CN101905040A - Method for preparing three-dimensional cell scaffold from elastic microsphere pore-forming agent - Google Patents
Method for preparing three-dimensional cell scaffold from elastic microsphere pore-forming agent Download PDFInfo
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Abstract
The invention discloses a method for preparing a three-dimensional cell scaffold from an elastic microsphere pore-forming agent. The method is characterized by comprising the following steps of: 1) preparing the elastic microsphere pore-forming agent and preparing the solution of a scaffold material; 2) placing the elastic microsphere pore-forming agent into a mould, adding the solution of the scaffold material to immerse the pore-forming agent, uniformly mixing the elastic microsphere pore-forming agent and the solution of the scaffold material to obtain a scaffold/elastic microsphere pore-forming agent complex, filter-pressing the complex by using a filter press plate with distributed microspheres to uniformly distribute the elastic microsphere pore-forming agent in the solution and contact with each other, and pre-freezing the complex for 3 hours at the temperature of -10 DEG C or lower; 3) demolding the complex and freeze-drying the complex for 6 to 8 hours; 4) rehydrating the complex in distilled water for 1 hour; and 5) removing the pore-forming agent. The scaffold preparation method of the invention can realize manual controllability of the pore structure of the scaffold and the preparation process is simple, convenient and easy.
Description
Technical field
The invention belongs to technical field of biological material, particularly a kind of elastic microsphere pore-forming agent is combined with the pressure filtration molding technology prepares the method for three-dimensional cell support.
Background technology
In organizational project, cell relies on support as growth templates, and cytoskeleton is guide tissue regeneration both, and control is knitted or the character of organ again, so three-dimensional porous rack plays pivotal role in tissue engineering, is one of key areas of Tissue Engineering Study.Studies show that in a large number cytoskeletal pore shape, size, porosity etc. directly influence the formation of migration, differentiation, propagation and the tissue of repopulating cell, ideal cytoskeleton need have certain aperture ranges and porosity communication passage.There is report to point out, influencing the grow into principal element of material hole of osseous tissue is the diameter of connection rate and pore interconnection between hole, the hole that be communicated with, the communication passage size is suitable is more conducive to blood vessel and osteoblast enters (Chang BS, Lee CK, Hong KS, etal.Osteoconduction at porous hydroxyapatite with various sporeconfiguration.Biomaterials, 2000,21 (12): 1291-8).And in the cytoskeletal various technology of traditional preparation, fibre cementing method can obtain connective good support between hole, but pore structure is difficult to control; Though gas foaming and phase detachment technique can be prepared the porous support of large volume but can not guarantee between hole to be communicated with fully, channel sized is uncontrollable between pore size and hole; The support connectedness of solvent cast/granule percolation preparation preparation is relatively poor, and pore morphology is coarse.
Summary of the invention
Purpose of the present invention is exactly in order to overcome above-mentioned the deficiencies in the prior art, provide a kind of elastic microsphere pore-forming agent and pressure filtration molding technology are combined to prepare the method for three-dimensional cell support, this method can obtain hole and be the artificial controlled three-dimensional cell support of communication passage between sphere, pore size and hole, and preparation process is simple and easy to do.
Method with preparing three-dimensional cell scaffold from elastic microsphere pore-forming agent involved in the present invention, its cardinal principle is: the pressure filtration molding technology is applied in the preparation of three-dimensional cell support, and combines with elastic microsphere pore-forming agent, can have both advantages concurrently.The present invention is uniformly mixed into complex with timbering material solution and elastic microsphere pore-forming agent, and the morphotropism of utilizing the flowability of timbering material solution and porogen is to the complex pressure filtration molding.When this complex was applied certain pressure, elastic microsphere pore-forming agent generation elastic deformation and be in contact with one another extruding caused the complex systems volume to reduce, and unnecessary polymer solution filtering is gone out.This moment is freeze settled, and the degree of microsphere shape and extruding each other all can be fixed.This method can be controlled the size in support aperture by the size of control microsphere, can become surface contact state by a contact condition after the stressed mutual extrusion of elastic microsphere, after removing porogen, the microsphere position has promptly formed hole, the face that is in contact with one another of microsphere can be converted into the brace aperture communication passage, can control brace aperture communication passage size by the contact area of adjusting between microsphere.
A kind of method involved in the present invention with preparing three-dimensional cell scaffold from elastic microsphere pore-forming agent, its step is as follows:
1) preparation elastic microsphere pore-forming agent, preparation timbering material solution;
2) elastic microsphere pore-forming agent is placed mould, add timbering material solution submergence porogen, mix homogeneously obtains support/elastic microsphere pore-forming agent complex, with the filter pressing plate that is distributed with micropore to this complex filter pressing, elastic microsphere pore-forming agent is uniformly distributed in the solution and can be in contact with one another, complex is placed pre-freeze 3h under-10 ℃ or the lower temperature;
3) with the complex demoulding, lyophilization 6～8h;
4) complex is inserted rehydration 1h in the distilled water;
5) remove porogen.
The above-mentioned the 1st) preparation of elastic microsphere pore-forming agent has following two kinds of methods in the step:
First method adopts the polyvinyl alcohol preparation, and its step is as follows:
(a) the preparation mass fraction is 7%～12% poly-vinyl alcohol solution;
(b) the high-pressure electrostatic legal system is equipped with the polyvinyl alcohol elastic microsphere, and high voltage range is 10～15KV, and interelectrode distance is 2～3cm; With the volume ratio that is lower than-25 ℃ is that 2: 1 chloroform and acetone mixed solution are as the consolidation liquid solidified microsphere; Preserve moisture at the logical a large amount of steam of poly-vinyl alcohol solution outflow port in the preparation process;
(c) with freezing repeatedly 4～6 times of the polyvinyl alcohol elastic microsphere in the consolidation liquid, cryogenic temperature is-25 ℃ or low temperature more, and each cooling time is 10h, and thawing time is 1h;
(d) the polyvinyl alcohol elastic microsphere is placed saturated sodium-chloride or saturated potassium chloride solution under the room temperature soak 20～60min; With distilled water flushing elastic microsphere 2～3 times, constant pressure and dry is removed its surface moisture;
(e) screening obtains diameter and is 100～1000 microns elastic microsphere pore-forming agent.
Second method adopts the sodium alginate preparation, and its step is as follows:
(a) the preparation mass fraction is 3%～5% sodium alginate soln;
(b) be equipped with the calcium alginate elastic microsphere with the high-pressure electrostatic legal system, high voltage range is 10～13KV, and interelectrode distance is 2～3cm; With mass fraction under the room temperature is that 20% calcium chloride solution is solidified into calcium alginate microsphere as consolidation liquid with sodium alginate micro ball; Preserve moisture at the logical a large amount of steam of sodium alginate soln outflow port in the preparation process;
(c) place consolidation liquid to soak 30～90min at normal temperatures the calcium alginate elastic microsphere; With distilled water flushing elastic microsphere 2～3 times, constant pressure and dry is removed its surface moisture;
(d) screening obtains diameter and is 100～1000 microns calcium alginate elastic microsphere pore-forming agent.
Major advantage of the present invention is: (1) this method can obtain hole and be the artificial controlled three-dimensional cell support of communication passage between sphere, pore size and hole; (2) elastic microsphere pore-forming agent and pressure filtration molding technology are combined, can have both advantages concurrently.By elastic microsphere pore-forming agent size control brace aperture size, control communication passage size between brace aperture by the contact area between the microsphere after the filter pressing; (3) preparation process is simple and easy to do.
Description of drawings
Fig. 1 is elastic microsphere pore-forming agent and the pressure filtration molding technology and the preparation three-dimensional cell support schematic diagram that combines.
Fig. 2 is that diameter is the pattern microscope picture of 100～1000 microns polyvinyl alcohol elastic microsphere pore-forming agent.
Fig. 3 is a three-dimensional cell support outward appearance picture.
Fig. 4 is the microscope picture of three-dimensional cell brace aperture structure, and amplification is 4 times.
Fig. 5 is the scanning electron microscope picture of three-dimensional cell support.
The specific embodiment
The present invention is further described as follows below in conjunction with drawings and Examples:
1) preparation polyvinyl alcohol elastic microsphere pore-forming agent, preparation mass fraction are that the dioxane solution of 3% polylactic acid is as timbering material solution; Polyvinyl alcohol elastic microsphere pore-forming agent preparation process is as follows:
(a) the preparation mass fraction is 7% poly-vinyl alcohol solution;
(b) the high-pressure electrostatic legal system is equipped with the polyvinyl alcohol elastic microsphere, voltage is 10KV, interelectrode distance is 3cm, preserves moisture at the logical a large amount of steam of poly-vinyl alcohol solution outflow port in the preparation process, and be that 2: 1 chloroform and acetone mixed solution are as the consolidation liquid solidified microsphere with-25 ℃ volume ratios;
(c) freezing repeatedly microsphere is 6 times, and cryogenic temperature is-25 ℃, and each cooling time is 10h, and thawing time is 1h;
(d) microsphere is inserted soak 20min in the saturated salt solution, with distilled water flushing 2～3 times, constant pressure and dry;
(e) screening obtains diameter and is 100～1000 microns polyvinyl alcohol elastic microsphere pore-forming agent.
2) with the 1st) the polyvinyl alcohol elastic microsphere pore-forming agent that makes of step inserts in the mould, add timbering material solution submergence porogen, mix homogeneously obtains support/polyvinyl alcohol elastic microsphere pore-forming agent complex, with the filter pressing plate that is distributed with micropore to this complex filter pressing, filter pressing is to 90% of the complex initial volume, the complex after the filter pressing is transferred to pre-freeze 3h in-10 ℃ the medical cryostat rapidly;
3) with the complex demoulding, lyophilization 8h;
4) complex was inserted in the distilled water rehydration 1 hour;
5) complex is inserted carry out the microwave heating repeated treatments in the distilled water 5～10 times, each 2～3min; Constant pressure and dry is removed excessive moisture, thereby obtains the three-dimensional rack of molding.
Embodiment 2
1) preparation polyvinyl alcohol elastic microsphere pore-forming agent, preparation mass fraction be the dioxane/chloroform soln of 3.5% polylactic acid as timbering material solution, dioxane and chloroform volume ratio are 5: 2; Polyvinyl alcohol elastic microsphere pore-forming agent preparation process is as follows:
(a) the preparation mass fraction is 10% poly-vinyl alcohol solution;
(b) the high-pressure electrostatic legal system is equipped with elastic microsphere, voltage is 12KV, interelectrode distance is 2.5cm, preserves moisture at the logical a large amount of steam of poly-vinyl alcohol solution outflow port in the preparation process, and be that 2: 1 chloroform and acetone mixed solution are as the consolidation liquid solidified microsphere with-25 ℃ volume ratios;
(c) freezing repeatedly microsphere is 5 times, and cryogenic temperature is-25 ℃, and each cooling time is 10h, and thawing time is 1h;
(d) microsphere is inserted soak 40min in the saturated salt solution, with distilled water flushing 2～3 times, constant pressure and dry;
(e) screening obtains diameter and is 100～1000 microns elastic microsphere pore-forming agent.
2) with the 1st) the polyvinyl alcohol elastic microsphere pore-forming agent that makes of step inserts in the mould, add timbering material solution submergence porogen, mix homogeneously obtains support/polyvinyl alcohol elastic microsphere pore-forming agent complex, with the filter pressing plate that is distributed with micropore to this complex filter pressing, filter pressing is to 80% of the complex initial volume, the complex after the filter pressing is transferred to pre-freeze 3h in-25 ℃ the medical cryostat rapidly;
3) with the complex demoulding, lyophilization 7h;
4) complex was inserted in the distilled water rehydration 1 hour;
5) complex is inserted carry out the microwave heating repeated treatments in the distilled water 5～10 times, each 2～3min; Constant pressure and dry is removed excessive moisture, thereby obtains the three-dimensional rack of molding.
Embodiment 3
1) preparation polyvinyl alcohol elastic microsphere pore-forming agent, preparation mass fraction be the dioxane/chloroform soln of 4% polylactic acid as timbering material solution, dioxane and chloroform volume ratio are 5: 2; Polyvinyl alcohol elastic microsphere pore-forming agent preparation process is as follows:
(a) the preparation mass fraction is 12% poly-vinyl alcohol solution;
(b) the high-pressure electrostatic legal system is equipped with elastic microsphere, voltage is 15KV, interelectrode distance is 2cm, preserves moisture at the logical a large amount of steam of poly-vinyl alcohol solution outflow port in the preparation process, and be that 2: 1 chloroform and acetone mixed solution are as the consolidation liquid solidified microsphere with-25 ℃ volume ratios;
(c) freezing repeatedly microsphere is 4 times, and cryogenic temperature is-25 ℃, and each cooling time is 10h, and thawing time is 1h;
(d) microsphere is inserted soak 60min in the saturated salt solution, with distilled water flushing 2～3 times, constant pressure and dry;
(e) screening obtains diameter and is 100～1000 microns elastic microsphere pore-forming agent.
2) with the 1st) the polyvinyl alcohol elastic microsphere pore-forming agent that makes of step inserts in the mould, add timbering material solution submergence porogen, mix homogeneously obtains support/polyvinyl alcohol elastic microsphere pore-forming agent complex, with the filter pressing plate that is distributed with micropore to this complex filter pressing, filter pressing is to 70% of the complex initial volume, the complex after the filter pressing is transferred to pre-freeze 3h in-25 ℃ the medical cryostat rapidly;
3) with the complex demoulding, lyophilization 6h;
4) complex was inserted in the distilled water rehydration 1 hour;
5) complex is inserted carry out the microwave heating repeated treatments in the distilled water 5～10 times, each 2～3min; Constant pressure and dry is removed excessive moisture, thereby obtains the three-dimensional rack of molding.
Embodiment 4
1) preparation calcium alginate elastic microsphere pore-forming agent, preparation mass fraction are that the dioxane solution of 3% polylactic acid is as timbering material solution; Calcium alginate elastic microsphere pore-forming agent preparation process is as follows:
(a) the preparation mass fraction is 3% sodium alginate soln;
(b) be equipped with the calcium alginate elastic microsphere with the high-pressure electrostatic legal system, voltage is 10KV, and interelectrode distance is 3cm; With mass fraction under the room temperature is that 20% calcium chloride solution is solidified into calcium alginate microsphere as consolidation liquid with sodium alginate micro ball; Preserve moisture at the logical a large amount of steam of sodium alginate soln outflow port in the preparation process;
(c) place consolidation liquid to soak 30min at normal temperatures the calcium alginate elastic microsphere; With distilled water flushing elastic microsphere 2～3 times, constant pressure and dry is removed its surface moisture;
(d) screening obtains diameter and is 100～1000 microns calcium alginate elastic microsphere pore-forming agent.
2) with the 1st) the calcium alginate elastic microsphere pore-forming agent that makes of step inserts in the mould, add timbering material solution submergence porogen, mix homogeneously obtains support/calcium alginate elastic microsphere pore-forming agent complex, with the filter pressing plate that is distributed with micropore to this complex filter pressing, filter pressing is to 90% of the complex initial volume, the complex after the filter pressing is transferred to pre-freeze 3h in-10 ℃ the medical cryostat rapidly;
3) with the complex demoulding, lyophilization 8h;
4) complex was inserted in the distilled water rehydration 1 hour;
5) complex is inserted in the EDTA solution of PH=7 and soak 2h, again support/porogen complex is inserted in the distilled water microwave heating repeated treatments 3～5 times, each 2～3min.
Embodiment 5
1) preparation calcium alginate elastic microsphere pore-forming agent, preparation mass fraction be the dioxane/chloroform soln of 3.5% polylactic acid as timbering material solution, dioxane and chloroform volume ratio are 5: 2; Calcium alginate elastic microsphere pore-forming agent preparation process is as follows:
(a) the preparation mass fraction is 4% sodium alginate soln;
(b) be equipped with the calcium alginate elastic microsphere with the high-pressure electrostatic legal system, voltage is 12KV, and interelectrode distance is 2.5cm; With mass fraction under the room temperature is that 20% calcium chloride solution is solidified into calcium alginate microsphere as consolidation liquid with sodium alginate micro ball; Preserve moisture at the logical a large amount of steam of sodium alginate soln outflow port in the preparation process;
(c) place consolidation liquid to soak 60min at normal temperatures the calcium alginate elastic microsphere; With distilled water flushing elastic microsphere 2～3 times, constant pressure and dry is removed its surface moisture;
(d) screening obtains diameter and is 100～1000 microns calcium alginate elastic microsphere pore-forming agent.
2) with the 1st) the calcium alginate elastic microsphere pore-forming agent that makes of step inserts in the mould, add timbering material solution submergence porogen, mix homogeneously obtains support/calcium alginate elastic microsphere pore-forming agent complex, with the filter pressing plate that is distributed with micropore to this complex filter pressing, filter pressing is to 80% of the complex initial volume, the complex after the filter pressing is transferred to pre-freeze 3h in-25 ℃ the medical cryostat rapidly;
3) with the complex demoulding, lyophilization 7h;
4) complex was inserted in the distilled water rehydration 1 hour;
5) complex is inserted in the EDTA solution of PH=7 and soak 2.5h, again support/porogen complex is inserted in the distilled water microwave heating repeated treatments 3～5 times, each 2～3min.
Embodiment 6
1) preparation calcium alginate elastic microsphere pore-forming agent, preparation mass fraction be the dioxane/chloroform soln of 4% polylactic acid as timbering material solution, dioxane and chloroform volume ratio are 5: 2; Calcium alginate elastic microsphere pore-forming agent preparation process is as follows:
(a) the preparation mass fraction is 5% sodium alginate soln;
(b) be equipped with the calcium alginate elastic microsphere with the high-pressure electrostatic legal system, voltage is 13KV, and interelectrode distance is 2cm; With mass fraction under the room temperature is that 20% calcium chloride solution is solidified into calcium alginate microsphere as consolidation liquid with sodium alginate micro ball; Preserve moisture at the logical a large amount of steam of sodium alginate soln outflow port in the preparation process;
(c) place consolidation liquid to soak 90min at normal temperatures the calcium alginate elastic microsphere; With distilled water flushing elastic microsphere 2～3 times, constant pressure and dry is removed its surface moisture;
(d) screening obtains diameter and is 100～1000 microns calcium alginate elastic microsphere pore-forming agent.
2) with the 1st) the calcium alginate elastic microsphere pore-forming agent that makes of step inserts in the mould, add timbering material solution submergence porogen, mix homogeneously obtains support/calcium alginate elastic microsphere pore-forming agent complex, with the filter pressing plate that is distributed with micropore to this complex filter pressing, filter pressing is to 70% of the complex initial volume, the complex after the filter pressing is transferred to pre-freeze 3h in-25 ℃ the medical cryostat rapidly;
3) with the complex demoulding, lyophilization 6h;
4) complex was inserted in the distilled water rehydration 1 hour;
5) complex is inserted in the EDTA solution of PH=7 and soak 3.5h, again support/porogen complex is inserted in the distilled water microwave heating repeated treatments 3～5 times, each 2～3min.
Fig. 2 is that diameter is the pattern microscope picture of 100～1000 microns polyvinyl alcohol elastic microsphere pore-forming agent.By preparation experiment interpretation of result to elastic microsphere pore-forming agent, the influence factor of elastic microsphere pore-forming agent particle diameter mainly contains: voltage and interelectrode distance, voltage is big more, interelectrode distance is more little, and the high voltage electric field intensity of formation is big more, and the microsphere average grain diameter that makes is more little; The influence factor of porogen elasticity and mechanical property mainly contains: solution concentration, saline solution soak time, and solution concentration is big more, the saline solution soak time is long more, and microsphere mechanical strength and elastic performance are good more.In addition, the freezing repeatedly mechanical strength and the elastic performance that can strengthen the polyvinyl alcohol elastic microsphere, when freezing number of times reached 5～6 times, microsphere mechanical strength and elastic performance can reach the experiment necessary requirement substantially; The polyvinyl alcohol elastic microsphere can not be long with the saturated salt solution soak time, can not reach the experiment necessary requirement on the contrary otherwise the elastic microsphere dehydration too much can make elasticity reduce.
Fig. 3 is a three-dimensional cell support outward appearance picture, has shown the outward appearance uniformity of support.
Fig. 4 is the microscope picture of three-dimensional cell brace aperture structure, has shown the pore structure and the interpore connectedness of support.
Fig. 5 is the scanning electron microscope picture of three-dimensional cell support.From picture, can clearly see, having because of removing the spherical void that elastic microsphere pore-forming agent forms (is the macropore on Fig. 5 medium-height trestle cross section, abbreviate macropore as), have many intercommunicating pores (be the aperture on the big hole wall among Fig. 5, abbreviate intercommunicating pore as) between the macropore.The existence of intercommunicating pore makes and can be interconnected between each macropore, illustrates that the contact surface of elastic microsphere pore-forming agent mutual extrusion in pressure-filtering process has become communication passage between brace aperture in better conversion after the filter pressing, has guaranteed the porous of three-dimensional cell support.
With the support of embodiment 1～6 preparation, carry out outward appearance and microstructure and observe, and with its pore size of tape measure and porosity communication channel sized, result such as table 1:
Table 1
| The filter pressing degree | Brace aperture size (μ m) | Channel size between brace aperture (μ m) |
| 70％ | 639 | ?326±26.6(8) |
| 80％ | 539 | ?283±52.0(8) |
| 90％ | 593 | ?257±36.2(8) |
By above-mentioned experimental result as can be known: (1) elastic microsphere pore-forming agent size is to the influence of brace aperture size.The porogen particle diameter is big more, and prepared brace aperture is big more; The suffered filter pressing degree of porogen/scaffold complex is big more, and the porogen elastic deformation is big more, and prepared brace aperture size is generally also big more.Can control the size of brace aperture size by the size of control elastic microsphere.(2) filter pressing degree is to the influence of communication passage size between brace aperture.As can be known from the above table, porogen/scaffold complex filter pressing degree is big more, and it is big more that microsphere is in contact with one another area, and it is big more to remove between the hole that forms behind the porogen communication passage size.By regulating the filter pressing degree, can control communication passage size between brace aperture.
Claims (3)
1. the method with preparing three-dimensional cell scaffold from elastic microsphere pore-forming agent is characterized in that step is as follows
1) preparation elastic microsphere pore-forming agent, preparation timbering material solution;
2) elastic microsphere pore-forming agent is placed mould, add timbering material solution submergence porogen, mix homogeneously obtains support/elastic microsphere pore-forming agent complex, with the filter pressing plate that is distributed with micropore to this complex filter pressing, elastic microsphere pore-forming agent is uniformly distributed in the solution and can be in contact with one another, complex is placed pre-freeze 3h under-10 ℃ or the lower temperature;
3) with the complex demoulding, lyophilization 6～8h;
4) complex is inserted rehydration 1h in the distilled water;
5) remove porogen.
2. a kind of method with preparing three-dimensional cell scaffold from elastic microsphere pore-forming agent according to claim 1 is characterized in that: the 1st) elastic microsphere pore-forming agent adopts the polyvinyl alcohol preparation in the step, and its step is as follows:
(a) the preparation mass fraction is 7%～12% poly-vinyl alcohol solution;
(b) the high-pressure electrostatic legal system is equipped with the polyvinyl alcohol elastic microsphere, and high voltage range is 10～15KV, and interelectrode distance is 2～3cm; With the volume ratio that is lower than-25 ℃ is that 2: 1 chloroform and acetone mixed solution are as the consolidation liquid solidified microsphere; Preserve moisture at the logical a large amount of steam of poly-vinyl alcohol solution outflow port in the preparation process;
(c) with freezing repeatedly 4～6 times of the polyvinyl alcohol elastic microsphere in the consolidation liquid, cryogenic temperature is-25 ℃ or low temperature more, and each cooling time is 10h, and thawing time is 1h;
(d) the polyvinyl alcohol elastic microsphere is placed saturated sodium-chloride or saturated potassium chloride solution under the room temperature soak 20～60min; With distilled water flushing elastic microsphere 2～3 times, constant pressure and dry is removed its surface moisture;
(e) screening obtains diameter and is 100～1000 microns elastic microsphere pore-forming agent;
The 5th) going on foot the porogen method of removing is support/polyvinyl alcohol elastic microsphere pore-forming agent complex to be inserted in the distilled water microwave heating repeated treatments 5～10 times, each 2～3min.
3. a kind of method with preparing three-dimensional cell scaffold from elastic microsphere pore-forming agent according to claim 1 is characterized in that: the 1st) elastic microsphere pore-forming agent adopts the sodium alginate preparation in the step, and its step is as follows:
(a) the preparation mass fraction is 3%～5% sodium alginate soln;
(b) be equipped with the calcium alginate elastic microsphere with the high-pressure electrostatic legal system, voltage range is 10～13KV, and interelectrode distance is 2～3cm; With mass fraction under the room temperature is that 20% calcium chloride solution is solidified into calcium alginate microsphere as consolidation liquid with sodium alginate micro ball; Preserve moisture at the logical a large amount of steam of sodium alginate soln outflow port in the preparation process;
(c) place consolidation liquid to soak 30～90min at normal temperatures the calcium alginate elastic microsphere; With distilled water flushing elastic microsphere 2～3 times, constant pressure and dry is removed its surface moisture;
(d) screening obtains diameter and is 100～1000 microns calcium alginate elastic microsphere pore-forming agent;
The 5th) the step porogen method of removing is support/calcium alginate elastic microsphere pore-forming agent complex to be inserted in the EDTA solution of PH=7 soak 2～3.5h, again support/sodium alginate elastic microsphere pore-forming agent complex is inserted in the distilled water, microwave heating repeated treatments 3～5 times, each 2～3min.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102631680A (en) * | 2012-04-19 | 2012-08-15 | 浙江大学 | Method of regulating microsphere drug-load rate by treating calcium alginate microspheres with sodium chloride solution |
| CN102698667A (en) * | 2012-06-19 | 2012-10-03 | 重庆大学 | Spherical pore foaming agent with nuclear shell structure and three-dimensional cytoskeleton prepared by same |
| CN102977224A (en) * | 2012-12-21 | 2013-03-20 | 青岛明月海藻集团有限公司 | Method for preparing calcium alginate |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1651099A (en) * | 2004-02-06 | 2005-08-10 | 上海组织工程研究与开发中心 | Water soluble binder and its application |
| CN101176799A (en) * | 2007-12-06 | 2008-05-14 | 同济大学 | Method for preparing polyalcohol stephanoporate bracket for tissue project by poragen agglutinating filtering off method |
-
2010
- 2010-07-23 CN CN 201010234842 patent/CN101905040B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1651099A (en) * | 2004-02-06 | 2005-08-10 | 上海组织工程研究与开发中心 | Water soluble binder and its application |
| CN101176799A (en) * | 2007-12-06 | 2008-05-14 | 同济大学 | Method for preparing polyalcohol stephanoporate bracket for tissue project by poragen agglutinating filtering off method |
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|---|---|---|---|---|
| CN102631680A (en) * | 2012-04-19 | 2012-08-15 | 浙江大学 | Method of regulating microsphere drug-load rate by treating calcium alginate microspheres with sodium chloride solution |
| CN102631680B (en) * | 2012-04-19 | 2013-05-29 | 浙江大学 | Method of regulating microsphere drug-load rate by treating calcium alginate microspheres with sodium chloride solution |
| CN102698667A (en) * | 2012-06-19 | 2012-10-03 | 重庆大学 | Spherical pore foaming agent with nuclear shell structure and three-dimensional cytoskeleton prepared by same |
| CN102698667B (en) * | 2012-06-19 | 2014-05-28 | 重庆大学 | Spherical pore foaming agent with nuclear shell structure and three-dimensional cytoskeleton prepared by same |
| CN102977224A (en) * | 2012-12-21 | 2013-03-20 | 青岛明月海藻集团有限公司 | Method for preparing calcium alginate |
| CN104711302A (en) * | 2015-03-17 | 2015-06-17 | 南京荣之盛生物科技有限公司 | Method for adjusting bacterial cellulose structure with calcium alginate bead |
| WO2018032664A1 (en) * | 2016-08-19 | 2018-02-22 | 北京派尔特医疗科技股份有限公司 | Titanium clip with drug-loading feature, drug-loaded titanium clip, and manufacturing method thereof |
| US10286117B2 (en) | 2016-08-19 | 2019-05-14 | B.J. Zh. F. Panther Medical Equipment Co. Ltd. | Titanium nail capable of loading drug and drug-loaded titanium nail and preparing method of the same |
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