CN106567151A - Calcium fluoride collagen micro-fiber composite and preparation method thereof - Google Patents
Calcium fluoride collagen micro-fiber composite and preparation method thereof Download PDFInfo
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- CN106567151A CN106567151A CN201610959141.5A CN201610959141A CN106567151A CN 106567151 A CN106567151 A CN 106567151A CN 201610959141 A CN201610959141 A CN 201610959141A CN 106567151 A CN106567151 A CN 106567151A
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Abstract
The invention relates to a calcium fluoride collagen micro-fiber composite and a preparation method thereof. The preparation method includes the steps of: (1) adding collagen protein fiber powder to an acetic acid solution to prepare a collagen protein solution, and uniformly coating a surface of a substrate with the collagen protein solution and diffusing the collagen protein solution with ammonia water to produce a collagen substrate; (2) uniformly mixing and stirring sodium fluoride, sodium chloride and water to prepare a solution A, uniformly mixing and stirring calcium chloride, a polyacrylic acid solution, 4-hydroxyethyl piperazine ethanesulfonic acid and water to obtain a solution B, slowly adding the solution A dropwise to the solution B and regulating the pH of the system to prepare a calcium fluoride precursor solution; and (3) soaking the collagen substrate in the calcium fluoride precursor solution for mineralization to prepare the calcium fluoride collagen micro-fiber composite. The method for producing the calcium fluoride collagen micro-fiber composite is carried out through internal mineralization of the collagen micro-fibers at normal temperature and under normal pressure, thereby achieving internal mineralization of the collagen micro-fibers.
Description
Technical field
The present invention relates to biomineralization field, mineralising prepares calcium fluoride collagen fento in more particularly to a kind of collagen microfibrilss
The method of dimension complex.
Background technology
Biomineralization refers to the process of the biological highly fine hybrid inorganic-organic structure of synthesis in vivo, such as skeleton, tooth
Tooth, shell etc., its essence is the process of biological mineral growth in vivo.Biogenic mineral then refer to produced by said process comprising ore deposit
Material and organic substrate, and the compound substance being assembled into according to certain ordered structure.
Although there is a problem of in terms of the research of mineralising mechanism many to be solved, mineralization model but has pervasive
Property：Matrix is made up of a dominant cross-linked structure framework of hydrophobicity, and the fixed hydrophilic macromole in surface is hydrophilic
The forming core surface that macromole can provide activity for external solution.Several typical biogenic minerals, such as skeleton, enamel, tooth
Essence, shell etc., its mineralization process meets above-mentioned model.Inorganic constituentss are mainly hydroxyapatite in skeleton, account for total matter
The 70% of amount；NTx fiber accounts for total protein content 90% in organic principle, and remaining is noncollagen protein.Its mineralising is general
Think to be assembled into supporting structure by collagen protein, noncollagen protein induces the formation of amorphous calcium phosphate, and in collagen protein
Growth.
Skeleton is that nature uses top-down methods in nanoscale self assembly molecule so as to obtain high in macro-scale
The Typical Representative of the orderly macromolecular structure of degree.Collagen-apatite complex that synthesis has similar nature bone chemical composition exists
It was widely studied in past 20 years.The successful part of these researchs is apatite mineral is deposited in collagen stroma,
And to understanding that bionical mineralized collagen has contributed much.But, related scholar does not obtain nanometer in collagen microfibrilss level
The labyrinth of crystal grain assembling.
The content of the invention
The technical problem to be solved is to be directed to above-mentioned deficiency present in prior art, there is provided a kind of collagen is micro-
Calcium fluoride collagen microfibrilss complex that in fiber prepared by mineralising and preparation method thereof.
To solve above-mentioned technical problem, the technical scheme that the present invention is provided is：
A kind of calcium fluoride collagen microfibrilss complex is provided, its preparation method is comprised the following steps：
1) pretreatment of collagen protein：Collagen fabric powder body is added in acetic acid solution, is sufficiently stirred for obtaining collagen
Protein solution, by gained collagen solution cryopreservation 24h, is then coated uniformly on base by the collagen solution for preparing
Piece surface, and substrate and ammonia spirit are respectively placed in 2 little glass dishes, then 2 little glass dishes are positioned over greatly jointly
Seal in glass dish and with preservative film, make collagen solution form collagen microfibrilss in substrate surface diffusion and assembling, obtain
Collagen substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained；Will
Calcium chloride, polyacrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B；Solution A is slow
In being added drop-wise to solution B, calcium fluoride forerunner is obtained to 7.0-7.4 with the pH value of sodium hydroxide solution regulation system after completion of dropwise addition
Liquid solution；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride presoma
Mineralising is carried out in solution, then substrate rinsing is taken out, is dried to obtain calcium fluoride collagen microfibrilss complex.
By such scheme, step 1) the acetic acid solution concentration be 0.1-0.2mol/L；The collagen solution concentration
For 2-3mg/mL；The cryopreservation temperature is 4-6 DEG C.
By such scheme, step 2) concentration of sodium fluoride is 10-20mM in the solution A, the concentration of Sodium Chloride in solution A
For 50-150mM, the concentration of calcium chloride is 5-10mM in the solution B, and polyacrylic acid solution is by number-average molecular weight in solution B
The polyacrylic acid of 40000-50000 is dissolved in concentration that water obtains for the aqueous solution of 25wt%, and polyacrylic acid concentration is in solution B
100-125 μ g/mL, 4- hydroxyethyl piperazine ethanesulfonic acid concentration is 0.5-1mM in solution B.
By such scheme, step 2) sodium fluoride and Sodium Chloride mol ratio are 1 in the solution A:10-1:15；The solution B
Middle calcium chloride, the mol ratio of 4- hydroxyethyl piperazine ethanesulfonic acid are 5:1-5:2；Sodium fluoride and calcium chloride rubs in solution A and solution B
You are than being 2:1.
By such scheme, step 2) it is described by solution A be slowly dropped to during solution B drip speed control in 3-6s/ drops, hydrogen
Sodium hydroxide solution concentration is 1M.
By such scheme, step 3) the mineralising time be 6-72h.
The present invention also provides the preparation method of above-mentioned calcium fluoride collagen microfibrilss complex, and it is comprised the following steps：
1) pretreatment of collagen protein：Collagen fabric powder body is added in acetic acid solution, is sufficiently stirred for obtaining collagen
Protein solution, by gained collagen solution cryopreservation 24h, is then coated uniformly on base by the collagen solution for preparing
Piece surface, and substrate and ammonia spirit are respectively placed in 2 little glass dishes, then 2 little glass dishes are positioned over greatly jointly
Seal in glass dish and with preservative film, make collagen solution form collagen microfibrilss in substrate surface diffusion and assembling, obtain
Collagen substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained；Will
Calcium chloride, polyacrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B；Solution A is slow
In being added drop-wise to solution B, calcium fluoride forerunner is obtained to 6.4-8.4 with the pH value of sodium hydroxide solution regulation system after completion of dropwise addition
Liquid solution；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride presoma
Mineralising is carried out in solution, then substrate rinsing is taken out, is dried to obtain calcium fluoride collagen microfibrilss complex.
The beneficial effects of the present invention is：The present invention prepares calcium fluoride collagen microfibrilss and answers using mineralising in collagen microfibrilss
Compound, compared with prior art, the inventive method is synthesized under normal temperature and pressure conditionses using biological organic native template, energy
Consumption is low, and pollution is few, and the calcium fluoride amorphous phase presoma of synthesis is combined by electrostatic interaction with collagen fiber, by capillarity
Penetrate into inside collagen, then mineralising growth is carried out inside collagen, it is achieved thereby that the inside mineralising of collagen microfibrilss, and
Internal mineral can also be promoted to grow, realized fluorination calcium nano and arranged in collagen microfibrilss internal order.
Description of the drawings
Fig. 1 is the SEM photograph of the calcium fluoride collagen microfibrilss complex prepared by comparative example of the present invention 1；
Fig. 2 is the SEM photograph of the calcium fluoride collagen microfibrilss complex prepared by comparative example of the present invention 2；
Fig. 3 is the SEM photograph of the calcium fluoride collagen microfibrilss complex prepared by comparative example of the present invention 3；
Fig. 4 is the SEM photograph of the calcium fluoride collagen microfibrilss complex prepared by comparative example of the present invention 4；
Fig. 5 is the SEM photograph of the calcium fluoride collagen microfibrilss complex prepared by comparative example of the present invention 5；
Fig. 6 is the SEM photograph of the calcium fluoride collagen microfibrilss complex prepared by the embodiment of the present invention 1；
Fig. 7 is the SEM photograph of the calcium fluoride collagen microfibrilss complex prepared by the embodiment of the present invention 2；
Fig. 8 is the SEM photograph of the calcium fluoride collagen microfibrilss complex prepared by the embodiment of the present invention 3.
Specific embodiment
To make those skilled in the art more fully understand technical scheme, the present invention is made below in conjunction with the accompanying drawings into
One step is described in detail.
Polyacrylic acid of the polyacrylic acid solution used by the embodiment of the present invention by number-average molecular weight for 40000-50000 is dissolved in water
The concentration for obtaining is the aqueous solution of 25wt%.
Comparative example 1
Calcium fluoride collagen microfibrilss complex is prepared, step is as follows：
1) pretreatment of collagen protein：During 20mg collagen fabrics powder body is added into concentration for the acetic acid solution of 0.1M,
It is sufficiently stirred for 24h and obtains the collagen solution that concentration is 2mg/mL, by gained collagen solution in 4 DEG C of cryopreservation 24h,
Then (glass chip size is 18 × 18mm, and thickness is the collagen solution that 20 μ L are prepared to be coated uniformly on into substrate surface
0.13-0.17mm) and by the ammonia spirit that substrate and volume fraction are 25% it is respectively placed in 2 little glass dishes, then by 2
Little glass dish is positioned in big glass dish and with preservative film and seals jointly, stands 12h, collagen solution is expanded in substrate surface
Dissipate and assembling forms collagen microfibrilss, obtain collagen substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained (molten
The concentration of sodium fluoride and Sodium Chloride is respectively 5mM and 50mM in liquid A, and mol ratio is 1:10, volume is 25mL)；By calcium chloride, gather
Acrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B (calcium chloride, polypropylene in solution B
Acid, the concentration of 4- hydroxyethyl piperazine ethanesulfonic acid are respectively 2.5mM, 100 μ g/mL, 0.5mM, calcium chloride and polyacrylic mole
Than for 5:2, volume is 25mL)；Solution A is slowly dropped in solution B (two kinds of liquor capacity ratios are 1:1) it is 6s/, to drip speed
Drop, with the pH value of sodium hydroxide solution (concentration is 1M) regulation system to 7.4 after completion of dropwise addition, obtains concentration for 2.5mM, volume
For the calcium fluoride precursor solution of 50mL；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride presoma
Mineralising 24h in solution, substrate then taken out rinsing, be dried to obtain calcium fluoride collagen microfibrilss complex.
Fig. 1 is the electron scanning micrograph of the calcium fluoride collagen microfibrilss complex prepared by this comparative example, by scheming
Understand, collagen microfibrilss pattern is flat, is all attached to substrate surface, do not show the threadiness of solid, calcium fluoride is not in glue
Former microfibre inside mineralising, illustrates system concentration relatively low (calcium fluoride precursor solution concentration is 2.5mM) and the mineralising time is shorter
(24h) calcium fluoride collagen microfibrilss complex is not formed under conditions of.
Comparative example 2
Calcium fluoride collagen microfibrilss complex is prepared, step is as follows：
1) pretreatment of collagen protein：During 20mg collagen fabrics powder body is added into concentration for the acetic acid solution of 0.1M,
It is sufficiently stirred for 24h and obtains the collagen solution that concentration is 2mg/mL, by gained collagen solution in 4 DEG C of cryopreservation 24h,
Then (glass chip size is 18 × 18mm, and thickness is the collagen solution that 20 μ L are prepared to be coated uniformly on into substrate surface
0.13-0.17mm) and by the ammonia spirit that substrate and volume fraction are 25% it is respectively placed in 2 little glass dishes, then by 2
Little glass dish is positioned in big glass dish and with preservative film and seals jointly, stands 12h, collagen solution is expanded in substrate surface
Dissipate and assembling forms collagen microfibrilss, obtain collagen substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained (molten
The concentration of sodium fluoride and Sodium Chloride is respectively 5mM and 50mM in liquid A, and mol ratio is 1:10, volume is 25mL)；By calcium chloride, gather
Acrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B (calcium chloride, polypropylene in solution B
Acid, the concentration of 4- hydroxyethyl piperazine ethanesulfonic acid are respectively 2.5mM, 100 μ g/mL, 0.5mM, calcium chloride and polyacrylic mole
Than for 5:2, volume is 25mL)；Solution A is slowly dropped in solution B (two kinds of liquor capacity ratios are 1:1), after completion of dropwise addition
With the pH value of sodium hydroxide solution (concentration is 1M) regulation system to 7.4, concentration is obtained for 2.5mM, the fluorination that volume is 50mL
Calcium precursor solution；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride presoma
Mineralising 72h in solution, substrate then taken out rinsing, be dried to obtain calcium fluoride collagen microfibrilss complex.
Fig. 2 is the electron scanning micrograph of the calcium fluoride collagen microfibrilss complex prepared by this comparative example, by scheming
Understand, partially fluorinated calcium crystal forms more piece fiber morphology, with non-growth site collagen fento in collagen microfibrilss growth inside
Dimension (Fig. 1) forms sharp contrast, when illustrating mineralising time longer (72h), there is certain remineralization efficacy, but due to system concentration compared with
Low (calcium fluoride precursor solution concentration is 2.5mM), calcium fluoride does not become single continuous fiber structure in collagen growth inside.
Comparative example 3
Calcium fluoride collagen microfibrilss complex is prepared, step is as follows：
1) pretreatment of collagen protein：During 30mg collagen fabrics powder body is added into concentration for the acetic acid solution of 0.1M,
It is sufficiently stirred for 24h and obtains the collagen solution that concentration is 3mg/mL, by gained collagen solution in 6 DEG C of cryopreservation 24h,
Then (glass chip size is 18 × 18mm, and thickness is the collagen solution that 30 μ L are prepared to be coated uniformly on into substrate surface
0.13-0.17mm) and by the ammonia spirit that substrate and volume fraction are 25% it is respectively placed in 2 little glass dishes, then by 2
Little glass dish is positioned in big glass dish and with preservative film and seals jointly, stands 12h, collagen solution is expanded in substrate surface
Dissipate and assembling forms collagen microfibrilss, obtain collagen substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained (molten
The concentration of sodium fluoride and Sodium Chloride is respectively 10mM and 150mM in liquid A, and mol ratio is 1:15, volume is 25mL)；By calcium chloride,
Polyacrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B (calcium chloride in solution B, poly- third
Olefin(e) acid, the concentration of 4- hydroxyethyl piperazine ethanesulfonic acid are respectively 5mM, 100 μ g/mL, 1mM, calcium chloride and polyacrylic mol ratio
For 5:1, volume is 25mL)；Solution A is slowly dropped in solution B (two kinds of liquor capacity ratios are 1:1), use after completion of dropwise addition
The pH value of sodium hydroxide solution (concentration is 1M) regulation system to 6.4, obtain concentration for 5mM, volume for the calcium fluoride of 50mL before
Drive liquid solution；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride presoma
Mineralising 72h in solution, substrate then taken out rinsing, be dried to obtain calcium fluoride collagen microfibrilss complex.
Fig. 3 is the electron scanning micrograph of the calcium fluoride collagen microfibrilss complex prepared by this comparative example, by scheming
Understand, collagen microfibrilss surface attachment grown fluorination calcium nano, and calcium fluoride mineralising outside collagen microfibrilss illustrates body
When being slant acidity (calcium fluoride precursor solution pH value be 6.4) calcium fluoride cannot inside collagen microfibrilss mineralising, have to
The calcium fluoride collagen microfibrilss complex of outside mineralising.
Comparative example 4
Calcium fluoride collagen microfibrilss complex is prepared, step is as follows：
1) pretreatment of collagen protein：During 30mg collagen fabrics powder body is added into concentration for the acetic acid solution of 0.1M,
It is sufficiently stirred for 24h and obtains the collagen solution that concentration is 3mg/mL, by gained collagen solution in 4 DEG C of cryopreservation 24h,
Then (glass chip size is 18 × 18mm, and thickness is the collagen solution that 30 μ L are prepared to be coated uniformly on into substrate surface
0.13-0.17mm) and by the ammonia spirit that substrate and volume fraction are 25% it is respectively placed in 2 little glass dishes, then by 2
Little glass dish is positioned in big glass dish and with preservative film and seals jointly, stands 12h, collagen solution is expanded in substrate surface
Dissipate and assembling forms collagen microfibrilss, obtain collagen substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained (molten
The concentration of sodium fluoride and Sodium Chloride is respectively 10mM and 150mM in liquid A, and mol ratio is 1:15, volume is 25mL)；By calcium chloride,
Polyacrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B (calcium chloride in solution B, poly- third
Olefin(e) acid, the concentration of 4- hydroxyethyl piperazine ethanesulfonic acid are respectively 5mM, 100 μ g/mL, 1mM, calcium chloride and polyacrylic mol ratio
For 5:1, volume is 25mL)；Solution A is slowly dropped in solution B (two kinds of liquor capacity ratios are 1:1), use after completion of dropwise addition
The pH value of sodium hydroxide solution (concentration is 1M) regulation system to 8.4, obtain concentration for 5mM, volume for the calcium fluoride of 50mL before
Drive liquid solution；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride presoma
Mineralising 72h in solution, substrate then taken out rinsing, be dried to obtain calcium fluoride collagen microfibrilss complex.
Fig. 4 is the electron scanning micrograph of the calcium fluoride collagen microfibrilss complex prepared by this comparative example, by scheming
Understand, calcium fluoride crystal collagen microfibrilss inside mineralising and show periodic structure, have rupture existing between each cycle
As can be achieved on calcium fluoride in collagen fento when (calcium fluoride precursor solution concentration is 5mM) when illustrating that system concentration is suitable
The internal mineralising of dimension, but due to system meta-alkalescence (when calcium fluoride precursor solution pH value is 8.4), collagen microfibrilss structure is broken
Bad, crystal result in the rupture of collagen after mineralising growth inside collagen microfibrilss, have to external structure rupture and inside
The discontinuous calcium fluoride collagen microfibrilss complex of crystal.
Comparative example 5
Calcium fluoride collagen microfibrilss complex is prepared, step is as follows：
1) pretreatment of collagen protein：During 30mg collagen fabrics powder body is added into concentration for the acetic acid solution of 0.1M,
It is sufficiently stirred for 24h and obtains the collagen solution that concentration is 3mg/mL, by gained collagen solution in 4 DEG C of cryopreservation 24h,
Then (glass chip size is 18 × 18mm, and thickness is the collagen solution that 30 μ L are prepared to be coated uniformly on into substrate surface
0.13-0.17mm) and by the ammonia spirit that substrate and volume fraction are 25% it is respectively placed in 2 little glass dishes, then by 2
Little glass dish is positioned in big glass dish and with preservative film and seals jointly, stands 12h, collagen solution is expanded in substrate surface
Dissipate and assembling forms collagen microfibrilss, obtain collagen substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained (molten
The concentration of sodium fluoride and Sodium Chloride is respectively 10mM and 150mM in liquid A, and mol ratio is 1:15, volume is 25mL)；By calcium chloride
With 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, solution B (calcium chloride, 4- hydroxyethyl piperazine second sulphurs in solution B are obtained
It is 5 that the concentration of acid is respectively the mol ratio of 5mM, 1mM, calcium chloride and 4- hydroxyethyl piperazine ethanesulfonic acid:1, volume is 25mL)；Will
Solution A is slowly dropped in solution B that (two kinds of liquor capacity ratios are 1:1), with sodium hydroxide solution, (concentration is after completion of dropwise addition
1M) pH value of regulation system obtains concentration for 5mM, the calcium fluoride precursor solution that volume is 50mL to 7.4；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride presoma
Mineralising 72h in solution, substrate then taken out rinsing, be dried to obtain calcium fluoride collagen microfibrilss complex.
Fig. 5 is the electron scanning micrograph of the calcium fluoride collagen microfibrilss complex prepared by this comparative example, by scheming
Understand, calcium fluoride crystal is directly generated in the solution, and collagen microfibrilss are not observed, and is being not added with this composition of polyacrylic acid
When, calcium fluoride cannot in the solution form amorphous phase so as to penetrate into the mineralising growth of collagen microfibrilss inside, explanation
Additive polyacrylic acid serves the effect for suppressing crystal nucleation rate, during without polyacrylic acid, it is impossible to synthesize calcium fluoride glue
Former microfibre complex.
Embodiment 1
Calcium fluoride collagen microfibrilss complex is prepared, step is as follows：
1) pretreatment of collagen protein：During 20mg collagen fabrics powder body is added into concentration for the acetic acid solution of 0.1M,
It is sufficiently stirred for 24h and obtains the collagen solution that concentration is 2mg/mL, by gained collagen solution in 4 DEG C of cryopreservation 24h,
Then (glass chip size is 18 × 18mm, and thickness is the collagen solution that 20 μ L are prepared to be coated uniformly on into substrate surface
0.13-0.17mm) and by the ammonia spirit that substrate and volume fraction are 25% it is respectively placed in 2 little glass dishes, then by 2
Little glass dish is positioned in big glass dish and with preservative film and seals jointly, stands 12h, collagen solution is expanded in substrate surface
Dissipate and assembling forms collagen microfibrilss, obtain collagen substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained (molten
The concentration of sodium fluoride and Sodium Chloride is respectively 10mM and 150mM in liquid A, and mol ratio is 1:15, volume is 25mL)；By calcium chloride,
Polyacrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B (calcium chloride in solution B, poly- third
Olefin(e) acid, the concentration of 4- hydroxyethyl piperazine ethanesulfonic acid are respectively 5mM, 100 μ g/mL, 1mM, calcium chloride and polyacrylic mol ratio
For 5:1, volume is 25mL)；Solution A is slowly dropped in solution B (two kinds of liquor capacity ratios are 1:1), use after completion of dropwise addition
The pH value of sodium hydroxide solution (concentration is 1M) regulation system to 7.4, obtain concentration for 5mM, volume for the calcium fluoride of 50mL before
Drive liquid solution；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride presoma
Mineralising 72h in solution, substrate then taken out rinsing, be dried to obtain calcium fluoride collagen microfibrilss complex.
Fig. 6 is the electron scanning micrograph of the calcium fluoride collagen microfibrilss complex prepared by the present embodiment, by scheming
Understand, form internal full single calcium fluoride collagen microfibrilss, illustrate to add additive polyacrylic acid, concentration in system
Moderate (calcium fluoride precursor solution concentration is 5mM), pH (pH value is 7.4) and mineralising time longer (72h) in neutral range
Under the conditions of remineralization efficacy it is good, calcium fluoride crystal mineralising growth inside collagen microfibrilss obtains internal full and continuous fluorine
Change calcium collagen microfibrilss complex.
Embodiment 2
Calcium fluoride collagen microfibrilss complex is prepared, step is as follows：
1) pretreatment of collagen protein：During 20mg collagen fabrics powder body is added into concentration for the acetic acid solution of 0.1M,
It is sufficiently stirred for 24h and obtains the collagen solution that concentration is 2mg/mL, by gained collagen solution in 4 DEG C of cryopreservation 24h,
Then (glass chip size is 18 × 18mm, and thickness is the collagen solution that 20 μ L are prepared to be coated uniformly on into substrate surface
0.13-0.17mm) and by the ammonia spirit that substrate and volume fraction are 25% it is respectively placed in 2 little glass dishes, then by 2
Little glass dish is positioned in big glass dish and with preservative film and seals jointly, stands 12h, collagen solution is expanded in substrate surface
Dissipate and assembling forms collagen microfibrilss, obtain collagen substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained (molten
The concentration of sodium fluoride and Sodium Chloride is respectively 10mM and 150mM in liquid A, and mol ratio is 1:15, volume is 25mL)；By calcium chloride,
Polyacrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B (calcium chloride in solution B, poly- third
Olefin(e) acid, the concentration of 4- hydroxyethyl piperazine ethanesulfonic acid are respectively 5mM, 100 μ g/mL, 1mM, calcium chloride and polyacrylic mol ratio
For 5:1, volume is 25mL)；Solution A is slowly dropped in solution B (two kinds of liquor capacity ratios are 1:1), use after completion of dropwise addition
The pH value of sodium hydroxide solution (concentration is 1M) regulation system to 7.2, obtain concentration for 5mM, volume for the calcium fluoride of 50mL before
Drive liquid solution；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride presoma
Mineralising 24h in solution, substrate then taken out rinsing, be dried to obtain calcium fluoride collagen microfibrilss complex.
Fig. 7 is the electron scanning micrograph of the calcium fluoride collagen microfibrilss complex prepared by the present embodiment, by scheming
Understand, form internal full single calcium fluoride collagen microfibrilss, and collagen microfibrilss surface is fluorinated completely calcium nano
Cover, illustrate to add additive polyacrylic acid, moderate concentration (calcium fluoride precursor solution concentration is 5mM), pH in system
In neutral range when (pH value is 7.2), under conditions of the mineralising time moderate (24h) preferable remineralization efficacy, portion are also reached
Fluorination calcium nano is divided to adsorb on collagen microfibrilss surface, partial crystals mineralising growth inside collagen microfibrilss obtains interior
The full and continuous calcium fluoride collagen microfibrilss complex in portion.
Embodiment 3
Calcium fluoride collagen microfibrilss complex is prepared, step is as follows：
1) pretreatment of collagen protein：During 20mg collagen fabrics powder body is added into concentration for the acetic acid solution of 0.1M,
It is sufficiently stirred for 24h and obtains the collagen solution that concentration is 2mg/mL, by gained collagen solution in 4 DEG C of cryopreservation 24h,
Then (glass chip size is 18 × 18mm, and thickness is the collagen solution that 20 μ L are prepared to be coated uniformly on into substrate surface
0.13-0.17mm) and by the ammonia spirit that substrate and volume fraction are 25% it is respectively placed in 2 little glass dishes, then by 2
Little glass dish is positioned in big glass dish and with preservative film and seals jointly, stands 12h, collagen solution is expanded in substrate surface
Dissipate and assembling forms collagen microfibrilss, obtain collagen substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained (molten
The concentration of sodium fluoride and Sodium Chloride is respectively 10mM and 150mM in liquid A, and mol ratio is 1:15, volume is 25mL)；By calcium chloride,
Polyacrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B (calcium chloride in solution B, poly- third
Olefin(e) acid, the concentration of 4- hydroxyethyl piperazine ethanesulfonic acid are respectively 5mM, 100 μ g/mL, 1mM, calcium chloride and polyacrylic mol ratio
For 5:1, volume is 25mL)；Solution A is slowly dropped in solution B (two kinds of liquor capacity ratios are 1:1), use after completion of dropwise addition
The pH value of sodium hydroxide solution (concentration is 1M) regulation system to 7.0, obtain concentration for 5mM, volume for the calcium fluoride of 50mL before
Drive liquid solution；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride presoma
Mineralising 6h in solution, substrate then taken out rinsing, be dried to obtain calcium fluoride collagen microfibrilss complex.
Fig. 8 is the electron scanning micrograph of the calcium fluoride collagen microfibrilss complex prepared by the present embodiment, by scheming
Understand, form internal full single calcium fluoride collagen microfibrilss, and collagen microfibrilss surface is fluorinated calcium nano covering,
Illustrate to be added in system additive polyacrylic acid, moderate concentration (calcium fluoride precursor solution concentration is 5mM), pH in neutrality
In the range of (pH value is 7.0) when, mineralising can also reach effect, partially fluorinated calcium nanometer under conditions of the mineralising time shorter (6h)
Granular absorption is obtained on collagen microfibrilss surface, and as the increase of mineralising time enters the mineralising growth of collagen microfibrilss inside
Internal full and continuous calcium fluoride collagen microfibrilss complex.
Claims (7)
1. a kind of calcium fluoride collagen microfibrilss complex, it is characterised in that its preparation method is comprised the following steps：
1) pretreatment of collagen protein：Collagen fabric powder body is added in acetic acid solution, is sufficiently stirred for obtaining collagen protein
Solution, by gained collagen solution cryopreservation 24h, is then coated uniformly on substrate table by the collagen solution for preparing
Face, and substrate and ammonia spirit are respectively placed in 2 little glass dishes, then 2 little glass dishes are positioned over into big glass jointly
Seal in ware and with preservative film, make collagen solution form collagen microfibrilss in substrate surface diffusion and assembling, obtain collagen
Substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained；By chlorination
Calcium, polyacrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B；Solution A is slowly added dropwise
To in solution B, calcium fluoride presoma is obtained molten to 7.0-7.4 with the pH value of sodium hydroxide solution regulation system after completion of dropwise addition
Liquid；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride precursor solution
In carry out mineralising, then by substrate take out rinsing, be dried to obtain calcium fluoride collagen microfibrilss complex.
2. calcium fluoride collagen microfibrilss complex according to claim 1, it is characterised in that：Step 1) acetic acid solution
Concentration is 0.1-0.2mol/L；The collagen solution concentration is 2-3mg/mL；The cryopreservation temperature is 4-6 DEG C.
3. calcium fluoride collagen microfibrilss complex according to claim 1, it is characterised in that：Step 2) in the solution A
The concentration of sodium fluoride is 10-20mM, and the concentration of Sodium Chloride is 50-150mM in solution A, and the concentration of calcium chloride is in the solution B
5-10mM, polyacrylic acid of the polyacrylic acid solution by number-average molecular weight for 40000-50000 is dissolved in the concentration that water is obtained in solution B
For the aqueous solution of 25wt%, polyacrylic acid concentration is 100-125 μ g/mL in solution B, 4- hydroxyethyl piperazine ethanesulfonic acid in solution B
Concentration is 0.5-1mM.
4. calcium fluoride collagen microfibrilss complex according to claim 1, it is characterised in that：Step 2) in the solution A
Sodium fluoride and Sodium Chloride mol ratio are 1:10-1:15；Calcium chloride, the mol ratio of 4- hydroxyethyl piperazine ethanesulfonic acid in the solution B
For 5:1-5:2；Sodium fluoride and the mol ratio of calcium chloride are 2 in solution A and solution B:1.
5. calcium fluoride collagen microfibrilss complex according to claim 1, it is characterised in that：Step 2) it is described by solution A
It is slowly dropped to during solution B and drips speed control in 3-6s/ drops, concentration of sodium hydroxide solution is 1M.
6. calcium fluoride collagen microfibrilss complex according to claim 1, it is characterised in that：Step 3) the mineralising time
For 6-72h.
7. the preparation method of the arbitrary calcium fluoride collagen microfibrilss complex of a kind of claim 1-6, it is characterised in that include
Following steps：
1) pretreatment of collagen protein：Collagen fabric powder body is added in acetic acid solution, is sufficiently stirred for obtaining collagen protein
Solution, by gained collagen solution cryopreservation 24h, is then coated uniformly on substrate table by the collagen solution for preparing
Face, and substrate and ammonia spirit are respectively placed in 2 little glass dishes, then 2 little glass dishes are positioned over into big glass jointly
Seal in ware and with preservative film, make collagen solution form collagen microfibrilss in substrate surface diffusion and assembling, obtain collagen
Substrate；
2) synthesis of calcium fluoride precursor solution：By sodium fluoride, Sodium Chloride and water mixing and stirring, solution A is obtained；By chlorination
Calcium, polyacrylic acid solution, 4- hydroxyethyl piperazine ethanesulfonic acid and water mixing and stirring, obtain solution B；Solution A is slowly added dropwise
To in solution B, calcium fluoride presoma is obtained molten to 7.0-7.4 with the pH value of sodium hydroxide solution regulation system after completion of dropwise addition
Liquid；
3) mineralising of collagen：By step 1 under room temperature) the collagen substrate handled well immersion step 2) gained calcium fluoride precursor solution
In carry out mineralising, then by substrate take out rinsing, be dried to obtain calcium fluoride collagen microfibrilss complex.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110426356A (en) * | 2019-08-06 | 2019-11-08 | 武汉轻工大学 | A kind of natural collagen protein/nano-Au composite and the method for synchronization monitoring natural collagen protein self assembly |
| CN116005361A (en) * | 2022-12-29 | 2023-04-25 | 东莞盛翔新材料技术有限公司 | Preparation method of cellulose-collagen composite nanofiber membrane |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1160187A (en) * | 1995-11-21 | 1997-09-24 | 赫德逊产品有限公司 | Heat pipe heat exchanger tubesheet |
| CN101327341A (en) * | 2008-08-01 | 2008-12-24 | 浙江理工大学 | Method for preparing nano-scale fibroin-hydroxylapatite mineralized complex fiber |
| CN101474429A (en) * | 2009-01-22 | 2009-07-08 | 浙江理工大学 | Method for preparing hydroxylapatite-silk fibroin compound stent material using two-step method |
| CN101693774A (en) * | 2009-09-28 | 2010-04-14 | 暨南大学 | Nanometer hydroxyapatite/natural polymer composite, preparation method and application thereof |
| CN102491298A (en) * | 2011-11-14 | 2012-06-13 | 苏州大学 | Preparation method for linear nanometre hydroxylapatite |
| CN104353118A (en) * | 2014-11-05 | 2015-02-18 | 上海师范大学 | Collagenous fiber non-woven hydroxyapatite biological scaffold material and preparation method thereof |
-
2016
- 2016-11-03 CN CN201610959141.5A patent/CN106567151B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1160187A (en) * | 1995-11-21 | 1997-09-24 | 赫德逊产品有限公司 | Heat pipe heat exchanger tubesheet |
| CN101327341A (en) * | 2008-08-01 | 2008-12-24 | 浙江理工大学 | Method for preparing nano-scale fibroin-hydroxylapatite mineralized complex fiber |
| CN101474429A (en) * | 2009-01-22 | 2009-07-08 | 浙江理工大学 | Method for preparing hydroxylapatite-silk fibroin compound stent material using two-step method |
| CN101693774A (en) * | 2009-09-28 | 2010-04-14 | 暨南大学 | Nanometer hydroxyapatite/natural polymer composite, preparation method and application thereof |
| CN102491298A (en) * | 2011-11-14 | 2012-06-13 | 苏州大学 | Preparation method for linear nanometre hydroxylapatite |
| CN104353118A (en) * | 2014-11-05 | 2015-02-18 | 上海师范大学 | Collagenous fiber non-woven hydroxyapatite biological scaffold material and preparation method thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110426356A (en) * | 2019-08-06 | 2019-11-08 | 武汉轻工大学 | A kind of natural collagen protein/nano-Au composite and the method for synchronization monitoring natural collagen protein self assembly |
| CN110426356B (en) * | 2019-08-06 | 2022-09-06 | 武汉轻工大学 | Natural collagen/nanogold composite and method for synchronously monitoring self-assembly of natural collagen |
| CN116005361A (en) * | 2022-12-29 | 2023-04-25 | 东莞盛翔新材料技术有限公司 | Preparation method of cellulose-collagen composite nanofiber membrane |
| CN116005361B (en) * | 2022-12-29 | 2023-07-21 | 东莞盛翔新材料技术有限公司 | Preparation method of cellulose-collagen composite nanofiber membrane |
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