CN109432495B - Mineralized bone tissue engineering scaffold and preparation method thereof - Google Patents
Mineralized bone tissue engineering scaffold and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a mineralized bone tissue engineering scaffold, which comprises a porous polyester/zwitter-ion hydrogel composite scaffold and a calcium-phosphorus mineralized substance formed in situ in the porous polyester/zwitter-ion hydrogel composite scaffold; the porous polyester/zwitterionic hydrogel composite scaffold comprises a porous polyester framework and a zwitterionic hydrogel compounded with the porous polyester framework. The invention also discloses a preparation method of the mineralized bone tissue engineering scaffold. The composite scaffold can realize the high-efficiency slow release of bone growth factors, endow the surface of the whole scaffold with high-efficiency bone conduction and bone induction performances, promote the functional repair of bone defect under the ultra-low rhBMP-2 dosage and improve the biosafety of the scaffold material.
Description
Technical Field
The invention belongs to the field of biomedical materials, and particularly relates to a mineralized composite bone tissue engineering scaffold capable of forming efficient slow release on growth factors and a preparation method thereof.
Background
Worldwide, bone defects caused by traumatic injuries, tumors, diseases, infections, etc. afflict a large number of patients each year. Over 2,000,000 patients are currently undergoing bone graft surgery annually worldwide. Autologous bone and allogeneic bone are the main means for repairing bone defects in clinical practice at present. However, due to the limited source of autologous bone, the defect of potential immunological rejection of allogeneic bone exists, and the search for alternative high-efficiency bone repair materials has great scientific and social significance.
The addition of exogenous growth factors such as BMP-2 to artificial bone repair materials is a clinically proven effective method for accelerating the repair of bone defects. However, the use of high-dose growth factors required for effective treatment in clinic poses risks in terms of biological safety, and therefore, how to reduce the dose of growth factors to improve the biological safety of treatment on the premise of ensuring the curative effect is a hot spot and a difficult point in the research of artificial bone repair materials at present. Embedding growth factors in scaffold materials (Biomaterials2013, 34, (6), 1644-. With the gradual degradation of the scaffold material, the growth factors are gradually released, thereby guiding the differentiation of the stem cells to osteoblasts and finally functionally repairing the bone defect. However, embedding growth factors in the scaffold, the degradation of the scaffold material is not completely controllable; the growth factors are immobilized on the scaffold material, and the activity of the growth factors is inevitably adversely affected due to the change in the conformation of the growth factors. Therefore, finding a strategy to load efficiently without affecting growth factor activity is critical to achieving efficient sustained release of growth factors.
Disclosure of Invention
The invention aims to provide the mineralized bone tissue engineering scaffold which has the advantages of simple preparation method, mild conditions, simple and convenient operation, capability of loading growth factors, high-efficiency slow release performance of the growth factors and high-efficiency promotion of osteogenesis.
The invention also aims to provide a method for preparing the mineralized bone tissue engineering scaffold.
The technical scheme is as follows: the invention provides a mineralized bone tissue engineering scaffold, which comprises a porous polyester/zwitter-ion hydrogel composite scaffold and a calcium-phosphorus mineralized substance formed in situ in the porous polyester/zwitter-ion hydrogel composite scaffold; the porous polyester/zwitterionic hydrogel composite scaffold comprises a porous polyester framework and a zwitterionic hydrogel compounded with the porous polyester framework.
The invention utilizes the interaction of the special positive and negative charge structures of zwitterions of the zwitterions hydrogel and calcium ions and phosphate ions of the mineralized precursor of the calcium phosphate, efficiently drives the deposition of the calcium phosphate on the surface and in the scaffold, forms a novel organic/inorganic composite porous bone tissue engineering scaffold, and has the efficient slow release function on growth factors and the efficient bone conduction and bone induction performance. Under the condition of ultra-low growth factor dosage, the functional repair of bone defect can be promoted, and the biological safety performance of the stent material is improved. The biomineralization bracket can be prepared into required size and shape according to different clinical requirements, and the shape can be columnar, cake-shaped, cuboid and the like.
Preferably, the porous polyester/zwitterionic hydrogel scaffold is internally provided with interconnected pores with the pore diameter of 10-200 μm.
The polyester is a biodegradable polyester material with good biological safety performance, and preferably, the polyester can be polylactic acid, polyglycolic acid, polycaprolactone or a copolymer of more than two monomers of lactic acid, glycolic acid and caprolactone.
The above-mentioned zwitterionic hydrogel should be a material with good biosafety, for example, it can be formed by crosslinking one or more than two zwitterionic monomers of ammonium phosphate, sulfanilamide and carboxyl ammonium zwitterionic monomers containing unsaturated chemical bonds, more specifically, the zwitterionic monomer of ammonium phosphate, sulfanilamide or carboxyl ammonium containing unsaturated chemical bonds can be one or more than two of methacrylate type zwitterionic monomer, acrylate type zwitterionic monomer, methacrylamide type zwitterionic monomer, acrylamide type zwitterionic monomer, styrene type zwitterionic monomer and vinyl type zwitterionic monomer.
In the porous polyester/zwitterionic hydrogel composite scaffold, the zwitterionic hydrogel accounts for 1-99% by mass, and preferably the zwitterionic hydrogel accounts for 5-90% by mass.
In the mineralized bone tissue engineering scaffold, calcium and phosphorus mineralized substances are distributed on the surface and in the internal pores of the porous polyester/zwitterionic hydrogel composite scaffold; preferably, the preparation method of the mineralized bone tissue engineering scaffold comprises the following steps: depositing a precursor containing calcium ions and a precursor containing phosphate ions in situ in the porous polyester/zwitterion hydrogel composite support to form a calcium-phosphorus mineralizer; calcium phosphorus mineralizationThe substance comprises one or more of calcium phosphate, hydroxyapatite and octacalcium phosphate; in the precursor containing calcium ions and the precursor containing phosphate radical ions, the molar ratio of calcium to phosphorus is 1: 1.0-2.0; the calcium ion-containing precursor may be any compound that can release calcium ions in water, and as an example of the present invention, is calcium chloride (CaCl)2) The precursor containing phosphate ions can be any compound that can release phosphate ions in water, and is phosphoric acid (H) as one example of the present invention3PO4) (ii) a The mass proportion of the calcium phosphate mineralized substance in the mineralized bone tissue engineering scaffold is 20-80%.
The invention also provides a preparation method of the mineralized bone tissue engineering scaffold, which comprises the following steps:
1) preparing a zwitterionic hydrogel pre-polymerization solution containing a zwitterionic monomer, an initiator and a cross-linking agent, dropwise adding the zwitterionic hydrogel pre-polymerization solution on the surface of a porous polyester framework, crosslinking a prepolymer in the zwitterionic hydrogel pre-polymerization solution after the pre-polymerization solution uniformly permeates into the porous polyester framework, standing in a sterile phosphate buffer solution to remove impurities, and finally freeze-drying to obtain a polyester/zwitterionic hydrogel composite bracket;
2) soaking the polyester/zwitter-ion hydrogel composite scaffold prepared in the step 1) in a solution containing calcium ions and phosphate ions, then soaking in ammonia water to enable the calcium ions and the phosphate ions to react, forming a calcium-phosphorus mineralized substance in the polyester/zwitter-ion hydrogel composite scaffold, finally standing in a sterile phosphate buffer solution, and performing vacuum drying to obtain the mineralized bone tissue engineering scaffold.
In the step 1), the crosslinking method only needs to crosslink the prepolymer to obtain a polymer, and as an example of the invention, the crosslinking method can adopt ultraviolet crosslinking, and the time of the ultraviolet crosslinking is preferably 1-300 min; preferably, in the step 2), the molar amount of the calcium ions in the solution containing the calcium ions and the phosphate ions is 0.1-5 mol/L, and PO is4 3-The molar weight of (a) is 0.1-5 mol/L; soaking the composite polyester/amphoteric ion hydrogel support in solution containing calcium ion and phosphate radical ionThe middle time is 0.1-96 h; soaking the composite stent in ammonia water to ensure that the time for forming the calcium phosphate mineralized substance in the composite stent is 0.05-96 hours; the mass concentration of the ammonia water is 1-25%.
Has the advantages that: the mineralized composite bone tissue engineering scaffold takes biodegradable polyester as a framework, introduces zwitter-ion hydrogel as a mineralized template, and effectively drives the uniform in-situ formation and uniform deposition of the calcium phosphate mineralized substance on the surface and inside of the scaffold by utilizing the interaction of the special positive and negative charge structures of zwitter-ions and calcium ions and phosphate ions of the calcium phosphate mineralized precursor, so that an excellent mineralized effect is realized. The surface lattice structure of the calcium-phosphorus mineral substance on the surface of the bracket can generate electrostatic interaction with protein, so that the rhBMP-2 is stably adsorbed on the surface, the efficient slow release function is realized, the bone conduction and bone induction performance of the whole bracket are endowed, and the treatment cost and the potential risk are greatly reduced. The calcium phosphate mineralization bracket provided by the invention is simple in preparation method, mild in reaction condition and high in mineralization speed, can be prepared into a calcium phosphate bone tissue engineering bracket applied to wound orthopedic repair and reconstruction, and has wide application market prospect in the field of tissue engineering.
Drawings
Fig. 1 is macro-morphology and micro-morphology of a cake-shaped mineralized composite bone tissue engineering scaffold and calcium-phosphorus element analysis, wherein a is a test chart of calcium-phosphorus mineral morphology structure and calcium-phosphorus element on the surface of the mineralized composite bone tissue engineering scaffold, b is a test chart of calcium-phosphorus mineral morphology structure and calcium-phosphorus element inside the mineralized composite bone tissue engineering scaffold, c is a statistic result of molar ratio of calcium-phosphorus elements on the surface and inside the mineralized composite bone tissue engineering scaffold, and d is macro-morphology and size of the surface and side surface of the mineralized composite bone tissue engineering scaffold; the detected elements in the insert at the lower left corner of the graph a are mainly C, O, Ca and P, and S elements in the hydrogel are not shown, so that the thick calcium phosphate mineral layer on the surface of the stent is illustrated; in the lower left hand insert of panel b, elements C, O, Ca, P and S are detected, indicating that the interior of the stent is thin in the layer of calcium phosphate mineral.
FIG. 2 is the slow release effect curve of the polyester/zwitterionic hydrogel composite scaffold and the polyester/zwitterionic hydrogel bone tissue engineering scaffold on bone morphogenetic protein-2 (rhBMP-2).
FIG. 3 is a mu-CT image of a calcium phosphate mineralized scaffold loaded with rhBMP-2 implanted at 4 weeks and 8 weeks after implantation into a skull defect of a mouse.
Detailed Description
The present invention is described in detail below by way of examples, it should be noted that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention.
Example 1
(1) Preparation of PCL/zwitterion hydrogel composite scaffold
0.5mmol of 3- (2-methacryloyloxyethyl dimethylamino) propanesulfonate (SBMA) was dissolved in 1.75mL of Phosphate Buffered Saline (PBS), and 50. mu.L of azo initiator VA-086(10mg/mL) and 25. mu.L of crosslinker polyethylene glycol diacrylate were added to form a sulfonamide zwitterionic hydrogel pre-polymerization solution. Dissolving a Polycaprolactone (PCL) polymer in 1, 4-dioxane, and preparing a porous polycaprolactone scaffold by a freeze-drying method; 50 mu L of sulfanilamide zwitterionic hydrogel pre-polymerization solution prepared in the embodiment is dripped on the surface of the porous PCL support, after the pre-polymerization solution is completely soaked into the support, ultraviolet crosslinking is carried out, after the front side and the back side are respectively irradiated for 5min, the support is soaked in sterile phosphate buffer solution at 37 ℃ for 12h, and after freeze drying and water removal, the PCL/zwitterionic hydrogel composite support with the zwitterionic hydrogel mass ratio of 10% is obtained, wherein the aperture is about 150 mu m.
(2) Mineralization of PCL/zwitterionic hydrogel composite scaffolds
Adding 0.041mol CaCl2And 0.024mol H3PO4Adding the solution into a centrifugal tube, adding 30mL of deionized water to completely dissolve the solution to prepare calcium phosphate mineralized liquid, then soaking the PCL/zwitter-ion hydrogel composite scaffold prepared in the step 1) in the calcium phosphate mineralized liquid for 6 hours, then soaking the PCL/zwitter-ion hydrogel composite scaffold in 15% ammonia water for 3 hours to separate out calcium phosphate crystals, finally standing the PCL/zwitter-ion hydrogel mineralized bone tissue engineering scaffold in a sterile phosphate buffer solution at 37 ℃ for 12 hours, and performing vacuum drying to obtain the PCL/zwitter-ion hydrogel mineralized bone tissue engineering scaffold.
Example 2
(1) Preparation of PLGA/zwitterionic hydrogel composite scaffold
1mmol of 3- (2-methacryloyloxyethyl dimethylamino) propanesulfonate (SBMA) was dissolved in 3.5mL of Phosphate Buffered Saline (PBS), and 100. mu.L of azo initiator VA-086(10mg/mL) and 50. mu.L of crosslinker polyethylene glycol diacrylate were added to form a sulfoammonium zwitterionic hydrogel pre-polymerization solution. Dissolving polylactic acid-glycolic acid copolymer (PLGA) in 1, 4-dioxane, and preparing a porous polycaprolactone stent by a freeze-drying method; 150 mu L of sulfanilamide zwitterionic hydrogel pre-polymerization solution prepared in the embodiment is dripped on the surface of the porous PLGA stent, after the pre-polymerization solution is completely infiltrated into the stent, ultraviolet crosslinking is carried out, after the front side and the back side are respectively irradiated for 12min, the stent is soaked in sterile phosphate buffer solution at 37 ℃ for 12h, and after freeze drying and water removal, the PLGA/zwitterionic hydrogel composite stent with the zwitterionic hydrogel mass ratio of 50% is obtained, wherein the aperture is about 100 mu m.
(2) Mineralization of PLGA/zwitterionic hydrogel composite scaffolds
Adding 0.075mol of CaCl2And 0.045molH3PO4Adding the PLGA/zwitter-ion hydrogel composite scaffold into a centrifugal tube, adding 40mL of deionized water to completely dissolve the PLGA/zwitter-ion hydrogel composite scaffold to prepare calcium phosphate mineralized liquid, soaking the PLGA/zwitter-ion hydrogel composite scaffold prepared in the step 1) in the calcium phosphate mineralized liquid for 10 hours, then soaking the PLGA/zwitter-ion hydrogel composite scaffold in 5% ammonia water for 5 hours to separate out calcium phosphate crystals, finally standing the PLGA/zwitter-ion hydrogel mineralized bone tissue engineering scaffold in a sterile phosphate buffer solution at 37 ℃ for 8 hours, and performing vacuum.
Example 3
(1) Preparation of PLA/zwitterion hydrogel composite scaffold
7mmol of 3- [ [2- (methacryloyloxy) ethyl ] dimethylammonium ] propionate (CBMA) was dissolved in 9mL Phosphate Buffered Saline (PBS), and 300. mu.L of azo initiator VA-086(10mg/mL) and 70. mu.L of crosslinker polyethylene glycol diacrylate were added to form a carboxyammonium zwitterionic hydrogel pre-polymerization solution. Dissolving polylactic acid (PLA) polymer in 1, 4-dioxane, and preparing a porous polycaprolactone stent by a freeze-drying method; 200 mu L of the prepared carboxamide zwitterionic hydrogel pre-polymerization solution is dripped on the surface of a porous PLA bracket, after the liquid is completely infiltrated into the bracket, ultraviolet crosslinking is carried out, after the front side and the back side are respectively irradiated for 15min, the bracket is soaked in sterile phosphate buffer solution at 37 ℃ for 12h, and after freeze drying and water removal, the PLA/zwitterionic hydrogel composite bracket with the zwitterionic hydrogel mass ratio of 65% is obtained, wherein the aperture is about 65 mu m.
(2) Mineralization of PLA/zwitterionic hydrogel composite scaffolds
Adding 0.09mol of CaCl2And 0.07molH3PO4Adding the mixture into a centrifugal tube, adding 45mL of deionized water to completely dissolve the mixture to prepare a calcium-phosphorus mineralized liquid, soaking the PLA/zwitter-ion hydrogel composite scaffold prepared in the step 1) in the calcium-phosphorus mineralized liquid for 8 hours, then soaking the calcium-phosphorus mineralized liquid in 10% ammonia water for 7 hours to separate out calcium-phosphorus crystals, finally standing the calcium-phosphorus mineralized liquid in a sterile phosphate buffer solution for 15 hours at 37 ℃, and performing vacuum drying to obtain the PLA/zwitter-ion hydrogel mineralized bone tissue engineering scaffold.
Example 4
(1) Preparation of PLGA/zwitterionic hydrogel composite scaffold
3mmol of 2-Methacryloyloxyethyl Phosphorylcholine (MPC) was dissolved in 7mL of Phosphate Buffered Saline (PBS), and 120. mu.L of azo initiator VA-086(10mg/mL) and 55. mu.L of crosslinker polyethylene glycol diacrylate were added to form a phosphamine zwitterionic hydrogel pre-polymerization solution. Dissolving polylactic acid-glycolic acid copolymer (PLGA) in 1, 4-dioxane, and preparing a porous polycaprolactone stent by a freeze-drying method; and (2) dropwise adding 100 mu L of the phosphamidon zwitterionic hydrogel pre-polymerization solution prepared in the embodiment on the surface of the porous PLGA stent, after the liquid is completely infiltrated into the interior of the stent, carrying out ultraviolet crosslinking, after the front side and the back side are respectively irradiated for 20min, soaking in sterile phosphate buffer solution at 37 ℃ for 11h, and carrying out freeze drying to remove water, thus obtaining the PLGA/zwitterionic hydrogel composite stent with the zwitterionic hydrogel mass ratio of 45%, wherein the pore diameter is about 80 mu m.
(2) Mineralization of PLGA/zwitterionic hydrogel composite scaffolds
Adding 0.04mol of CaCl2And 0.025mol H3PO4Adding the mixture into a centrifugal tube, adding 80mL of deionized water to completely dissolve the mixture to prepare calcium-phosphorus mineralized liquid, then soaking the PLGA/zwitter-ion hydrogel composite scaffold prepared in the step 1) in the calcium-phosphorus mineralized liquid for 8 hours, then soaking the PLGA/zwitter-ion hydrogel composite scaffold in 1% ammonia water for 4.5 hours to separate out calcium-phosphorus crystals, and finally, placing the calcium-phosphorus crystals in a sterile phosphate buffer solutionStanding for 20h at 25 ℃, and drying in vacuum to obtain the PLGA/zwitter-ion hydrogel mineralized bone tissue engineering scaffold.
Example 5
(1) Preparation of PLGA/zwitterionic hydrogel composite scaffold
3mmol of 3- [ [2- (methacryloyloxy) ethyl ] dimethylammonium ] propionate (CBMA) was dissolved in 7mL Phosphate Buffered Saline (PBS), and 120. mu.L of azo initiator VA-086(10mg/mL) and 55. mu.L of crosslinker polyethylene glycol diacrylate were added to form a carboxyamide zwitterionic hydrogel pre-polymerization solution. Dissolving polylactic acid-glycolic acid copolymer (PLGA) in 1, 4-dioxane, and preparing a porous polycaprolactone stent by a freeze-drying method; and (2) dropwise adding 100 mu L of the carboxyl amine zwitterionic hydrogel pre-polymerization solution prepared in the embodiment on the surface of the porous PLGA stent, after the liquid is completely infiltrated into the interior of the stent, carrying out ultraviolet crosslinking, after the front side and the back side are respectively irradiated for 20min, soaking in sterile phosphate buffer solution at 37 ℃ for 11h, and carrying out freeze drying to remove water, thus obtaining the PLGA/zwitterionic hydrogel composite stent with the zwitterionic hydrogel mass ratio of 45%, wherein the pore diameter is about 80 mu m.
(2) Mineralization of PLGA/zwitterionic hydrogel composite scaffolds
Adding 0.04mol of CaCl2And 0.025mol H3PO4Adding the mixture into a centrifugal tube, adding 80mL of deionized water to completely dissolve the mixture to prepare calcium phosphate mineralized liquid, then soaking the PLGA/zwitter-ion hydrogel composite scaffold prepared in the step 1) in the calcium phosphate mineralized liquid for 8 hours, then soaking the calcium phosphate mineralized liquid in 25% ammonia water for 4.5 hours to separate out calcium phosphate crystals, finally standing the calcium phosphate crystals in a sterile phosphate buffer solution for 20 hours at 25 ℃, and performing vacuum drying to obtain the PLGA/zwitter-ion hydrogel mineralized bone tissue engineering scaffold.
Example 6
(1) Preparation of PLGA/zwitterionic hydrogel composite scaffold
4mmol of 2-Methacryloyloxyethyl Phosphorylcholine (MPC) was dissolved in 6mL of Phosphate Buffered Saline (PBS), and 85. mu.L of azo initiator VA-086(10mg/mL) and 45. mu.L of crosslinker polyethylene glycol diacrylate were added to form a phosphamine zwitterionic hydrogel pre-polymerization solution. Dissolving polylactic acid-glycolic acid copolymer (PLGA) in 1, 4-dioxane, and preparing a porous polycaprolactone stent by a freeze-drying method; 125 mu L of the phosphamidon zwitterionic hydrogel pre-polymerization solution prepared in the embodiment is dripped on the surface of the porous PLGA stent, after the liquid is completely infiltrated into the stent, ultraviolet crosslinking is carried out, after the front side and the back side are respectively irradiated for 18min, the stent is soaked in sterile phosphate buffer solution at 37 ℃ for 8h, and after freeze drying and water removal, the PLGA/zwitterionic hydrogel composite stent with the zwitterionic hydrogel mass ratio of 48% is obtained, wherein the aperture is about 75 mu m.
(2) Mineralization of PLGA/zwitterionic hydrogel composite scaffolds
Adding 0.05mol of CaCl2 and 0.016mol of H3PO4 into a centrifuge tube, adding 35mL of deionized water to completely dissolve the materials to prepare calcium-phosphorus mineralized liquid, then soaking the PLGA/zwitterionic hydrogel composite scaffold prepared in the step 1) in the calcium-phosphorus mineralized liquid for 8h, then soaking the calcium-phosphorus mineralized liquid in 20% ammonia water for 6h to separate out calcium-phosphorus crystals, finally standing the calcium-phosphorus mineralized liquid in a sterile phosphate buffer solution at 37 ℃ for 15h, and performing vacuum drying to obtain the PLGA/zwitterionic hydrogel mineralized bone tissue engineering scaffold.
Example 7
(1) Preparation of PLGA/zwitterionic hydrogel composite scaffold
4mmol of 2-Methacryloyloxyethyl Phosphorylcholine (MPC) was dissolved in 6mL of Phosphate Buffered Saline (PBS), and 85. mu.L of azo initiator VA-086(10mg/mL) and 45. mu.L of crosslinker polyethylene glycol diacrylate were added to form a phosphamine zwitterionic hydrogel pre-polymerization solution. Dissolving polylactic acid-glycolic acid copolymer (PLGA) in 1, 4-dioxane, and preparing a porous polycaprolactone stent by a freeze-drying method; 125 mu L of the phosphamidon zwitterionic hydrogel pre-polymerization solution prepared in the embodiment is dripped on the surface of the porous PLGA stent, after the liquid is completely infiltrated into the stent, ultraviolet crosslinking is carried out, after the front side and the back side are respectively irradiated for 18min, the stent is soaked in sterile phosphate buffer solution at 37 ℃ for 8h, and after freeze drying and water removal, the PLGA/zwitterionic hydrogel composite stent with the zwitterionic hydrogel mass ratio of 48% is obtained, wherein the aperture is about 75 mu m.
(2) Mineralization of PLGA/zwitterionic hydrogel composite scaffolds
Adding 0.05mol of CaCl2And 0.016mol of H3PO4Adding into a centrifuge tube, adding 35mL deionized waterCompletely dissolving the PLGA/zwitter-ion hydrogel composite scaffold to prepare a calcium-phosphorus mineralized liquid, soaking the PLGA/zwitter-ion hydrogel composite scaffold prepared in the step 1) in the calcium-phosphorus mineralized liquid for 8 hours, then soaking in 15% ammonia water for 6 hours to separate out calcium-phosphorus crystals, finally standing in a sterile phosphate buffer solution at 37 ℃ for 15 hours, and performing vacuum drying to obtain the PLGA/zwitter-ion hydrogel mineralized bone tissue engineering scaffold.
Example 8
(1) Preparation of PCL/zwitterion hydrogel composite scaffold
5mmol 2-Methacryloyloxyethyl Phosphorylcholine (MPC) was dissolved in 10mL Phosphate Buffered Saline (PBS) and 100. mu.L of azo initiator VA-086(10mg/mL) and 70. mu.L of crosslinker polyethylene glycol diacrylate were added to form a phosphamine zwitterionic hydrogel pre-polymerization solution. Dissolving polylactic acid-glycolic acid copolymer (PLGA) in 1, 4-dioxane, and preparing a porous polycaprolactone stent by a freeze-drying method; and (3) dropwise adding 180 mu L of the phosphamidon zwitterionic hydrogel pre-polymerization solution prepared in the embodiment on the surface of the porous PLGA stent, after the liquid is completely infiltrated into the stent, carrying out ultraviolet crosslinking, respectively irradiating the front side and the back side for 25min, soaking in a sterile phosphate buffer solution at 25 ℃ for 10h, and carrying out freeze drying to remove water, thus obtaining the PCL/zwitterionic hydrogel composite stent with the zwitterionic hydrogel mass ratio of 52% and the pore diameter of about 66 mu m.
(2) Mineralization of PCL/zwitterionic hydrogel composite scaffolds
Adding 0.035mol CaCl2And 0.038mol H3PO4Adding the solution into a centrifugal tube, adding 44mL of deionized water to completely dissolve the solution to prepare calcium phosphate mineralized liquid, then soaking the PCL/zwitter-ion hydrogel composite scaffold prepared in the step 1) in the calcium phosphate mineralized liquid for 16h, then soaking the PCL/zwitter-ion hydrogel composite scaffold in 15% ammonia water for 12h to separate out calcium phosphate crystals, finally standing the PCL/zwitter-ion hydrogel mineralized bone tissue engineering scaffold in a sterile phosphate buffer solution at 37 ℃ for 12h, and performing vacuum drying to obtain the PCL/zwitter-ion hydrogel mineralized bone tissue engineering scaffold.
Structural and performance characterization
The mineralized bone tissue engineering scaffold prepared in example 2 was subjected to structural and performance characterization. Fig. 1 is macro and micro morphology of a cake-shaped mineralized bone tissue engineering scaffold and calcium and phosphorus element analysis, wherein a is a test chart of morphology structure of calcium and phosphorus mineral substances and calcium and phosphorus elements on the surface of the mineralized bone tissue engineering scaffold, b is a test chart of morphology structure of calcium and phosphorus mineral substances and calcium and phosphorus elements inside the mineralized bone tissue engineering scaffold, c is a statistical result of molar ratio of calcium and phosphorus elements on the surface and inside the mineralized bone tissue engineering scaffold, and d is macro morphology and size of the surface and side surface of the mineralized bone tissue engineering scaffold; the detected elements in the insert at the lower left corner of the graph a are mainly C, O, Ca and P, and S elements in the hydrogel are not shown, so that the thick calcium phosphate mineral layer on the surface of the stent is illustrated; in the lower left hand insert of panel b, elements C, O, Ca, P and S are detected, indicating that the interior of the stent is thin in the layer of calcium phosphate mineral.
FIG. 2 is a slow release effect curve of PLGA/ammonium sulfonate zwitterionic hydrogel composite scaffold and PLGA/ammonium sulfonate zwitterionic hydrogel mineralized bone tissue engineering scaffold on bone morphogenetic protein-2 (rhBMP-2). The PLGA/ammonium sulfonate zwitterionic hydrogel composite scaffold releases about 8% of rhBMP-2 within 2h, the total release amount within 72h reaches about 14%, the special lattice structure on the surface of the PLGA/ammonium sulfonate zwitterionic hydrogel mineralized bone tissue engineering scaffold can generate electrostatic interaction with protein, the rhBMP-2 is stably adsorbed on the surface, only 0.04% of the total release amount within 2h is released, and only 0.5% of the total loading amount is released within 72h, so that the combination of the calcium-phosphorus mineral substance and the PLGA/ammonium sulfonate zwitterionic hydrogel composite scaffold endows the overall scaffold with a high-efficiency slow release function for growth factors.
FIG. 3 is the mu-CT images of the mineralized bone tissue engineering scaffold loaded with rhBMP-2 at 4 weeks and 8 weeks after being implanted into the skull defect of a mouse. After the mineralized bone tissue engineering scaffold loaded with 400ng of rhBMP-2 is implanted into a skull defect of a mouse, obvious new bone generation already occurs 4 weeks later, new bones grow inwards from the edge, after 8 weeks after operation, the new bone amount at the defect is obviously increased, and the new bones almost completely cover the defect position, so that the mineralized bone tissue engineering scaffold loaded with low dose (400ng) of rhBMP-2 can realize obvious bone repair effect in a short period.
The structural and performance characterization of examples 1, 3-8 are similar to example 2.
Claims (9)
1. The mineralized bone tissue engineering scaffold is characterized by comprising a porous polyester/zwitter-ion hydrogel composite scaffold and a calcium-phosphorus mineralized substance formed in situ in the porous polyester/zwitter-ion hydrogel composite scaffold; the porous polyester/zwitterionic hydrogel composite scaffold comprises a porous polyester framework and a zwitterionic hydrogel compounded with the porous polyester framework; the calcium phosphate mineralized substances are distributed on the surface and in the internal pores of the porous polyester/zwitterionic hydrogel composite bracket; the mineralized bone tissue engineering scaffold is used for loading bone growth factors, can realize efficient slow release of the bone growth factors and endows the surface of the whole scaffold with efficient bone conduction and bone induction performances.
2. The mineralized bone tissue engineering scaffold according to claim 1, wherein interconnected pores with a pore size of 10-200 μm penetrate through the porous polyester/zwitterionic hydrogel scaffold.
3. The mineralized bone tissue engineering scaffold according to claim 1, wherein the polyester is a homopolymer of one monomer or a copolymer of two or more monomers selected from lactic acid, glycolic acid, and caprolactone; the zwitterionic hydrogel is formed by crosslinking one or more than two zwitterionic monomers of ammonium phosphate, sulfanilamide and carboxyl ammonium zwitterionic monomers containing unsaturated chemical bonds; the weight proportion of the zwitterionic hydrogel in the porous polyester/zwitterionic hydrogel composite support is 1-99%.
4. The mineralized bone tissue engineering scaffold according to claim 3, wherein the ammonium phosphate, sulfonamide, or carboxylate zwitterionic monomers containing unsaturated chemical bonds comprise one or more of methacrylate zwitterionic monomers, acrylate zwitterionic monomers, methacrylamide zwitterionic monomers, acrylamide zwitterionic monomers, styrene zwitterionic monomers, and vinyl zwitterionic monomers.
5. The mineralized bone tissue engineering scaffold according to claim 1, wherein the preparation method of the mineralized bone tissue engineering scaffold comprises: depositing a precursor containing calcium ions and a precursor containing phosphate ions in situ in the porous polyester/zwitterion hydrogel composite support to form a calcium-phosphorus mineralizer; the calcium phosphate mineralizer comprises one or more of calcium phosphate, hydroxyapatite and octacalcium phosphate; in the calcium-phosphorus mineralizer, the molar ratio of calcium to phosphorus is 1: 1.0 to 2.0.
6. The mineralized bone tissue engineering scaffold according to claim 5, wherein the precursor containing calcium ions is calcium chloride, and the precursor containing phosphate ions is phosphoric acid.
7. The mineralized bone tissue engineering scaffold according to claim 1, wherein the calcium phosphate mineralized substance accounts for 20-80% by mass of the mineralized bone tissue engineering scaffold.
8. The method for preparing the mineralized bone tissue engineering scaffold according to any one of claims 1 to 7, wherein the method comprises the following steps:
1) preparing a zwitterionic hydrogel pre-polymerization solution containing a zwitterionic monomer, an initiator and a cross-linking agent, dropwise adding the zwitterionic hydrogel pre-polymerization solution on the surface of a porous polyester framework, crosslinking a prepolymer in the zwitterionic hydrogel pre-polymerization solution when the pre-polymerization solution uniformly permeates into the porous polyester framework, standing in a sterile phosphate buffer solution to remove impurities, and finally freeze-drying to obtain the polyester/zwitterionic hydrogel composite bracket;
2) soaking the polyester/zwitter-ion hydrogel composite scaffold prepared in the step 1) in a solution containing calcium ions and phosphate ions, then soaking in ammonia water to enable the calcium ions and the phosphate ions to react, forming a calcium-phosphorus mineralized substance in the polyester/zwitter-ion hydrogel composite scaffold, finally standing in a sterile phosphate buffer solution, and performing vacuum drying to obtain the mineralized bone tissue engineering scaffold.
9. The preparation method according to claim 8, wherein in the step 1), the crosslinking method is ultraviolet crosslinking, and the time of the ultraviolet crosslinking is 1-300 min; in the step 2), the concentration of calcium ions in the solution containing calcium ions and phosphate ions is 0.1-5 mol/L, and PO is4 3-The concentration of (A) is 0.1-5 mol/L; the polyester/zwitter-ion hydrogel composite bracket is soaked in a solution containing calcium ions and phosphate ions for 0.1-96 h; the mass concentration of the ammonia water is 1-25%, and the time for soaking the composite bracket in the ammonia water to form the calcium phosphate mineralized substance is 0.05-96 hours.
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| CN108690309A (en) * | 2017-04-06 | 2018-10-23 | 天津大学 | High intensity mineralising hydrogel and its application in Bone Defect Repari |
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