CN103768658A - Hydroxyapatite-loading polylactic acid porous microsphere and preparation method thereof - Google Patents
Hydroxyapatite-loading polylactic acid porous microsphere and preparation method thereof Download PDFInfo
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- CN103768658A CN103768658A CN201210396694.6A CN201210396694A CN103768658A CN 103768658 A CN103768658 A CN 103768658A CN 201210396694 A CN201210396694 A CN 201210396694A CN 103768658 A CN103768658 A CN 103768658A
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Abstract
The invention provides a hydroxyapatite-loading polylactic acid composite porous microsphere which is characterized in that: the composite porous microsphere has an obvious pore appearance structure, the particle size is 50-500 mum, the pore diameter is 5-20 mum, the inside of the microsphere and external environment are communicated, and the water absorption rate is 900-3200%. The invention also provides a preparation method of the hydroxyapatite-loading polylactic acid composite porous microsphere. The prepared hydroxyapatite-loading polylactic acid composite porous microsphere can be used as an adult cell growth scaffold, also can be used as an attachment scaffold for stem cell growth and differentiation, and can be used in tissue engineering. The preparation method of the composite porous microsphere is simple and convenient in operation, low in cost, safe in use, and is favorable for market promotion.
Description
Technical field
The present invention relates to a kind of porous microsphere and preparation method thereof, particularly relate to porous polylactic acid microball of a kind of load hydroxyapatite and preparation method thereof, belong to field of tissue engineering technology.
Background technology
Porous polymer bead, refers to that material is made up of macromolecular material, and particle size range is between 1-1000 micron, and surface has the microsphere of all vesicular textures.Can be used at present the fields such as medicament slow release, cytoskeleton, cell injection.
The macromolecular material of preparing at present porous microsphere mainly contains polylactic acid, the polyesters such as blend or the lactic acid-caprolactone copolymer macromolecular material of polylactic acid and polycaprolactone.Above-mentioned material has good biological degradability and biological safety and is widely used as tissue engineering bracket and pharmaceutical carrier field (Lin Sun, 2009; Hyun Jung Chung, 2006; Stefano Giovagnoli, 2007).But prepared porous microsphere also exists variety of issue, main cause is because catabolite is acid, and pH value reduces and easily causes inflammation.
The light base apatite of inorganic material, is widely used as organizational project because having good biocompatibility, the particularly relevant research of orthopaedics.Hydroxyapatite discharges weakly alkaline ion in aqueous solution, just in time can be used for neutralizing the acid problem of above-mentioned Degradation of Polymer Materials product, can also improve in addition the biological activity of material.
Jointly less as carrier report about polylactic acid and hydroxyapatite at present, both compound porous microspheres of preparing be there is not yet to report at home and abroad.
Summary of the invention
One of the object of the invention is to provide a kind of polylactic acid composite porous microspheres of load hydroxyapatite, can be used as the support of cell adhesion in organizational project.
A polylactic acid composite porous microspheres for load hydroxyapatite, is characterized in that, the outward appearance of composite porous microspheres has obvious cavernous structure, its particle diameter is 50～500 μ m,, aperture is 5～20 μ m, be connected with external environment inner microsphere, its water absorption rate is 900～3200%.
The present invention also provides a kind of preparation method of polylactic acid composite porous microspheres of load hydroxyapatite, it is characterized in that comprising the steps:
Macromolecular material and hydroxyapatite are dissolved in organic solvent simultaneously, and heating condensing reflux dissolve, and its macromolecular material and hydroxyapatite are dispersed in organic solvent, form organic facies;
(2) ammonium bicarbonate is dissolved in the water, and normal-temperature dissolution forms water I;
(3) polyethylene alcoholysis, in water, when high-temperature digestion, forms water II;
(4) take out above-mentioned organic facies and be placed in dry small beaker, add above-mentioned water I, carry out subsequently homogenized, form emulsion;
(5) at once emulsion even speed is added drop-wise in water II, magnetic stir bar after 4 hours, by centrifugal microsphere 5 minutes, then is used ultra-pure water Eddy diffusion with stirring at low speed, repeatedly washes away unnecessary PVA 3 times, then by microsphere lyophilization processing.
Described macromolecular material is the blend of polylactic acid and polycaprolactone, or lactic acid-caprolactone copolymer.
Described macromolecular material is 2～8g/100ml at the final concentration of organic facies, and described hydroxyapatite is 2～4g/100ml at the final concentration of organic facies.
The final concentration that described ammonium bicarbonate is dissolved in water I is 5～20 g/100ml.
Described polyvinyl alcohol is 0.1～1 g/100ml at the final concentration of water II.
The volume ratio of described water I and organic facies is 3:1～5:1.
Polylactic acid composite porous microspheres of load hydroxyapatite disclosed by the invention and preparation method thereof, can be used as the support of cell adhesion in organizational project.The polylactic acid composite porous microspheres particle diameter of prepared load hydroxyapatite is 50-500 μ m, its outward appearance has obvious cavernous structure, and aperture is 5-20 μ m, is connected with external environment inner microsphere, increase its hydrophilic by load hydroxyapatite, water absorption rate is 900-3200%.Prepared composite porous microspheres, for the support of adult Growth of Cells, also can be used as the support that stem cell growth and differentiation phase adhere to, and is applied to organizational project.In the present invention, the preparation method of composite porous microspheres, simple to operation, with low cost, uses safety, is conducive to marketing.
accompanying drawing explanation:
Fig. 1 is the schematic diagram of the polylactic acid composite porous microspheres of load hydroxyapatite of the present invention.
Wherein 1 polylactic acid component, 2 is hydroxyapatite component, 3 is cavernous structure.
Fig. 2 is the schematic diagram scanning electron microscope diagram of the polylactic acid composite porous microspheres of load hydroxyapatite of the present invention.
The specific embodiment
Embodiment 1:
Take the polylactic acid of 200 mg, the hydroxyapatite of 200mg is dissolved in 5ml dichloromethane, and heating condensing reflux dissolve 1 hour, and formation polylactic acid and hydroxyapatite are dispersed in the organic facies of dichloromethane.Take 10g ammonium bicarbonate and be dissolved in 100ml water, normal-temperature dissolution 1 hour, forms water I.Take the alcoholysis of 2g polyethylene in 200ml water, high-temperature digestion 1 hour, forms water II.Take out above-mentioned organic facies 4ml and be placed in dry small beaker, add above-mentioned 1ml water I, carry out subsequently homogenized, form emulsion.At once emulsion even speed is added drop-wise in the water II of 200ml.3cm magnetic stir bar stirs after 4 hours with the speed of 200rpm, by the centrifugal 5min of speed of microsphere 50 rpm, then uses the ultra-pure water Eddy diffusion of 200ml, 3 times repeatedly.By microsphere lyophilization processing, both obtained the load hydroxyapatite/polylactic acid porous microsphere of white appearance.
Through sem observation, the mean diameter of hydroxyapatite/polylactic acid porous microsphere is 201.4 ± 51.6 μ m, and its outward appearance has obvious cavernous structure, is connected with external environment inner microsphere, and the average diameter of cavernous structure is 23.4 ± 5.2 μ m.Water absorption rate is 2045.7%.
Embodiment 2:
Take the polylactic acid of 200 mg, the hydroxyapatite of 400mg is dissolved in 5ml dichloromethane, and heating condensing reflux dissolve 1 hour, and formation polylactic acid and hydroxyapatite are dispersed in the organic facies of dichloromethane.Take 10g ammonium bicarbonate and be dissolved in 100ml water, normal-temperature dissolution 1 hour, forms water I.Take the alcoholysis of 2g polyethylene in 200ml water, high-temperature digestion 1 hour, forms water II.Take out above-mentioned organic facies 4ml and be placed in dry small beaker, add above-mentioned 1ml water I, carry out subsequently homogenized, form emulsion.At once emulsion even speed is added drop-wise in the water II of 200ml.3cm magnetic stir bar stirs after 4 hours with the speed of 200rpm, by the centrifugal 5min of speed of microsphere 50 rpm, then uses the ultra-pure water Eddy diffusion of 200ml, 3 times repeatedly.By microsphere lyophilization processing, both obtained the load hydroxyapatite/polylactic acid porous microsphere of white appearance.Through sem observation, the mean diameter of hydroxyapatite/polylactic acid porous microsphere is 164.5 ± 31.1 μ m, and its outward appearance has obvious cavernous structure, is connected with external environment inner microsphere, and the average diameter of cavernous structure is 11.5 ± 3.1 μ m.Water absorption rate is 1743.0%.
Embodiment 3:
Take the polylactic acid of 400 mg, the hydroxyapatite of 200mg is dissolved in 5ml dichloromethane, and heating condensing reflux dissolve 1 hour, and formation polylactic acid and hydroxyapatite are dispersed in the organic facies of dichloromethane.Take 5g ammonium bicarbonate and be dissolved in 100ml water, normal-temperature dissolution 1 hour, forms water I.Take the alcoholysis of 2g polyethylene in 200ml water, high-temperature digestion 1 hour, forms water II.Take out above-mentioned organic facies 4ml and be placed in dry small beaker, add above-mentioned 1ml water I, carry out subsequently homogenized, form emulsion.At once emulsion even speed is added drop-wise in the water II of 200ml.3cm magnetic stir bar stirs after 4 hours with the speed of 200rpm, by the centrifugal 5min of speed of microsphere 50 rpm, then uses the ultra-pure water Eddy diffusion of 200ml, 3 times repeatedly.By microsphere lyophilization processing, both obtained the load hydroxyapatite/polylactic acid porous microsphere of white appearance.
Through sem observation, the mean diameter of hydroxyapatite/polylactic acid porous microsphere is 274.2 ± 52.1 μ m, and its outward appearance has obvious cavernous structure, is connected with external environment inner microsphere, and the average diameter of cavernous structure is 14.3 ± 3.1 μ m.Water absorption rate is 913.9%.
Take the polylactic acid of 200 mg, the hydroxyapatite of 200mg is dissolved in 5ml dichloromethane, and heating condensing reflux dissolve 1 hour, and formation polylactic acid and hydroxyapatite are dispersed in the organic facies of dichloromethane.Take 20g ammonium bicarbonate and be dissolved in 100ml water, normal-temperature dissolution 1 hour, forms water I.Take the alcoholysis of 2g polyethylene in 200ml water, high-temperature digestion 1 hour, forms water II.Take out above-mentioned organic facies 4ml and be placed in dry small beaker, add above-mentioned 1ml water I, carry out subsequently homogenized, form emulsion.At once emulsion even speed is added drop-wise in the water II of 200ml.3cm magnetic stir bar stirs after 4 hours with the speed of 200rpm, by the centrifugal 5min of speed of microsphere 50 rpm, then uses the ultra-pure water Eddy diffusion of 200ml, 3 times repeatedly.By microsphere lyophilization processing, both obtained the load hydroxyapatite/polylactic acid porous microsphere of white appearance.
Through sem observation, the mean diameter of hydroxyapatite/polylactic acid porous microsphere is 300.2 ± 25.7 μ m, and its outward appearance has obvious cavernous structure, is connected with external environment inner microsphere, and the average diameter of cavernous structure is 21.5 ± 5.2 μ m.Water absorption rate is 2532.2%.
Embodiment 5:
Take the polylactic acid of 200 mg, the hydroxyapatite of 200mg is dissolved in 5ml dichloromethane, and heating condensing reflux dissolve 1 hour, and formation polylactic acid and hydroxyapatite are dispersed in the organic facies of dichloromethane.Take 10g ammonium bicarbonate and be dissolved in 100ml water, normal-temperature dissolution 1 hour, forms water I.Take the alcoholysis of 0.2g polyethylene in 200ml water, high-temperature digestion 1 hour, forms water II.Take out above-mentioned organic facies 4ml and be placed in dry small beaker, add above-mentioned 1ml water I, carry out subsequently homogenized, form emulsion.At once emulsion even speed is added drop-wise in the water II of 200ml.3cm magnetic stir bar stirs after 4 hours with the speed of 200rpm, by the centrifugal 5min of speed of microsphere 50 rpm, then uses the ultra-pure water Eddy diffusion of 200ml, 3 times repeatedly.By microsphere lyophilization processing, both obtained the load hydroxyapatite/polylactic acid porous microsphere of white appearance.
Through sem observation, the mean diameter of hydroxyapatite/polylactic acid porous microsphere is 321.7 ± 16.4 μ m, and its outward appearance has obvious cavernous structure, is connected with external environment inner microsphere, and the average diameter of cavernous structure is 35.3 ± 5.4 μ m.Water absorption rate is 3190.5%.
Embodiment 6:
Take the polylactic acid of 200 mg, the hydroxyapatite of 200mg is dissolved in 5ml dichloromethane, and heating condensing reflux dissolve 1 hour, and formation polylactic acid and hydroxyapatite are dispersed in the organic facies of dichloromethane.Take 10g ammonium bicarbonate and be dissolved in 100ml water, normal-temperature dissolution 1 hour, forms water I.Take the alcoholysis of 2g polyethylene in 200ml water, high-temperature digestion 1 hour, forms water II.Take out above-mentioned organic facies 4ml and be placed in dry small beaker, add above-mentioned 1ml water I, carry out subsequently homogenized, form emulsion.At once emulsion even speed is added drop-wise in the water II of 200ml.3cm magnetic stir bar stirs after 4 hours with the speed of 100rpm, by the centrifugal 5min of speed of microsphere 50 rpm, then uses the ultra-pure water Eddy diffusion of 200ml, 3 times repeatedly.By microsphere lyophilization processing, both obtained the load hydroxyapatite/polylactic acid porous microsphere of white appearance.
Through sem observation, the mean diameter of hydroxyapatite/polylactic acid porous microsphere is 241.9 ± 13.8 μ m, and its outward appearance has obvious cavernous structure, is connected with external environment inner microsphere, and the average diameter of cavernous structure is 32.1 ± 6.6 μ m.Water absorption rate is 1969.1%.
Embodiment 7:
Take the polylactic acid of 200 mg, the hydroxyapatite of 200mg is dissolved in 5ml dichloromethane, and heating condensing reflux dissolve 1 hour, and formation polylactic acid and hydroxyapatite are dispersed in the organic facies of dichloromethane.Take 10g ammonium bicarbonate and be dissolved in 100ml water, normal-temperature dissolution 1 hour, forms water I.Take the alcoholysis of 2g polyethylene in 200ml water, high-temperature digestion 1 hour, forms water II.Take out above-mentioned organic facies 4ml and be placed in dry small beaker, add above-mentioned 1ml water I, carry out subsequently homogenized, form emulsion.At once emulsion even speed is added drop-wise in the water II of 200ml.3cm magnetic stir bar stirs after 4 hours with the speed of 300rpm, by the centrifugal 5min of speed of microsphere 50 rpm, then uses the ultra-pure water Eddy diffusion of 200ml, 3 times repeatedly.By microsphere lyophilization processing, both obtained the load hydroxyapatite/polylactic acid porous microsphere of white appearance.
Through sem observation, the mean diameter of hydroxyapatite/polylactic acid porous microsphere is 149.2 ± 8.1 μ m, and its outward appearance has obvious cavernous structure, is connected with external environment inner microsphere, and the average diameter of cavernous structure is 12.2 ± 4.1 μ m.Water absorption rate is 2285.2%.
Embodiment 8:
Take the polylactic acid of 200 mg, the hydroxyapatite of 200mg is dissolved in 5ml dichloromethane, and heating condensing reflux dissolve 1 hour, and formation polylactic acid and hydroxyapatite are dispersed in the organic facies of dichloromethane.Take 10g ammonium bicarbonate and be dissolved in 100ml water, normal-temperature dissolution 1 hour, forms water I.Take the alcoholysis of 2g polyethylene in 200ml water, high-temperature digestion 1 hour, forms water II.Take out above-mentioned organic facies 4ml and be placed in dry small beaker, add above-mentioned 1ml water I, carry out subsequently homogenized, form emulsion.At once emulsion even speed is added drop-wise in the water II of 200ml.3cm magnetic stir bar stirs after 4 hours with the speed of 400rpm, by the centrifugal 5min of speed of microsphere 50 rpm, then uses the ultra-pure water Eddy diffusion of 200ml, 3 times repeatedly.By microsphere lyophilization processing, both obtained the load hydroxyapatite/polylactic acid porous microsphere of white appearance.
Through sem observation, the mean diameter of hydroxyapatite/polylactic acid porous microsphere is 111.7 ± 6.9 μ m, and its outward appearance has obvious cavernous structure, is connected with external environment inner microsphere, and the average diameter of cavernous structure is 13.8 ± 8.1 μ m.Water absorption rate is 1522.9%.
Claims (7)
1. a polylactic acid composite porous microspheres for load hydroxyapatite, is characterized in that, the outward appearance of composite porous microspheres has obvious cavernous structure, its particle diameter is 50～500 μ m,, aperture is 5～20 μ m, be connected with external environment inner microsphere, its water absorption rate is 900～3200%.
2. a kind of preparation method of polylactic acid composite porous microspheres of load hydroxyapatite according to claim 1, is characterized in that comprising the steps:
(1) macromolecular material and hydroxyapatite are dissolved in organic solvent simultaneously, heating condensing reflux dissolve, and its macromolecular material and hydroxyapatite are dispersed in organic solvent, form organic facies;
(2) ammonium bicarbonate is dissolved in the water, and normal-temperature dissolution forms water I;
(3) polyvinyl alcohol is dissolved in the water, and when high-temperature digestion, forms water II;
(4) take out above-mentioned organic facies and be placed in dry small beaker, add above-mentioned water I, carry out subsequently homogenized, form emulsion;
(5) at once emulsion even speed is added drop-wise in water II, magnetic stir bar after 4 hours, by centrifugal microsphere 5 minutes, then is used ultra-pure water Eddy diffusion with stirring at low speed, repeatedly washes away unnecessary PVA 3 times, then by microsphere lyophilization processing.
3. a kind of preparation method of polylactic acid composite porous microspheres of load hydroxyapatite according to claim 2, is characterized in that, described macromolecular material is the blend of polylactic acid and polycaprolactone, or lactic acid-caprolactone copolymer.
4. a kind of preparation method of polylactic acid composite porous microspheres of load hydroxyapatite according to claim 2, it is characterized in that, described macromolecular material is 2～8g/100ml at the final concentration of organic facies, and described hydroxyapatite is 2～4g/100ml at the final concentration of organic facies.
5. a kind of preparation method of polylactic acid composite porous microspheres of load hydroxyapatite according to claim 2, is characterized in that, the final concentration that described ammonium bicarbonate is dissolved in water I is 5～20 g/100ml.
6. a kind of preparation method of polylactic acid composite porous microspheres of load hydroxyapatite according to claim 2, is characterized in that, described polyvinyl alcohol is 0.1～1 g/100ml at the final concentration of water II.
7. a kind of preparation method of polylactic acid composite porous microspheres of load hydroxyapatite according to claim 2, is characterized in that, the volume ratio of described water I and organic facies is 3:1～5:1.
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