CN112220965A - Janus porous biological patch for wound repair and preparation method thereof - Google Patents
Janus porous biological patch for wound repair and preparation method thereof Download PDFInfo
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- 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
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A61L27/025—Other specific inorganic materials not covered by A61L27/04 - A61L27/12
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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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- 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
- A61L27/56—Porous materials, e.g. foams or sponges
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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
- A61L27/60—Materials for use in artificial skin
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a Janus porous biological patch for wound repair and a preparation method thereof. The Janus porous biological patch for wound repair, which is prepared by the invention, is constructed based on two colloid droplet templates with different surface wettability, so that the patch has controllable surface wettability, a hydrophobic surface is directly contacted with a wound to avoid secondary wound caused by tissue adhesion, and a hydrophilic surface can adsorb exudate at the wound, so that the wound repair is accelerated. The preparation method is simple and easy to implement, strong in controllability, good in repeatability and wide in application prospect.
Description
Technical Field
The invention relates to the technical field of biomedical materials, in particular to a Janus porous biological patch for wound repair and a preparation method thereof.
Background
Skin wounds are not only very common in life, but also very easy to cause in surgical treatment, so wound repair has attracted high attention from the medical field. The ideal wound repair method is to promote cell adhesion and proliferation, directionally migrate to the wound area, secrete extracellular matrix and restore the original functions of tissues. To accomplish these processes, biological patches are produced. The biological patch is a film patch prepared from one or more biocompatible materials, and can meet the requirements of wound repair of different parts of a human body.
Generally, an anti-adhesion biological patch made of hydrophobic materials with low surface energy and small surface roughness can effectively isolate a wound area from adjacent tissues, but the promotion effect of the anti-adhesion biological patch on tissue regeneration is not ideal. On the other hand, the adhesive biological patch, such as acellular tissue matrix, polymer gauze and the like, has the function of promoting cell adhesion and infiltration, but also has unnecessary wound surface adhesion, and is easy to cause secondary wound.
Disclosure of Invention
The invention aims to solve the technical problem of the prior art and provides a Janus porous biological patch for wound repair and a preparation method thereof.
The invention designs a Janus porous biological patch for wound repair based on surface wettability, the biological patch has a Janus structure, the wettability of the surfaces on two sides of the biological patch is completely different, a hydrophobic surface is directly contacted with a wound to avoid secondary wound caused by tissue adhesion, and a hydrophilic surface can adsorb exudate at the wound, so that the wound repair is accelerated. The biological patch is an ideal tissue repair material and has wide clinical application value.
In order to achieve the technical purpose, the technical scheme adopted by the invention is as follows:
a Janus porous biological patch for wound repair is prepared by the following steps:
(1) constructing a microfluidic single emulsion device: breaking a glass capillary tube by using a capillary tube drawing instrument, and then polishing the broken glass capillary tube into a tip end by using abrasive paper to serve as an internal phase tube; taking another glass capillary tube with the same size, and polishing one end of the glass capillary tube smoothly by using sand paper to serve as an outer phase tube; assembling the inner phase tube and the outer phase tube on a cover glass, keeping the central lines horizontally aligned, and fixing; connecting and sealing the interface of the two glass capillary tubes for later use;
(2) preparing a colloid liquid drop template: preparing a colloid liquid drop template with hydrophilic silicon dioxide nano particles and hydrophobic magnetic nano particles wrapped on the surface by using the microfluidic single emulsion device built in the step (1); the inner phase solution is n-hexadecane dispersed with hydrophilic silicon dioxide nanoparticles and hydrophobic magnetic nanoparticles, the outer phase solution is a photo-polymerization aqueous polyurethane solution added with a surfactant, the inner phase solution is sheared by the outer phase solution at the tip of an inner phase tube to generate colloid droplets, and the colloid droplets are automatically layered under the action of a magnetic field and assembled in a hexagonal close-packed structure to form a colloid droplet template;
(3) preparing a Janus porous biological patch for wound repair: and (3) taking the colloid liquid drop template prepared in the step (2) as a positive template, carrying out in-situ ultraviolet photopolymerization on the external phase solution-the photopolymerization aqueous polyurethane solution, and then removing the positive template to obtain the Janus porous biological patch for wound repair.
The outer diameter of the glass capillary tube adopted in the step (1) is 1mm, the inner diameter is 0.8mm, and the inner phase tube is polished by sand paper until the inner diameter of the tip is 0.4 mm; and ultrasonically cleaning the polished inner phase tube and the polished outer phase tube for 2min by using absolute ethyl alcohol, and naturally drying for later use.
Furthermore, in the external phase solution, the surfactant is a mixed solution of sodium dodecyl sulfate and a polyether surfactant F108, and the mass concentration of the mixed solution is 2% m/v.
Further, the curing method of the photo-polymerization aqueous polyurethane solution is to add 2% m/v of 2-hydroxy-2-methyl propiophenone, and carry out ultraviolet crosslinking curing: wherein the photopolymerisable aqueous polyurethane solution needs to be processed in a dark place before photocuring.
Further, the particle size of the hydrophilic silica nano-particles is 220 nm; the particle size of the hydrophobic magnetic nano particles is 240 nm;
further, the diameter of the colloid droplet is 300-500 μm.
Further, the colloid liquid drop template removing method is that normal hexane is used for soaking and cleaning.
In the step (2), the colloid droplet template coated with the hydrophilic silica nanoparticles and the hydrophobic magnetic nanoparticles on the surface is automatically divided into two layers under the action of a magnetic field, wherein the lower layer is a single-layer colloid droplet coated with the hydrophobic magnetic nanoparticles, and the upper layer is a double-layer colloid droplet coated with the hydrophilic silica nanoparticles.
Furthermore, the colloid liquid drop template coated with the hydrophilic silicon dioxide nano particles and the hydrophobic magnetic nano particles on the surface is automatically divided into two layers under the action of a magnetic field, and the proportion of the hydrophilic colloid liquid drops in the hydrophobic colloid liquid drop layer can be adjusted according to the action time and the distance of the magnetic field.
Furthermore, the lower layer of the Janus porous biological patch for wound repair is a hydrophobic surface doped with hydrophilic holes, the upper layer of the Janus porous biological patch for wound repair is a hydrophilic surface, the hydrophobic surface is in direct contact with the wound to avoid secondary trauma caused by tissue adhesion, and the hydrophilic surface can adsorb exudate at the wound, so that the wound repair is accelerated.
Compared with the prior art, the invention has the beneficial effects that:
1) the preparation method of the Janus porous biological patch for wound repair provided by the invention utilizes a microfluidic single emulsion droplet generation technology to prepare the colloid droplet template coated with the hydrophilic silicon dioxide nanoparticles and the hydrophobic magnetic nanoparticles on the surface, and the preparation method is simple, convenient and feasible, has good controllability and high accuracy, and can adjust the diameter of the droplet on line;
2) the lower layer of the Janus porous biological patch for wound repair provided by the invention is a hydrophobic surface doped with hydrophilic holes, and the upper layer is a hydrophilic surface, wherein the hydrophobic surface is directly contacted with the wound to avoid secondary wound caused by tissue adhesion, and the hydrophilic surface can adsorb exudate at the wound, so that the wound repair is accelerated;
3) the Janus porous biological patch for wound repair provided by the invention has an inverse opal porous structure, uniform pore structure and high connectivity, is beneficial to the transfer of oxygen and nutrient substances, and is capable of abreast removing biological effusion and waste generated by cell metabolism, thereby effectively accelerating wound repair.
Drawings
Fig. 1 is a schematic diagram of a colloidal droplet template for a Janus porous biological patch for wound repair according to the present invention.
Fig. 2 is a schematic diagram of the preparation of the Janus porous biological patch for wound repair according to the present invention.
FIG. 3 is electron microscope images of the hydrophilic side and the hydrophobic side of the Janus porous biological patch for wound repair of the invention; wherein, the figure a is a hydrophilic side electron microscope picture with a ruler of 5 μm, the figure b is a hydrophobic side electron microscope picture with a ruler of 2.5 μm.
FIG. 4 shows the application of Janus porous biological patch in wound repair according to the embodiment of the present invention; wherein a is the H & E staining result with scale of 1mm, b is the Masson staining result with scale of 100 μm.
Detailed Description
In order to make those skilled in the art better understand the technical solution of the present invention, the following detailed description of the embodiments of the present invention is provided with reference to the accompanying drawings.
The experimental procedures used in the examples below are, unless otherwise specified, conventional procedures and the reagents, methods and equipment used are, unless otherwise specified, conventional in the art.
The invention provides a preparation method of a Janus porous biological patch for wound repair, which is characterized in that a colloid droplet template coated with hydrophilic silicon dioxide nanoparticles and hydrophobic magnetic nanoparticles on the surface is prepared by utilizing a microfluidic single emulsion droplet generation technology.
The Janus porous biological patch for wound repair is characterized in that the lower layer is a hydrophobic surface doped with hydrophilic holes, the upper layer is a hydrophilic surface, the hole structure is uniform and high in connectivity, the transfer of oxygen and nutrient substances is facilitated, biological effusion and waste generated by cell metabolism are arranged side by side, the hydrophobic surface is in direct contact with the wound to avoid secondary wound caused by tissue adhesion, and the hydrophilic surface can adsorb exudate at the wound to accelerate wound repair.
The following are examples:
example 1
A Janus porous biological patch with complete anisotropic wettability and used for wound repair is prepared according to the following method:
(1) construction of microfluidic single emulsion devices
A glass capillary (the outer diameter is 1mm, the inner diameter is 0.8mm) is broken by using a capillary tube drawing instrument, and then the tip of the broken glass capillary is polished by abrasive paper until the inner diameter is 0.4mm for later use, so that the glass capillary is an inner phase tube; and taking another glass capillary tube with the same size, polishing one end of the glass capillary tube with sand paper to be smooth to form an outer phase tube, ultrasonically cleaning the inner phase tube and the outer phase tube for 2min by using absolute ethyl alcohol, and naturally drying for later use. Assembling the inner phase tube and the outer phase tube on a cover glass, keeping the central lines horizontally aligned, fixing by using AB glue, connecting the interface of the two glass capillary tubes by using a flat-mouth needle head, finally sealing by using the AB glue, and using after the glue is dried.
(2) Preparation of colloid droplet template
N-hexadecane containing 7% hydrophobic magnetic nanoparticles (m/v) was prepared as the internal phase solution 1. The aqueous solution containing 7% hydrophilic silica nanoparticles (m/v) was replaced with anhydrous ethanol, and this was repeated 3 times to ensure complete removal of water, and then the solution was dispersed in n-hexadecane, and placed in an oven at 40 ℃ overnight to remove the anhydrous ethanol, to obtain a solution of n-hexadecane containing 7% hydrophilic silica nanoparticles as an internal phase solution 2. Preparing an aqueous polyurethane solution containing 2% of sodium dodecyl sulfate and a polyether surfactant F108(m/v), and then adding 2% of a photoinitiator 2-hydroxy-2-methyl propiophenone (m/v) to serve as an external phase solution. Taking two microfluidic single emulsion devices, wherein an inner phase pipe of one device is communicated with an inner phase solution 1, an outer phase pipe is communicated with an outer phase solution, an inner phase pipe of the other device is communicated with an inner phase solution 2, an outer phase pipe is communicated with an outer phase solution, and colloid liquid drops with different sizes are obtained by adjusting the flow rates of the inner phase solution and the outer phase solution of the two devices.
(3) Preparation of Janus porous biological patch for wound repair
And (3) the colloid drops prepared in the step (2) are automatically and completely layered under the action of a magnetic field and assembled in a hexagonal close-packed structure to form a colloid drop positive template, the external phase solution-the photopolymerization aqueous polyurethane solution are subjected to in-situ ultraviolet photopolymerization, and then n-hexane is used for cleaning n-hexadecane to remove the positive template, so that the Janus porous biological patch for wound repair is obtained.
(4) Characterization of Janus porous biological patches for wound repair
The Janus porous biological patch structure for wound repair is observed and characterized under a body type microscope and a scanning electron microscope, and the measured diameter of the holes of the Janus porous biological patch for wound repair is about 350 mu m, and the connectivity among the holes is good.
Example 2
A Janus porous biological patch with partial anisotropic wettability and used for wound repair is prepared according to the following method:
(1) construction of microfluidic single emulsion devices
A glass capillary (the outer diameter is 1mm, the inner diameter is 0.8mm) is broken by using a capillary tube drawing instrument, and then the tip of the broken glass capillary is polished by abrasive paper until the inner diameter is 0.4mm for later use, so that the glass capillary is an inner phase tube; and taking another glass capillary tube with the same size, polishing one end of the glass capillary tube with sand paper to be smooth to form an outer phase tube, ultrasonically cleaning the inner phase tube and the outer phase tube for 2min by using absolute ethyl alcohol, and naturally drying for later use. Assembling the inner phase tube and the outer phase tube on a cover glass, keeping the central lines horizontally aligned, fixing by using AB glue, connecting the interface of the two glass capillary tubes by using a flat-mouth needle head, finally sealing by using the AB glue, and using after the glue is dried.
(2) Preparation of colloid droplet template
N-hexadecane containing 7% hydrophobic magnetic nanoparticles (m/v) was prepared as the internal phase solution 1. The aqueous solution containing 7% hydrophilic silica nanoparticles (m/v) was replaced with anhydrous ethanol, and this was repeated 3 times to ensure complete removal of water, and then the solution was dispersed in n-hexadecane, and placed in an oven at 40 ℃ overnight to remove the anhydrous ethanol, to obtain a solution of n-hexadecane containing 7% hydrophilic silica nanoparticles as an internal phase solution 2. Preparing an aqueous polyurethane solution containing 2% of sodium dodecyl sulfate and a polyether surfactant F108(m/v), and then adding 2% of a photoinitiator 2-hydroxy-2-methyl propiophenone (m/v) to serve as an external phase solution. Taking two microfluidic single emulsion devices, wherein an inner phase pipe of one device is communicated with an inner phase solution 1, an outer phase pipe is communicated with an outer phase solution, an inner phase pipe of the other device is communicated with an inner phase solution 2, an outer phase pipe is communicated with an outer phase solution, and colloid liquid drops with different sizes are obtained by adjusting the flow rates of the inner phase solution and the outer phase solution of the two devices.
(3) Preparation of Janus porous biological patch for wound repair
And (3) automatically layering the colloid drops prepared in the step (2) under the action of a magnetic field and assembling the colloid drops in a hexagonal close-packed structure to form a colloid drop positive template, adjusting the proportion of hydrophilic colloid drops in a hydrophobic colloid drop layer according to the action time and distance of the magnetic field in the process, then carrying out in-situ ultraviolet polymerization on the external phase solution-photopolymerization aqueous polyurethane solution, and then cleaning n-hexadecane with n-hexane to remove the positive template to obtain the Janus porous biological patch for wound repair.
(4) Characterization of Janus porous biological patches for wound repair
The Janus porous biological patch structure for wound repair is observed and characterized under a body type microscope and a scanning electron microscope, the diameter of the holes of the Janus porous biological patch for wound repair is measured to be about 350 micrometers, a certain proportion of hydrophilic holes are dispersed on the hydrophobic side, and the connectivity among the holes is good.
Example 3
Janus porous biological patches were used in rat wound models:
taking the Janus porous biological patch for wound repair prepared in the example 2 as an example, randomly dividing 10 male SD rats weighing about 200g into 2 groups, namely a control group and an experimental group, wherein each group comprises 5 male SD rats, injecting 10% (w/v) chloral hydrate solution into an abdominal cavity to anaesthetize the male SD rats, establishing a wound model with the diameter of 1cm on the back of the male SD rats, wherein the control group does not receive any treatment, the experimental group is attached to the Janus porous biological patch prepared in the example 2, the hydrophobic side of the Janus porous biological patch directly contacts the wound, and after 9 days of treatment, taking down a tissue sample of the wound and carrying out Masson and H & E staining;
the results are shown in fig. 4, and the H & E staining results show that the thickness of the granulation tissue of the experimental group is significantly higher than that of the control group, which indicates that the tissue regeneration condition of the experimental group is better than that of the control group. According to the Masson color-staining image result, the collagen deposition amount of the experimental group is obviously larger than that of the control group, and the Janus porous biological patch for wound repair can effectively promote wound collagen regeneration. The above results show that the Janus porous biological patch for wound repair can effectively promote wound repair, wherein the lower layer of the Janus porous biological patch is a hydrophobic surface doped with hydrophilic holes, the upper layer of the Janus porous biological patch is a hydrophilic surface, the hole structures are uniform and high in connectivity, transfer of oxygen and nutrients is facilitated, biological effusion and waste generated by cell metabolism are arranged side by side, in addition, the hydrophobic surface and the wound are in direct contact, secondary wound caused by tissue adhesion can be avoided, and the hydrophilic surface can adsorb exudate at the wound, so that wound repair is accelerated.
The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention in any way, and any person skilled in the art can make any simple modification, equivalent replacement, and improvement on the above embodiment without departing from the technical spirit of the present invention, and still fall within the protection scope of the technical solution of the present invention.
Claims (10)
1. A Janus porous biological patch for wound repair, which is characterized in that: the preparation method comprises the following steps:
(1) constructing a microfluidic single emulsion device: breaking a glass capillary tube by using a capillary tube drawing instrument, and then polishing the broken glass capillary tube into a tip end by using abrasive paper to serve as an internal phase tube; taking another glass capillary tube with the same size, and polishing one end of the glass capillary tube smoothly by using sand paper to serve as an outer phase tube; assembling the inner phase tube and the outer phase tube on a cover glass, keeping the central lines horizontally aligned, and fixing; connecting and sealing the interface of the two glass capillary tubes for later use;
(2) preparing a colloid liquid drop template: preparing a colloid liquid drop template with hydrophilic silicon dioxide nano particles and hydrophobic magnetic nano particles wrapped on the surface by using the microfluidic single emulsion device built in the step (1); the inner phase solution is n-hexadecane dispersed with hydrophilic silicon dioxide nanoparticles and hydrophobic magnetic nanoparticles, the outer phase solution is a photo-polymerization aqueous polyurethane solution added with a surfactant, the inner phase solution is sheared by the outer phase solution at the tip of an inner phase tube to generate colloid droplets, and the colloid droplets are automatically layered under the action of a magnetic field and assembled in a hexagonal close-packed structure to form a colloid droplet template;
(3) preparing a Janus porous biological patch for wound repair: and (3) taking the colloid liquid drop template prepared in the step (2) as a positive template, carrying out in-situ ultraviolet photopolymerization on the external phase solution-the photopolymerization aqueous polyurethane solution, and then removing the positive template to obtain the Janus porous biological patch for wound repair.
2. The Janus porous biological patch for wound repair of claim 1, wherein: the outer diameter of the glass capillary tube adopted in the step (1) is 1mm, the inner diameter is 0.8mm, and the inner phase tube is polished by sand paper until the inner diameter of the tip is 0.4 mm; and ultrasonically cleaning the polished inner phase tube and the polished outer phase tube for 2min by using absolute ethyl alcohol, and naturally drying for later use.
3. The Janus porous biological patch for wound repair of claim 1, wherein: in the external phase solution, the surfactant is a mixed solution of sodium dodecyl sulfate and a polyether surfactant F108, and the mass concentration of the mixed solution is 2% m/v.
4. The Janus porous biological patch for wound repair of claim 1, wherein: the curing method of the photo-polymerization aqueous polyurethane solution is to add 2% m/v of 2-hydroxy-2-methyl propiophenone, and to carry out ultraviolet crosslinking curing: wherein the photopolymerisable aqueous polyurethane solution needs to be processed in a dark place before photocuring.
5. The Janus porous biological patch for wound repair of claim 1, wherein: the particle size of the hydrophilic silicon dioxide nano particles is 220 nm; the particle size of the hydrophobic magnetic nanoparticles is 240 nm.
6. The Janus porous biological patch for wound repair of claim 1, wherein: the diameter of the colloid liquid drop is 300-500 mu m.
7. The Janus porous biological patch for wound repair of claim 1, wherein: the method for removing the colloid liquid drop template comprises the step of soaking and cleaning by using normal hexane.
8. The Janus porous biological patch for wound repair of claim 1, wherein: in the step (2), the colloid droplet template coated with the hydrophilic silica nanoparticles and the hydrophobic magnetic nanoparticles on the surface is automatically divided into two layers under the action of a magnetic field, wherein the lower layer is a single-layer colloid droplet coated with the hydrophobic magnetic nanoparticles, and the upper layer is a double-layer colloid droplet coated with the hydrophilic silica nanoparticles.
9. The Janus porous biological patch for wound repair of claim 1, wherein: the colloid liquid drop template coated with the hydrophilic silicon dioxide nano particles and the hydrophobic magnetic nano particles on the surface is automatically divided into two layers under the action of a magnetic field, and the proportion of the hydrophilic colloid liquid drops in the hydrophobic colloid liquid drop layer can be adjusted according to the action time and distance of the magnetic field.
10. The Janus porous biological patch for wound repair of claim 1, wherein: the Janus porous biological patch for wound repair has the advantages that the lower layer is a hydrophobic surface doped with hydrophilic holes, the upper layer is a hydrophilic surface, the hydrophobic surface is in direct contact with the wound to avoid secondary wound caused by tissue adhesion, and the hydrophilic surface can adsorb exudate at the wound to accelerate wound repair.
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