Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a drive response composite material and a preparation method and application thereof. The invention can enable the wound dressing to have the effects of controllable and drug release according to requirements, has excellent antibacterial, anti-inflammatory and healing promotion functions, has excellent air permeability and moisture retention and can maintain the mechanical property of the dressing, reduce the replacement and use of the dressing and effectively solve the problems of the wound dressing.
In order to achieve the purpose, the invention adopts the following scheme:
a drive-responsive composite material characterized by: comprises a nanofiber layer and functional substances in the nanofiber layer;
the material of the nanofiber in the nanofiber layer is a mixed material of a hydrophilic high molecular polymer I and a hydrophobic high molecular polymer;
the functional substance can be driven under the applied external condition; the addition conditions were aqueous ethanol (75% ethanol).
As a preferred technical scheme:
in an actuated response composite as described above, the nanofibers in the nanofiber layer comprise hydrophilic high molecular polymer I nanofibers and hydrophobic high molecular polymer nanofibers.
A drive-responsive composite as described above, further comprising a gel layer bonded to the nanofiber layer; the main material of the gel layer is hydrophilic high molecular polymer II; the functional substance is capable of being driven into the gel layer under applied external conditions.
In a drive-responsive composite material as described above, the functional substance is dispersed in the form of fine particles (or powder) in the nanofibers.
The drive response composite material is characterized in that the functional substance is an extract of salvia miltiorrhiza, a monomer modifier of salvia miltiorrhiza, an antibacterial drug or a growth factor (the former three substances have antibacterial and anti-inflammatory effects, and the growth factor has a tissue repair promoting effect); the loading amount of the functional substance in the nanofiber layer is 0.3-29.7 ug/cm 2 。
The drive response composite material is prepared from PLGA, PGA, PLA or PLLA-CL (PLGA: polylactic acid-glycolic acid; PGA: polyglycolic acid; PLA: polylactic acid; PLLA-CL (block copolymer, copolymer of PLLA and PCL with the molar ratio of 1: 1) polylactic acid-caprolactone; PCL: polycaprolactone); the number average molecular weight Mn of the hydrophobic high molecular polymer is 8-15 ten thousand;
the hydrophilic high molecular polymer I is silk fibroin, collagen or collagen denaturant (such as gelatin);
the hydrophilic high molecular polymer II is hyaluronic acid or alginic acid polysaccharide; the number average molecular weight Mn of the hydrophilic high molecular polymer II is 20-100 ten thousand;
the mass ratio of the hydrophobic high-molecular polymer to the hydrophilic high-molecular polymer I is 0.8-1.2: 1.
The invention also provides a wound dressing made of a drive responsive composite material as described above, characterised in that: the gel layer directly contacts the biological tissue; the functional substance can be driven and seeped into the gel layer to act on biological tissues under the applied condition; the functional material does not exude when no external condition is applied; and the single release amount of the functional substance in the wound dressing is less than 33.2 wt%.
The invention also provides a preparation method of the drive response composite material, which is characterized in that a functional substance, a hydrophilic high molecular polymer I and a hydrophobic high molecular polymer are used as raw materials to prepare spinning solution, and an electrostatic spinning process is adopted to prepare the nanofiber layer.
The electrostatic spinning conditions can be as follows: the mass concentration (Mv) of the spinning solution is 4-20 percent; the spinning voltage is 10-30 kV, the spinning advancing speed is 0.5-2.5 mL/h, the receiving distance is 10-25 cm, and the receiver is conductive tinfoil paper. Wherein, the size of the fiber membrane obtained by electrostatic spinning of every 10mL of spinning solution is 18cmX30 cm.
As a preferred technical scheme:
a method of making a drive-responsive composite as described above, a gel layer is formed on one side of the nanofiber layer.
The preparation method of the drive response composite material comprises the following steps:
(1) dissolving a hydrophilic high molecular polymer II in an aqueous solution containing methanol or ethanol, and adjusting to obtain a pre-gel;
(2) applying a pre-gel to a single-sided surface of the nanofiber layer; the coating thickness is 10-40 μm.
The preparation method of the drive response composite material comprises the following steps:
(1) dissolving a hydrophilic high molecular polymer II in an aqueous solution containing methanol or ethanol, and adjusting to obtain a pre-gel;
(2) adding a cross-linking agent, quickly stirring for 1-2 minutes, and immediately coating the pre-gel on the surface of one side of the nanofiber layer;
the coating thickness is 10-40 mu m, and the cross-linking agent is EDC/NHS or CaCl 2 The mass ratio of the cross-linking agent to the hydrophilic high molecular polymer II is 0.2-1: 1;
(3) standing at room temperature for crosslinking reaction, then ventilating and drying, and soaking the gel surface of the dried composite membrane into deionized water to remove the residues of the crosslinking agent and then drying;
the temperature of the crosslinking reaction is 4-20 ℃, and the crosslinking time is 2-4 hours.
The specific process of coating is as follows: and (2) pressing the polytetrafluoroethylene rectangular frame (with the size of 7cm, the length and the width of 15 cm) on the electrostatic spinning fiber membrane prepared in the step (1), quickly injecting 20mL of gel liquid and 1mL of cross-linking agent mixed solution into the mold frame to coat the fiber membrane, standing at 4-20 ℃ for cross-linking reaction for 2-4 hours, and then ventilating and drying at room temperature. And finally, immersing the gel surface of the composite membrane into deionized water for 6 hours to remove the residues of the cross-linking agent, sucking the surface moisture, and then placing the composite membrane in a vacuum drying oven to dry for 1 week at room temperature.
In the preparation method of the driving response composite material, the solvent of the spinning solution is hexafluoroisopropanol;
in the aqueous solution containing methanol or ethanol, the volume of the methanol or ethanol accounts for 5-10% of the total volume;
the mass content of the hydrophilic high molecular polymer II in the pre-gel is 1 to 5 percent;
the principle of the invention is as follows:
in the prior art, functional substances are embedded in gel and release drugs through gel degradation and other modes, the situation of drug burst release inevitably occurs, the release of the drugs is uninterrupted and cannot be forcibly stopped, the use period of validity is short when the drug is used as a wound dressing, and the utilization rate of the drugs is low; in the invention, the designed driving response composite material for the wound dressing is of a fiber and gel composite double-layer structure, and the gel layer directly contacting the tissue does not contain functional substances (in the preparation process, because the gel solution contains ethanol solution, a small amount of medicaments are enabled to be permeated into the gel layer closest to the fiber layer in a gradient manner, so that the aim of the driving response composite material is that the amount of the functional substances released in the first spraying is larger than the subsequent spraying release amount because the firstly required medicament amount of the damaged tissue is generally larger), and the fiber layer as the outer layer is formed by fibers of high molecular polymer loaded with the functional substances (namely, the fiber layer is a medicament reservoir); because the fibers in the fiber layer are mutually entangled and have better mechanical property, and the fibers in the fiber layer are a mixed material of hydrophilic high molecular polymer I and hydrophobic high molecular polymer, when the hydrophilic substance acts on the fiber layer, the functional substance is dissolved in the hydrophilic substance and permeates into the gel layer under the guiding action of the hydrophilic high molecular polymer I in the fiber layer, so that the transfer of the functional substance is more effective compared with the pure hydrophobic high molecular polymer fibers; compared with pure hydrophilic polymer fibers, the hydrophobic fibers can enable the fat-soluble medicine to be better dispersed in the fibers; further, when the solution formed by the functional substance enters the gel layer, the gel layer serves as a drug release passage, and since the permeation rate of the functional substance is related to the degree of crosslinking in the gel layer, the method of the present invention can reasonably design the degree of crosslinking in the gel layer according to the needs to adjust the permeation rate and dosage effect of the functional substance. The invention effectively controls the transfer of the functional substance by designing a unique fiber layer structure, controls the release of the functional substance by utilizing the structure of the gel layer and effectively avoids the burst release of the functional substance.
In addition, because the functional substance is positioned in the fiber layer and is continuously guided to the gel layer by applying external factors, the functional substance is dissolved by water or ethanol water solution and is guided to the gel layer; this "guided" actuation allows the release of the functional substance to be an external factor-responsive actuation rather than an uninterrupted or terminated actuation process, i.e.: when the external factor is present, the functional substance is driven into the gel layer, and when the external factor disappears, it is not driven. The controllable driving mode can avoid the burst release of the functional substances on one hand, and also control the release amount of the functional substances on the other hand, so that the utilization rate of the functional substances is improved, and the effective application time of the composite material is greatly prolonged.
Advantageous effects
(1) The preparation method of the drive response composite material is simple in process and wide in application range;
(2) according to the driving response composite material, functional substances can firstly permeate into the gel coating and then reach biological tissues by spraying the alcohol-containing aqueous solution according to needs, the wound healing is promoted, the antibacterial and anti-inflammatory effects are achieved, and the dressing does not need to be frequently replaced or even not replaced (the gel layer has a good moisturizing effect, the wound is not torn during replacement, and the anti-adhesion effect is achieved).
Detailed Description
The invention will be further illustrated with reference to specific embodiments. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.
Example 1
A method for preparing a drive response composite material, as shown in fig. 1, comprises the following specific steps:
(1) cryptotanshinone, silk fibroin with molecular weight of more than 14000 and PLGA with molecular weight of 10 ten thousand are taken as raw materials and added into hexafluoroisopropanol to prepare spinning solution with mass concentration (Mv) of 10 percent, and an electrostatic spinning process is adopted to prepare a nanofiber layer; wherein the content of cryptotanshinone in the spinning solution is 0.4 percent of the total mass of PLGA and silk fibroin; the mass ratio of PLGA to silk fibroin is 1: 1;
the conditions of electrostatic spinning are as follows: the spinning voltage is 18kV, the spinning advancing speed is 1.5mL/h, the receiving distance is 15cm, and the receiver is conductive tinfoil paper; wherein, the size of the fiber membrane obtained by electrostatic spinning of every 10mL of spinning solution is 18cmx30 cm;
(2) dissolving hyaluronic acid with molecular weight of 40 ten thousand in an aqueous solution containing ethanol (the volume of the ethanol accounts for 10% of the total volume), and adjusting to obtain a pre-gel; wherein the mass content of hyaluronic acid in the pre-gel is 3%;
(3) and coating the pregel on the surface of one side of the nanofiber layer, wherein the thickness of the coating is 28 microns, and thus the driving response composite material is prepared.
The driving response composite material is used as a wound dressing and comprises a nanofiber layer and cryptotanshinone (the load is 6.6 ug/cm) dispersed in the nanofiber layer 2 ) The gel layer is attached to the nanofiber layer; wherein the gel layer directly contacts with biological tissue, and cryptotanshinone can be driven and permeate into the gel layer under the action of ethanol water solution to act on biological tissue; the invention carries out cryptotanshinone driving test on the wound dressing, and the specific test process is as follows: spraying 500uL of 75% ethanol to one side of the nanofiber in the wound dressing in a spraying manner, and standing for 10min, and testing the release amount of cryptotanshinone driven for the first time; after 30min, spraying 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing in a spraying mode again, staying for 10min, and testing the release amount of cryptotanshinone driven for the second time; after 30min, spraying 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing in a spraying mode again, staying for 10min, and testing the release amount of cryptotanshinone driven for the third time; the results of three tests are as follows:
example 2
A preparation method of a drive response composite material comprises the following specific steps:
(1) cryptotanshinone, silk fibroin with molecular weight of more than 14,000 and PLGA with molecular weight of 10 ten thousand are taken as raw materials and added into hexafluoroisopropanol to prepare spinning solution with mass concentration (Mv) of 10%, and an electrostatic spinning process is adopted to prepare a nanofiber layer, namely the driving response composite material;
wherein the content of cryptotanshinone in the spinning solution is 0.4 percent of the total mass of PLGA and silk fibroin; the mass ratio of PLGA to silk fibroin is 1: 1; the conditions of electrostatic spinning are as follows: the spinning voltage is 18kV, the spinning advancing speed is 1.5mL/h, the receiving distance is 15cm, and the receiver is conductive tinfoil paper; wherein, the size of the fiber membrane obtained by electrostatic spinning of every 10mL of spinning solution is 18cmx30 cm;
the driving response composite material is used as a wound dressing and comprises a nanofiber layer and cryptotanshinone (the load is 6.9 ug/cm) dispersed in the nanofiber 2 ) (ii) a And the cryptotanshinone can be driven to act on biological tissues; the wound dressing is subjected to cryptotanshinone driving test, and the specific test process comprises the following steps: spraying 500uL of 75% ethanol to one side of the nanofiber in the wound dressing in a spraying mode, and after staying for 10min, testing the release amount of the first drive; after 30min, spraying 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing in a spraying mode again, and after staying for 10min, testing the release amount of the second driving; after 30min, spraying 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing in a spraying mode again, and after staying for 10min, testing the release amount of the third driving; the results of three tests are given in the following table:
example 3
A preparation method of a drive response composite material comprises the following specific steps:
(1) adding cryptotanshinone, silk fibroin with the number average molecular weight Mn of more than 14,000 and PLGA with the number average molecular weight of 10 ten thousand serving as raw materials into hexafluoroisopropanol to prepare spinning solution with the mass concentration (Mv) of 10%, and preparing a nanofiber layer by adopting an electrostatic spinning process;
wherein, the content of cryptotanshinone in the spinning solution is 0.4 percent of the total mass of PLGA and silk fibroin; the mass ratio of PLGA to silk fibroin is 1: 1; the conditions of electrostatic spinning are as follows: the spinning voltage is 18kV, the spinning advancing speed is 1.5mL/h, the receiving distance is 15cm, and the receiver is conductive tinfoil paper. Wherein, the size of the fiber membrane obtained by electrostatic spinning of every 10mL of spinning solution is 18cmX30 cm.
(2) Dissolving hyaluronic acid with the molecular weight of 40 ten thousand in an aqueous solution containing ethanol (the volume of the ethanol accounts for 10 percent of the total volume) to obtain a pre-gel; wherein the mass content of the hyaluronic acid in the pre-gel is 3%;
(3) adding a cross-linking agent, quickly stirring for 1-2 minutes, and immediately coating the pre-gel on the surface of one side of the nanofiber layer, wherein the thickness of the coating is 26 microns; the cross-linking agent is an EDC/NHS aqueous solution with the mass concentration of 20%, the mass ratio of EDC to NHS is 3:2, and the mass ratio of the cross-linking agent to hyaluronic acid is 1: 1;
(4) standing at 20 ℃, carrying out crosslinking reaction for 2 hours, then carrying out ventilation drying, immersing the gel surface of the dried composite membrane into deionized water to remove the residues of the crosslinking agent, and then drying to obtain the drive response composite material.
The driving response composite material is used as a wound dressing and comprises a nanofiber layer and cryptotanshinone (the load is 6.7 ug/cm) mainly dispersed in the nanofiber 2 ) The gel layer is attached to the nanofiber layer; wherein the gel layer directly contacts with biological tissue, and cryptotanshinone can be driven and permeated into the gel layer under the action of ethanol water solution to act on the biological tissue; the invention carries out cryptotanshinone driving test on the wound dressing, and the specific test process is as follows: spraying 500uL of 75% ethanol to one side of the nanofiber in the wound dressing in a spraying manner, and standing for 10min, and testing the release amount of cryptotanshinone driven for the first time; after 30min, spraying 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing in a spraying mode again, staying for 10min, and testing the release amount of cryptotanshinone driven for the second time; spraying in form of spray again after 30minDischarging 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing, staying for 10min, and testing the release amount of cryptotanshinone driven for the third time; the results of three tests are as follows:
example 4
A preparation method of a drive response composite material comprises the following specific steps:
(1) cryptotanshinone, silk fibroin with the number average molecular weight Mn of more than 14,000 and PLGA with the number average molecular weight of 10 ten thousand are taken as raw materials, added into hexafluoroisopropanol to prepare spinning solution with the mass concentration (Mv) of 12%, and an electrostatic spinning process is adopted to prepare a nanofiber layer;
wherein the content of cryptotanshinone in the spinning solution is 1.5 percent of the total mass of PLGA and silk fibroin; the mass ratio of PLGA to silk fibroin is 0.8: 1;
the conditions of electrostatic spinning are as follows: the spinning voltage is 12kV, the spinning advancing speed is 0.5mL/h, the receiving distance is 10cm, and the receiver is conductive tinfoil paper. Wherein, the size of the fiber membrane obtained by electrostatic spinning of every 10mL of spinning solution is 18cmX30 cm.
(2) Dissolving hyaluronic acid with the molecular weight of 60 ten thousand in an aqueous solution containing ethanol (the volume of the ethanol accounts for 5 percent of the total volume) to obtain a pre-gel; wherein the mass content of hyaluronic acid in the pre-gel is 1%;
(3) adding an EDC/NHS aqueous solution (a cross-linking agent and the mass ratio of EDC to NHS is 3:2) with the mass concentration of 10% into the pre-gel, quickly stirring for 1 minute, immediately coating the pre-gel on the single-side surface of the nanofiber layer, wherein the thickness of the coating is 14 mu m; the mass ratio of the cross-linking agent to the hyaluronic acid is 0.4: 1;
(4) standing at 4 ℃, carrying out crosslinking reaction for 4 hours, then carrying out ventilation drying, immersing the gel surface of the dried composite membrane into deionized water to remove the residues of the crosslinking agent, and then drying to obtain the drive response composite material.
With drive-responsive composite as a wound dressing, whichComprises a nanometer fiber layer, cryptotanshinone (loading amount is 29.7 ug/cm) dispersed in the nanometer fiber layer 2 ) The gel layer is attached to the nanofiber layer; wherein the gel layer directly contacts with biological tissue, and cryptotanshinone can be driven and permeated into the gel layer under the action of ethanol to act on the biological tissue; the invention carries out cryptotanshinone driving test on the wound dressing, and the specific test process is as follows: spraying 500uL of 75% ethanol to one side of the nanofiber in the wound dressing in a spraying manner, and standing for 10min, and testing the release amount of cryptotanshinone driven for the first time; after 30min, spraying 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing in a spraying mode again, staying for 10min, and testing the release amount of cryptotanshinone driven for the second time; after 30min, spraying 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing in a spraying mode again, staying for 10min, and testing the release amount of cryptotanshinone driven for the third time; the results of three tests are as follows:
example 5
A preparation method of a drive response composite material comprises the following specific steps:
(1) adding tanshinone II B, gelatin with number average molecular weight Mn of 80,000 and PLA with number average molecular weight of 10.5 ten thousand serving as raw materials into hexafluoroisopropanol to prepare spinning solution with mass concentration (Mv) of 15%, and preparing a nanofiber layer by adopting an electrostatic spinning process;
wherein the content of tanshinone II B in the spinning solution is 1 percent of the total mass of PLA and gelatin; the mass ratio of PLA to gelatin is 1.2: 1;
the conditions of electrostatic spinning are as follows: the spinning voltage is 30kV, the spinning advancing speed is 2.0mL/h, the receiving distance is 25cm, and the receiver is conductive tinfoil paper. Wherein, the size of the fiber membrane obtained by electrostatic spinning of every 10mL of spinning solution is 18cmX30 cm.
(2) Dissolving alginic acid polysaccharide with molecular weight of 100 ten thousand in an aqueous solution containing ethanol (the volume of the ethanol accounts for 8% of the total volume), and adjusting to obtain a pre-gel; wherein the mass content of alginic acid polysaccharide in the pre-gel is 4%;
(3) adding a cross-linking agent, quickly stirring for 2 minutes, and immediately coating the pre-gel on the surface of one side of the nanofiber layer, wherein the thickness of the coating is 36 microns; the cross-linking agent is CaCl with the mass concentration of 15 percent 2 The mass ratio of the cross-linking agent to the alginic acid polysaccharide is 0.8: 1;
(4) standing at 20 ℃, carrying out crosslinking reaction for 2 hours, then carrying out ventilation drying, immersing the gel surface of the dried composite membrane into deionized water to remove the residues of the crosslinking agent, and then drying to obtain the drive response composite material.
The driving response composite material is used as a wound dressing and comprises a nanofiber layer, tanshinone II B (with the loading amount of 24.2 ug/cm) dispersed in the nanofiber in a particle form 2 ) The gel layer is attached to the nanofiber layer; wherein the gel layer directly contacts with biological tissue, and tanshinone II B can be driven and permeated into the gel layer under the action of ethanol to act on the biological tissue; the driving test of the tanshinone II B is carried out on the wound dressing, and the specific test process comprises the following steps: spraying 500uL of 75% ethanol to one side of the nanofiber in the wound dressing in a spraying manner, and standing for 10min, and testing the release amount of the first-driven tanshinone II B; after 30min, spraying 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing in a spraying mode again, staying for 10min, and testing the release amount of the tanshinone II B driven for the second time; after 30min, spraying 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing in a spraying mode again, staying for 10min, and testing the release amount of the third driven tanshinone II B; the results of three tests are as follows:
example 6
A preparation method of a drive response composite material comprises the following specific steps:
(1) adding growth factor (VEGF), collagen with the number average molecular weight Mn of 100,000 and PLLA-CL (the molar ratio is 1: 1) with the number average molecular weight of 11 ten thousand serving as raw materials into hexafluoroisopropanol to prepare spinning solution with the mass concentration (Mv) of 10%, and preparing a nanofiber layer by adopting an electrostatic spinning process;
wherein, the content of the growth factor (VEGF) in the spinning solution is 0.02 percent of the total mass of the PLLA-CL and the collagen; the mass ratio of PLLA-CL to collagen is 0.9: 1; the conditions of electrostatic spinning are as follows: the spinning voltage is 20kV, the spinning advancing speed is 2mL/h, the receiving distance is 15cm, and the receiver is conductive tinfoil paper. Wherein, the size of the fiber membrane obtained by electrostatic spinning of every 10mL of spinning solution is 18cm x30 cm.
(2) Dissolving alginic acid polysaccharide with molecular weight of 80 ten thousand in aqueous solution containing methanol (the volume of the methanol accounts for 8% of the total volume), and adjusting to obtain pre-gel; wherein the mass content of alginic acid polysaccharide in the pre-gel is 2 percent;
(3) adding a cross-linking agent, quickly stirring for 1 minute, and immediately coating the pre-gel on the surface of one side of the nanofiber layer, wherein the thickness of the coating is 23 mu m; the cross-linking agent is CaCl with the mass concentration of 10 percent 2 The mass ratio of the cross-linking agent to the alginic acid polysaccharide is 0.4: 1;
(4) standing at 10 ℃, carrying out crosslinking reaction for 3 hours, then carrying out ventilation drying, immersing the gel surface of the dried composite membrane into deionized water to remove the residues of the crosslinking agent, and then drying to obtain the drive response composite material.
The driving response composite material is used as a wound dressing and comprises a nanofiber layer, and growth factors (VEGF) (the loading amount is 0.3 ug/cm) mainly dispersed in the nanofibers 2 ) The gel layer is attached to the nanofiber layer; wherein the gel layer directly contacts with biological tissue, and growth factor (VEGF) can be driven by ethanol and permeate into the gel layer to act on biological tissue; the wound dressing is subjected to a driving test of a growth factor (VEGF), and the specific test process comprises the following steps: spraying 500uL of 75% ethanol to one side of the nanofiber in the wound dressing in a spraying mode, and after staying for 10min, testing the release amount of the growth factor (VEGF) driven for the first time; after 30min, spraying 500uL of 75% ethanol again to the same woundTesting the release amount of a second-driven growth factor (VEGF) after the side of the nanofiber in the mouth dressing stays for 10 min; after 30min, spraying 500uL of 75% ethanol to one side of the nanofiber in the same wound dressing in a spraying mode again, and after staying for 10min, testing the release amount of growth factor (VEGF) driven for the third time; the results of three tests are as follows: