CN105348545A - Preparation method of ultra-high-strength gelatin/collagen-based hydrogel - Google Patents
Preparation method of ultra-high-strength gelatin/collagen-based hydrogel Download PDFInfo
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Abstract
The invention relates to a preparation method of ultra-high-strength gelatin/collagen-based hydrogel. The preparation method comprises the following steps: first, performing microfibrillar treatment on cellulose to obtain an MFC suspension; then, oxidizing the MFC suspension to obtain dialdehyde MFC; finally, mixing dialdehyde MFC with a gelatin/collagen solution, so as to prepare dialdehyde MFC/gelatin compound hydrogel or dialdehyde MFC/collagen compound hydrogel. MFC has very high strength and is a good reinforcing agent; a chemical reaction can be performed between an aldehyde group on the molecular chain of dialdehyde MFC and an amino group on the gelatin/collagen molecular chain, so as to from a huge network structure in a whole dialdehyde MFC/gelatin system or dialdehyde MFC/collagen system; through adoption of the preparation method, the strength of the gelatin/collagen compound hydrogel can be greatly improved.
Description
Technical field
The invention belongs to composite aquogel technical field, be specifically related to a kind of method preparing superstrength gelatin or collagen-based composite aquogel.
Background technology
Hydrogel is that a class has strongly hydrophilic, but can not dissolve in water swelling material can only occur, and is the ubiquitous a kind of physical form of nature.The multicomponent system that macromolecule hydrogel is made up of high molecular three-dimensional crosslinked network and solution, can keep definite shape and three-dimensional space network structure in water.From micro-scale, macromolecule hydrogel has three-dimensional net structure, and due to hydration, water molecules is strapped in three-dimensional net structure by the hydrophilic radical in macromolecular chain, thus imparting the good water retention property of macromolecule hydrogel, shipwreck to squeeze out from hydrogel network simultaneously.Human body, except tooth and bone, is all made up of biologic soft tissue, studies the macromolecule hydrogel material similar to the stuctures and properties of biologic soft tissue and has important scientific meaning and using value.Such as, macromolecule hydrogel has important application prospect in tissue engineering bracket.When hydrogel is used as tissue engineering bracket material, need bear enough application and not be destroyed.But most of hydrogel is all more crisp, its energy-to-break is 10J/m
2left and right, and the energy-to-break of cartilage is 1000J/m
2left and right, remains between the two in larger gap.In recent years, the intensity how improving macromolecule hydrogel becomes the problem of many investigators concern.
Summary of the invention
The object of the invention is to overcome prior art defect, provides the preparation method of a kind of high strength gelatin or collagen-based composite aquogel.
For achieving the above object, the present invention adopts following technical scheme:
Prepare a method for superstrength gelatin or collagen-based composite aquogel, it comprises the steps:
1) fibrillation process is carried out to Mierocrystalline cellulose, obtain MFC suspension (i.e. fibrillation cellulose suspension);
2) with periodate, oxide treatment is carried out to MFC suspension, after oxide treatment terminates, dialyse 1-3 days to remove the complete periodate of unreacted with deionized water, obtain dialdehyde MFC suspension;
3) the gelatin-compounded hydrogel of dialdehyde MFC/ or dialdehyde MFC/ collagen composite hydrogel is prepared:
Dialdehyde MFC suspension and gelatin are mixed, then in 30-50 DEG C, react 1-3h under pH7-9, obtain the gelatin-compounded hydrogel of dialdehyde MFC/;
Or dialdehyde MFC suspension is carried out lyophilize and obtains dialdehyde MFC sponge, then dialdehyde MFC sponge is joined in collagen acetum, 37 DEG C, react 1-3h under the condition of pH7.4, obtain dialdehyde MFC/ collagen composite hydrogel.
Further preferably, in step 1), fibrillation process is specially: by Mierocrystalline cellulose and water in mass ratio (1-10): after (90-99) mixes, add in high pressure homogenizer, homogeneous 10-30 time, then centrifugal concentrating under 500-1500bar, obtain the MFC suspension that MFC content is 1-6wt%.
Step 2) in, oxide treatment is specially: joined by sodium periodate in MFC suspension, and under room temperature, lucifuge stirs 36-48h; Wherein, MFC suspension is with MFC dry weight basis, and the mass ratio of sodium periodate and MFC is 1-9:10.Mass ratio as sodium periodate and MFC can be 1:10,3:10,5:10,7:10,9:10 etc.
In step 3), dialdehyde MFC suspension is with dialdehyde MFC dry weight basis, and dialdehyde MFC addition accounts for the 0.1-40% of dialdehyde MFC and gelatin or collagen quality summation.
The gelatin-compounded hydrogel of above-mentioned dialdehyde MFC/ is slightly different from the preparation process of dialdehyde MFC/ collagen composite hydrogel:
Because gelatin can be dissolved in warm water, therefore when preparing the gelatin-compounded hydrogel of dialdehyde MFC/, gelatin particle directly can be joined in dialdehyde MFC suspension, reacting with dialdehyde MFC after making Gelatin.But collagen but can not be dissolved in water, therefore be first collagen is dissolved in acetum obtain collagen acetum, again dialdehyde MFC suspension is made dialdehyde MFC sponge through lyophilize, then dialdehyde MFC sponge is added in collagen acetum, dialdehyde MFC and collagen are reacted.Because collagen sex change can occur more than 40 DEG C, lose as the necessary life of biomaterial active, therefore react and carry out at 37 DEG C.Also for ease of the biological activity retaining collagen, the pH of reaction system is defined as 7.4.
MFC suspension can be prepared by mechanical process (as high pressure homogenization method, speed lapping method, high-speed mixing method etc.).Dialdehyde MFC be by Microcrystalline Cellulose through high-pressure homogeneous method (pressure of employing be 500-1500bar, homogenization cycles be 10-30 time) first obtained diameter be nano level and there is the fibrillation Mierocrystalline cellulose MFC(nano-cellulose of three-dimensional network-like structure), and then with periodate, it is oxidized, obtained dialdehyde MFC.Dialdehyde MFC and gelatin or collagen generation chemical reaction, make whole composite aquogel System forming have the hydrogel of huge network structure.
Mierocrystalline cellulose is the abundantest renewable polymer resource of nature, its abundance and cheap, has the advantages such as pollution-free, readily biodegradable.Mierocrystalline cellulose has multistage aggregated structure.A fiber is made up of some primitive fibers, and primitive fiber is again form by some cellulose molecular chains are arranged in parallel.Employing appropriate means (as acid-hydrolysis method, enzyme hydrolysis method and mechanical phonograph recorder separation) can isolate the nano level primitive fiber in Mierocrystalline cellulose, and wherein, mechanical phonograph recorder separation does not introduce contaminative chemical substance, is eco-friendly physical method.With the high pressure homogenization method in mechanical phonograph recorder separation, fibrillation is carried out to Mierocrystalline cellulose, diameter can be prepared in tens nanometers, the very large fibrillation Mierocrystalline cellulose of length-to-diameter ratio (also referred to as nano-cellulose, MFC), and these MFC do not rupture, but form interconnective network structure.Huge surface-area and meticulous network structure make nano-cellulose can give full play of the mechanical property of Mierocrystalline cellulose excellence, become a kind of excellent toughener.The cellulose molecular chain of MFC there is the oh group of One's name is legion, can easily to its modification.Such as, can be oxidized MFC, be aldehyde groups by part of hydroxyl group transformations wherein, thus prepare the dialdehyde MFC with new chemical functional group.
Self is just had the dialdehyde MFC of network structure and gelatin or collagen and carry out compound, prepare the gelatin-compounded hydrogel of dialdehyde MFC/ or dialdehyde MFC/ collagen composite hydrogel, in the process, aldehyde groups on dialdehyde MFC can with the amino group generation chemical reaction on gelatin or tropocollagen molecule chain, make MFC while playing toughener effect, play again the effect of linking agent, the gelatin-compounded hydrogel of whole dialdehyde MFC/ or dialdehyde MFC/ collagen composite hydrogel is made to become a huge network, thus the physical strength of gelatin-based or collagen-based composite aquogel is greatly improved.
Compared to the prior art, beneficial effect of the present invention:
Main raw material (Mierocrystalline cellulose, gelatin and collagen) in the inventive method all derives from natural macromolecular material, have wide material sources, good biocompatibility, the advantage such as biodegradable, and preparation process is simple, nontoxic, security is high.MFC has three-dimensional network-like structure and higher physical strength, on nanometer MFC molecular chain, aldehyde groups is introduced after oxidation, aldehyde radical and gelatin or collagen generation chemical reaction, make dialdehyde MFC not only play toughener effect but also play linking agent effect, the physical strength of gelatin or collagen-based composite aquogel is greatly improved.
Accompanying drawing explanation
Fig. 1 is the compound support frame material scanning electron microscope (SEM) photograph of different moiety; In figure, a is pure gelatin, b is MFC/ gelatin, and c is dialdehyde MFC/ gelatin; As can be seen from the figure: three all can form vesicular structure; In pure gelatin, bore dia is 50-200 μm; After adding MFC and dialdehyde MFC, the hole in system is flats, and diameter reduces, and hole becomes intensive;
Fig. 2 is the digital photograph figure of different moiety composite aquogel; As can be seen from the figure: pure gelatin hydrogel is lower due to intensity, easily fragmentation occurs when peeling off from mould, gel surface is caused to occur slight crack; Adding of MFC and dialdehyde MFC makes system elasticity increase, and fragility reduces, and therefore composite aquogel smooth surface, does not have slight crack;
Fig. 3 is the compressive strength of different moiety hydrogel;
Fig. 4 is the compressive strength of the dialdehyde MFC/ gelatin-based composite aquogel of the different dialdehyde MFC content adopting the inventive method to prepare;
Fig. 5 is the compressive strength of the dialdehyde MFC/ gelatin-based composite aquogel of the different oxidisabilities adopting the inventive method to prepare.
Embodiment
Do to introduce in detail further to technical scheme of the present invention below in conjunction with embodiment, but protection scope of the present invention is not limited thereto.
In following embodiment, the Microcrystalline Cellulose used is all through following pre-treatment: get 20g Microcrystalline Cellulose, 40gNaOH and 340g water adds beaker, 2h is stirred under 60 DEG C of water-baths, object is to make that fiber is swelling under the effect of NaOH, activation, Mierocrystalline cellulose generation fibrillation when being beneficial to high-pressure homogeneous.Suction filtration after alkaline purification, then fully wash to neutrality with deionized water, suspension can be used for high-pressure homogeneous.
embodiment 1
Prepare a method for superstrength gelatin-based composite aquogel, it comprises the steps:
1) get 20g Microcrystalline Cellulose, adding distil water is diluted to 1000mL, is transferred in high pressure homogenizer, and under 1000bar, homogeneous 25 times, centrifugal, after removing part supernatant liquor, obtains the MFC suspension that MFC content is 2.56wt%;
2) join in 500gMFC suspension by 8.96g sodium periodate, under room temperature, lucifuge stirs 36h, then dialyses 2 days with deionized water, leaves standstill after removing part supernatant liquid, obtains the dialdehyde MFC suspension that solid content is 3.49%;
3) get 25.27g dialdehyde MFC suspension, add 25.61g distilled water and supply as 50g to make the quality of water, stir under 40 DEG C of water-baths; Then 5g gelatin (to make the mass concentration of gelatin for 10%) is added, stirring reaction 2h under 40 DEG C and pH are the condition of 7.4, ultrasonator is adopted to be exhausted bubble to mixed solution, then mixed solution is poured in mould, room temperature cooling 12h forms gel, obtain the gelatin-compounded hydrogel of dialdehyde MFC/, the content of dialdehyde MFC is 15wt%.
The computation process of dialdehyde MFC content: the dry weight first calculating dialdehyde MFC, then by its quality sum divided by dialdehyde MFC and gelatin.Be specially: 25.27g × 3.49%=0.88g, 0.88g/(0.88g+5g) × 100%=15%.
The compression performance of the gelatin-compounded hydrogel of above-mentioned gained dialdehyde MFC/ is measured with CMT5140 electronic universal mechanics tester.Sample is cylindrical shape, is highly 20mm, and diameter is 22mm.During compression, the loading rate of pressure head is 1.5mm/min.Each sample repeated test 5 times, gets its mean value.After tested, the compressive strength of the gelatin-compounded hydrogel of this dialdehyde MFC/ is 1.074MPa.Lower same.
embodiment 2
Prepare a method for superstrength gelatin-based composite aquogel, it comprises the steps:
1) get 20g Microcrystalline Cellulose, adding distil water is diluted to 1000mL, is transferred in high pressure homogenizer, and under 1250bar, homogeneous 20 times, centrifugal, after removing part supernatant liquor, obtains the MFC suspension that MFC content is 2wt%;
2) add in 500gMFC suspension by 5g sodium periodate, under room temperature, lucifuge stirs 36h, then dialyses 2 days with deionized water, leaves standstill after removing part supernatant liquid, obtains the dialdehyde MFC suspension that solid content is 3.75%;
3) get 23.52g dialdehyde MFC suspension, add 27.36g distilled water, stir under 40 DEG C of water-baths; Then add 5g gelatin, stirring reaction 2h under 40 DEG C and pH are the condition of 7.4, adopt ultrasonator to be exhausted bubble to mixed solution, then pour in mould by mixed solution, room temperature cooling 12h forms gel, obtains the gelatin-compounded hydrogel of dialdehyde MFC/.
After tested, the compressive strength of the gelatin-compounded hydrogel of above-mentioned dialdehyde MFC/ is 0.805MPa.
embodiment 3
Prepare a method for superstrength collagen base composite aquogel, it comprises the steps:
1) get 20g Microcrystalline Cellulose, adding distil water is diluted to 1000mL, is transferred in high pressure homogenizer, and under 1500bar, homogeneous 15 times, centrifugal, after removing part supernatant liquor, obtains the MFC suspension that MFC content is 4.6wt%;
2) add in 500gMFC suspension by 20.7g sodium periodate, under room temperature, lucifuge stirs 36h, then dialyses 2 days with deionized water, centrifugal rear removal part supernatant liquid, obtain the dialdehyde MFC suspension that solid content is 6%, lyophilize 24h at-50 DEG C, makes dialdehyde MFC sponge;
3) being in the acetic acid of 3 by being dissolved in 100mL, pH at 1.0g collagen 4 DEG C, being made into the collagen acetum of 10mg/mL, using Na
2hPO
4buffered soln adjust ph is 7.4;
4) getting 0.25g dialdehyde MFC sponge joins in 50mL collagen acetum, and at 37 DEG C of reaction 2h, room temperature cooling 12h forms gel, obtains dialdehyde MFC/ collagen composite hydrogel.
After tested, the compressive strength of above-mentioned dialdehyde MFC/ collagen composite hydrogel is 1.352MPa.
embodiment 4
Prepare a method for superstrength collagen base composite aquogel, it comprises the steps:
1) get 20g Microcrystalline Cellulose, adding distil water is diluted to 1000mL, is transferred in high pressure homogenizer, and under 500bar, homogeneous 30 times, centrifugal, after removing part supernatant liquor, obtains the MFC suspension that MFC content is 5wt%;
2) add in 300gMFC suspension by 4.5g sodium periodate, under room temperature, lucifuge stirs 36h, then dialyses 2 days with deionized water, centrifugal rear removal part supernatant liquid, obtain the MFC suspension that solid content is 6%, lyophilize 24h at-50 DEG C, makes dialdehyde MFC sponge;
3) being in the acetic acid of 3 by being dissolved in 100mL, pH at 1.5g collagen 4 DEG C, being made into the collagen acetum of 15mg/mL, using Na
2hPO
4buffered soln adjust ph is 7.4;
4) getting 0.5g dialdehyde MFC sponge joins in 50mL collagen solution, and at 37 DEG C of reaction 2h, room temperature cooling 12h forms gel, obtains dialdehyde MFC/ collagen composite hydrogel.
After tested, the compressive strength of above-mentioned dialdehyde MFC/ collagen composite hydrogel is 1.680MPa.
Fig. 3 gives the compression testing figure of pure gelatin, MFC/ gelatin and the gelatin-compounded hydrogel of dialdehyde MFC/.Can find out in figure: compressive strength when pure gelatin hydrogel is broken only has 0.039MPa; The compressive strength of the gelatin-compounded hydrogel of MFC/ increases to some extent, is 1.10 times of pure gelatin, illustrates that the network structure of nano-cellulose is conducive to strengthening the mechanical property of hydrogel; The mechanical property of the gelatin-compounded hydrogel of dialdehyde MFC/ is 27.54 times of pure gelatin, reach 1.074MPa, illustrate that the aldehyde radical on dialdehyde MFC cellulose molecular chain and the amino in gelatin chains there occurs crosslinked, the molecular chain that the network structure of dialdehyde MFC and gelatin are disperseed forms a huge three-dimensional network, more be conducive to the propagation of stress, substantially increase its mechanical property, make it have huge potential using value at cartilage tissue engineering rack.
Fig. 4 gives the impact of dialdehyde MFC content on the gelatin-compounded hydrogel compressive strength of dialdehyde MFC/, and wherein, the oxidisability of dialdehyde MFC is 1.471mmol/100g.Can find out in figure: along with the increase of dialdehyde MFC content, the compressive strength of composite aquogel enlarges markedly.The compressive strength of pure gelatin is 0.039MPa, and the dialdehyde MFC only adding 5wt% just makes its compressive strength be increased to 0.182MPa, is 4.67 times of pure gelatin.After adding 10wt% dialdehyde MFC, the compressive strength of composite aquogel is significantly improved, and is 20.56 times of pure gelatin.When dialdehyde MFC content is increased to 25wt%, the compressive strength of composite aquogel is up to 1.631MPa, and be 41.82 times of pure gelatin, its reinforced effects is very outstanding.
Fig. 5 gives the oxidisability of dialdehyde MFC to the impact of the gelatin-compounded hydrogel compressive strength of dialdehyde MFC/, and wherein, the content of dialdehyde MFC is 15wt%.Can find out in figure: as oxidisability less (lower than 1.069mmol/100g), the reinforced effects of dialdehyde MFC to system is limited; But after the oxidisability of dialdehyde MFC is more than 1.302mmol/100g, the compressive strength increase of system is very remarkable; When oxidisability is 1.725mmol/100g, the dialdehyde MFC adding 15wt% makes the compressive strength of system be increased to 1.532MPa, is 35.62 times of MFC/ gelatin hydrogel.This shows after the hydroxyl in MFC Mierocrystalline cellulose is oxidized to aldehyde radical in a large number, amino in aldehyde radical and gelatin chains fully reacts, make the network structure with certain density that dialdehyde MFC and gelatin are formed, can effectively bear external force load, the physical strength of system is significantly increased.
Claims (4)
1. prepare a method for superstrength gelatin or collagen-based composite aquogel, it is characterized in that, comprise the steps:
1) fibrillation process is carried out to Mierocrystalline cellulose, obtain MFC suspension;
2) with periodate, oxide treatment is carried out to MFC suspension, after oxide treatment terminates, dialyse 1-3 days to remove the complete periodate of unreacted with deionized water, obtain dialdehyde MFC suspension;
3) the gelatin-compounded hydrogel of dialdehyde MFC/ or dialdehyde MFC/ collagen composite hydrogel is prepared:
Dialdehyde MFC suspension and gelatin are mixed, then in 30-50 DEG C, react 1-3h under pH7-9, obtain the gelatin-compounded hydrogel of dialdehyde MFC/;
Or dialdehyde MFC suspension is carried out lyophilize and obtains dialdehyde MFC sponge, then dialdehyde MFC sponge is joined in collagen acetum, 37 DEG C, react 1-3h under the condition of pH7.4, obtain dialdehyde MFC/ collagen composite hydrogel.
2. prepare the method for superstrength gelatin or collagen-based composite aquogel as claimed in claim 1, it is characterized in that, in step 1), fibrillation process is specially: by Mierocrystalline cellulose and water in mass ratio (1-10): after (90-99) mixes, to add in high pressure homogenizer under 500-1500bar homogeneous 10-30 time, then centrifugal concentrating, obtains the MFC suspension that MFC content is 1-6wt%.
3. prepare the method for superstrength gelatin or collagen-based composite aquogel as claimed in claim 1, it is characterized in that, step 2) in, oxide treatment is specially: joined by sodium periodate in MFC suspension, and under room temperature, lucifuge stirs 36-48h; Wherein, MFC suspension is with MFC dry weight basis, and the mass ratio of sodium periodate and MFC is 1-9:10.
4. prepare the method for superstrength gelatin or collagen-based composite aquogel as claimed in claim 1, it is characterized in that, in step 3), dialdehyde MFC suspension is with dialdehyde MFC dry weight basis, and dialdehyde MFC addition accounts for the 0.1-40% of dialdehyde MFC and gelatin or collagen quality summation.
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