CN103157143A - Bone cement porous support with inner surface provided with stamen-type laminated-sheet-shaped micro-nano topology shape and preparation method thereof - Google Patents
Bone cement porous support with inner surface provided with stamen-type laminated-sheet-shaped micro-nano topology shape and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a bone cement porous support with an inner surface provided with a stamen-type laminated-sheet-shaped micro-nano topology shape and a preparation method of the bone cement porous support with the inner surface provided with the stamen-type laminated-sheet-shaped micro-nano topology shape. The interior of the bone cement porous support is in an irregular porous structure. A hole diameter is 100-1000 micrometers. The porosity is 5-95%. Holes are mutually communicated. An inner surface of each hole is an unsmooth surface, and grains with diameters of 200-800 nanometers are continuously distributed on the inner surface of each hole, wherein protrusion structures of a micrometer order are discontinuously distributed on the inner surface of each hole, a diameter of each protrusion structure is 2-8 micrometers, and a height of each protrusion structure is 1-5 micrometers. The bone cement porous support with a special shape and prepared with the method can promote the adhesion of cells on the inner surface of the support, and other behaviors of the cells and repair for defect parts are further regulated and controlled, and the bone cement porous support with the inner surface provided with the stamen-type laminated-sheet-shaped micro-nano topology shape and the preparation method of the bone cement porous support with the inner surface provided with the stamen-type laminated-sheet-shaped micro-nano topology shape can be used for repair of sclerosis tissues, in particular to repair of bone tissues.
Description
Technical field
The present invention relates to a kind of bone cement porous support and preparation method thereof, particularly a kind of inner surface has bone cement porous support of the stacked lamellar micron-nano topological of pistil type pattern and preparation method thereof.
Background technology
Utilize tissue engineering to repair the damaged three elements of osseous tissue and be seed cell, signal factor and porous support materials.Wherein, porous support materials can provide three-dimensional space environment that cell and blood vessel grow into and the transmission channel of matter transportation, and for the generation of cambium provides physical support, desirable porous support materials also should have good biocompatibility, biological degradability, cell-material and stick interface and certain characteristics such as mechanical strength.Calcium phosphate cement bracket has voluntarily and solidifies, facilitates that figuration, osteoinductive and bone conductibility are good, the mechanical strength advantages of higher, although enbrittle large shortcoming, still becomes one of important branch of bone tissue engineer area research, and wide market prospect is arranged.
According to the literature, the nanometer of two-dimensional surface and micron topological structure have important regulating action [Kulangara K and Leong KW, 2009, Soft Matter to cell adhesion and cell differentiation; Geblinger D, et al, 2010, Journal of Cell Science; Pan Z, et al, 2012, Biomaterials.].Smooth surface with surface ratio same material of suitable micron, nanoscale topological structure more is conducive to the tactophily of cell and the regulation and control of cell differentiation direction.Utilize the porous support of Biodegradable polymer material preparation, its inner surface is often relatively smooth, is not easy to carry out the modification of topological structure.And the surface after calcium phosphate bone cement curing generally forms micron or nano level topological structure voluntarily, common are the structures such as lamellar, pit, projection.
(biphasic calcium phosphate (BCP) of β-TCP/HA) form comes across nineteen twenty (thinking at that time tricalcium phosphate) the earliest by bata-tricalcium phosphate/hydroxyapatite, when having the good biocompatibility of general synthos, also combine the advantage that the hydroxyapatite osteoinductive is good, mechanical strength is high and the bata-tricalcium phosphate degraded is fast, bone resorption is good.β-TCP and HA ratio also can be regulated, and the BCP pottery of proper ratio and pore structure can have osteoinductive.Therefore, BCP by large quantity research, has entered clinical in early 1980s in dentistry and medicinal application with the form of BCP pottery in the application of the aspects such as hard tissue repair, displacement.[LeGeros?RZ,?et?a1,?Journal?of?Materials?Science:?Materials?in?Medicine,?2003;?Kurashina?K,?et?al,?Biomaterials,?2002;?Gerber?T,?et?al,?Journal?of?Sol-Gel?Science?and?Technology,?2003.?]
In existing document and patent report about the bone cement porous support, the inner surface in hole mostly is the topological structure of particle packing, or there is no obvious height fluctuating, and the tactophily of cell is better than common high score subclass tissue engineering bracket.The present invention is directed to above background, a kind of Bone Colonizable Cements In Rabbits porous support with topology of multistage yardstick is provided, further improve the tactophily of cell.
Summary of the invention
One of purpose of the present invention is to provide a kind of inner surface to have the bone cement porous support of the stacked lamellar micron-nano topological of pistil type pattern, to improve cell sticking at the support inner surface, and then other behaviors of regulating cell and promote tissue repair, its inner surface is distributed with the stacked lamellar projection of pistil type of discontinuous micro nano structure, with the grain structure of piling up continuously in the background area.
Two of purpose of the present invention is to provide the preparation method of this porous support.
For achieving the above object, the design that the present invention adopts is: bata-tricalcium phosphate/hydroxyapatite biphasic calcium phosphate mixes calcium carbonate as foaming agent as the primary raw material of bone cement powder by physical mixed; Take the aqueous citric acid solution system as consolidation liquid, add chitosan as reinforcing agent.Bone cement powder and consolidation liquid are reconciled into the thickness paste according to certain liquid-solid ratio, and reaction generates the carbon dioxide pore of foaming, and injects mould and carries out plastotype, wait solidifying the rear demoulding.Then maintenance at a certain temperature and humidity conditions, basicly stable to intensity, again through polishing processing, remove the part that directly contacts with mould, the linked hole of internal stent is exposed, finally make the bone cement porous support materials that inner surface has the complex topology pattern, inner surface has the grain structure of the stacked laminated structure of discontinuous micro-nano pistil type and continuous hundreds of nanometers size.
According to above-mentioned design, the present invention adopts following technical scheme:
A kind of inner surface has the bone cement porous support of the stacked lamellar micron-nano topological of pistil type pattern, the inside that it is characterized in that this bone cement porous support is irregular loose structure, the aperture is that 100～1000 μ m, porosity are 5～95%, mutual UNICOM between Kong Yukong, the inner surface in hole is rough surface, and it is the granule of 200～800 nm that continuous distribution size, wherein the discontinuous micron-sized bulge-structure that distributing, the size of this bulge-structure is 2～8 μ m, is highly 1～5 μ m.
Above-mentioned micron order bulge-structure is the micron and nanometer composite structure of the stacked lamellar of pistil type, and the thickness of its petal-shaped monolithic is less than 10 nm.
A kind ofly prepare the preparation method that above-mentioned inner surface has the bone cement porous support of the stacked lamellar micron-nano topological of pistil type pattern, it is characterized in that the concrete steps of the method are:
A. mix calcium carbonate in the powder of bata-tricalcium phosphate/hydroxyapatite biphasic calcium phosphate, as the bone cement powder powder stock; The quality of calcium carbonate is the 0.5-50% of the quality of biphasic calcium phosphate.
B. preparing mass percent concentration is the Fructus Citri Limoniae aqueous acid of 0.05-50%, and adds chitosan, stirs and makes its dissolving, obtains the acid consolidation liquid of pH＜4.0, and to make the mass percent concentration of chitosan be 0.05～3%.
C. step a gained bone cement powder is mixed according to the liquid-solid ratio of 0.2～1.0 ml:1g with subsidy b gained consolidation liquid, reconcile into the thickness paste, the injection molding plastotype solidifies the rear demoulding until bone cement;
D. the bone cement after the demoulding is processed through maintenance, polishing, finally made the bone cement porous support that inner surface has the stacked lamellar micron-nano topological of pistil type pattern.
The size of the biphasic calcium phosphate described in above-mentioned step a is 150 nm～1 μ m, and wherein the crystalline phase ratio of bata-tricalcium phosphate and hydroxyapatite is at 99:1～30:70.
The particle diameter of above-mentioned calcium carbonate is no more than 5 μ m.
The technological parameter of the above-mentioned described maintenance processes of steps d is: 15～50 ℃ of temperature, damp condition are relative humidity 5～80%, and curing time is more than 18 h.
Polishing mode described in above-mentioned steps d is the wet method polishing of mechanical grinding, sand paper dry grinding or non-aqueous media.。
The present invention with bata-tricalcium phosphate/hydroxyapatite biphasic calcium phosphate as the bone cement pressed powder, with the calcium carbonate pressed powder as foaming agent, as consolidation liquid, has the bone cement porous support of the stacked lamellar micron-nano topological of pistil type pattern with citric acid/chitosan aqueous solution through the chemical blowing preparation.The bone cement porous support of this method preparation has suitable aperture (150～1000 μ m), porosity (5～95%) and comprcssive strength (reaching as high as 40 MPa); Its inner surface has the micro-nano projection of the discontinuously arranged stacked laminated structure of pistil type, and the background area is piled up continuously by the granule of hundreds of nanometers and formed.The projection of pistil type is 1～20 μ m, high 0.5～15 μ m, and its petal-shaped single-sheet thickness is less than 10 nm, and granular size is 150～1000 nm.The size of pistil type projection and distribution density can be regulated by changing the parameters such as calcium carbonate content, consolidation liquid pH value, liquid-solid ratio; The background area particle size is mainly determined by the initial particle size of bone cement powder.
The bone cement porous support with special appearance of this method preparation can promote cell in the sticking of support inner surface, and then other behaviors of regulating cell and to the reparation of defect, can be used for the especially reparation of osseous tissue of sclerous tissues.
Description of drawings
Fig. 1 is the irregular loose structure that the prepared inner surface of the embodiment of the present invention one has the bone cement porous support of the micro-nano pattern of the stacked lamellar of pistil type, and aperture size is 100～500 μ m, mutual UNICOM between micropore and micropore.
Fig. 2 is the surface topology that the prepared inner surface of the embodiment of the present invention one has the bone cement porous support of the micro-nano pattern of the stacked lamellar of pistil type, and the micropore inner surface is rough surface.
Fig. 3 is the enlarged drawing of Fig. 2, shows the structure chart of micron order projection discontinuously arranged on the micropore inner surface, and its size is 2-8 μ m, is highly 1-5 μ m.
The prepared inner surface of Fig. 4 embodiment of the present invention one has the protruding fine structure at nanoscale of micron of the bone cement porous support of the micro-nano pattern of the stacked lamellar of pistil type, is the stacked laminated structure of pistil type, and the thickness of its petal-shaped monolithic is less than 10 nm.
Fig. 5 be the micron projection of the bone cement porous support of the prepared inner surface of inventive embodiments one with the micro-nano pattern of the stacked lamellar of pistil type at the fine structure of nanoscale, be the stacked laminated structure of pistil type, the thickness of its petal-shaped monolithic is less than 10 nm.
Fig. 6 is the enlarged drawing of Fig. 3.Show that the prepared inner surface of inventive embodiments one has the appearance structure on the rough surface in the micropore of bone cement porous support of the micro-nano pattern of the stacked lamellar of pistil type, consisted of by the granule of 200-800 nm size.
The specific embodiment
Following examples are implemented as prerequisite take the invention technical scheme, provided detailed embodiment and concrete operating process, but protection scope of the present invention are not limited to following embodiment.
Embodiment one: choose bata-tricalcium phosphate/hydroxyapatite that particle size range is 200-800 nm (65:35) biphasic calcium phosphate powder, take 9.4 g, add calcium carbonate 0.6 g(mass fraction 6.0%), take dehydrated alcohol as medium, through ball milling, centrifugal, dry, smash to pieces after, obtain the bone cement powder.Take respectively monohydrate potassium, the some grams of chitosan, make it be dissolved in water by mechanical agitation, make the consolidation liquid of pH=2.0.The bone cement powder is mixed in the ratio of 0.5 ml/g with consolidation liquid, reconcile into the paste of thickness, inject cylindrical stainless steel mould, an end sealing, an end is open, makes the bone cement free wxpansion in foaming under room temperature condition, solidifies the rear demoulding.After maintenance 48 h, with the sand papering surface, linked hole is communicated with atmosphere under room temperature environment.The gained inner surface has the scanning electron microscope picture of bone cement porous support of the stacked lamellar micron-nano topological of pistil type pattern as shown in Fig. 1-6.This cement bracket has linked hole, and pore size is 200-600 μ m, and referring to Fig. 1, porosity is 58%, and mean compressive strength is 18.8 MPa.The micropore inner surface is distributed with the micron order projection, and its size is 2-8 μ m, is highly 1-5 μ m, and the distribution density scope is 1 ~ 2/100 μ m
2(Fig. 2 and Fig. 3).This projection is the micron and nanometer composite structure of the stacked lamellar of pistil type, and the petal-shaped single-sheet thickness is less than 10 nm(Fig. 4 and Fig. 5); The background area of micron projection consists of (Fig. 6) by the granule of 200-800 nm size.The cell culture result shows, cell adhesion is all right, and a little higher than bore area of cell proliferation speed is the bone cement porous support of simple particle packing.
Embodiment two: choose bata-tricalcium phosphate/hydroxyapatite that particle size range is 200-600 nm (63:37) biphasic calcium phosphate powder, take 9.5 g, add calcium carbonate 0.5 g(mass fraction 5.0%), take dehydrated alcohol as medium, through ball milling, centrifugal, dry, smash to pieces after, obtain the bone cement powder.Take respectively monohydrate potassium, the some grams of chitosan, make it be dissolved in water by mechanical agitation, make the consolidation liquid of pH=2.0.The bone cement powder is mixed in the ratio of 0.5 ml/g with consolidation liquid, reconcile into the paste of thickness, inject cylindrical stainless steel mould, an end sealing, an end is open, makes the bone cement free wxpansion in foaming under room temperature condition, solidifies the rear demoulding.After maintenance 48 h, with the sand papering surface, linked hole is communicated with atmosphere under room temperature environment.The internal surface shape of gained bone cement porous support is similar to Fig. 1-6.This cement bracket has linked hole, and pore size is 150-550 μ m, and porosity is 53%, and mean compressive strength is 24.5 MPa.The micropore inner surface is distributed with the micron order projection, and its size is 2-7 μ m, is highly 1-4 μ m, and the distribution density scope is 1 ~ 2/100 μ m
2This projection is the micron and nanometer composite structure of the stacked lamellar of pistil type, and the petal-shaped single-sheet thickness is less than 10 nm; The background area of micron projection is made of the granule of 200-600 nm size.The cell culture result shows, cell adhesion is all right, and cell proliferation speed is high.
Embodiment three: choose bata-tricalcium phosphate/hydroxyapatite that particle size range is 200-700 nm (55:45) biphasic calcium phosphate powder, take 9.38 g, add calcium carbonate 0.62 g(mass fraction 6.2%), take dehydrated alcohol as medium, through ball milling, centrifugal, dry, smash to pieces after, obtain the bone cement powder.Take respectively monohydrate potassium, the some grams of chitosan, make it be dissolved in water by mechanical agitation, make the consolidation liquid of pH=2.0.The bone cement powder is mixed in the ratio of 0.5 ml/g with consolidation liquid, reconcile into the paste of thickness, inject cylindrical stainless steel mould, an end sealing, an end is open, makes the bone cement free wxpansion in foaming under room temperature condition, solidifies the rear demoulding.After maintenance 48 h, with the sand papering surface, linked hole is communicated with atmosphere under room temperature environment.The internal surface shape of gained bone cement porous support is similar to Fig. 1-6.This cement bracket has linked hole, and pore size is 250-600 μ m, and porosity is 65%, and mean compressive strength is 16.5 MPa.The micropore inner surface is distributed with the micron order projection, and its size is 2-7 μ m, is highly 1-4 μ m, and the distribution density scope is 1 ~ 2/100 μ m
2This projection is the micron and nanometer composite structure of the stacked lamellar of pistil type, and the petal-shaped single-sheet thickness is less than 10 nm; The background area of micron projection is made of the granule of 200-700 nm size.The cell culture result shows, cell adhesion is all right, and cell proliferation speed is slightly high.
Embodiment four: choose bata-tricalcium phosphate/hydroxyapatite that particle size range is 150-700 nm (60:40) biphasic calcium phosphate powder, take 9.4 g, add calcium carbonate 0.6 g(mass fraction 6%), take dehydrated alcohol as medium, through ball milling, centrifugal, dry, smash to pieces after, obtain the bone cement powder.Take respectively monohydrate potassium, the some grams of chitosan, make it be dissolved in water by mechanical agitation, make the consolidation liquid of pH=2.0.The bone cement powder is mixed in the ratio of 0.5 ml/g with consolidation liquid, reconcile into the paste of thickness, inject cylindrical stainless steel mould, an end sealing, an end is open, makes the bone cement free wxpansion in foaming under room temperature condition, solidifies the rear demoulding.After maintenance 48 h, with the sand papering surface, linked hole is communicated with atmosphere under room temperature environment.The internal surface shape of gained bone cement porous support is similar to Fig. 1-6.This cement bracket has linked hole, and pore size is 250-600 μ m, and porosity is 60%, and mean compressive strength is 18.3 MPa.The micropore inner surface is distributed with the micron order projection, and its size is 2-7 μ m, is highly 1-4 μ m, and the distribution density scope is 1 ~ 2/100 μ m
2This projection is the micron and nanometer composite structure of the stacked lamellar of pistil type, and the petal-shaped single-sheet thickness is less than 10 nm; The background area of micron projection is made of the granule of 150-700 nm size.The cell culture result shows, cell adhesion is all right, and cell proliferation speed is slightly high.
Embodiment five:
Choose bata-tricalcium phosphate/hydroxyapatite that particle size range is 200-1000 nm (60:40) biphasic calcium phosphate powder, take 9.4 g, add calcium carbonate 0.6 g(mass fraction 8%), take dehydrated alcohol as medium, through ball milling, centrifugal, dry, smash to pieces after, obtain the bone cement powder.Take respectively monohydrate potassium, the some grams of chitosan, make it be dissolved in water by mechanical agitation, make the consolidation liquid of pH=2.0.The bone cement powder is mixed in the ratio of 0.5 ml/g with consolidation liquid, reconcile into the paste of thickness, inject cylindrical stainless steel mould, an end sealing, an end is open, makes the bone cement free wxpansion in foaming under room temperature condition, solidifies the rear demoulding.After maintenance 48 h, with the sand papering surface, linked hole is communicated with atmosphere under room temperature environment.The internal surface shape of gained bone cement porous support is similar to Fig. 1-6.This cement bracket has linked hole, and pore size is 300-700 μ m, and porosity is 68%, and mean compressive strength is 17.1 MPa.The micropore inner surface is distributed with the micron order projection, and its size is 3-10 μ m, is highly 1.5-5 μ m, and distribution density is about 1/100 μ m 2 This projection is the micron and nanometer composite structure of the stacked lamellar of pistil type, and the petal-shaped single-sheet thickness is less than 10 nm; The background area of micron projection is made of the granule of 200-1000 nm size.The cell culture result shows, cell adhesion is all right, and cell proliferation speed is slightly high
Claims (7)
1. an inner surface has the bone cement porous support of the stacked lamellar micron-nano topological of pistil type pattern, the inside that it is characterized in that this bone cement porous support is irregular loose structure, the aperture is that 100～1000 μ m, porosity are 5～95%, mutual UNICOM between Kong Yukong, the inner surface in hole is rough surface, it is the granule of 200～800 nm that continuous distribution size, wherein discontinuously be distributed with micron-sized bulge-structure, the size of this bulge-structure is 2～8 μ m, is highly 1～5 μ m.
2. inner surface according to claim 1 has the bone cement porous support of the stacked lamellar micron-nano topological of pistil type pattern, it is characterized in that described micron order bulge-structure is the micron and nanometer composite structure of the stacked lamellar of pistil type, the thickness of its petal-shaped monolithic is less than 10 nm.
3. one kind prepares the preparation method that inner surface according to claim 1 and 2 has the bone cement porous support of the stacked lamellar micron-nano topological of pistil type pattern, it is characterized in that the concrete steps of the method are:
A. mix calcium carbonate in the powder of bata-tricalcium phosphate/hydroxyapatite biphasic calcium phosphate, as the bone cement powder powder stock; The quality of calcium carbonate is the 0.5-50% of the quality of biphasic calcium phosphate.
B. preparing mass percent concentration is the Fructus Citri Limoniae aqueous acid of 0.05-50%, and adds chitosan, stirs and makes its dissolving, obtains the acid consolidation liquid of pH＜4.0, and to make the mass percent concentration of chitosan be 0.05～3%.
C. step a gained bone cement powder is mixed according to the liquid-solid ratio of 0.2～1.0 ml:1g with subsidy b gained consolidation liquid, reconcile into the thickness paste, the injection molding plastotype solidifies the rear demoulding until bone cement;
D. the bone cement after the demoulding is processed through maintenance, polishing, finally made the bone cement porous support that inner surface has the stacked lamellar micron-nano topological of pistil type pattern.
4. method according to claim 3, the size that it is characterized in that the biphasic calcium phosphate described in step a is 150 nm～1 μ m, wherein the crystalline phase ratio of bata-tricalcium phosphate and hydroxyapatite is at 99:1～30:70.
5. method according to claim 3, is characterized in that the particle diameter of described calcium carbonate is no more than 5 μ m.
6. method according to claim 3, it is characterized in that the technological parameter of the described maintenance processes of steps d is: 15～50 ℃ of temperature, damp condition are relative humidity 5～80%, curing time is more than 18 h.
7. method according to claim 3, the polishing mode described in steps d that it is characterized in that are the wet method polishing of mechanical grinding, sand paper dry grinding or non-aqueous media.
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