CN109897387A - Application, porous gel and its preparation of a kind of modified gelatin in water packet air lotion - Google Patents
Application, porous gel and its preparation of a kind of modified gelatin in water packet air lotion Download PDFInfo
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- CN109897387A CN109897387A CN201910072576.1A CN201910072576A CN109897387A CN 109897387 A CN109897387 A CN 109897387A CN 201910072576 A CN201910072576 A CN 201910072576A CN 109897387 A CN109897387 A CN 109897387A
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Abstract
The invention belongs to the technical field of medical macromolecular materials, application, porous gel and its preparation of a kind of modified gelatin in water packet air lotion are disclosed.Application of the modified gelatin in lotion, the surfactant as stable emulsion；Modified gelatin is methacrylic anhydride modified gelatin；Lotion is water packet air lotion.The water packet air lotion, is mainly obtained by methacrylic anhydride modified gelatin and water by high speed homogenization；It further include inorganic particulate, photoinitiator.The porous gel obtains by the following method: 1) methacrylic anhydride modified gelatin, photoinitiator and inorganic particulate high speed homogenization in water obtains lotion；2) lotion illumination is crosslinked, obtains porous gel.The present invention can form stable water packet air lotion and chemical cross-linking agent is not added when preparing porous gel using methacrylic anhydride modified gelatin as surfactant, environmental protection, safety；Prepared porous gel has preferable mechanical property, good biocompatibility.
Description
Technical field
The invention belongs to biological medical polymer material technical fields, and in particular to a kind of modified gelatin is newborn in water packet air
Application in liquid, the surfactant as stable emulsion, and utilize the porous aquagel and preparation method thereof of lotion preparation.
Background technique
Hydrogel is used as drug conveying, cell encapsulation and tissue engineering bracket, can effectively simulate natural extracellular matrix
(ECM).If the adherency and amplification of cell will be limited by space however, being wrapped in closely knit hydrogel, so that it is raw to influence cell
Long and regeneration.In recent years, porous aquagel is used as cell delivery vehicle and dimensional culture is concerned.Draw in hydrogel
Enter porous structure, can not only improve the permeability of hydrogel, can also quickly conduct nutrition and oxygen, for cell growth and
ECM deposition creates space.
Currently, the methods of emulsion template, freeze-drying, salt grain template and 3D printing have applied to hydrogel pore-creating to manufacture
Super porous hydrogel.In these methods, emulsion template has unique advantage, and the interconnected aperture knot of hole can be prepared for hydrogel
Structure is very beneficial for nutrition exchange and cell migration growth.Emulsion template is divided into oil-in-water type and water-in-oil type, works as internal phase volume
Higher than 74%, can get High Internal Phase Emulsion (HIPEs).Monomer or hydrogel precursor and crosslinking agent are typically dissolved in continuous aqueous phase
In, after organic solvent emulsifying, by aqueous polymerization or solidify available porous gel.
For aperture and the porosity for regulating and controlling such hydrogel, surfactant is extremely crucial.Emulsion template can be by big
Molecular surface active agent or colloidal particle are stablized, such as synthetic polymer, protein, inorganic oxide particles, graphene oxide
(GO), Cellulose nanocrystal body, microgel etc..However, the porous aquagel for the emulsion template method preparation reported at present is directed to
Using organic solvent, it is used as dispersed phase, needs more purification steps to remove organic solvent, to reduce cytotoxicity.In addition, most of
Surfactant non-degradable influences the biologic applications of prepared porous gel to a certain degree.
Summary of the invention
In order to overcome the shortcomings and deficiencies of the prior art, it is empty in water packet that the purpose of the present invention is to provide a kind of modified gelatins
Application in gas emulsions.The present invention is by the grafting modification to gelatin, and the surface for being used as water packet air lotion is living
Property agent so that water packet air lotion have preferable stability.
Another object of the present invention is to provide a kind of porous aquagels and preparation method thereof.Porous aquagel of the invention
It is to be prepared using above-mentioned water packet air lotion.
The purpose of the present invention is achieved through the following technical solutions:
A kind of application of modified gelatin in lotion, the surfactant as stable emulsion；The modified gelatin is first
Base acrylic anhydride modified gelatin.
The grafting rate of methacrylic anhydride modified gelatin (GelMA) is 50%~85%.
Methacrylic anhydride modified gelatin is to be modified to obtain to gelatin by methacrylic anhydride.
Methacrylic anhydride modified gelatin is prepared especially by following methods:
(S1) Gelatin is obtained by gelatin solution using PBS solution；(S2) methyl-prop is slowly added into gelatin solution
Olefin(e) acid acid anhydride, reaction, subsequent processing obtain methacrylic anhydride modified gelatin.
The amount ratio of the gelatin and methacrylic anhydride is (2~10) g:(5~20) mL.The gelatin and PBS solution
Amount ratio be (2~10) g:(20~100) mL.
The methacrylic anhydride is added in a manner of being added dropwise, and the speed of dropwise addition is 1-3mL/min；The temperature of the reaction
It is 40~60 DEG C, the time of reaction is 4~10h.After reaction, with PBS diluting reaction to terminate reaction.The subsequent processing
Refer to and the solution after reaction is dialysed, be centrifuged, take upper liquid, is freeze-dried.The bag filter of dialysis is bag filter (8-
14KDa), it is dialysed with water.The revolving speed of centrifugation is 3000~6000rpm.
The lotion is water packet air lotion.
The water packet air lotion is mainly obtained by methacrylic anhydride modified gelatin and water by high speed homogenization；Methyl-prop
The mass volume ratio of olefin(e) acid anhydride modification gelatin and (solution that methacrylic anhydride modified gelatin and water are formed) be (0.01~
0.15) g:1mL.
Further, the water packet air lotion further includes inorganic particulate, and water packet air lotion is mainly by methyl-prop at this time
Olefin(e) acid anhydride modification gelatin is obtained by high speed homogenization in water with inorganic particulate.
Further, the water packet air lotion further includes photoinitiator, and water packet air lotion is mainly by methyl at this time
Acrylic anhydride modified gelatin, photoinitiator and inorganic particulate are obtained by high speed homogenization in water.
The mass ratio of the methacrylic anhydride modified gelatin, photoinitiator and inorganic particulate is (0.01~0.15) g:
(0.002~0.01) g:(0.004~0.03) g, preferably (0.05~0.15) g:(0.002~0.01) g:(0.004~
0.03)g。
The concentration of the inorganic particulate in water is 0.5%~2.5%, i.e., contains in the inorganic particulate solution of every 1mL
The inorganic particulate of 0.005~0.025g.
The inorganic particulate is more than one in lithium diatomaceous earth, hydroxyapatite or calcium carbonate.
The water packet air lotion is preparing the application in porous gel.
The porous gel is prepared by the following method to obtain:
1) methacrylic anhydride modified gelatin, photoinitiator and inorganic particulate carry out high speed homogenization in water, obtain lotion；
2) lotion is subjected to illumination crosslinking, obtains porous gel.
The mass ratio of the methacrylic anhydride modified gelatin, photoinitiator and inorganic particulate is (0.01~0.15) g:
(0.002~0.01) g:(0.004~0.03) g, preferably (0.05~0.15) g:(0.002~0.01) g:(0.004~
0.03)g。
The concentration of the inorganic particulate in water is 0.5%~2.5% (w/v), i.e., contains in the inorganic particulate solution of every 1mL
There is the inorganic particulate of 0.005~0.025g.
The revolving speed of the high speed homogenization is 8000~16000rpm, and the time of homogeneous is 30~120s.
The light irradiation is UV illumination, and the time of illumination is 8~15min, and ultraviolet lamp power is 5-20mw/cm2, cream
Liquid and lamp source distance 5-20cm.
The photoinitiator is 2- hydroxyl -4'- (2- hydroxy ethoxy) -2- methyl phenyl ketone (I2959).
Application of the porous gel in the timbering material of Bone Defect Repari.
Gelatin modified object GelMA is prepared water packet air lotion by the present invention, and is prepared by template of water packet air lotion
Porous aquagel timbering material.Gelatin chains contain a large amount of hydrophilic and hydrophobic functional groups, gelatin amphiphilic irregular due to being distributed
Property is poor.Gelatin by with methacrylic acid anhydride reactant, introduce hydrophobic grouping, obtain modified gelatin product gelatin
Methacrylate (GelMA), can have excellent amphipathic, be used as surfactant to prepare lotion.The present invention utilizes
The double bond of GelMA side chain carries out chemical crosslinking and prepares hydrogel, improves the mechanical strength of gelatin materials.Therefore, in conjunction with GelMA
Stable lotion and double bond cross-linking method, can prepare the good porous hydrogel material of biocompatibility, widen gelatin in life
The application of object medical field.
Compared with the domestic and international prior art, the present invention has the advantage that
(1) present invention is using methacrylic anhydride modified gelatin as surfactant, and method is simple, in low concentration
(1%) it can be formed when and stablize water packet air lotion, and lotion can be mixed to form with other inorganic particulates, be greatly broadened bright
The application range of glue；
(2) porous gel that the present invention is prepared using methacrylic anhydride modified gelatin as surfactant, is not added
Any chemical cross-linking agent is conducive to the safety for improving material；
(3) porous gel that the present invention is prepared using methacrylic anhydride modified gelatin as surfactant has phase
Intercommunicated open-celled structure is conducive to medium and quickly spreads；
(4) porous gel that is prepared using methacrylic anhydride modified gelatin as surfactant of the present invention, wherein lithium
The addition of diatomaceous earth can effectively improve the mechanical property of gelatin materials, can be used as the timbering material of Bone Defect Repari.
Detailed description of the invention
Fig. 1 is the nuclear magnetic spectrogram that embodiment 1 prepares GelMA；
Fig. 2 is that modified gelatin GelMA prepares lotion (corresponding to GelMA) in embodiment 2 and the lotion of gelatin preparation is (corresponding
Gelatin photo)；(a) freshly prepared lotion after high speed shear；(b) lotion stands 3 hours, the lotion of unmodified gelatin preparation
Demulsification；
Fig. 3 is the photo (a) and water packet air lotion of the water packet air lotion prepared under various concentration in embodiment 2
Fluorescent microscopy images (b-e)；
Fig. 4 is the fluorescent microscopy images of lotion prepared by embodiment 3, and (a) is lithium diatomaceous earth solution, that is, Laponite solution
The photo that concentration is 0 corresponds to 0clay, and (b) photo that lithium diatomaceous earth solution, that is, Laponite solution concentration is 1% corresponds to 1clay,
(c) photo that lithium diatomaceous earth solution, that is, Laponite solution concentration is 2.5% corresponds to 2.5clay；
Fig. 5 is the stereoscan photograph of porous gel prepared by embodiment 4, and (a) is that lithium diatomaceous earth solution, that is, Laponite is molten
The photo that liquid concentration is 0, (b) lithium diatomaceous earth solution, that is, Laponite solution concentration is 1% photo, and (c) lithium diatomaceous earth solution is
The photo that Laponite solution concentration is 2.5%；The lower left corner is the figure of different multiples in each figure；
Fig. 6 is Proliferation data histogram of the mesenchymal stem cell in porous gel prepared by embodiment 4；0 refers to
The gel that lithium diatomaceous earth solution, that is, Laponite solution concentration is prepared when being 0,1% refers to that the i.e. Laponite solution of lithium diatomaceous earth solution is dense
The porous gel prepared when degree is 1%, 2.5% refers to and prepares when lithium diatomaceous earth solution i.e. Laponite solution concentration is 2.5%
Porous gel.
Specific embodiment
The present invention is specifically described below with reference to embodiment, but protection scope of the present invention is not limited to following implementation
Example.
Embodiment 1
The gelatin (Gelatin) for weighing 5g is added in PBS (pH=7.4) solution of 50mL, the heating stirring at 50 DEG C
Half an hour, until gelatin is completely dissolved；It is slowly added to the methacrylic anhydride (MA) of 10mL, rate of addition 1mL/min；At 50 DEG C
Continue to be stirred to react 4h, after reaction, with the PBS diluting reaction of 200mL to terminate reaction；Gained liquid is placed in bag filter
In (8-14KDa), to be used deionized water dialysis 7 days at 40 DEG C, change water daily twice, the solution after dialysis pours into centrifuge tube,
It is centrifuged 10min at 3,000 rpm, takes supernatant liquor, is lyophilized with freeze drier to get product GelMA, yield is arrived
91%, acid anhydrides grafting rate 83%.Fig. 1 is the nuclear magnetic spectrogram that embodiment 1 prepares GelMA.
Embodiment 2
The GelMA aqueous solution that concentration is 1% (w/v), 3% (w/v), 5% (w/v), 10% (w/v) is respectively configured, respectively takes
0.2mL stirs 30s in 8000rpm revolving speed using high speed agitator respectively, then stirs 1min under 20000rpm revolving speed,
Obtain water packet air lotion.
Fig. 2 is that modified gelatin GelMA prepares the lotion (cream prepared under the conditions of corresponding GelMA, 10% (w/v) in embodiment 2
Liquid) with gelatin preparation lotion (lotion prepared under the conditions of corresponding Gelatin, 10% (w/v)) photo；(a) high speed shear
Freshly prepared lotion afterwards；(b) lotion stands 3 hours, the emulsion breaking of unmodified gelatin preparation；
Fig. 3 is the photo (a) and water packet air lotion of the water packet air lotion prepared under various concentration in embodiment 2
Fluorescent microscopy images (b-e).
Embodiment 3
Weigh 0.1g GelMA, be added 0.9mL prepare in advance concentration be 0% (w/v), 1% (w/v), 2.5% (w/v's)
Laponite solution (lithium diatomaceous earth) is stirred 30 seconds in 8000rpm revolving speed using high speed agitator, then stirred under 20000rpm revolving speed
It mixes 1 minute.Fig. 4 is the fluorescent microscopy images of lotion prepared by embodiment 3, and (a) is lithium diatomaceous earth solution, that is, Laponite solution
The photo that concentration is 0 corresponds to 0clay, and (b) photo that lithium diatomaceous earth solution, that is, Laponite solution concentration is 1% corresponds to 1clay,
(c) photo that lithium diatomaceous earth solution, that is, Laponite solution concentration is 2.5% corresponds to 2.5clay.
Embodiment 4
0.1g GelMA is weighed, it is respectively 0% (w/v), 1% (w/v), 2.5% (w/ that 0.9mL, which is added, and prepares concentration in advance
V) 0.002g photoinitiator is added in Laponite solution, stirs 30 in 8000rpm revolving speed using high speed agitator after dissolution
Second, then stirred 1 minute under 20000rpm revolving speed, the stable emulsion of formation is placed under ultraviolet lamp and is irradiated 15 minutes, is obtained more
Hole gel rubber material.
Fig. 5 is the stereoscan photograph of porous gel prepared by embodiment 4, and (a) is that lithium diatomaceous earth solution, that is, Laponite is molten
The photo that liquid concentration is 0, (b) lithium diatomaceous earth solution, that is, Laponite solution concentration is 1% photo, and (c) tripolite solution is
The photo that Laponite solution concentration is 2.5%.
Fig. 6 is Proliferation data histogram of the mesenchymal stem cell in porous gel prepared by embodiment 4；0 refers to
The gel that lithium diatomaceous earth solution, that is, Laponite solution concentration is prepared when being 0,1% refers to that the i.e. Laponite solution of lithium diatomaceous earth solution is dense
The porous gel prepared when degree is 1%, 2.5% refers to and prepares when lithium diatomaceous earth solution i.e. Laponite solution concentration is 2.5%
Porous gel.Abscissa is the time that cell is grown in porous gel, and ordinate is the proliferative amount of cell.
The explanation of raw material sources used by the above implementation method: methacrylic anhydride, photoinitiator, is purchased from west at gelatin
Chinese (Shanghai) trade Co., Ltd of lattice Ma aldrich, lithium diatomaceous earth model Laponite-XLG.
Claims (10)
1. a kind of application of modified gelatin in lotion, it is characterised in that: the surfactant as stable emulsion；The modification
Gelatin is methacrylic anhydride modified gelatin；The lotion is water packet air lotion.
2. a kind of water packet air lotion, it is characterised in that: mainly pass through high speed homogenization by methacrylic anhydride modified gelatin and water
It obtains.
3. water packet air lotion according to claim 2, it is characterised in that: methacrylic anhydride modified gelatin and (methyl-prop
The solution that olefin(e) acid anhydride modification gelatin and water are formed) mass volume ratio be (0.01~0.15) g:1mL.
4. water packet air lotion according to claim 2, it is characterised in that: further include inorganic particulate, water packet air is newborn at this time
Liquid is mainly obtained by high speed homogenization by methacrylic anhydride modified gelatin with inorganic particulate in water.
5. water packet air lotion according to claim 4, it is characterised in that: further include photoinitiator, water packet air is newborn at this time
Liquid is mainly obtained by high speed homogenization by methacrylic anhydride modified gelatin, photoinitiator and inorganic particulate in water.
6. water packet air lotion according to claim 5, it is characterised in that: the methacrylic anhydride modified gelatin, light draw
The mass ratio for sending out agent and inorganic particulate is (0.01~0.15): (0.002~0.01): (0.004~0.03)；
The concentration of the inorganic particulate in water is 0.5%~2.5% (w/v), i.e., contains in the inorganic particulate solution of every 1mL
The inorganic particulate of 0.005~0.025g；
The inorganic particulate is more than one in lithium diatomaceous earth, hydroxyapatite or calcium carbonate.
7. a kind of porous gel, it is characterised in that: be mainly prepared by any one of the claim 2~6 water packet air lotion
It arrives.
8. porous gel according to claim 7, it is characterised in that: be prepared by the following method to obtain:
1) methacrylic anhydride modified gelatin, photoinitiator and inorganic particulate carry out high speed homogenization in water, obtain lotion；
2) lotion is subjected to illumination crosslinking, obtains porous gel.
9. porous gel according to claim 8, it is characterised in that: the methacrylic anhydride modified gelatin, photoinitiator
Mass ratio with inorganic particulate is (0.01~0.15): (0.002~0.01): (0.004~0.03)；
The concentration of the inorganic particulate in water is 0.5%~2.5% (w/v), i.e., contains in the inorganic particulate solution of every 1mL
The inorganic particulate of 0.005~0.025g；
The inorganic particulate is more than one in lithium diatomaceous earth, hydroxyapatite or calcium carbonate；
The revolving speed of the high speed homogenization is 8000~16000rpm, and the time of homogeneous is 30~120s；
The light irradiation is UV illumination, and the condition of light irradiation: the time of illumination is 8~15min, and ultraviolet lamp power is 5-
20mw/cm2, lotion and lamp source distance 5-20cm；
The photoinitiator is 2- hydroxyl -4'- (2- hydroxy ethoxy) -2- methyl phenyl ketone.
10. according to application of any one of claim 7~9 porous gel in the timbering material of Bone Defect Repari.
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Cited By (8)
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| CN111040199A (en) * | 2019-12-31 | 2020-04-21 | 华南理工大学 | Photo-crosslinking porous hydrogel based on two water-phase immiscible emulsions and preparation method and application thereof |
| CN111621032A (en) * | 2020-04-27 | 2020-09-04 | 上海海洋大学 | Octenylsuccinic anhydride modified gelatin and preparation method and application thereof |
| CN112980009A (en) * | 2021-03-16 | 2021-06-18 | 华南理工大学 | Nano composite porous gel scaffold and construction method and application thereof |
| CN113527605A (en) * | 2021-07-22 | 2021-10-22 | 华南理工大学 | Tissue adhesion conductive porous hydrogel and preparation method thereof |
| CN113563609A (en) * | 2021-07-28 | 2021-10-29 | 华南理工大学 | Nano composite porous hydrogel and preparation and application thereof |
| CN114712566A (en) * | 2022-04-22 | 2022-07-08 | 华南理工大学 | PCL/GelMA biological porous scaffold based on Pickering emulsion and preparation method thereof |
| CN115850730A (en) * | 2021-09-23 | 2023-03-28 | 四川大学 | Use of photo-cured porous hydrogel material as hemostatic material |
| TWI797630B (en) * | 2021-05-27 | 2023-04-01 | 台灣創新材料股份有限公司 | Microcarriers with scaffold structure and continuous outer wall for culturing cells |
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Cited By (9)
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|---|---|---|---|---|
| CN111040199A (en) * | 2019-12-31 | 2020-04-21 | 华南理工大学 | Photo-crosslinking porous hydrogel based on two water-phase immiscible emulsions and preparation method and application thereof |
| CN111621032A (en) * | 2020-04-27 | 2020-09-04 | 上海海洋大学 | Octenylsuccinic anhydride modified gelatin and preparation method and application thereof |
| CN111621032B (en) * | 2020-04-27 | 2022-04-05 | 上海海洋大学 | Octenylsuccinic anhydride modified gelatin and preparation method and application thereof |
| CN112980009A (en) * | 2021-03-16 | 2021-06-18 | 华南理工大学 | Nano composite porous gel scaffold and construction method and application thereof |
| TWI797630B (en) * | 2021-05-27 | 2023-04-01 | 台灣創新材料股份有限公司 | Microcarriers with scaffold structure and continuous outer wall for culturing cells |
| CN113527605A (en) * | 2021-07-22 | 2021-10-22 | 华南理工大学 | Tissue adhesion conductive porous hydrogel and preparation method thereof |
| CN113563609A (en) * | 2021-07-28 | 2021-10-29 | 华南理工大学 | Nano composite porous hydrogel and preparation and application thereof |
| CN115850730A (en) * | 2021-09-23 | 2023-03-28 | 四川大学 | Use of photo-cured porous hydrogel material as hemostatic material |
| CN114712566A (en) * | 2022-04-22 | 2022-07-08 | 华南理工大学 | PCL/GelMA biological porous scaffold based on Pickering emulsion and preparation method thereof |
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Application publication date: 20190618 |