, dropwise adding 1-3vt% acetic acid solution into the mixed solution obtained in the step S8, and fully stirring to obtain a spinning solution C;
s10, preparing an upper layer membrane from the spinning solution C obtained in the step S9 on the basis of the middle layer membrane by an electrostatic spinning method to obtain a three-layer biological membrane;
s11, drying the three-layer biomembrane obtained in the step S10, placing the dried three-layer biomembrane into a crosslinking solution for crosslinking for 0.5-72h, and cleaning and drying the dried three-layer biomembrane to obtain the biomembrane;
wherein the crosslinking temperature in the step S11 is 0-5 ℃, the crosslinking solution is an ethanol mixed solution with the concentration of the crosslinking agent of 5-100mM, and the crosslinking agent is at least one of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide, genipin, glutaraldehyde and vanillin.
2. A multilayer gradient biofilm according to claim 1, wherein: the gelatin is pig skin gelatin or fish skin gelatin.
3. A multilayer gradient biofilm according to claim 1, wherein: the apatite is one of hydroxyapatite, tricalcium phosphate, calcium hydrogen phosphate, calcium pyrophosphate and octacalcium phosphate, and the particle size of the apatite is 20-200 nm.
4. A multilayer gradient biofilm according to claim 1, wherein: the organic solvent is trifluoroethanol or hexafluoroisopropanol.
5. A multilayer gradient biofilm according to claim 1, wherein: the polycaprolactone can be replaced by one of polylactic acid, polylactic-glycolic acid and polyurethane.
6. A multilayer gradient biofilm according to claim 1, wherein: the voltage of the electrostatic spinning is 4-12kV, the distance of a receiving plate is 10-20cm, and the injection speed is 0.1-1 mL/h.
7. A multilayer gradient biofilm according to claim 1, wherein: the ultrasonic power in the step S1 is 200-400W, and the ultrasonic time is 30-120 min.
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| CN111472093A (en) * | 2020-04-08 | 2020-07-31 | 武汉理工大学 | Nano-hydroxyapatite/polylactic acid composite fiber membrane and preparation method thereof |
| CN111803713A (en) * | 2020-05-29 | 2020-10-23 | 广州新诚生物科技有限公司 | Woven film, woven bag and weaving method |
| CN113026199A (en) * | 2021-03-11 | 2021-06-25 | 北京化工大学 | Component-gradient tendon adhesion-prevention fibrous membrane and preparation method thereof |
| CN113332497B (en) * | 2021-04-30 | 2022-04-22 | 国家纳米科学中心 | Double-sided bracket and preparation method and application thereof |
| CN113230460A (en) * | 2021-05-07 | 2021-08-10 | 四川大学 | Porous scaffold for guiding integrated restoration of soft and hard tissues of oral cavity and preparation method thereof |
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6869701B1 (en) * | 1999-08-16 | 2005-03-22 | Carolyn Aita | Self-repairing ceramic coatings |
| CN1943801A (en) * | 2006-11-01 | 2007-04-11 | 华中科技大学 | A gradient laminated composite supporting frame material based on bionic structures and its preparation method |
| CN101584885A (en) * | 2009-06-25 | 2009-11-25 | 同济大学 | Preparation method of three-layer lead tissue regenerating velum with gradient |
| CN103736153A (en) * | 2013-12-30 | 2014-04-23 | 北京市创伤骨科研究所 | Single-layer and double-layer polycaprolactone-based guided tissue regeneration membranes and preparation method thereof |
| CN104414772A (en) * | 2013-09-06 | 2015-03-18 | 山东百多安医疗器械有限公司 | In-vivo degradable and absorbable artificial medical tissue repairing film |
| CN105031739A (en) * | 2015-06-25 | 2015-11-11 | 中国人民解放军第四军医大学 | Doxycycline-hyclate-carried GTR/GBR composite membrane and preparation method thereof |
| CN106110407A (en) * | 2016-08-12 | 2016-11-16 | 上海交通大学医学院附属第九人民医院 | A kind of inductive bone regeneration composite film material and preparation method thereof |
| CN107456607A (en) * | 2017-07-03 | 2017-12-12 | 广州医科大学附属口腔医院 | Guide Periodontal Tissue Regeneration film of new " sandwich " structure a kind of of difunctionalization and its preparation method and application |
| CN107596448A (en) * | 2017-11-14 | 2018-01-19 | 四川大学 | Can gradient degradation biomembrane timbering material and preparation method thereof |
| CN109876192A (en) * | 2019-03-13 | 2019-06-14 | 东华大学 | A kind of Bone Defect Repari film and preparation method thereof |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102166378B (en) * | 2011-01-13 | 2013-08-21 | 北京化工大学 | Tissue regeneration guiding membrane and preparation method thereof |
| US10227568B2 (en) * | 2011-03-22 | 2019-03-12 | Nanofiber Solutions, Llc | Fiber scaffolds for use in esophageal prostheses |
| US10195022B2 (en) * | 2015-04-30 | 2019-02-05 | Mayo Foundation For Medical Education And Research | Nanofibrous biologic heart valve leaflets and fibrosa layer of a leaflet |
| CN106267341A (en) * | 2016-08-30 | 2017-01-04 | 圆容生物医药无锡有限公司 | One can organize induction bio-medical material |
| US20180064853A1 (en) * | 2016-09-06 | 2018-03-08 | Roohollah Bagherzadeh | Hierarchically Structured and Multifunctional Nanofibrous Composite Structure for Soft-Tissue Engineering Applications |
| IN201741001087A (en) * | 2017-01-11 | 2018-07-13 | ||
| CN106975106A (en) * | 2017-03-31 | 2017-07-25 | 北京化工大学 | A kind of double-deck Bone Defect Repari membrane material and preparation method thereof |
| CN109513046B (en) * | 2018-12-18 | 2021-06-22 | 诺一迈尔（苏州）医学科技有限公司 | Absorbable oral cavity repairing film and preparation method thereof |
-
2019
- 2019-06-21 CN CN201910542360.7A patent/CN110193098B/en active Active
- 2019-07-12 WO PCT/CN2019/095664 patent/WO2020252825A1/en active Application Filing
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6869701B1 (en) * | 1999-08-16 | 2005-03-22 | Carolyn Aita | Self-repairing ceramic coatings |
| CN1943801A (en) * | 2006-11-01 | 2007-04-11 | 华中科技大学 | A gradient laminated composite supporting frame material based on bionic structures and its preparation method |
| CN101584885A (en) * | 2009-06-25 | 2009-11-25 | 同济大学 | Preparation method of three-layer lead tissue regenerating velum with gradient |
| CN104414772A (en) * | 2013-09-06 | 2015-03-18 | 山东百多安医疗器械有限公司 | In-vivo degradable and absorbable artificial medical tissue repairing film |
| CN103736153A (en) * | 2013-12-30 | 2014-04-23 | 北京市创伤骨科研究所 | Single-layer and double-layer polycaprolactone-based guided tissue regeneration membranes and preparation method thereof |
| CN105031739A (en) * | 2015-06-25 | 2015-11-11 | 中国人民解放军第四军医大学 | Doxycycline-hyclate-carried GTR/GBR composite membrane and preparation method thereof |
| CN106110407A (en) * | 2016-08-12 | 2016-11-16 | 上海交通大学医学院附属第九人民医院 | A kind of inductive bone regeneration composite film material and preparation method thereof |
| CN107456607A (en) * | 2017-07-03 | 2017-12-12 | 广州医科大学附属口腔医院 | Guide Periodontal Tissue Regeneration film of new " sandwich " structure a kind of of difunctionalization and its preparation method and application |
| CN107596448A (en) * | 2017-11-14 | 2018-01-19 | 四川大学 | Can gradient degradation biomembrane timbering material and preparation method thereof |
| CN109876192A (en) * | 2019-03-13 | 2019-06-14 | 东华大学 | A kind of Bone Defect Repari film and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| A novel spatially designed and functionally graded electrospun membrane for periodontal regeneration;Marco C. Bottino et al;《Acta Biomaterialia》;20100827;第216–224页 * |
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