CN101716331B - Method for embedding protein biological macromolecules by using amylopectin-p-nitrocinnamic acid graft in situ - Google Patents
Method for embedding protein biological macromolecules by using amylopectin-p-nitrocinnamic acid graft in situ Download PDFInfo
- Publication number
- CN101716331B CN101716331B CN2009102139408A CN200910213940A CN101716331B CN 101716331 B CN101716331 B CN 101716331B CN 2009102139408 A CN2009102139408 A CN 2009102139408A CN 200910213940 A CN200910213940 A CN 200910213940A CN 101716331 B CN101716331 B CN 101716331B
- Authority
- CN
- China
- Prior art keywords
- amylopectin
- nitrocinnamic
- graft
- protide
- situ
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Medicinal Preparation (AREA)
- Materials For Medical Uses (AREA)
Abstract
The invention discloses a method for embedding protein biological macromolecules by using amylopectin-p-nitrocinnamic acid graft in situ. The method comprises the steps of: evenly mixing the amylopectin-p-nitrocinnamic acid graft solution with the mass percent concentration of 5-20% and the protein biological macromolecule solution with the mass percent concentration of 4-8%; pouring the mixed solution into a mould; and under the illumination of a UV lamp, obtaining amylopectin gel embedded with the protein biological macromolecules in situ. The method takes p-nitrocinnamic acid as photoreactive group which is crosslinked by UV light, so that the amylopectin gel embedded with the protein biological macromolecules in situ can be prepared. The method does not relate to other chemical cross-linking agent and photoinitiator, has the advantages of simple preparation technique, easy control, mild reaction and room temperature formation, takes amylopectin as main raw material, and is wide in the material source. Furthermore, the obtained amylopectin gel has excellent biocompatibility and biodegradability, so that the method can be expected to be applied to the field of biomedical engineering.
Description
Technical field
The invention belongs to the bio-medical engineering material field, particularly a kind of amylopectin-of utilizing to the method for nitrocinnamic graft embedding in situ protide biomacromolecule.
Background technology
In recent years, photocrosslinkable hydrogel has obtained in fields such as medicine controlled releasing and organizational projects to use widely as biomaterial.Compare with other crosslinking method; The photo-crosslinking method prepares hydrogel and has following advantage (Amsden B G; Sukarto A, Knight D K and Shapka S N.Biomacromolecules, 2007; 8:3758-3766): (1) can make the polymer precursor aqueous solution in-situ cross-linked, thereby can be used for preparing injection aquagel; (2) rapid polymerization forms hydrogel under room temperature or physiological temp; (3) the product geometry is easy to control; (4) produce lower heat etc. in the in-situ polymerization process.These advantages make photocrosslinkable hydrogel enjoy favor aspect the embedding of protide biopharmaceutical macromolecular drug and the sustained release.
The photocrosslinkable hydrogel that is used for protide biopharmaceutical macromolecular drug controlled release carrier; Can be divided into synthetic high polymer class photocrosslinkable hydrogel and natural polymer subclass photocrosslinkable hydrogel according to its primary raw material source, general optical active group is the two keys in the macromonomer terminal acrylate.The synthetic presoma of common chemical has Polyethylene Glycol (methyl) acrylate derivative, Polyethylene Glycol polylactide block copolymer (methyl) acrylate derivative etc.People such as Elisseeff (Elisseeff J, Anseth K, Langer R.Plastic andReconstructive Surgery, 1999,104:1014-1022) utilize the transdermal light polymerization technique of injection aquagel, realized that the dystopy of cartilage forms; They are suspended in the cattle articular chondrocytes in the aqueous solution of Polyethylene Glycol double methacrylate and Polyethylene Glycol, then with its subcutaneous injection in the Mus back, polymerization under external light source irradiation is taken out after week through 4-7, finds to have newborn type cartilage structure to form.Liu etc. (Liu C C, Sawicki S M, Metters A T.Biomacromolecules, 2008, be big monomer 9:75-83), in the presence of light trigger, through photo-crosslinking embedding in situ lysozyme with the Polyethylene Glycol double methacrylate.But this type chemosynthesis presoma does not possess favorable biological degradability, and its application receives certain restriction.
Compare with synthetic high polymer class hydrogel, natural polymer subclass photocrosslinkable hydrogel has advantages such as nontoxic easy degraded, has more development potentiality as protide biopharmaceutical macromolecular drug carrier.(LeachaJ B such as Leacha; Schmidt C E.Biomaterials; 2005,26:125-135) hyaluronic acid and GMA reaction are introduced two keys, mix with bovine serum albumen solution again; Effect at light trigger issues third contact of a total solar or lunar eclipse cross-linking reaction, the embedding in situ bovine serum albumin.(Jeon O, Bouhadir K H, Mansour J M such as Jeon; Alsberg E.Biomaterials; 2009, be light trigger 30:2724-2734) with Irgacure 2959, the methacrylate alginate is mixed with the cattle chondrocyte; Polymerization under the ultraviolet lighting of 365nm, embedding in situ cattle chondrocyte.Dyeing and MTT experiment through to work/dead cell find that the cattle chondrocyte of embedding in situ in photo-crosslinking alginate gel has certain biological activity.But, all having used certain Cytotoxic light trigger when relevant synthetic, and in the photopolymerization process, light trigger maybe induced protein class biomacromolecule generation radical polymerization and cause the protide biomacromolecule to lose activity.Therefore, how not adding light trigger condition diarrhea, just become an important topic in bio-medical engineering material field with the natural polymer subclass hydrogel that the preparation of light crosslinking technological is suitable for.
Summary of the invention
In order to overcome the shortcoming and defect of prior art, it is a kind of in room temperature with do not add under the light trigger condition that primary and foremost purpose of the present invention is to provide, and utilizes amylopectin-to the method for nitrocinnamic graft embedding in situ protide biomacromolecule.
Another object of the present invention is to the amylopectin gel that provides a kind of said method to obtain.
A purpose more of the present invention is to provide the application of above-mentioned amylopectin gel.
The object of the invention is realized through following technical proposals: a kind of amylopectin-to the method for nitrocinnamic graft embedding in situ protide biomacromolecule, comprise following operating procedure of utilizing:
(1) under 60～90 ℃ of conditions, the 1g amylopectin is dissolved in 20～30mL anhydrous dimethyl sulphoxide; Cool the temperature to 40～50 ℃, add 0.23～0.32gN, N '-dicyclohexylcarbodiimide and 0.07～0.09g 4-dimethylamino naphthyridine obtain amylopectin solution after the dissolving fully; 0.22～0.30g is dissolved in 5～10mL dimethyl sulfoxide to nitrocinnamic, obtains nitrocinnamic solution; To be added in the amylopectin solution lucifuge reaction to the nitrocinnamic drips of solution; After reaction finishes, remove by filter precipitate, will filtrate with ethanol precipitation, washing, drying obtains amylopectin-to the nitrocinnamic graft;
(2) be 5～20% amylopectin-with step (1) gained amylopectin-nitrocinnamic graft is mixed with mass percent concentration to nitrocinnamic graft solution; Be 4～8% protide biological macromolecule solns mix homogeneously then with mass percent concentration, obtain mixed solution; Mixed solution is poured in the mould, and illumination 5～50min under uviol lamp obtains the amylopectin gel of embedding in situ protide biomacromolecule.
The temperature of the said lucifuge reaction of step (1) is 40～50 ℃, and the response time is 24h.
The said drying of step (1) is a vacuum drying, and baking temperature is 40～50 ℃, and be 48h drying time.
The said amylopectin of step (2)-to nitrocinnamic graft solution is with the amylopectin-nitrocinnamic graft is dissolved in deionized water, buffer reagent or the normal saline; Said buffer reagent is phosphate, carbonate or citrate.
Protide biomacromolecule and amylopectin in the said mixed solution of step (2)-to the mass ratio of nitrocinnamic graft are 1: 10～1: 100.
The said protide biomacromolecule of step (2) is bovine serum albumin, egg white powder, lysozyme, horseradish peroxidase or trypsin; Said protide biological macromolecule solns is that the protide biomacromolecule is dissolved in deionized water, buffer reagent or the normal saline; Said buffer reagent is phosphate, carbonate or citrate.
The said mould of step (2) is politef mould, glass mold or silicon rubber mould.
The wavelength of the said uviol lamp of step (2) is 365nm, and power is 4～15W, and light intensity is 0.4～1mW/cm
2
A kind of amylopectin gel according to method for preparing.
Above-mentioned amylopectin gel can be used for preparing injectable protide biopharmaceutical macromolecular drug carrier or oral protein class biopharmaceutical macromolecular drug carrier.
Principle of the present invention is: the present invention is a basic raw material with the amylopectin with good biological degraded and biocompatibility; At first synthesized amylopectin-to the nitrocinnamic graft; Being photoreactive group to nitrocinnamic; Through the ultraviolet light cross-linking photoreactive group, the graft mixed solution forms the hydrogel of embedding protide biopharmaceutical macromolecular drug fast; The present invention is through control amylopectin-to the concentration and the light application time of nitrocinnamic graft; In room temperature with do not add under the light trigger condition, preparation series is biodegradable and have an amylopectin gel that is loaded with the protide biopharmaceutical macromolecular drug of good biocompatibility.
The present invention compared with prior art; Have following advantage and beneficial effect: (1) the present invention is being photoreactive group to nitrocinnamic; Adopt optical cross-linking method to prepare the amylopectin gel of embedding in situ protide biomacromolecule; Do not relate to other chemical cross-linking agent and light trigger, preparation technology is simple, and control easily; (2) reaction condition is gentle, at room temperature forms gel fast, can effectively avoid protide biomacromolecule inactivation; (3) gelation process carries out in aqueous medium; (4) intensity of gel can be through the amylopectin-concentration and the light application time of nitrocinnamic graft are regulated and control; (5) primary raw material that is adopted is an amylopectin, and raw material sources are extensive, and formed hydrogel has favorable biological degradability and biocompatibility.
Description of drawings
Fig. 1 is the cumulative in vitro releasing curve diagram of bovine serum albumin in the embedding in situ bovine serum albumin amylopectin gel.
Fig. 2 is the circular dichroism spectrogram of natural bovine serum albumin and embedding bovine serum albumin.
Fig. 3 is the cumulative in vitro releasing curve diagram of lysozyme in the embedding in situ lysozyme amylopectin gel.
Fig. 4 is the circular dichroism spectrogram of natural lysozyme and embedding lysozyme.
The specific embodiment
Below in conjunction with embodiment and accompanying drawing the present invention is described in further detail, but embodiment of the present invention is not limited thereto.
Embodiment 1
Amylopectin-to the preparation of nitrocinnamic graft: under 70 ℃ of conditions, the 1g amylopectin is dissolved in the 20mL anhydrous dimethyl sulphoxide; Reduce temperature to 40 ℃, add 0.25gN, N '-dicyclohexylcarbodiimide and 0.074g 4-dimethylamino naphthyridine obtain amylopectin solution after the stirring and dissolving; 0.24g is dissolved in the 10mL dimethyl sulfoxide to nitrocinnamic, obtains nitrocinnamic solution; To be added in the amylopectin solution lucifuge reaction 24h under 40 ℃ of conditions to the nitrocinnamic drips of solution; After reaction finishes; Remove by filter precipitate; To filtrate and dropwise be added drop-wise in 6 times of volume of ethanol; To with the UV, visible light sub-ray spectrometer detection absworption peak less than cinnamic acid, 40 ℃ of vacuum drying 48h then obtain amylopectin-to the nitrocinnamic graft to the deposition that filtration obtains with the ethanol cyclic washing.Record this amylopectin-in the nitrocinnamic graft with ultraviolet spectroscopy, containing nitrocinnamic ester group number on per 100 sugar rings is 1.6.
Embodiment 2
Amylopectin-to the preparation of nitrocinnamic graft: under 60 ℃ of conditions, the 1g amylopectin is dissolved in the 25mL anhydrous dimethyl sulphoxide; Reduce temperature to 50 ℃, add 0.23g N, N '-dicyclohexylcarbodiimide and 0.074g 4-dimethylamino naphthyridine obtain amylopectin solution after the stirring and dissolving; 0.22g is dissolved in the 5mL dimethyl sulfoxide to nitrocinnamic, obtains nitrocinnamic solution; To be added in the amylopectin solution lucifuge reaction 24h under 50 ℃ of conditions to the nitrocinnamic drips of solution; After reaction finishes; Remove by filter precipitate; To filtrate and dropwise be added drop-wise in 6 times of volume of ethanol; To with the UV, visible light sub-ray spectrometer detection absworption peak less than cinnamic acid, 45 ℃ of vacuum drying 48h then obtain amylopectin-to the nitrocinnamic graft to the deposition that filtration obtains with the ethanol cyclic washing.Record this amylopectin-in the nitrocinnamic graft with ultraviolet spectroscopy, containing nitrocinnamic ester group number on per 100 sugar rings is 1.5.
Embodiment 3
Amylopectin-to the preparation of nitrocinnamic graft: under 90 ℃ of conditions, the 1g amylopectin is dissolved in the 30mL anhydrous dimethyl sulphoxide; Reduce temperature to 45 ℃, add 0.32g N, N '-dicyclohexylcarbodiimide and 0.09g 4-dimethylamino naphthyridine obtain amylopectin solution after the stirring and dissolving; 0.30g is dissolved in the 8mL dimethyl sulfoxide to nitrocinnamic, obtains nitrocinnamic solution; To be added in the amylopectin solution lucifuge reaction 24h under 45 ℃ of conditions to the nitrocinnamic drips of solution; After reaction finishes; Remove by filter precipitate; To filtrate and dropwise be added drop-wise in 6 times of volume of ethanol; To with the UV, visible light sub-ray spectrometer detection absworption peak less than cinnamic acid, 50 ℃ of vacuum drying 48h then obtain amylopectin-to the nitrocinnamic graft to the deposition that filtration obtains with the ethanol cyclic washing.Record this amylopectin-in the nitrocinnamic graft with ultraviolet spectroscopy, containing nitrocinnamic ester group number on per 100 sugar rings is 1.9.
Embodiment 4
With the embodiment 1 gained amylopectin-nitrocinnamic graft is mixed with the 1mL mass percent concentration is 10% amylopectin-to the PBS of nitrocinnamic graft; Then to wherein adding the PBS that 25 μ L mass percent concentrations are 4% bovine serum albumin; Fully mixing obtains mixed solution; Mixed solution is poured in the politef mould into (die size is 20mm * 20mm * 1mm), is that (power of uviol lamp is 6W, and light intensity is 0.6mW/cm for the uviol lamp of 365nm at wavelength
2) descend illumination 10min, obtain the amylopectin gel of embedding in situ bovine serum albumin.
The amylopectin gel of the embedding in situ bovine serum albumin that makes is placed the PBS of 10mL; In 37 ℃ shaking baths (concussion speed is 50 rev/mins), carry out vitro drug release; The content that discharges bovine serum albumin in the liquid adopts the Coomassie brilliant blue method to measure, and the release liquid with 24h carries out the circular dichroism spectra analysis again.This amylopectin gel that is loaded with bovine serum albumin continues to discharge in PBS, and no tangible initial stage burst release (as shown in Figure 1) in dispose procedure.
Through the circular dichroism spectrometry conformation of bovine serum albumin before and after the embedding is measured; The circular dichroism spectrogram of the natural bovine serum albumin buffer solution of (●) expression among Fig. 2, (zero) expression embedding in situ bovine serum albumin amylopectin gel of the present invention discharges the circular dichroism spectrogram of the release liquid of 24h.As can beappreciated from fig. 2, bovine serum albumin is behind the amylopectin gel embedding of photo-crosslinking, and its circular dichroism spectrogram is consistent with the spectrogram of natural bovine serum albumin, bovine serum albumin is described in the process of embedding and release, and its conformation has obtained keeping preferably.
Embodiment 5
With the embodiment 1 gained amylopectin-nitrocinnamic graft is mixed with the 1mL mass percent concentration is 10% amylopectin-to the carbonate buffer solution of nitrocinnamic graft; Then to wherein adding the carbonate buffer solution that 50 μ L mass percent concentrations are 4% bovine serum albumin; Fully mixing obtains mixed solution; Mixed solution is poured in the politef mould into (die size is 20mm * 20mm * 1mm), is that (power of uviol lamp is 8W, and light intensity is 0.6mW/cm for the uviol lamp of 365nm at wavelength
2) descend illumination 20min, obtain the amylopectin gel of embedding in situ bovine serum albumin.
The amylopectin gel of the embedding in situ bovine serum albumin that makes is placed the PBS of 10mL; In 37 ℃ shaking baths (concussion speed is 50 rev/mins), carry out vitro drug release, the content that discharges bovine serum albumin in the liquid adopts the Coomassie brilliant blue method to measure.The amylopectin gel of this embedding bovine serum albumin continues to discharge in PBS, and no tangible initial stage burst release in dispose procedure.
Embodiment 6
With the embodiment 1 gained amylopectin-nitrocinnamic graft is mixed with the 1mL mass percent concentration is 5% amylopectin-to the citrate buffer solution of nitrocinnamic graft; Then to wherein adding the citrate buffer solution that 25 μ L mass percent concentrations are 4% bovine serum albumin; Fully mixing obtains mixed solution; Mixed solution is poured in the politef mould into (die size is 20mm * 20mm * 1mm), is that (power of uviol lamp is 4W, and light intensity is 0.4mW/cm for the uviol lamp of 365nm at wavelength
2) descend illumination 50min, obtain the amylopectin gel of embedding in situ bovine serum albumin.
The amylopectin gel of the embedding in situ bovine serum albumin that makes is placed the PBS of 10mL; In 37 ℃ shaking baths (concussion speed is 50 rev/mins), carry out vitro drug release, the content that discharges bovine serum albumin in the liquid adopts the Coomassie brilliant blue method to measure.The amylopectin gel of this embedding bovine serum albumin continues to discharge in PBS, and no tangible initial stage burst release in dispose procedure.
Embodiment 7
With the embodiment 1 gained amylopectin-nitrocinnamic graft is mixed with the 1mL mass percent concentration is 15% amylopectin-to nitrocinnamic graft aqueous solution; Be 4% bovine serum albumin aqueous solution to wherein adding 50 μ L mass percent concentrations then; Fully mixing obtains mixed solution; Mixed solution is poured in the politef mould into (die size is 20mm * 20mm * 1mm), is that (power of uviol lamp is 15W, and light intensity is 1mW/cm for the uviol lamp of 365nm at wavelength
2) descend illumination 5min, obtain the amylopectin gel of embedding in situ bovine serum albumin.
The amylopectin gel of the embedding in situ bovine serum albumin that makes is placed the PBS of 10mL; In 37 ℃ shaking baths (concussion speed is 50 rev/mins), carry out vitro drug release, the content that discharges bovine serum albumin in the liquid adopts the Coomassie brilliant blue method to measure.The amylopectin gel of this embedding bovine serum albumin continues to discharge in PBS, and no tangible initial stage burst release in dispose procedure.
Embodiment 8
With the embodiment 1 gained amylopectin-nitrocinnamic graft is mixed with the 1mL mass percent concentration is 10% amylopectin-to the normal saline solution of nitrocinnamic graft; Then to wherein adding the normal saline solution that 50 μ L mass percent concentrations are 8% lysozyme; Fully mixing obtains mixed solution; Mixed solution is poured in the silicon rubber mould into (die size is 20mm * 20mm * 1mm), is that (power of uviol lamp is 6W, and light intensity is 0.6mW/cm for the uviol lamp of 365nm at wavelength
2) descend illumination 10min, obtain the amylopectin gel of embedding in situ lysozyme.
The amylopectin gel of the embedding in situ lysozyme that makes is placed the PBS of 10mL; In 37 ℃ shaking baths (concussion speed is 50 rev/mins), carry out vitro drug release; The content that discharges lysozyme in the liquid adopts the Coomassie brilliant blue method to measure, and the release liquid with 24h carries out the circular dichroism spectra analysis again.The amylopectin gel of this embedding lysozyme continues to discharge in PBS, and no tangible initial stage burst release (as shown in Figure 3) in dispose procedure.Compare with the release in vitro situation of bovine serum albumin among the embodiment 2, the rate of release of lysozyme is faster, and the cumulative release amount of 48h is higher.
Through the circular dichroism spectrometry conformation of lysozyme before and after the embedding is measured, the circular dichroism spectrogram of the natural lysozyme buffer solution of (●) expression among Fig. 4, (zero) expression embedding in situ lysozyme amylopectin gel of the present invention discharges the circular dichroism spectrogram of the release liquid of 24h.As can beappreciated from fig. 4, lysozyme is behind the amylopectin gel embedding of photo-crosslinking, and its circular dichroism spectrogram is consistent with the spectrogram of natural lysozyme, lysozyme is described in the process of embedding and release, and its conformation has obtained keeping preferably.
Embodiment 9
With the embodiment 1 gained amylopectin-nitrocinnamic graft is mixed with the 1mL mass percent concentration is 10% amylopectin-to the PBS of nitrocinnamic graft; Then to wherein adding the PBS that 100 μ L mass percent concentrations are 8% lysozyme; Fully mixing obtains mixed solution; Mixed solution is poured in the glass mold into (die size is 20mm * 20mm * 1mm), is that (power of uviol lamp is 12W, and light intensity is 0.8mW/cm for the uviol lamp of 365nm at wavelength
2) descend illumination 20min, obtain the amylopectin gel of embedding in situ lysozyme.
The amylopectin gel of the embedding in situ lysozyme that makes is placed the PBS of 10mL; In 37 ℃ shaking baths (concussion speed is 50 rev/mins), carry out vitro drug release, the content that discharges lysozyme in the liquid adopts the Coomassie brilliant blue method to measure.The amylopectin gel of this embedding lysozyme continues to discharge in PBS, and no tangible initial stage burst release in dispose procedure.
The foregoing description is a preferred implementation of the present invention; But embodiment of the present invention is not restricted to the described embodiments; Other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; All should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. one kind is utilized amylopectin-to the method for nitrocinnamic graft embedding in situ protide biomacromolecule, it is characterized in that comprising following operating procedure:
(1) under 60～90 ℃ of conditions, the 1g amylopectin is dissolved in 20～30mL anhydrous dimethyl sulphoxide; Cool the temperature to 40～50 ℃, add 0.23～0.32g N, N '-dicyclohexylcarbodiimide and 0.07～0.09g 4-dimethylamino naphthyridine obtain amylopectin solution after the dissolving fully; 0.22～0.30g is dissolved in 5～10mL dimethyl sulfoxide to nitrocinnamic, obtains nitrocinnamic solution; To be added in the amylopectin solution lucifuge reaction to the nitrocinnamic drips of solution; After reaction finishes, remove by filter precipitate, will filtrate with ethanol precipitation, washing, drying obtains amylopectin-to the nitrocinnamic graft;
(2) be 5～20% amylopectin-with step (1) gained amylopectin-nitrocinnamic graft is mixed with mass percent concentration to nitrocinnamic graft solution; Be 4～8% protide biological macromolecule solns mix homogeneously then with mass percent concentration, obtain mixed solution; Mixed solution is poured in the mould, and illumination 5～50min under uviol lamp obtains the amylopectin gel of embedding in situ protide biomacromolecule.
2. a kind of amylopectin-to the method for nitrocinnamic graft embedding in situ protide biomacromolecule, it is characterized in that of utilizing according to claim 1: the temperature of the said lucifuge reaction of step (1) is 40～50 ℃, and the response time is 24h.
3. a kind of amylopectin-to the method for nitrocinnamic graft embedding in situ protide biomacromolecule, it is characterized in that of utilizing according to claim 1: the said drying of step (1) is a vacuum drying, and baking temperature is 40～50 ℃, and be 48h drying time.
4. a kind of amylopectin-to the method for nitrocinnamic graft embedding in situ protide biomacromolecule, it is characterized in that of utilizing according to claim 1: the said amylopectin of step (2)-to nitrocinnamic graft solution is with the amylopectin-nitrocinnamic graft is dissolved in deionized water, buffer reagent solution or the normal saline; Said buffer reagent solution is phosphate, carbonate or citrate.
5. a kind of amylopectin-to the method for nitrocinnamic graft embedding in situ protide biomacromolecule, it is characterized in that of utilizing according to claim 1: protide biomacromolecule and amylopectin in the said mixed solution of step (2)-to the mass ratio of nitrocinnamic graft are 1: 10～1: 100.
6. a kind of amylopectin-to the method for nitrocinnamic graft embedding in situ protide biomacromolecule, it is characterized in that of utilizing according to claim 1: the said protide biomacromolecule of step (2) is bovine serum albumin, egg white powder, lysozyme, horseradish peroxidase or trypsin; Said protide biological macromolecule solns is that the protide biomacromolecule is dissolved in deionized water, buffer reagent solution or the normal saline; Said buffer reagent solution is phosphate, carbonate or citrate.
7. a kind of amylopectin-to the method for nitrocinnamic graft embedding in situ protide biomacromolecule, it is characterized in that of utilizing according to claim 1: the said mould of step (2) is politef mould, glass mold or silicon rubber mould.
8. a kind of amylopectin-of utilizing according to claim 1 to the method for nitrocinnamic graft embedding in situ protide biomacromolecule; It is characterized in that: the wavelength of the said uviol lamp of step (2) is 365nm; Power is 4～15W, and light intensity is 0.4～1mW/cm
2
9. amylopectin gel according to each said method preparation of claim 1～8.
10. the application of amylopectin gel according to claim 9 in preparation injectable protide biopharmaceutical macromolecular drug carrier or oral protein class biopharmaceutical macromolecular drug carrier.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102139408A CN101716331B (en) | 2009-12-16 | 2009-12-16 | Method for embedding protein biological macromolecules by using amylopectin-p-nitrocinnamic acid graft in situ |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102139408A CN101716331B (en) | 2009-12-16 | 2009-12-16 | Method for embedding protein biological macromolecules by using amylopectin-p-nitrocinnamic acid graft in situ |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101716331A CN101716331A (en) | 2010-06-02 |
| CN101716331B true CN101716331B (en) | 2012-03-07 |
Family
ID=42431028
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009102139408A Expired - Fee Related CN101716331B (en) | 2009-12-16 | 2009-12-16 | Method for embedding protein biological macromolecules by using amylopectin-p-nitrocinnamic acid graft in situ |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101716331B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1207744A (en) * | 1995-11-15 | 1999-02-10 | 生化学株式会社 | Photocured cross-linked-hyaluronic acid gel and method of preparation thereof |
-
2009
- 2009-12-16 CN CN2009102139408A patent/CN101716331B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1207744A (en) * | 1995-11-15 | 1999-02-10 | 生化学株式会社 | Photocured cross-linked-hyaluronic acid gel and method of preparation thereof |
Non-Patent Citations (1)
| Title |
|---|
| 李振中等.紫外光辐照交联EVA的制备及表征.《研究与开发》.2007,第24卷(第4期),16-19. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101716331A (en) | 2010-06-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Three‐dimensional extrusion bioprinting of single‐and double‐network hydrogels containing dynamic covalent crosslinks | |
| Bryant et al. | Synthesis and characterization of photopolymerized multifunctional hydrogels: water-soluble poly (vinyl alcohol) and chondroitin sulfate macromers for chondrocyte encapsulation | |
| Sun et al. | Current research progress of photopolymerized hydrogels in tissue engineering | |
| US9987393B2 (en) | Covalently cross linked hydrogels and methods of making and using same | |
| Truong et al. | Photodegradable gelatin-based hydrogels prepared by bioorthogonal click chemistry for cell encapsulation and release | |
| Zustiak et al. | Hydrolytically degradable poly (ethylene glycol) hydrogel scaffolds with tunable degradation and mechanical properties | |
| Jin et al. | Synthesis and characterization of hyaluronic acid–poly (ethylene glycol) hydrogels via Michael addition: An injectable biomaterial for cartilage repair | |
| Martens et al. | Tailoring the degradation of hydrogels formed from multivinyl poly (ethylene glycol) and poly (vinyl alcohol) macromers for cartilage tissue engineering | |
| Bartnikowski et al. | Protective effects of reactive functional groups on chondrocytes in photocrosslinkable hydrogel systems | |
| Uliniuc et al. | New approaches in hydrogel synthesis—Click chemistry: A review | |
| CN112341640B (en) | Bio-based self-repairing hydrogel and preparation method and application thereof | |
| Bae et al. | Fabrication of hyaluronic acid hydrogel beads for cell encapsulation | |
| JP2008508959A5 (en) | ||
| Hong et al. | Complex tuning of physical properties of hyperbranched polyglycerol‐based bioink for microfabrication of cell‐laden hydrogels | |
| Wells et al. | Photosensitive controlled release with polyethylene glycol–anthracene modified alginate | |
| KR101898229B1 (en) | Method for producing photo-crosslinked silk fibroin, and photo-crosslinked silk fibroin produced by the same | |
| CN109125808A (en) | A kind of biodegradable collagen-based cornea substitute and preparation method thereof | |
| Hamilton et al. | Hyaluronic acid hydrogel microspheres for slow release stem cell delivery | |
| Zanon et al. | Microwave-assisted methacrylation of chitosan for 3D printable hydrogels in tissue engineering | |
| Pei et al. | Photocrosslinkable chitosan hydrogels and their biomedical applications | |
| CN1348470A (en) | Hydrophilic macromolecular compounds | |
| Jiang et al. | Comparative cytocompatibility of multiple candidate cell types to photoencapsulation in PEGNB/PEGDA macroscale or microscale hydrogels | |
| Declercq et al. | Encapsulation of osteoblast seeded microcarriers into injectable, photopolymerizable three-dimensional scaffolds based on D, L-lactide and ε-caprolactone | |
| Swilem et al. | Development of carboxymethyl cellulose/gelatin hybrid hydrogels via radiation-induced cross-linking as novel anti-adhesion barriers | |
| Lee et al. | Green chemistry for crosslinking biopolymers: Recent advances in riboflavin-mediated photochemistry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120307 Termination date: 20141216 |
|
| EXPY | Termination of patent right or utility model |