CN103920187A - Bone repair material prepared by combining silk fibroin and decalcified bone matrix - Google Patents
Bone repair material prepared by combining silk fibroin and decalcified bone matrix Download PDFInfo
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- CN103920187A CN103920187A CN201310130923.4A CN201310130923A CN103920187A CN 103920187 A CN103920187 A CN 103920187A CN 201310130923 A CN201310130923 A CN 201310130923A CN 103920187 A CN103920187 A CN 103920187A
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- fibroin albumen
- decalcified
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Abstract
The invention relates to the technical field of biomaterials, and relates to a preparation method for a bone repair material. The bond repair material is composed of silk fibroin and decalcified bone matrix. A salting-out process is employed for preparing the bone repair material by combining silk fibroin and decalcified bone matrix. The preparation method helps to keep the bone induction and bone conduction capabilities of decalcified bone matrix, and helps to improve the mechanic properties of the bone repair material. The prepared bone repair material has good promotion capability on bone regeneration and excellent mechanical properties, and is substantially improved in shape keeping capability and implanting capability.
Description
(1) technical field
The present invention relates to biomedical materials field, particularly relate to a kind of have good bone induction and bone conduction effect, there is fibroin albumen and the compound bone renovating material of decalcified bone matrix of outstanding mechanical property simultaneously.
(2) background technology
As everyone knows, the damaged and bone does not connect of bone is a great problem that orthopedist faces always, and how repairing bone defect and bone does not connect are the thorny problems of orthopedics circle always.In recent years, bone graft substitute becomes study hotspot.At present conventional repair materials comprises three kinds of autologous bone, allograph bone and synthetic substitutes.Autologous bone is owing to containing bone-inducing factor, bone conduction substrate and Osteogenic Cells, induced multipotent stem cells is to osteoblast differentiation, be beneficial to growing into and osteanagenesis of blood vessel, therefore, autologous bone is people's desirable bone grafting material, but it has following shortcoming: 1. source is limited, particularly can not meet the needs of large segmental bone defect operation; 2. needs of patients bears the extra misery of bringing for district's complication; 3. extend operating time, increased accordingly amount of bleeding.Allogenic bone transplantation has good bone conductibility, can also retain its osteoinductive, but it easily causes immunological rejection, and the cost that samples, processes, stores is high through ad hoc approach after processing, and its application is very limited.Synthetic substitute, as polymethyl methacrylate, poly-sough, sulphurated siliastic etc., be all non-degradable material, although wide material sources, due to these material non-degradables, cause defective region not rebuild Regenerated Bone by bone.In addition, the stress in daily life, bone filling position being produced is also a major reason that finally causes repairing failure.So, desirable bone renovating material except should there is bone conduction and bone inducibility and in surgical operation easy operating, also should there is certain mechanical strength.
The decalcified bone matrix (DBM) of being made through special handling by homogeneous allogenic bone, not only has the constituent of nature bone, and has bone conduction and bone inductive effect.DBM is proved to be effective bone-grafting material, is day by day applied in bone grafting surgical operation.Due to the good bone regeneration capability of DBM, it is widely used in orthopedics, dentistry, neurosurgery, is also used to treat osteoma, bone injury and fracture.Because the mechanical property of DBM is poor, be not suitable for bearing the position of gravity and other stress, this greatly reduces the scope of application of DBM.In addition, because DBM has lower water solublity, when DBM can not maintain separately the shape needing when transplanting preferably, be included in the also digested hydrolysis in vivo of osteogenesis albumen in DBM.In addition, because the incomplete shape hold facility of DBM causes the low implantation capability of DBM, make to find and a kind ofly there is good biocompatibility and there is material and the DBM of better mechanical property compound, become one of effective ways of dealing with problems.
In recent years, a kind of natural macromolecular material-silkworm silk, due to its good mechanical property, is better than again traditional the absorbable macromolecule material simultaneously aspect biocompatibility, becomes the special biomaterial of a class that field of tissue engineering technology attracts people's attention.Fibroin albumen is the natural polymer fibrin extracting from silkworm silk, content accounts for 70%～80% of silkworm silk, contain 18 seed amino acids, wherein glycine (gly), alanine (ala) and serine (ser) account for the more than 80% of total composition.Much research shows, fibroin albumen has outstanding mechanical property and good biocompatibility.
Therefore, the present invention utilizes salting out method that fibroin albumen and decalcified bone matrix are carried out to the compound bone renovating material of preparing, and on the one hand, has retained bone conduction and the bone inducibility of decalcified bone matrix; On the other hand, mechanical property and the biocompatibility of bone renovating material be can improve, and shape-retaining force and the implantation capability of bone renovating material significantly improved.
(3) summary of the invention
The object of the invention is to prepare a kind of have good bone induction and bone conduction ability, there is fibroin albumen and the compound bone renovating material of decalcified bone matrix of excellent mechanical performances simultaneously.
To achieve the above object of the invention, the technical solution used in the present invention is as follows:
A) preparation of fibroin albumen:
Domestic silkworm silk is placed in to 0.1% Na
2cO
3in solution, in 98～100 ° of C, process 30 minutes, in triplicate, to slough the sericin on silk fiber surface, in air at room temperature, dry.Silk fiber is put into CaCl
2-CH
3cH
2oH-H
2o(mol ratio is 1:2:8) dissolve in solution, maintain the temperature at 78 ± 2 ℃ and constantly stir, obtain silk fibroin protein solution, with Dialysis tubing, in distilled water, dialyse subsequently.Finally, silk fibroin protein solution is carried out in freezer dryer to lyophilization;
B) preparation of blend solution:
Cryodesiccated fibroin albumen and decalcified bone matrix are placed in to beaker by a certain percentage, add hexafluoroisopropanol, concussion, until fibroin albumen all dissolves, obtains blend solution;
C) preparation of blend:
Sodium chloride particle with standard molecule sieve screening 200～300um, joins in above-mentioned solution, stirs and obtains blend;
D) proceed to mould:
Above-mentioned blend is proceeded in mould, and note compacting;
E) forming materials:
Mould is placed in fume hood, until composite presents curing tendency.From mould, take out blend, soaked in absolute ethyl alcohol 0.5 hour.Then be placed in distilled water, make sodium chloride be dissolved in distilled water and obtain porous decalcified bone matrix and the compound bone renovating material of fibroin albumen.Dry in last vacuum drying oven.
Advantage of the present invention is:
(1) prepared fibroin albumen and the compound bone renovating material of decalcified bone matrix of the present invention not only has good bone induction and bone conduction ability, and has good mechanical property.
(2) technique of the present invention is simple, and consumption is few, consuming time short, easily realizes commodity production.
(3) shape of adjustable material and size as required.
(4) prepared fibroin albumen and the compound bone renovating material of decalcified bone matrix of the present invention has good shape-retaining force and implantation capability.
(5) the prepared fibroin albumen of the present invention and the compound bone renovating material of decalcified bone matrix be expected to be applied to clinical, repairing bone defect and bone does not connect.
(4) specific embodiment
Specifically in conjunction with the embodiments, the present invention is further elaborated.These embodiment are only not used in and limit the scope of the invention for the present invention is described.In addition should be understood that those skilled in the art can make various changes or modification to invention after having read the content of the present invention's instruction, these equivalent form of values fall within the application's appended claims limited range equally.
Embodiment 1
The preparation method of the bone renovating material that fibroin albumen and decalcified bone matrix are compound is as follows: the bone renovating material that it is comprised of following weight proportioning, decalcified bone matrix: fibroin albumen=1:5.And the concentration of silk fibroin protein solution is 17% (g/ml).
A) preparation of fibroin albumen:
Domestic silkworm silk is placed in to 0.1% Na
2cO
3in solution, in 98～100 ° of C, process 30 minutes, in triplicate, to slough the sericin on silk fiber surface, in air at room temperature, dry.Silk fiber is put into CaCl
2-CH
3cH
2oH-H
2o(mol ratio is 1:2:8) dissolve in solution, maintain the temperature at 78 ± 2 ℃ and constantly stir, obtain silk fibroin protein solution, with Dialysis tubing, in distilled water, dialyse subsequently.Finally, silk fibroin protein solution is carried out to lyophilization;
B) preparation of blend solution:
Just cryodesiccated fibroin albumen 0.85g and decalcified bone matrix 0.17g are placed in beaker, add hexafluoroisopropanol 5ml, and concussion, until fibroin albumen all dissolves, obtains blend solution;
C) preparation of blend:
Sodium chloride particle with standard molecule sieve screening 200～300um, joins in above-mentioned solution, stirs and obtains blend;
D) proceed to mould:
Above-mentioned blend is proceeded in mould, and note compacting;
E) forming materials:
Mould is placed in fume hood, until composite presents curing tendency.From mould, take out blend, soaked in absolute ethyl alcohol 0.5 hour.Then be placed in distilled water, make sodium chloride be dissolved in distilled water and obtain porous decalcified bone matrix and the compound bone renovating material of fibroin albumen.Dry in last vacuum drying oven.
Embodiment 2
The preparation method of the bone renovating material that fibroin albumen and decalcified bone matrix are compound is as follows: the bone renovating material that it is comprised of following weight proportioning, decalcified bone matrix: fibroin albumen=2:5.And the concentration of silk fibroin protein solution is 17% (g/ml).
A) preparation of fibroin albumen:
Domestic silkworm silk is placed in to 0.1% Na
2cO
3in solution, in 98～100 ° of C, process 30 minutes, in triplicate, to slough the sericin on silk fiber surface, in air at room temperature, dry.Silk fiber is put into CaCl
2-CH
3cH
2oH-H
2o(mol ratio is 1:2:8) dissolve in solution, maintain the temperature at 78 ± 2 ℃ and constantly stir, obtain silk fibroin protein solution, with Dialysis tubing, in distilled water, dialyse subsequently.Finally, silk fibroin protein solution is carried out in freezer dryer to lyophilization;
B) preparation of blend solution:
Just cryodesiccated fibroin albumen 0.85g and decalcified bone matrix 0.34g are placed in beaker, add hexafluoroisopropanol 5ml, and concussion, until fibroin albumen all dissolves, obtains blend solution;
C) preparation of blend:
Sodium chloride particle with standard molecule sieve screening 200～300um, joins in above-mentioned solution, stirs and obtains blend;
D) proceed to mould:
Above-mentioned blend is proceeded in mould, and note compacting;
E) forming materials:
Mould is placed in fume hood, until composite presents curing tendency.From mould, take out blend, soaked in absolute ethyl alcohol 0.5 hour.Then be placed in distilled water, make sodium chloride be dissolved in distilled water and obtain porous decalcified bone matrix and the compound bone renovating material of fibroin albumen.Dry in last vacuum drying oven.
Embodiment 3
The preparation method of the bone renovating material that fibroin albumen and decalcified bone matrix are compound is as follows: the bone renovating material that it is comprised of following weight proportioning, decalcified bone matrix: fibroin albumen=4:5.And the concentration of silk fibroin protein solution is 17% (g/ml).
A) preparation of fibroin albumen:
Domestic silkworm silk is placed in to 0.1% Na
2cO
3in solution, in 98～100 ° of C, process 30 minutes, in triplicate, to slough the sericin on silk fiber surface, in air at room temperature, dry.Silk fiber is put into CaCl
2-CH
3cH
2oH-H
2o(mol ratio is 1:2:8) dissolve in solution, maintain the temperature at 78 ± 2 ℃ and constantly stir, obtain silk fibroin protein solution, with Dialysis tubing, in distilled water, dialyse subsequently.Finally, silk fibroin protein solution is carried out in freezer dryer to lyophilization;
B) preparation of blend solution:
Just cryodesiccated fibroin albumen 0.85g and decalcified bone matrix 0.68g are placed in beaker, add hexafluoroisopropanol 5ml, and concussion, until fibroin albumen all dissolves, obtains blend solution;
C) preparation of blend:
Sodium chloride particle with standard molecule sieve screening 200～300um, joins in above-mentioned solution, stirs and obtains blend;
D) proceed to mould:
Above-mentioned blend is proceeded in mould, and note compacting;
E) forming materials:
Mould is placed in fume hood, until composite presents curing tendency.From mould, take out blend, soaked in absolute ethyl alcohol 0.5 hour.Then be placed in distilled water, make sodium chloride be dissolved in distilled water and obtain porous decalcified bone matrix and the compound bone renovating material of fibroin albumen.Dry in last vacuum drying oven.
Claims (5)
1. the preparation method of decalcified bone matrix and fibroin albumen composite bone repairing material, is characterized in that the method carries out as follows: a) preparation of fibroin albumen:
Domestic silkworm silk is placed in to 0.1% Na
2cO
3in solution, in 98～100 ° of C, process 30 minutes, in triplicate, to slough the sericin on silk fiber surface, in air at room temperature, dry.Silk fiber is put into CaCl
2-CH
3cH
2oH-H
2o(mol ratio is 1:2:8) dissolve in solution, maintain the temperature at 78 ± 2 ℃ and constantly stir, obtain silk fibroin protein solution, with Dialysis tubing, in distilled water, dialyse subsequently.Finally, silk fibroin protein solution is carried out in freezer dryer to lyophilization;
B) preparation of blend solution:
Cryodesiccated fibroin albumen and decalcified bone matrix are placed in to beaker by a certain percentage, add hexafluoroisopropanol, concussion, until fibroin albumen all dissolves, obtains blend solution;
C) preparation of blend:
Sodium chloride particle with standard molecule sieve screening 200～300um, joins in above-mentioned solution, stirs and obtains blend.
D) proceed to mould:
Above-mentioned blend is proceeded in mould, and note compacting.
E) forming materials:
Mould is placed in fume hood, until composite presents curing tendency.From mould, take out blend, soaked in absolute ethyl alcohol 0.5 hour.Then be placed in distilled water, make sodium chloride be dissolved in distilled water and obtain porous decalcified bone matrix and the compound bone renovating material of fibroin albumen.Dry in last vacuum drying oven.
2. the compound bone renovating material of decalcified bone matrix as claimed in claim 1 and fibroin albumen, is characterized in that: decalcified bone matrix: fibroin albumen=1:10～4:5.
3. the compound bone renovating material of decalcified bone matrix as claimed in claim 1 and fibroin albumen, is characterized in that: the shape of this bone renovating material can be selected corresponding mould as required.
4. the compound bone renovating material of decalcified bone matrix as claimed in claim 1 and fibroin albumen, is characterized in that: the concentration range of silk fibroin protein solution is 8%～30%.
5. the compound bone renovating material of decalcified bone matrix as claimed in claim 1 and fibroin albumen, is characterized in that: be expected to for clinical repairing bone defect and bone does not connect.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310130923.4A CN103920187B (en) | 2013-04-16 | 2013-04-16 | The bone renovating material that a kind of fibroin albumen and decalcified bone matrix are compound |
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|---|---|---|---|
| CN201310130923.4A CN103920187B (en) | 2013-04-16 | 2013-04-16 | The bone renovating material that a kind of fibroin albumen and decalcified bone matrix are compound |
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| CN103920187A true CN103920187A (en) | 2014-07-16 |
| CN103920187B CN103920187B (en) | 2016-05-25 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105661041A (en) * | 2016-01-11 | 2016-06-15 | 浙江国光生化股份有限公司 | Method for preparing granular feed additives based on powdery L-shaped amino acids |
| CN106178117A (en) * | 2016-08-09 | 2016-12-07 | 中国人民解放军***武汉总医院 | A kind of PLL DBM porous composite bone graft material, its preparation method and application |
| CN113952512A (en) * | 2021-10-04 | 2022-01-21 | 重庆工商大学 | Composite porous gel microsphere and preparation method and application thereof |
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| WO2005011764A1 (en) * | 2003-07-25 | 2005-02-10 | Sdgi Holdings, Inc. | Crosslinked compositions comprising collagen and demineralized bone matrix |
| CN101401974A (en) * | 2008-11-11 | 2009-04-08 | 中国人民解放军第二军医大学 | Process for producing homogeneous decalcification bone matrix material nano-DBM |
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2013
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Patent Citations (5)
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| WO2005011764A1 (en) * | 2003-07-25 | 2005-02-10 | Sdgi Holdings, Inc. | Crosslinked compositions comprising collagen and demineralized bone matrix |
| CN101401974A (en) * | 2008-11-11 | 2009-04-08 | 中国人民解放军第二军医大学 | Process for producing homogeneous decalcification bone matrix material nano-DBM |
| CN101664346A (en) * | 2009-09-02 | 2010-03-10 | 南通大学 | Artificial nerve graft prepared by electrostatic spinning and preparation method and special device thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105661041A (en) * | 2016-01-11 | 2016-06-15 | 浙江国光生化股份有限公司 | Method for preparing granular feed additives based on powdery L-shaped amino acids |
| CN106178117A (en) * | 2016-08-09 | 2016-12-07 | 中国人民解放军***武汉总医院 | A kind of PLL DBM porous composite bone graft material, its preparation method and application |
| CN113952512A (en) * | 2021-10-04 | 2022-01-21 | 重庆工商大学 | Composite porous gel microsphere and preparation method and application thereof |
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|---|---|
| CN103920187B (en) | 2016-05-25 |
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