CN102475917A - Preparation method of injectable bracket material used for bone tissue engineering - Google Patents
Preparation method of injectable bracket material used for bone tissue engineering Download PDFInfo
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- CN102475917A CN102475917A CN2010105536159A CN201010553615A CN102475917A CN 102475917 A CN102475917 A CN 102475917A CN 2010105536159 A CN2010105536159 A CN 2010105536159A CN 201010553615 A CN201010553615 A CN 201010553615A CN 102475917 A CN102475917 A CN 102475917A
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- chitosan
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- calcium phosphate
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- bone tissue
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Abstract
The invention discloses a preparation method of an injectable bracket material used for the bone tissue engineering, belonging to a bone tissue engineering material technology in the biomedical engineering field. The method is characterized in that the injectable bracket material is prepared by chitosan hydrochloric acidification and lyophobic interaction of molecules in a solution by a ternary dispersoid composed of hydrochloric acidification chitosan, beta-glycerophosphate and calcium phosphate. The preparation method is characterized in that a prepared thermo-responsive injectable chitosan/ calcium phosphate sol conversion system realizes sol-gel conversion thermosensitivity by regulating interaction with water molecules. The optimization of components ensures that the components are suitable for injection application and are formed in situ in a body, and meanwhile, the bionic degree of the injectable bracket material to a tone tissue is enhanced. Therefore, the injectable bracket material is more suitable for bone reconstruction and is more convenient for clinic application.
Description
Technical field
The invention belongs to the engineering material of bone tissue technology of biomedical engineering field, the concrete method for preparing of a kind of bone tissue engineer that relate to the injectable type timbering material.
Background technology
Bone is the Biocomposite material that collagen fiber form through hydroxyapatite.Osseous tissue is through the osteoclast metabolism, and osteoblast is rebuild and constituted a kind of dynamic equilibrium system.The through engineering approaches bone utilizes seed cell to be planted in porous support exactly, behind amplification in vitro, cell/supporting structure thing is implanted, with repairing bone defect; Also can adopt the good biomaterial of biocompatibility to implant, bone rebuild through bioprocess.Support is molding in advance, and also original position forms in vivo.The linear polysaccharide that chitosan is made up of the D-glycosamine residue and the random N-acetyl-glucosamine of number different distributions of β (1 → 4) bonding, the N-acetyl-glucosamine in its structure also is present in the glycosaminoglycans (GAGs).Because chitosan structure is similar to GAGS, so have the multi-biological activity.It is the lysozyme degraded, and its biocompatibility is good.Chitosan-modified gathering (L, D-lactic acid) can be improved material to osteoblastic affinity.Chenite A etc. prepare chitosan/glycerophosphate water solublity constituent with glycerophosphate regulation and control chitosan and hydrone interphase interaction, and it can implant by injection system, gelatinizing-in-situ.
Summary of the invention
The present invention is directed to above-mentioned prior art state of development, the method for preparing of a kind of bone tissue engineer with the injectable type timbering material is provided.The timbering material of preparation original position formation in vivo according to said method, its clinical practice is convenient.
For achieving the above object, the present invention realizes through following technical proposals.Comprise with chitosan hydrochloride (CH) and β-phosphoglycerol disodium (GP) and calcium phosphate (TCP) forming the ternary dispersion, prepare the method for injectable type chitosan hydrochloride/calcium phosphate bone repair materials.It is characterized in that adopting molecular weight is 15-30 ten thousand; It is raw material that degree of deacetylation reaches the above chitosan of 85 %, after being dissolved in 0.5-lwt % acetic acid aqueous solution and filtering, dialyses with 0.2mol/L NaCl aqueous solution; The dialysis of reuse deionized water, the chitosan hydrochloride of obtained by freeze drying; Press 10 with β-phosphoglycerol disodium with calcium phosphate again: 3.0-3.5: the preparation of 5-15 mixed forms ternary solution; 30-39 ℃ of sol-gel transition temperature range; PH scope 7.00-7.40 obtains the sol gel transformsystem of injectable type chitosan hydrochloride/calcium phosphate.Above-mentioned chitosan comprises the methyl chitosan of completing of chitosan derivatives, phosphonized chitosan; Above-mentioned calcium phosphate comprises bata-tricalcium phosphate, type alpha tricalcium phosphate, hydroxyapatite bioactive pottery.
The invention has the advantages that the employing chitosan hydrochloride; The temperature-responsive injectable type chitosan/calcium phosphate sol gel transformsystem of β-phosphoglycerol disodium and type alpha tricalcium phosphate preparation; Through regulation and control and hydrone interphase interaction, realize temperature sensitiveization of sol-gel transition.Composite body inner gel characteristic is adjustable, can adapt to the damaged reparation of difformity bone.Optimizing of components makes it be suitable for infusion application, and according to said method the timbering material of preparation can form through being injected at internal in-situ, not only kept the chitosan/biocompatibility of calcium phosphate biomimetic biomaterial capable, biological degradability, and its clinical practice is convenient.Chitosan structure is similar to GAGs; So have the multi-biological activity; And calcium phosphate is the bone inorganic constituents, in chitosan hydrochloride solution, has added the sol gel transformsystem that calcium phosphate forms, and belongs to particle enhancement mode gel; Strengthen its bionical degree simultaneously, make this injection-type timbering material be more suitable for rebuilding in bone to osseous tissue.
Description of drawings
Accompanying drawing is the temperature dependency of chitosan hydrochloride-β of the present invention-phosphoglycerol disodium salt (CH-GP) and chitosan hydrochloride-β-phosphoglycerol disodium salt-bata-tricalcium phosphate (CH-GP-TCP) sol-gel transition time.
The specific embodiment
Embodiment one:
With molecular weight 200,000, the chitosan of deacetylation 87 % is dissolved in the acetic acid aqueous solution of 1 %, after the chitosan solution of formation lwt % filters; Dialyse with 0.2mol/L NaCl aqueous solution; The dialysis of reuse deionized water, obtained by freeze drying chitosan hydrochloride, productive rate 92 %.Confirm the primary amino radical (1599cm on the chitosan molecule pyranoid ring by infrared spectrum
-l) the characteristic peak disappearance, and be transformed into season by salt.The X-diffraction pattern shows that chitosan degree of crystallinity obviously reduces, and forms unformed shape.Chitosan hydrochloride is added in the 2T3 osteoblast culture medium by 1-20mg/mL concentration, and the MTT experimental result shows that it is good to osteoblastic affinity.
Embodiment two
Get the chitosan hydrochloride of the embodiment one method preparation of 0.10g; Add the 4.50mL deionized water, dissolving is cooled to 4 ℃ fully; Dropwise add 0.70g/ mL β-phosphoglycerol disodium salt solution 500uL under the stirring state; Add the 010g bata-tricalcium phosphate again, fully vibration makes its uniform mixing, forms the sol-gel conversion system.This system pH value is 7.15,37 ℃ following 10 minutes be that cocoa changes into gel, can stablize preservation under 4 ℃.
Embodiment three
Get the chitosan hydrochloride of the embodiment one method preparation of 0.10g; Add the 4.50mg deionized water, dissolving is cooled to 4 ℃ fully; Dropwise add 0.60g/mL β-phosphoglycerol disodium salt solution 500mL under the stirring state; Add the 0.10g bata-tricalcium phosphate again, fully vibration makes its uniform mixing, forms the sol-gel conversion system.This system pH value is 7.01, can change into sol system in following 13 minutes at 37 ℃, can stablize preservation under 4 ℃.
Embodiment four:
Prepare (the wherein chitosan hydrochloride Y x radiation x sterilization that obtains of bata-tricalcium phosphate and embodiment one of chitosan hydrochloride-β-phosphoglycerol disodium salt-bata-tricalcium phosphate dispersion in the ratio of embodiment two; β-phosphoglycerol disodium salt solution is through the syringe needle filter filtration sterilization of 0.20 μ m) pass through injector to inject to 2 monthly age Wistar subcutaneous rat; Draw materials after one month; The neutral formalin solution of 10 % is fixed, specimens paraffin embedding slices, Histological method's dyeing.It is good that optical microscope is observed biocompatibility down.
Claims (2)
1. a bone tissue engineer is with the method for preparing of injectable type timbering material, and it is characterized in that: its preparation method may further comprise the steps:
(1) be 15-30 ten thousand with molecular weight; Degree of deacetylation reaches the above chitosan of 85 %, is dissolved in mass ratio 0.5-1.0% acetic acid aqueous solution, filters the back with 0.2 mol/L NaCl aqueous solution dialysis; The dialysis of reuse deionized water, the chitosan hydrochloride of obtained by freeze drying;
(2) above-mentioned chitosan hydrochloride and β-phosphoglycerol disodium and press 10:3.0-3.5:5-15 mixed preparation formation ternary solution with calcium phosphate; 30-39 ℃ of sol-gel transition temperature range; PH scope 7.00-7.40 obtains the sol gel transformsystem of injectable type chitosan hydrochloride/calcium phosphate.
2. according to the method for preparing of the described a kind of bone tissue engineer of claim 1 with the injectable type timbering material, it is characterized in that: the chitosan of employing can be the derivant carboxymethyl chitosan or the phosphonized chitosan of chitosan; Said calcium phosphate can be bata-tricalcium phosphate, type alpha tricalcium phosphate or hydroxyapatite bioactive pottery.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2010105536159A CN102475917A (en) | 2010-11-22 | 2010-11-22 | Preparation method of injectable bracket material used for bone tissue engineering |
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| CN2010105536159A CN102475917A (en) | 2010-11-22 | 2010-11-22 | Preparation method of injectable bracket material used for bone tissue engineering |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103599567A (en) * | 2013-11-25 | 2014-02-26 | 四川大学华西医院 | Temperature-sensitive composite as well as preparation method and application thereof |
| CN104906637A (en) * | 2015-05-15 | 2015-09-16 | 武汉大学 | Injectable-porous-drug loaded polymethyl methacrylate-based composite scaffold bone transplant material and preparation method thereof |
| CN106421929A (en) * | 2016-09-22 | 2017-02-22 | 四川大学 | Injectable calcium phosphate/natural polymer composite material and preparation method and application thereof |
| CN111375067A (en) * | 2020-03-18 | 2020-07-07 | 四川大学 | Chitosan scaffold and preparation method and application thereof |
-
2010
- 2010-11-22 CN CN2010105536159A patent/CN102475917A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103599567A (en) * | 2013-11-25 | 2014-02-26 | 四川大学华西医院 | Temperature-sensitive composite as well as preparation method and application thereof |
| CN103599567B (en) * | 2013-11-25 | 2015-06-03 | 四川大学华西医院 | Temperature-sensitive composite as well as preparation method and application thereof |
| CN104906637A (en) * | 2015-05-15 | 2015-09-16 | 武汉大学 | Injectable-porous-drug loaded polymethyl methacrylate-based composite scaffold bone transplant material and preparation method thereof |
| CN104906637B (en) * | 2015-05-15 | 2018-01-12 | 武汉大学 | Porous polymethyl methacrylate base compound rest bone grafting material for carrying medicine of a kind of injectable and preparation method thereof |
| CN106421929A (en) * | 2016-09-22 | 2017-02-22 | 四川大学 | Injectable calcium phosphate/natural polymer composite material and preparation method and application thereof |
| CN106421929B (en) * | 2016-09-22 | 2019-11-15 | 四川大学 | A kind of injectable calcium phosphate/natural high molecular composite material and its preparation method and application |
| CN111375067A (en) * | 2020-03-18 | 2020-07-07 | 四川大学 | Chitosan scaffold and preparation method and application thereof |
| CN111375067B (en) * | 2020-03-18 | 2022-02-11 | 四川大学 | Chitosan scaffold and preparation method and application thereof |
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Application publication date: 20120530 |