CN111333824B - Process for preparing porous elastomeric materials and uses thereof - Google Patents

Process for preparing porous elastomeric materials and uses thereof Download PDF

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CN111333824B
CN111333824B CN202010206790.4A CN202010206790A CN111333824B CN 111333824 B CN111333824 B CN 111333824B CN 202010206790 A CN202010206790 A CN 202010206790A CN 111333824 B CN111333824 B CN 111333824B
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CN111333824A (en
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乔元昊
木仁
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Quanzhou Shengde Light Industry Co ltd
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Hangzhou Luyang Technology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G63/00—Macromolecular compounds obtained by reactions forming a carboxylic ester link in the main chain of the macromolecule
    • C08G63/02—Polyesters derived from hydroxycarboxylic acids or from polycarboxylic acids and polyhydroxy compounds
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    • 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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    • C08G63/02—Polyesters derived from hydroxycarboxylic acids or from polycarboxylic acids and polyhydroxy compounds
    • 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
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Abstract

The invention provides a preparation method of a porous elastomer material and application thereof, belonging to the technical field of tissue engineering, comprising the steps of taking a prepolymer of modified POC, adding the prepolymer into acetone to fully dissolve the prepolymer, pouring the prepolymer into a tetrafluoroethylene mold, adding NaCl particles and guanethidine sulfate, uniformly stirring, ventilating to completely volatilize the acetone, and polymerizing for 4-7d under the vacuum condition at 100-120 ℃; and taking out the sample, soaking the sample in deionized water, completely washing NaCl and guanethidine sulfate in the bracket, and drying. According to the invention, through modification of POC, the rigidity of the molecular main chain in a cross-linked network structure can be reduced while the internal cross-linking of the porous scaffold is increased, the strength and toughness of the porous scaffold can be improved, and the hydrophilicity can be increased; can inhibit the sedimentation of sodium chloride, so that the sodium chloride can be uniformly distributed in the polymer, the porosity and the distribution uniformity of the porous support are improved, and the connectivity of the support is improved.

Description

Process for preparing porous elastomeric materials and uses thereof
Technical Field
The invention belongs to the technical field of tissue engineering, and particularly relates to a preparation method and application of a porous elastomer material.
Background
The biological materials can be classified into three major categories, i.e., biomedical polymer materials, biomedical inorganic non-metallic materials (ceramics, composite materials, etc.) and biomedical metal materials according to the composition and properties of the biological materials. Compared with metal materials and inorganic non-metal materials, the biomedical polymer material has better physicochemical properties, and also has incomparable elasticity and flexibility, and can be better matched with human tissues and organs. With the rapid development of science, medical technology and medical materials, various medical technologies appear in succession, such as tissue engineering, drug release, gene therapy and the like, which puts higher requirements on biomedical polymer materials, and requires that the materials are only temporarily substituted after being implanted into human bodies, and can be gradually degraded along with the regeneration of tissues or organs and discharged out of the bodies along with the metabolism of the human bodies, thereby reducing the long-term influence of the materials on the bodies as much as possible.
The tissue engineering porous scaffold is a biodegradable material which can be combined with tissue living cells, can meet the requirements of different tissues in organisms and can be implanted into the organisms for tissue substitution. The porous scaffold plays a role of extracellular matrix, and meanwhile, the three-dimensional scaffold structure provides a good environment for the growth, proliferation and differentiation of cells. Specifically, tissue cells and a scaffold material are subjected to compound culture in vitro to form a cell-biomaterial complex, then the complex is implanted into an organism, and the cells continue to proliferate and secrete a matrix in the organism, so that new tissues and organs are generated. The porous scaffold can repair and reconstruct damaged tissues and even permanently replace the damaged tissues.
The prior art, such as the Chinese patent with the publication number of CN 102973984B, discloses a preparation method of a composite material porous scaffold. The method compounds silk fibroin, collagen and chitosan together to obtain the white spongy composite porous scaffold. The porous scaffold prepared by the method can be applied to the technical fields of tissue and wound surface filling and repairing, tissue engineering, drug release and the like. The porosity of the composite porous scaffold prepared by the invention is more than 98%, the size of the composite porous scaffold is distributed in a range of 50-400 mu m, and micropores are communicated with each other. The composite porous scaffold obtained by the invention can keep the original shape after being soaked in water again, has the function of sponge, and can recover the original shape after being extruded.
Disclosure of Invention
The invention aims to provide a poly citrate porous scaffold and a preparation method thereof, the method can reduce the rigidity of molecular main chains in a cross-linked network structure while increasing the internal cross-linking of the porous scaffold by modifying POC, can improve the strength and toughness of the porous scaffold, and can increase the hydrophilicity; can inhibit the sedimentation of sodium chloride, so that the sodium chloride can be uniformly distributed in the polymer, the porosity and the distribution uniformity of the porous support are improved, and the connectivity of the support is improved.
The technical scheme adopted by the invention for realizing the purpose is as follows:
the application of 4-hydroxy-2-butenoic acid in improving the toughness of the poly citrate is provided.
Provided is a preparation method of modified poly (1, 8-octanediol-citric acid), comprising the following steps:
s1, placing citric acid, 1, 8-octanediol and 4-hydroxy-2-butenoic acid in a container, placing in an oil bath at 160-165 ℃ under the protection of nitrogen, heating and stirring, cooling to 135-140 ℃ after the mixture is melted, adding 2-aminoethanol-1-phosphoric acid, and reacting for 5-7 hours at normal pressure to obtain a prepolymer;
s2, dissolving the prepolymer in acetone, precipitating in ethanol to remove unreacted monomers, pouring into a mold, standing until acetone is completely volatilized, and polymerizing for 4-7d under the vacuum condition of 100-120 ℃. Poly (1, 8-octanediol-citric acid) (POC) is a network type polyester bio-elastomer prepared by reacting citric acid with 1, 8-octanediol, and is widely used in the field of tissue engineering due to its good mechanical and biological properties. The mechanical properties of POC can be adjusted by adjusting the molar ratio of the reactive monomers and the degree of cure crosslinking, but such adjustment is limited. The POC tensile deformability is improved, and the crosslinking degree is inevitably sacrificed, so that the material viscosity is increased, and the use is not facilitated. Thermosetting resins generally have a very high crosslinking density and therefore are very brittle. In the presence of 2-aminoethanol-1-phosphoric acid, 4-hydroxy-2-butenoic acid is used for modifying POC, alcoholic hydroxyl on 4-hydroxy-2-butenoic acid can react with alcoholic hydroxyl on citric acid to form ether bond, so that a flexible chain segment containing carbon double bond and carboxyl bond is introduced to the side surface of a POC main chain, and carboxylic acid group is introduced.
Preferably, the molar ratio of citric acid, 1, 8-octanediol and 4-hydroxy-2-butenoic acid in the step S1 is 5-6:4-5: 2-3.
Provides a degradable biological elastomer which is prepared by the preparation method.
A preparation method of a poly citrate porous scaffold is provided, which comprises the following steps:
a. taking the prepolymer in the step S1, adding acetone to fully dissolve the prepolymer, pouring the prepolymer into a tetrafluoroethylene mold, adding NaCl particles and guanethidine sulfate, uniformly stirring, ventilating to completely volatilize the acetone, and polymerizing for 4-7d under the vacuum condition at 100-120 ℃;
b. and taking out the sample, soaking the sample in deionized water, completely washing NaCl and guanethidine sulfate in the stent, and drying to obtain the poly-citrate porous stent. When the porous scaffold is prepared by using the solution casting/particle leaching technology, a compact skin layer is easily formed on the surface of a material particularly under the influence of the distribution of a pore-forming agent, the pore-forming agent is easy to precipitate at the bottom of a container to form an uneven pore structure, the obtained closed pore structure is more, and the connectivity among pores is poor. Guanethidine sulfate is added when the porous scaffold is prepared, can penetrate through the modified POC network structure, can wrap sodium chloride, and inhibit sodium chloride from settling, so that the sodium chloride can be uniformly distributed in the polymer, the formation of an open pore structure by the sodium chloride is facilitated, the porosity and distribution uniformity of the porous scaffold can be improved, the connectivity of the scaffold is improved, and cell adhesion and proliferation are facilitated.
Preferably, the mass ratio of the performed polymer in the step a, NaCl and guanethidine sulfate is 1:4-6: 3-5.
Providing a porous elastomeric material characterized by: the preparation method of the poly citrate porous scaffold is adopted for preparation.
The use of guanethidine sulfate to improve the connectivity of a stent is provided.
Provides the application of a degradable biological elastomer in preparing a tissue scaffold and/or a drug controlled release carrier.
Provides the application of a porous elastic material in repairing bone defects.
The invention has the beneficial effects that:
1) according to the invention, in the presence of 2-aminoethanol-1-phosphoric acid, 4-hydroxy-2-butenoic acid is used for modifying POC, alcoholic hydroxyl on 4-hydroxy-2-butenoic acid can react with alcoholic hydroxyl on citric acid to form ether bond, so that a flexible chain segment containing carbon double bond and carboxyl bond is introduced to the side surface of a POC main chain, and carboxylic acid group is introduced, when the modified POC is used for preparing the porous scaffold, the rigidity of the molecular main chain in a cross-linked network structure can be reduced while cross-linking is increased, so that the porous scaffold has better strength and toughness, and meanwhile, the hydrophilicity of the scaffold can be increased, and cell adhesion and proliferation are promoted;
2) according to the invention, guanethidine sulfate is added when the porous scaffold is prepared, and can penetrate through the modified POC network structure, so that sodium chloride can be wrapped, sodium chloride sedimentation can be inhibited, the sodium chloride can be uniformly distributed in the polymer, the formation of an open pore structure by the sodium chloride is facilitated, the porosity and distribution uniformity of the porous scaffold can be improved, the connectivity of the scaffold is improved, and cell adhesion and proliferation are facilitated.
Drawings
FIG. 1 is a Fourier infrared spectrum of an unmodified and a modified POC prepolymer in example 1 of the present invention;
FIG. 2 is a graph showing the results of the measurement of the graft ratio in example 2 of the present invention;
FIG. 3 is a stress-strain curve of a porous scaffold in test example 1 of the present invention;
FIG. 4 is a graph showing the breaking strength, elongation at break and elastic modulus of the porous scaffold in test example 1 of the present invention;
FIG. 5 is a water contact angle of a porous support in test example 1 of the present invention;
FIG. 6 is an SEM photograph of a porous support in test example 2 of the present invention;
FIG. 7 is a result of measurement of porosity in test example 2 of the present invention;
FIG. 8 shows the results of measurement of the percentage reduction of alamarBlue in test example 3 of the present invention.
Detailed Description
Unless otherwise indicated, all publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety as if set forth in their entirety.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.
When an amount, concentration, or other value or parameter is given as either a range, preferred range, or a list of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any larger range limit or preferred value and any smaller range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when a range of "1 to 5" is described, the described range should be construed as including ranges of "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. Where numerical ranges are described herein, unless otherwise stated, the stated ranges are intended to include the endpoints of the ranges and all integers and fractions within the ranges.
In addition, the words "a" and "an" preceding an element or component of the invention are intended to mean no limitation on the number of times that the element or component appears (i.e., occurs). Thus, "a" or "an" should be understood to include one or at least one and the singular forms of an element or component also include the plural unless the singular is explicitly stated.
Embodiments of the present invention, including embodiments of the invention described in the summary section and any other embodiments described herein below, can be combined arbitrarily.
The present invention is described in detail below.
Provides the application of 4-hydroxy-2-butenoic acid in improving the toughness of the poly citrate.
Provided is a preparation method of modified poly (1, 8-octanediol-citric acid), comprising the following steps:
s1, placing citric acid, 1, 8-octanediol and 4-hydroxy-2-butenoic acid in a container, placing in an oil bath at 160-165 ℃ under the protection of nitrogen, heating and stirring, cooling to 135-140 ℃ after the mixture is melted, adding 2-aminoethanol-1-phosphoric acid, and reacting for 5-7 hours at normal pressure to obtain a prepolymer;
s2, dissolving the prepolymer in acetone, precipitating in ethanol to remove unreacted monomers, pouring into a mold, standing until acetone is completely volatilized, and polymerizing for 4-7d under the vacuum condition of 100-120 ℃. Poly (1, 8-octanediol-citric acid) (POC) is a network type polyester bio-elastomer prepared by reacting citric acid with 1, 8-octanediol, and is widely used in the field of tissue engineering due to its good mechanical and biological properties. The mechanical properties of POC can be adjusted by adjusting the molar ratio of the reactive monomers and the degree of cure crosslinking, but such adjustment is limited. The POC tensile deformability is improved, and the crosslinking degree is inevitably sacrificed, so that the material viscosity is increased, and the use is not facilitated. Thermosetting resins generally have a very high crosslinking density and therefore are very brittle. In the presence of 2-aminoethanol-1-phosphoric acid, 4-hydroxy-2-butenoic acid is used for modifying POC, alcoholic hydroxyl on 4-hydroxy-2-butenoic acid can react with alcoholic hydroxyl on citric acid to form ether bond, so that a flexible chain segment containing carbon double bond and carboxyl bond is introduced to the side surface of a POC main chain, and carboxylic acid group is introduced.
Preferably, the molar ratio of citric acid, 1, 8-octanediol and 4-hydroxy-2-butenoic acid in the step S1 is 5-6:4-5: 2-3.
Preferably, the mass ratio of the 2-aminoethanol-1-phosphoric acid to the 4-hydroxy-2-butenoic acid in the step S1 is 1: 2-3.
Provides a degradable biological elastomer which is prepared by the preparation method.
A preparation method of a poly citrate porous scaffold is provided, which comprises the following steps:
a. adding the prepolymer in the step S1 into acetone to fully dissolve the prepolymer, pouring the prepolymer into a tetrafluoroethylene mold, adding NaCl particles and guanethidine sulfate which are sieved by a 100-400-mesh sieve, uniformly stirring, ventilating to completely volatilize the acetone, and polymerizing for 4-7d under the vacuum condition at 100-120 ℃;
b. and taking out the sample, soaking the sample in deionized water, completely washing NaCl and guanethidine sulfate in the stent, and drying to obtain the poly-citrate porous stent. When the porous scaffold is prepared by using the solution casting/particle leaching technology, a compact skin layer is easily formed on the surface of a material particularly under the influence of the distribution of a pore-forming agent, the pore-forming agent is easy to precipitate at the bottom of a container to form an uneven pore structure, the obtained closed pore structure is more, and the connectivity among pores is poor. Guanethidine sulfate is added when the porous scaffold is prepared, can penetrate through the modified POC network structure, can wrap sodium chloride, and inhibit sodium chloride from settling, so that the sodium chloride can be uniformly distributed in the polymer, the formation of an open pore structure by the sodium chloride is facilitated, the porosity and distribution uniformity of the porous scaffold can be improved, the connectivity of the scaffold is improved, and cell adhesion and proliferation are facilitated.
Preferably, the mass ratio of the performed polymer in the step a, NaCl and guanethidine sulfate is 1:4-6: 3-5.
Providing a porous elastomeric material characterized by: the preparation method of the poly citrate porous scaffold is adopted for preparation.
The use of guanethidine sulfate to improve the connectivity of a stent is provided.
Provides the application of a degradable biological elastomer in preparing a tissue scaffold and/or a drug controlled release carrier.
Provides the application of a porous elastic material in repairing bone defects.
The present invention is further described in detail with reference to the following examples:
example 1:
1. a method for preparing modified poly (1, 8-octanediol-citric acid), comprising the steps of:
1.1115.3 g of citric acid, 58.5g of 1, 8-octanediol and 20.4g of 4-hydroxy-2-butenoic acid are put in a four-neck flask, stirred under the protection of nitrogen, put in an oil bath at 165 ℃ for heating and stirring, after the mixture is melted, the temperature is reduced to 140 ℃, 10g of 2-aminoethanol-1-phosphoric acid is added, and the reaction is carried out for 5 hours under normal pressure, thus obtaining the modified POC prepolymer. The Fourier IR spectra of the unmodified and modified POC prepolymers are shown in FIG. 1.
1.2 dissolving the prepolymer in acetone, precipitating in ethanol to remove unreacted monomers, repeating the purification process twice, pouring into a stainless steel mold to naturally cast the solution until the solution is flat, and polymerizing for 6d under vacuum at 120 ℃ after the acetone is completely volatilized.
2. A preparation method of a porous polycitrate scaffold comprises the following steps:
2.1 taking 10g of the prepolymer in the step 1.1, adding acetone to fully dissolve the prepolymer, pouring the mixture into a tetrafluoroethylene mold, adding 60g of NaCl particles which are sieved by a 400-mesh sieve and 30g of guanethidine sulfate, uniformly stirring, ventilating to completely volatilize the acetone, and then carrying out vacuum condition post-polymerization for 6d at 120 ℃;
2.2 taking out the sample, soaking the sample in deionized water, completely washing NaCl and guanethidine sulfate in the stent, and drying to obtain the multi-hole citrate stent.
Example 2:
poly (1, 8-octanediol-citric acid) was modified without adding 2-aminoethanol-1-phosphoric acid, and the remainder was completely the same as in example 1. The modified POC prepolymers obtained in examples 1 and 2 were subjected to graft ratio measurement by Fourier transform infrared spectroscopy, and the results of the graft ratio measurement are shown in FIG. 2.
As can be seen from FIG. 1, 3512cm of modified POC prepolymer-1The peak of O-H stretching vibration of hydroxyl group is weakened and is 1082cm-1A stretching vibration peak of C-O bond of the fatty ether appears at 1630cm-1A stretching vibration peak of-C ═ C-double bond appears at 2718cm-1The peak of stretching and contraction of O-H with carboxyl appears, which shows that the POC is modified by 4-hydroxy-2-butenoic acid, the alcoholic hydroxyl on the 4-hydroxy-2-butenoic acid can react with the alcoholic hydroxyl on the citric acid to form ether bond, a flexible chain segment containing carbon double bond and carboxyl bond is introduced at the side of the POC main chain, and carboxylic acid group is introduced, and as can be seen from figure 2, the grafting ratio of the 4-hydroxy-2-butenoic acid on the POC prepolymer in example 2 is almost 0, which shows that the 4-hydroxy-2-butenoic acid can be successfully grafted on the POC with higher grafting ratio in the presence of 2-aminoethanol-1-phosphoric acid.
Example 3:
poly (1, 8-octanediol-citric acid) was not modified, and the rest was completely the same as in example 1.
Example 4:
the preparation of the multicitrate scaffolds was completed in the same manner as in example 1 except that guanethidine sulfate was not added.
Example 5:
the preparation of the multicitrate scaffolds was completed in the same manner as in example 3 except that guanethidine sulfate was not added.
Test example 1:
and (3) mechanical property characterization: examples 1, 1 were processed through a die punch (2.67 mm in width at the narrowest and 7.49mm in length),
The porous scaffolds obtained in examples 3, 4 and 5 were cut into dumbbell-shaped specimens and tested on an HY-941 Universal materials tester (Shanghai Hengyu instruments, Inc.) at a tensile speed of 10 mm/min. And measuring for 5 times to obtain the data of breaking strength, breaking elongation and elastic modulus, and taking an average value. The stress-strain curve of the porous scaffold is shown in FIG. 3, and the breaking strength, elongation at break and elastic modulus of the porous scaffold are shown in FIG. 4.
And (3) detecting hydrophilicity: the contact angles of the porous scaffolds prepared in example 1, example 3, example 4 and example 5 were measured by a high-speed video optical contact angle measuring instrument of type Data Physics OCA 200. The sample is placed on a laboratory bench, the position of the sample is adjusted by a level regulator, 2 mu L of deionized water is dripped on the sample by a micro-injector, and then the size of the contact angle is measured under the shooting of an infrared thermal imager. And randomly selecting three points to test the sample, and taking the average value of the three points as the result of the hydrophilicity and hydrophobicity of the surface of the sample. The water contact angle of the porous scaffold is shown in figure 5.
As can be seen from fig. 3 and 4, the breaking strength, elongation at break and elastic modulus of the porous scaffold in example 1 are significantly higher than those of example 3, and the breaking strength, elongation at break and elastic modulus of the porous scaffold in example 4 are significantly higher than those of example 5, and as can be seen from fig. 5, the water contact angle of the porous scaffold in example 1 is significantly higher than that of example 3, and the water contact angle of the porous scaffold in example 4 is significantly higher than that of example 5, which shows that, after POC is modified with 4-hydroxy-2-butenoic acid in the presence of 2-aminoethanol-1-phosphoric acid, the rigidity of the molecular main chains in the cross-linked network structure can be reduced while the cross-linking is increased, so that the porous scaffold has better strength and toughness, and at the same time, the hydrophilicity of the scaffold can be increased.
Test example 2:
and (4) SEM observation: the porous scaffold pore structures in examples 3 and 5 were observed using a TM-100 type scanning electron microscope (Hitachi, Japan). Quenching in liquid nitrogen, and treating the cross section by low-temperature metal spraying. The SEM image of the porous scaffold is shown in figure 6.
And (3) measuring the porosity of the porous support material by adopting an ethanol substitution method. The mass of the pycnometer filled with ethanol is called m 1; immersing the porous support material with the mass of ms into ethanol, vacuumizing for many times to remove bubbles so that the pores of the porous support are completely filled with the ethanol, and then filling the pores with the ethanol to obtain a material with the mass of m 2; the sample of the porous support impregnated with ethanol was removed and the remaining ethanol and pycnometer were weighed again to mass m 3. Q is the density of ethanol.
Volume of the porous scaffold itself: vs ═ m1-m2+ ms)/Q;
porous scaffold pore volume: vp (m2-m 3-ms)/Q;
porosity of the porous scaffold: e ═ Vp/(Vp + Vs) ═ m2-m3-ms)/(m1-m 3);
and weighing 5 groups of the samples, calculating, and finally averaging. The results of the porosity measurements are shown in FIG. 7.
As can be seen from fig. 6, in example 5, the pores on the upper surface of the porous scaffold are less and denser, the pore diameter at the bottom is larger, and a non-uniform pore structure is formed, and in example 3, the pore structure of the porous scaffold is uniformly distributed and has good connectivity, as can be seen from fig. 7, the porosity of the porous scaffold in example 1 is significantly higher than that in example 4, and the porosity of the porous scaffold in example 3 is significantly higher than that in example 5, which indicates that guanethidine sulfate is added during the preparation of the porous scaffold, and the guanethidine sulfate can penetrate through the modified POC network structure, can wrap sodium chloride, and inhibit the settlement of sodium chloride, so that the sodium chloride can be uniformly distributed in the polymer, and is helpful for the sodium chloride to form an open pore structure, and can improve the porosity and distribution uniformity of the porous scaffold, and improve the connectivity.
Test example 3:
and (3) testing the cellular compatibility of the porous scaffold:
treating the porous scaffold: PLA porous scaffolds prepared by different molding techniques are respectively placed on polystyrene culture plates with 24 pores, and before use, the culture plates are soaked in alcohol and sterilized by an autoclave, then are rinsed with PBS (phosphate buffered saline), and then are soaked in a culture solution (DMEM, GIBCO). The culture solution used in the experiment is prepared by the laboratory.
And (3) cell planting: NIH-3T3 murine fibroblasts were treated with EDTA (Ethylene Diamine Tetraacetic Acid) for 5 minutes prior to implantation, washed with PBS, and then washed at 1.25X 105cells/cm2The concentration distribution of the culture medium is planted on a treated bracket for cell culture. The cell-seeded scaffolds were placed at 37 ℃ in CO2The culture solution was changed every day in an incubator at a concentration of 5% and a temperature of 95%. The cells were observed and characterized after 10 days of culture. Each set of scaffolds was replicated 3 times.
Preparing alamarBlue cell activity detection solution: in the experiment, 10% alamarBlue detection solution is adopted, namely 9 ml of cell culture solution (DMEM/F12+ 10% FBS) is taken, 1 ml of alamarBlue reagent solution is added, and the mixture is sufficiently and uniformly mixed and then kept in dark for later use.
And (3) detecting the activity of the cells: after NIH-3T3 mouse fibroblasts were planted on the sample surface and cultured for 1, 2 and 5 days, respectively, the cell culture solution in the wells containing each sample was aspirated, 200. mu.l of the prepared alamarBlue cell activity assay solution was added to each well, and the wells were returned to the cell culture chamber and cultured for 4 hours. Thereafter, 100. mu.l of liquid was aspirated from each well containing the sample, transferred to a new 96-well plate, and placed in a quantitative microplate reader to read and record at 570nm and 600nm, respectively.
And (3) processing cell activity data: the number of NIH-3T3 murine fibroblasts which have withered to grow on the surface of the sample is indirectly shown by the alamarBlue reduction ratio, and the calculation formula is as follows:
percent (%) reduction of alamarBlue { [ (O2 × A1) - (O1 × A2) ]/[ (R1 × N2) - (R2 × N1) ] } × 100%
O1 represents the molar absorption coefficient of alamarBlue at a wavelength of 570nm (equal to 80586);
o2 represents the molar absorption coefficient of alamarBlue at a wavelength of 600nm (equal to 117216);
r1 represents a reduced molar absorption coefficient of alamarBlue at a wavelength of 570nm (equal to 155677);
r2 represents a reduced molar absorption coefficient of alamarBlue at a wavelength of 600nm (equal to 14652); a1 represents the absorbance value of the plate hole of the detection sample at the wavelength of 570 nm;
a2 represents the absorbance value of the plate well of the test sample at a wavelength of 600 nm;
n1 represents the absorbance value at 570nm for negative control plate wells (cell culture medium added to alamarBlue, but no cells);
n2 represents the absorbance value at 600nm for negative control plate wells (cell culture medium added to alamarBlue, but no cells).
The results of the measurement of the percentage reduction of alamarBlue are shown in FIG. 8.
As can be seen from fig. 8, after culturing for 1 day, 2 days and 5 days, the reduction percentage of alamarBlue in example 1 is significantly higher than that in examples 3, 4 and 5, the reduction percentage of alamarBlue in example 3 is significantly higher than that in example 5, and guanethidine sulfate is added during preparation of the porous scaffold, so that the connectivity of the scaffold can be improved, and cell adhesion and proliferation are promoted; the percentage of alamarBlue reduction in example 4 is significantly higher than that in example 5, which shows that the modified POC can be used to prepare porous scaffolds after the modification of POC with 4-hydroxy-2-butenoic acid in the presence of 2-aminoethanol-1-phosphoric acid, thereby improving the hydrophilicity of the porous scaffolds and further promoting cell adhesion and proliferation.
Conventional techniques in the above embodiments are known to those skilled in the art, and therefore, will not be described in detail herein.
The above embodiments are merely illustrative, and not restrictive, and those skilled in the art can make various changes and modifications without departing from the spirit and scope of the invention. Therefore, all equivalent technical solutions also belong to the scope of the present invention, and the protection scope of the present invention should be defined by the claims.

Claims (1)

  1. The use of 4-hydroxy-2-butenoic acid for improving the toughness of a polycitrate ester.
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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2008311957A1 (en) * 2007-10-15 2009-04-23 Wake Forest University Methods and compositions for printing biologically compatible nanotube composites of autologous tissue
CN102246297A (en) * 2008-12-16 2011-11-16 横滨橡胶株式会社 Silanol condensation catalyst, heat-curable silicone resin composition for sealing photosemiconductors and sealed photosemiconductor using same
CN106310370A (en) * 2016-08-09 2017-01-11 东华大学 Preparing method of elastic degradable biomedical material
CN107349465A (en) * 2016-05-09 2017-11-17 香港大学深圳医院 A kind of implantable click chemistry is modified poly- citric acid 1,8 pungent two ester films and preparation method thereof
CN108624005A (en) * 2017-03-24 2018-10-09 华东理工大学 Double factor slow-released system based on POC and meso-porous nano microballoon
CN108939150A (en) * 2018-09-05 2018-12-07 华东理工大学 Osteoporosis defect repair bracket based on POFC/ β-TCP and Strontium Ranelate
CN110200963A (en) * 2011-10-19 2019-09-06 墨卡托医疗系统公司 Local modulation tissue and cell include kidney denervation to improve curative effect

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2008311957A1 (en) * 2007-10-15 2009-04-23 Wake Forest University Methods and compositions for printing biologically compatible nanotube composites of autologous tissue
CN102246297A (en) * 2008-12-16 2011-11-16 横滨橡胶株式会社 Silanol condensation catalyst, heat-curable silicone resin composition for sealing photosemiconductors and sealed photosemiconductor using same
CN110200963A (en) * 2011-10-19 2019-09-06 墨卡托医疗系统公司 Local modulation tissue and cell include kidney denervation to improve curative effect
CN107349465A (en) * 2016-05-09 2017-11-17 香港大学深圳医院 A kind of implantable click chemistry is modified poly- citric acid 1,8 pungent two ester films and preparation method thereof
CN106310370A (en) * 2016-08-09 2017-01-11 东华大学 Preparing method of elastic degradable biomedical material
CN108624005A (en) * 2017-03-24 2018-10-09 华东理工大学 Double factor slow-released system based on POC and meso-porous nano microballoon
CN108939150A (en) * 2018-09-05 2018-12-07 华东理工大学 Osteoporosis defect repair bracket based on POFC/ β-TCP and Strontium Ranelate

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