CN104027840A - Preparation method of porous scaffold for bone repair - Google Patents
Preparation method of porous scaffold for bone repair Download PDFInfo
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- CN104027840A CN104027840A CN201410267249.9A CN201410267249A CN104027840A CN 104027840 A CN104027840 A CN 104027840A CN 201410267249 A CN201410267249 A CN 201410267249A CN 104027840 A CN104027840 A CN 104027840A
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Abstract
The invention relates to a preparation method of a porous scaffold for bone repair, belonging to the technical field of bio-medical materials. The preparation method comprises the steps of firstly dissolving poly-L-lactic acid into a solvent to obtain a first solution; dissolving polycaprolactone into the solvent to obtain a second solution; mixing the first solution and the second solution to obtain a third solution; adding nano hydroxyapatite particles into the third solution to obtain a fourth solution; adding aragonite or vaterite pearl powder particles into the fourth solution to obtain a fifth solution; pouring the fifth solution into a scaffold mold, freezing, and then performing freeze drying to obtain a dried bone repair porous composite scaffold. The porous scaffold prepared by using the method disclosed by the invention has good mechanical performances; inorganic components of the scaffold are more close to the bone so as to ensure that a bone repair material has better bio-compatibility, is beneficial for the growth and proliferation of cells, and can be used for prolonging the degradation time of the bone repair scaffold.
Description
Technical field
The present invention relates to a kind of preparation method of the porous support of repairing for bone, relate in particular to a kind of preparation method of polylactic acid and caprolactone/Margarita powder/nanometer hydroxyapatite porous support of repairing for bone, belong to bio-medical material technical field.
Background technology
At present, the research of porous compound support frame mainly concentrates on that biodegradable macromolecule is compound has a bioactive ceramic particle.Biodegradable high-molecular-Poly-L-lactic acid (hereinafter to be referred as PLLA), polycaprolactone (hereinafter to be referred as PCL) have good biocompatibility, biological degradability and preferably mechanical property, controlled degradation property and machinability, its catabolite can participate in the metabolism of human body.This makes them become in current biomedical sector one of the most valued material, and by united states drug food control office (FDA) approval for human body.
Polylactide and its copolymer is mainly used in controlled drug delivery system, bone inside-fixture, tissue repair and cell culture material and medical operation suture thread etc.But there is the defect of some application aspect in Poly-L-lactic acid: (1) polylactic acid is lyophobic dust, reduced its cell compatibility and biocompatibility; (2) sour environment that its catabolite causes accelerates the degraded of PLLA self, also easily causes aseptic inflammation in body; (3), as porous support, its mechanical property can not meet the requirement of bone implant material.
Polycaprolactone (PCL) is a kind of biodegradable high polymer, has good biocompatibility, and its elongation at break is high, plasticity is good, but PCL degradation time is longer, degradable needs 24 months, the mechanical property and the degradation speed that are generally used for the compound controlled material of other materials.
In order to improve the mechanical property of porous support materials, in preparation process, often add having of some strong mechanical performances bioactive granule, particularly nanometer hydroxyapatite (nano-hydroxylapatite, hereinafter to be referred as nHA) and Margarita powder.NHA is the main inorganic composition of human bone, has good biocompatibility, and non-stimulated and rejection in vivo, can conduct osteogenesis.But there is the defect of some application aspect in HA: inanimate object is active, and the nHA degradation speed of sintering is slower.Nacreous layer has reduced immunogenicity, good biocompatibility, degradability, bone conductibility and ossification preferably, and self-contained bioactie agent, meets the design concept of desirable timbering material.Natural mussel pearl can be divided into aragonite Margarita and ball aragonite Margarita, all contains somatomedin in the organic substrate of two kinds of nacreous layers, and in organic substrate, the albumen of soluble micromolecule amount is for promoting that the Differentiation and proliferation of osteocyte is even more important.But the degradation speed of nacreous layer block is slower.
Summary of the invention
The preparation method that the object of the invention is to propose a kind of porous support of repairing for bone, to improve mechanical property and the degradability of existing pure PLLA porous support, adds bioactie agent, and makes preparation method simple.
The preparation method of the porous support of repairing for bone that the present invention proposes, comprises the following steps:
(1) Poly-L-lactic acid is dissolved in solvent 1-4-dioxane, the mass volume ratio of Poly-L-lactic acid and solvent 1-4-dioxane is: Poly-L-lactic acid: solvent 1-4-dioxane)=(0.03～0.08): 1, within 1～8 hour under magnetic agitation, dissolve, obtain the first solution;
(2) polycaprolactone is dissolved in solvent 1-4-dioxane, the mass volume ratio of polycaprolactone and solvent 1-4-dioxane is: polycaprolactone: solvent 1-4-dioxane=(0.03～0.08): 1, within 1～8 hour under magnetic agitation, dissolve, obtain the second solution;
(3) the first solution is mixed with the second solution, the volume ratio of mixing is: the first solution: the second solution=(0.8～1.2): 1, obtain the 3rd solution;
(4) at stirring, nano-hydroapatite particles is joined in the 3rd solution, obtain the 4th solution, in the 4th solution, the mass ratio of nano-hydroapatite particles, Poly-L-lactic acid and polycaprolactone is: nano-hydroapatite particles: Poly-L-lactic acid: polycaprolactone=1:(1～9): stirred 1～2 hour (1～9), with ultrasonic dispersion 1～2 hour;
(5) by aragonite or ball aragonite Margarita with alcohol-pickled 1～4 hour, after dry, pulverize after washing down with deionized water, obtain Margarita powder granule, the granularity of Margarita powder granule is less than 50 microns;
(6) at stirring, above-mentioned Margarita powder granule is joined in the 4th solution, obtain the 5th solution, in the 5th solution, the mass ratio of each composition is: Margarita powder granule: nano-hydroapatite particles: Poly-L-lactic acid: polycaprolactone=1:(1～2): (1～9): (1～9), stir 10～30 minutes;
(7) the 5th solution, is poured in bracket mould after 1～2 hour with ultrasonic dispersion, shaken up after 5～10 minutes, under-20 ℃ of environment freezing 10～24 hours;
(8) sample freezing is put into freezer dryer, lyophilization 48～75 hours, the bone repair porous compound rest that obtains being dried.
The preparation method of the porous support of repairing for bone that the present invention proposes, its advantage is:
(1) in porous support preparation method of the present invention, the PLLA selecting, PCL, nHA and Margarita powder, all have good biocompatibility, and four kinds of Material claddings are used, and makes the support of preparation have good biocompatibility.
(2) in preparation method of the present invention, PLLA wherein mixes by a certain percentage with PCL, can regulate and control the degradation speed of support integral body, increases the plasticity of support simultaneously, has therefore optimized the mechanical property of support.
(3) in preparation method of the present invention, appropriate after the Margarita powder and nHA adding, do not affect pattern, aperture and the porosity of PLLA support, and retaining that the porous pattern of support and certain aperture are conducive to that cell attaches, in propagation and differentiation, increased the mechanical property of support.
(4) bone repairing support that utilizes preparation method of the present invention to prepare, has good bone inductive effect.Add after Margarita powder, because physics, chemical property and biological characteristics and the osseous tissue of nacreous layer are very approaching, there is bone conductibility and good ossification, in stent area, be easy to skeletonization.The self-contained bioactie agent of Margarita powder, and signal factor is comparatively stable in nacreous layer, changeableness not, plays the effect of slow release.
(5) preparation method of the present invention is mixed nHA by a certain percentage with Margarita powder, increases on the one hand the dissolubility of granule, improves the degradation speed of support integral body; On the other hand, can control the calcium-phosphorus ratio in granule, make the inorganic constituents of support and bone more approaching, make bone renovating material there is better biocompatibility.
(6) preparation method process of the present invention is simple, and in prior art, the support that carries atopen or albumen is often in order to ensure the slow release of albumen and changeableness and make complicated process of preparation not, and PLLA/PCL/ prepared by the inventive method Margarita powder/nHA support, utilize signal factor stability in nacreous layer, only Margarita powder, nHA need be added into lyophilization in the solution of PLLA/PCL, preparation process is the stick signal factor more simply more intactly.
(7) preparation method of the present invention has well solved the acid problem causing owing to alleviating PLLA degraded, aragonite Margarita powder wherein or ball aragonite Margarita powder and nHA degraded are alkalescence, alkalescence when Margarita powder and nHA dissolve can in and the acidic materials that produce of PLLA degraded, therefore can stablize the pH value of microenvironment, the growth and the propagation that are conducive to cell, the degradation time of prolongation bone repairing support.
The specific embodiment
The preparation method of the porous support of repairing for bone that the present invention proposes, comprises the following steps:
(1) Poly-L-lactic acid is dissolved in solvent 1-4-dioxane, the mass volume ratio of Poly-L-lactic acid and solvent 1-4-dioxane is: Poly-L-lactic acid: solvent 1-4-dioxane)=(0.03～0.08): 1, within 1～8 hour under magnetic agitation, dissolve, obtain the first solution;
(2) polycaprolactone is dissolved in solvent 1-4-dioxane, the mass volume ratio of polycaprolactone and solvent 1-4-dioxane is: polycaprolactone: solvent 1-4-dioxane=(0.03～0.08): 1, within 1～8 hour under magnetic agitation, dissolve, obtain the second solution;
(3) the first solution is mixed with the second solution, the volume ratio of mixing is: the first solution: the second solution=(0.8～1.2): 1, obtain the 3rd solution;
(4) at stirring, nano-hydroapatite particles is joined in the 3rd solution, obtain the 4th solution, in the 4th solution, the mass ratio of nano-hydroapatite particles, Poly-L-lactic acid and polycaprolactone is: nano-hydroapatite particles: Poly-L-lactic acid: polycaprolactone=1:(1～9): stirred 1～2 hour (1～9), with ultrasonic dispersion 1～2 hour;
(5) by aragonite or ball aragonite Margarita with alcohol-pickled 1～4 hour, after dry, pulverize after washing down with deionized water, obtain Margarita powder granule, the granularity of Margarita powder granule is less than 50 microns;
(6) at stirring, above-mentioned Margarita powder granule is joined in the 4th solution, obtain the 5th solution, in the 5th solution, the mass ratio of each composition is: Margarita powder granule: nano-hydroapatite particles: Poly-L-lactic acid: polycaprolactone=1:(1～2): (1～9): (1～9), stir 10～30 minutes;
(7) the 5th solution, is poured in bracket mould after 1～2 hour with ultrasonic dispersion, shaken up after 5～10 minutes, under-20 ℃ of environment freezing 10～24 hours;
(8) sample freezing is put into freezer dryer, lyophilization 48～75 hours, the bone repair porous compound rest that obtains being dried.
Below introduce the embodiment of the inventive method.
Embodiment 1: prepare the bone repairing support that a kind of composition is PLLA/PCL/ aragonite Margarita powder/nHA.
(1) PLLA of 0.4g (mean molecule quantity 179kDa) is added in the 1-4-dioxane of 8mL to magnetic agitation 8h, the 1-4-dioxane solution of formation 5% (w/v) PLLA.
(2) PCL of 0.4g (mean molecule quantity 8kDa) is added in the 1-4-dioxane of 8mL to magnetic agitation 1h, the 1-4-dioxane solution of formation 5% (w/v) PCL.
(3) PLLA, the PCL solution of getting respectively 4mL mix by 1:1.
(4), at stirring, the nHA granule of getting 0.0225g adds in mixed solution, ultrasonic dispersion 1h, and ultrasound intensity is 60W.
(5) with pulverizer, crushing pearl is sieved, obtain the aragonite Margarita powder that particle diameter is less than 50 microns.
(6) 0.0225g, particle diameter are less than to the aragonite Margarita powder of 50 microns and add in solution, continue to stir 0.5h.Then ultrasonic dispersion 1h, ultrasound intensity is 60W, makes uniform mixed solution.
(7) the 5th solution, is poured in bracket mould after 1 hour with ultrasonic dispersion, shaken up after 10 minutes, under-20 ℃ of environment freezing 24 hours;
(8) sample freezing is put into freezer dryer.Wherein, freezer dryer is at-60 ℃ of pre-cooling 0.5h.Lyophilization 72 hours, the bone repair porous compound rest that obtains being dried.
Embodiment 2: PLLA/PCL/ ball aragonite Margarita powder/nHA support of preparing a kind of composition.
(1) PLLA of 0.64g (mean molecule quantity 179kDa) is added in the 1-4-dioxane of 8mL to magnetic agitation 8h, the 1-4-dioxane solution of formation 8% (w/v) PLLA.
(2) PCL of 0.64g (mean molecule quantity 8kDa) is added in the 1-4-dioxane of 8mL to magnetic agitation 1h, the 1-4-dioxane solution of formation 8% (w/v) PCL.
(3) get respectively the PLLA of 3.55mL and the PCL solution of 4.45mL mixes by 0.8:1.
(4), at stirring, the nHA granule of getting 0.213g adds in mixed solution, ultrasonic dispersion 1h, and ultrasound intensity is 60W.
(5) with pulverizer, crushing pearl is sieved, obtain the aragonite Margarita powder that particle diameter is less than 50 microns.
(6) 0.213g, particle diameter are less than to the ball aragonite Margarita powder of 50 microns and add in solution, continue to stir 0.5h.Then ultrasonic dispersion 1h, ultrasound intensity is 60W, makes uniform mixed solution.
(7) the 5th solution, is poured in bracket mould after 1 hour with ultrasonic dispersion, shaken up after 10 minutes, under-20 ℃ of environment freezing 24 hours;
(8) sample freezing is put into freezer dryer.Wherein, freezer dryer is at-60 ℃ of pre-cooling 0.5h.Lyophilization 48 hours, the bone repair porous compound rest that obtains being dried.
Embodiment 3: preparing a kind of composition is the bone repairing support of PLLA/PCL/ aragonite Margarita powder/ball aragonite Margarita powder/nHA.
(1) PLLA of 0.32g (mean molecule quantity 179kDa) is added in the 1-4-dioxane of 8mL to magnetic agitation 8h, the 1-4-dioxane solution of formation 4% (w/v) PLLA.
(2) PCL of 0.32g (mean molecule quantity 8kDa) is added in the 1-4-dioxane of 8mL to magnetic agitation 1h, the 1-4-dioxane solution of formation 4% (w/v) PCL.
(3) PLLA, the PCL solution of getting respectively 4mL mix by 1:1.
(4), at stirring, the nHA granule of getting 0.16g adds in mixed solution, ultrasonic dispersion 1h, and ultrasound intensity is 60W.
(5) with pulverizer, crushing pearl is sieved, obtain aragonite or ball aragonite Margarita powder that particle diameter is less than 50 microns.
(6) 0.08g, particle diameter are less than to the ball aragonite Margarita powder that the aragonite Margarita powder of 50 microns and 0.08g, particle diameter be less than 50 microns and add in solution, continue to stir 0.5h.Then ultrasonic dispersion 1h, ultrasound intensity is 60W, makes uniform mixed solution.
(7) the 5th solution, is poured in bracket mould after 1 hour with ultrasonic dispersion, shaken up after 10 minutes, under-20 ℃ of environment freezing 24 hours;
(8) sample freezing is put into freezer dryer.Wherein, freezer dryer is at-60 ℃ of pre-cooling 0.5h.Lyophilization 72 hours, the bone repair porous compound rest that obtains being dried.
In the above embodiment of the present invention, the reaction raw materials using, can buy from market, for example, aragonite Margarita, the ball aragonite Margarita Ke Zhejiang Province Zhuji City pearl market used buy, nanometer hydroxyapatite Ke Haitai Nano Material Co., Ltd., Nanjing buys, and Poly-L-lactic acid and polycaprolactone Ke Shandong medical apparatus corporation, Ltd buy.
Claims (1)
1. a preparation method for the porous support of repairing for bone, is characterized in that this preparation method comprises the following steps:
(1) Poly-L-lactic acid is dissolved in solvent 1-4-dioxane, the mass volume ratio of Poly-L-lactic acid and solvent 1-4-dioxane is: Poly-L-lactic acid: solvent 1-4-dioxane)=(0.03～0.08): 1, within 1-8 hour under magnetic agitation, dissolve, obtain the first solution;
(2) polycaprolactone is dissolved in solvent 1-4-dioxane, the mass volume ratio of polycaprolactone and solvent 1-4-dioxane is: polycaprolactone: solvent 1-4-dioxane=(0.03～0.08): 1, within 1-8 hour under magnetic agitation, dissolve, obtain the second solution;
(3) the first solution is mixed with the second solution, the volume ratio of mixing is: the first solution: the second solution=(0.8～1.2): 1, obtain the 3rd solution;
(4) at stirring, nano-hydroapatite particles is joined in the 3rd solution, obtain the 4th solution, in the 4th solution, the mass ratio of nano-hydroapatite particles, Poly-L-lactic acid and polycaprolactone is: nano-hydroapatite particles: Poly-L-lactic acid: polycaprolactone=1:(1～9): stirred 1～2 hour (1～9), with ultrasonic dispersion 1～2 hour;
(5) by aragonite or ball aragonite Margarita with alcohol-pickled 1～4 hour, after dry, pulverize after washing down with deionized water, obtain Margarita powder granule, the granularity of Margarita powder granule is less than 50 microns;
(6) at stirring, above-mentioned Margarita powder granule is joined in the 4th solution, obtain the 5th solution, in the 5th solution, the mass ratio of each composition is: Margarita powder granule: nano-hydroapatite particles: Poly-L-lactic acid: polycaprolactone=1:(1～2): (1～9): (1～9), stir 10～30 minutes;
(7) the 5th solution, is poured in bracket mould after 1～2 hour with ultrasonic dispersion, shaken up after 5～10 minutes, under-20 ℃ of environment freezing 10～24 hours;
(8) sample freezing is put into freezer dryer, lyophilization 48～75 hours, the bone repair porous compound rest that obtains being dried.
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Application publication date: 20140910 |