CN108478874A - A kind of preparation method of the nano combined bone holder material of hydroxyethyl chitosan - Google Patents
A kind of preparation method of the nano combined bone holder material of hydroxyethyl chitosan Download PDFInfo
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- CN108478874A CN108478874A CN201810378947.4A CN201810378947A CN108478874A CN 108478874 A CN108478874 A CN 108478874A CN 201810378947 A CN201810378947 A CN 201810378947A CN 108478874 A CN108478874 A CN 108478874A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
- A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
- A61L27/48—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with macromolecular fillers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
- A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
- A61L27/46—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with phosphorus-containing inorganic fillers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
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Abstract
The present invention relates to a kind of preparation methods of the nano combined bone holder material of hydroxyethyl chitosan, first phosphorus calcium powder is obtained using the precipitation method, after calcination process, obtain hydroxyapatite/β tricalcium phosphate two-phase phosphorus calcium nanometer powders, chitosan is subjected to hydroxyethylation again, hydroxyethyl chitosan is prepared, finally hydroxyethyl chitosan is dissolved in 1% acetum, according to mass ratio (1~4):1:Hydroxyethyl chitosan, gelatin and two-phase phosphorus calcium nanometer powder are mixed the mass ratio of (0.5~4), and degassing is uniformly dispersed, and solution is then carried out circulating frozen defrosting processing, finally dry to get.Whole preparation process is simple to operation, can prepare in batches, cost is relatively low, pollution-free, ternary material system of the invention makes the bone holder of preparation possess suitable degradation rate, improves the cell compatibility of bone holder, with better bioactivity, cell can be made preferably to adhere to, be proliferated.
Description
Technical field
The present invention relates to a kind of preparation methods of the nano combined bone holder material of hydroxyethyl chitosan, belong to biomedical material
Expect technical field.
Background technology
Chitosan is that the chitin being widely present by nature is obtained by deacetylation, this natural polymer tool
It has the excellent performances such as biocompatibility, blood compatibility, safety, microbic resolvability, is prepared one in bio-medical field
The preferable material of kind.But chitosan gel rubber is needed in preparation process through crosslinking, and crosslinking agent usually selects glutaraldehyde, crosslinking agent
Introducing can reduce the cell compatibility of holder, and mechanical property is also poor.
Contain a large amount of phosphorus calcium material, such as tricalcium phosphate, hydroxyapatite, calcium carbonate etc., therefore phosphorus calcium in bone tissue
Material is commonly used for preparing artificial bone scaffold.Due to the compositional similarity with bone, calcium orthophosphate base biomaterial such as hydroxyapatite
(HA, Ca10(PO4)6(OH)2), tricalcium phosphate (β-TCP, Ca3(PO4)2) and biphase calcium phosphor sour (BCP) etc. can be used for bone holder
It prepares.BCP is the mixture of HA and β-TCP, is made of a certain proportion of inorganic substances tricalcium phosphate and hydroxyapatite, two-phase
Phosphorus calcium can overcome tricalcium phosphate to be easy degradation and the not degradable disadvantage of hydroxyapatite, and have good osteoconductive
And osteoinductive.In the various Calcium phosphate biomaterials for bone surgery, BCP is combined with high mechanical properties, and HA's is excellent
The important candidate of the bone substitute of different bioactivity and faster β-TCP degradation rates.The bioceramic made of calcium phosphate
All there is poor mechanical performance, there is certain limitation in weight-bearing areas.
Invention content
Present invention aims to solve the deficiencies of the prior art, and provides a kind of a kind of nano combined bone holder materials of hydroxyethyl chitosan
The preparation method of material, simple process and low cost is easily operated, and bone holder material obtained can make cell more preferable on holder
Attachment and proliferation, activity is high, and mechanical property is good.
Technical solution
The present inventor can improve the cell compatibility and mechanical property of chitosan by carrying out hydroxyethylation to chitosan
Can, and the two-phase phosphorus calcium nanometer powder and cellulose of different proportion is added, nano combined bone holder material is made, makes bone branch
Frame possesses suitable degradation rate, and has better bioactivity, cell can be made preferably to adhere to, it is allowed preferably to increase
It grows.Concrete scheme is as follows：
A kind of preparation method of the nano combined bone holder material of hydroxyethyl chitosan, includes the following steps：
(1) two-phase phosphorus calcium nanometer powder is prepared：By Ca (NO3)2.4H2O solution and (NH4)2HPO4Solution is according to calcium phosphorus mole
Than for (1.55~1.70):1 is added in container, is uniformly mixed, and pH is then adjusted to 11, and the powder for obtaining white is heavy
It forms sediment, after filtering, white powder is cleaned with deionized water, is then dried, roasts, grinds successively, finally cross 200 mesh sieve,
Obtain two-phase phosphorus calcium nanometer powder；
(2) hydroxyethyl chitosan is prepared：Take 10g chitosans be added in the 50wt% sodium hydroxide solutions of 15~18g into
Row alkalization, obtains chitosan solution, after standing 1~3h under the conditions of -20 DEG C, 15~25ml solvent isopropanols is added, stirring rises
Temperature is stirred for being cooled to room temperature to 70~80 DEG C, and the mixed of 130~180ml isopropanols and 15~19ml ethylene chlorhydrins is then added
Solution is closed, after stirring evenly, is washed using hydrochloric acid solution tune pH7, then using absolute ethyl alcohol, then dialyses, finally freeze
Drying is to get hydroxyethyl chitosan；
(3) compound bone holder material is prepared：It takes hydroxyethyl chitosan to be dissolved in the acetum of 1wt%, obtains hydroxyl second
Cellulose and two-phase are added into hydroxyethyl chitosan solution by base chitosan solution, a concentration of 2~5wt% of hydroxyethyl chitosan
Phosphorus calcium nanometer powder, hydroxyethyl chitosan, cellulose, two-phase phosphorus calcium nanometer powder mass ratio be (1~4):1:(0.5~4),
After stirring evenly at room temperature, then deaerating makes it be uniformly dispersed, and obtained solution, which is then poured into circulating frozen in 24 orifice plates, thaws
Processing 3~9 times, be finally putting into freeze drier be dried to get.
Further, in step (1), Ca (NO3)2.4H2A concentration of 0.4M, (NH of O solution4)2HPO4Solution it is a concentration of
0.2M。
Further, in step (1), the roasting carries out in Muffle furnace, is warming up to first with the rate of 10 DEG C/min
1125 DEG C, 1h is then kept, finally drops to room temperature with stove.
Further, in step (2), the alkalization carries out in -20 DEG C of refrigerators, and alkalization time is more than for 24 hours.
Further, in step (2), concentration of hydrochloric acid solution 0.1M.
Further, in step (2), selection of dialysing is bag filter of the molecular cut off in 10000Da, dialysis time 2d
More than.
Further, in step (3), the hydroxyethyl chitosan, cellulose, two-phase phosphorus calcium nanometer powder mass ratio be 1:
1:2。
Further, in step (3), the circulating frozen defrosting processing refers to：Freezing 24 hours in -20 DEG C of refrigerators are put into,
Then 12 hours of defrosting under room temperature are taken out, freezing in -20 DEG C of refrigerators is then placed into.
Beneficial effects of the present invention：The present invention by the two-phase phosphorus calcium nanometer powder of hydroxyethyl chitosan and different proportion and
Cellulose mixes, and obtains Composite Bone holder.Compared with prior art, advantage of the invention is that：1. selecting hydroxyethyl chitosan
The cell compatibility of bone holder can be improved, phosphorus calcium nanometer powder is added in bone holder, bone holder is made to possess suitable degradation speed
Rate makes it when new bone is grown, and the degradation appropriate of bone holder does not interfere with the reparation of bone；2. selecting ternary material system, make new
The bone holder of preparation has better bioactivity, cell can be made preferably to adhere to, it is allowed preferably to be proliferated；3. entire prepare
Process is simple to operation, can largely be prepared, and the high molecular material cost for preparing holder is relatively low, pollution-free.
Description of the drawings
Fig. 1 is the scanning electron microscope (SEM) photograph of nano combined bone holder material made from embodiment 1-3；
Fig. 2 is the porosity of nano combined bone holder material made from embodiment 1-6；
Fig. 3 is the cell compatibility test result of nano combined bone holder material made from embodiment 1-6.
Specific implementation mode
The present invention will be illustrated by the drawings and specific embodiments below, but these specific embodiments are not with any side
Formula limits the protection domain of the invention.Raw material used in the experimental program can be bought in market.
Embodiment 1
(1) two-phase phosphorus calcium nanometer powder is prepared：By the Ca (NO of 0.4M3)2.4H2(the NH of O solution and 0.2M4)2HPO4Solution
It is 1.65 according to calcium phosphorus molar ratio:1 is added in three-neck flask, is uniformly mixed, and is then adjusted to pH using 25% ammonium hydroxide
11, the powder precipitation of white is obtained, after filtering, white powder is cleaned with deionized water, then 80 DEG C of dryings, place into Muffle
Kiln roasting is first warming up to 1125 DEG C with the rate of 10 DEG C/min, after keeping 1h, drops to room temperature with stove, then takes out grinding, most
200 mesh sieve is crossed afterwards, obtains two-phase phosphorus calcium nanometer powder；
(2) hydroxyethyl chitosan is prepared：It takes 10g chitosans to be added in the 50wt% sodium hydroxide solutions of 16.7g to carry out
Alkalization, obtains chitosan solution, and the alkalization carries out in -20 DEG C of refrigerators, and alkalization time is for 24 hours, to be stood under the conditions of -20 DEG C
After 1h, be added solvent isopropanol, stirring be warming up to 78 DEG C, be stirred for being cooled to room temperature, then be added 133ml isopropanols and
The mixed solution of 18.3ml ethylene chlorhydrins after stirring evenly, is washed using the hydrochloric acid solution tune pH7 of 0.1M, then using absolute ethyl alcohol
It washs 3 times, then selects molecular cut off in the bag filter dialysis 2d of 10000Da, be finally freeze-dried poly- to get ethoxy shell
Sugar；
(3) compound bone holder material is prepared：It takes hydroxyethyl chitosan to be dissolved in the acetum of 1wt%, obtains hydroxyl second
Cellulose and two-phase phosphorus calcium are added into hydroxyethyl chitosan solution by base chitosan solution, a concentration of 2wt% of hydroxyethyl chitosan
Nanometer powder, hydroxyethyl chitosan, cellulose, two-phase phosphorus calcium nanometer powder mass ratio be 1:1:1, it stirs evenly at room temperature
Afterwards, then deaerating makes it be uniformly dispersed, and obtained solution, which is then poured into circulating frozen defrosting processing in 24 orifice plates, (puts 28 orifice plates
Enter freezing 24 hours in -20 DEG C of refrigerators, then takes out 12 hours of defrosting under room temperature, then place into cold in -20 DEG C of refrigerators
Freeze) 9 times, be finally putting into freeze drier dry 48h to get.
The scanning electron microscope (SEM) photograph of compound bone holder material made from embodiment 1 is shown in Fig. 1.
Embodiment 2
Preparation process is substantially the same manner as Example 1, only in addition to hydroxyethyl chitosan, cellulose, two-phase phosphorus calcium in step (2)
Nanometer powder mass ratio is 2:1:1.
The scanning electron microscope (SEM) photograph of compound bone holder material made from embodiment 2 is shown in Fig. 1, wherein Figure 1A is the compound of embodiment 1
Bone holder material, Figure 1B are the compound bone holder material of embodiment 2.
Embodiment 3
Preparation process is substantially the same manner as Example 1, only in addition to hydroxyethyl chitosan, cellulose, two-phase phosphorus calcium in step (2)
Nanometer powder mass ratio is 1:1:2.
The scanning electron microscope (SEM) photograph of compound bone holder material made from embodiment 3 is shown in Fig. 1, wherein Figure 1A is the compound of embodiment 1
Bone holder material, Figure 1B are the compound bone holder material of embodiment 2, and Fig. 1 C are the compound bone holder material of embodiment 3.
Embodiment 4
Preparation process is substantially the same manner as Example 1, only in addition to hydroxyethyl chitosan, cellulose, two-phase phosphorus calcium in step (2)
Nanometer powder mass ratio is 2:1:2.
Embodiment 5
Preparation process is substantially the same manner as Example 1, only in addition to hydroxyethyl chitosan, cellulose, two-phase phosphorus calcium in step (2)
Nanometer powder mass ratio is 2:1:3.
Embodiment 6
Preparation process is substantially the same manner as Example 1, only in addition to hydroxyethyl chitosan, cellulose, two-phase phosphorus calcium in step (2)
Nanometer powder mass ratio is 3:1:3.
Performance test
1. the porosity of the compound bone holder material of testing example 1-6, is as a result shown in Fig. 2.
The porosity of these types of holder has all reached 65% or more as seen from Figure 2, and gap connected ratio meets cell
Attachment proliferation on holder.
2. the compound bone holder material of embodiment 1-6 is carried out cell compatibility detection.
Cell compatibility testing process is as follows：Sample is carried out disinfection with 75% alcohol, it is then dry thin with medulla mesenchyma
Born of the same parents cultivate, and are evaluated using CCK-8.Fig. 3 is the cell compatibility test of compound bone holder material prepared by the present invention
As a result.As seen from Figure 3 cell on holder as time increases, cell survival rate be in increase trend, for preparing answers
Closing bone holder material has good cell compatibility.
Claims (8)
1. a kind of preparation method of the nano combined bone holder material of hydroxyethyl chitosan, which is characterized in that include the following steps：
(1) two-phase phosphorus calcium nanometer powder is prepared：By Ca (NO3)2·4H2O solution and (NH4)2HPO4Solution is according to calcium phosphorus molar ratio
For (1.55~1.70):1 is added in container, is uniformly mixed, and pH is then adjusted to 11, obtains the powder precipitation of white,
After filtering, white powder is cleaned with deionized water, is then dried, roasts, grinds successively, finally crossed 200 mesh sieve, obtain
Two-phase phosphorus calcium nanometer powder；
(2) hydroxyethyl chitosan is prepared：It takes 10g chitosans to be added in the 50wt% sodium hydroxide solutions of 15~18g and carries out alkali
Change, obtain chitosan solution, after standing 1~3h under the conditions of -20 DEG C, 15~25ml solvent isopropanols is added, stirring is warming up to
It 70~80 DEG C, is stirred for being cooled to room temperature, 130~180ml isopropanols is then added and the mixing of 15~19ml ethylene chlorhydrins is molten
Liquid after stirring evenly, is washed using hydrochloric acid solution tune pH7, then using absolute ethyl alcohol, then dialyses, be finally freeze-dried,
Up to hydroxyethyl chitosan；
(3) compound bone holder material is prepared：It takes hydroxyethyl chitosan to be dissolved in the acetum of 1wt%, obtains ethoxy shell
Cellulose and two-phase phosphorus calcium are added into hydroxyethyl chitosan solution by glycan solution, a concentration of 2~5wt% of hydroxyethyl chitosan
Nanometer powder, hydroxyethyl chitosan, cellulose, two-phase phosphorus calcium nanometer powder mass ratio be (1~4):1:(0.5~4), room temperature
Under stir evenly after, then deaerating makes it be uniformly dispersed, and obtained solution, which is then poured into circulating frozen in 24 orifice plates, thaws processing 3
~9 times, be finally putting into freeze drier be dried to get.
2. the preparation method of the nano combined bone holder material of hydroxyethyl chitosan as described in claim 1, which is characterized in that step
Suddenly in (1), Ca (NO3)2·4H2A concentration of 0.4M, (NH of O solution4)2HPO4A concentration of 0.2M of solution.
3. the preparation method of the nano combined bone holder material of hydroxyethyl chitosan as described in claim 1, which is characterized in that step
Suddenly in (1), the roasting carries out in Muffle furnace, is warming up to 1125 DEG C with the rate of 10 DEG C/min first, then keeps 1h, most
Afterwards room temperature is dropped to stove.
4. the preparation method of the nano combined bone holder material of hydroxyethyl chitosan as described in claim 1, which is characterized in that step
Suddenly in (2), the alkalization carries out in -20 DEG C of refrigerators, and alkalization time is more than for 24 hours.
5. the preparation method of the nano combined bone holder material of hydroxyethyl chitosan as described in claim 1, which is characterized in that step
Suddenly in (2), concentration of hydrochloric acid solution 0.1M.
6. the preparation method of the nano combined bone holder material of hydroxyethyl chitosan as described in claim 1, which is characterized in that step
Suddenly in (2), selection of dialysing is bag filter of the molecular cut off in 10000Da, and dialysis time is 2d or more.
7. the preparation method of the nano combined bone holder material of hydroxyethyl chitosan as described in claim 1, which is characterized in that step
Suddenly in (3), the hydroxyethyl chitosan, cellulose, two-phase phosphorus calcium nanometer powder mass ratio be 2:1:1.
8. the preparation method of the nano combined bone holder material of hydroxyethyl chitosan as described in any one of claim 1 to 7, special
Sign is, in step (3), the circulating frozen defrosting processing refers to：Freezing 24 hours in -20 DEG C of refrigerators are put into, are then taken out
Carry out 12 hours of thawing under room temperature, then places into freezing in -20 DEG C of refrigerators.
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| CN201810378947.4A CN108478874B (en) | 2018-04-25 | 2018-04-25 | Preparation method of hydroxyethyl chitosan nano composite bone scaffold material |
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Cited By (2)
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| CN109749094A (en) * | 2018-12-07 | 2019-05-14 | 信阳师范学院 | A kind of bifunctional microgel PNIPAM/HECs/D-cGO and the preparation method and application thereof |
| CN110327493A (en) * | 2019-08-07 | 2019-10-15 | 重庆市畜牧科学院 | A kind of compound bio bracket and preparation method |
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| CN109749094A (en) * | 2018-12-07 | 2019-05-14 | 信阳师范学院 | A kind of bifunctional microgel PNIPAM/HECs/D-cGO and the preparation method and application thereof |
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