CN104548202B - Titanium dioxide/albumin/biogenic signalling molecular composite coating with photo-inducible cell detachment function and preparation method thereof - Google Patents
Titanium dioxide/albumin/biogenic signalling molecular composite coating with photo-inducible cell detachment function and preparation method thereof Download PDFInfo
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- CN104548202B CN104548202B CN201410769505.4A CN201410769505A CN104548202B CN 104548202 B CN104548202 B CN 104548202B CN 201410769505 A CN201410769505 A CN 201410769505A CN 104548202 B CN104548202 B CN 104548202B
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Abstract
The invention discloses a titanium dioxide/albumin/biogenic signalling molecular composite coating with a photo-inducible cell detachment function. The titanium dioxide/albumin/biogenic signalling molecular composite coating with the photo-inducible cell detachment function sequentially comprises a base, a titanium dioxide nano point layer and an albumin and biogenic signalling molecular layer from top to bottom, wherein dimension of a titanium dioxide nano point in the titanium dioxide nano point layer is 20-300nm, density is 1.0*10<9>-1.0*10<11>/cm<2>, and gaps between titanium dioxide nano points are filled with the alumin and biogenic signalling molecular layer which covers the titanium dioxide nano points. A preparation method of the titanium dioxide/albumin/biogenic signalling molecular composite coating with the photo-inducible cell detachment function comprises the following steps: preparing titanium dioxide precursor sol; and sequentially spin-coating the precursor sol, a mixture of alumin and biogenic signalling molecules on the base, and carrying out heat treatment. The obtained titanium dioxide/albumin/biogenic signalling molecular composite coating has good biocompatibility and is beneficial to initial attachment, multiplication and follow-up detachment of cells, so that the titanium dioxide/albumin/biogenic signalling molecular composite coating can be widely applied to biomedical engineering fields of in-vitro cell culture, tissue engineering and the like.
Description
Technical field
The invention belongs to bio-medical coatings art, and in particular to have promotion cell adhesion concurrently and desorption is titania-based multiple
Close coating and preparation method thereof.
Background technology
At present mainly with pancreatin process during Cell culture invitro harvesting, but comprehend at pancreatin and make cell with extracellular base
Matter is separated, and is unfavorable for the later use of cell；In recent years, OKANO etc. can using its hydrophobic water-wet behavior using temperature sensitive polymer
The characteristic for varying with temperature successfully passes through temperature change and realizes cell and separates from cell culture vessel and do not destroy cell
Connection [N. Matsuda, T. Shimizu, M. Yamato, T. Okano, the Tissue of extracellular matrix protein
Engineering Based on Cell Sheet Technology, Advanced Materials, 2007, 19,
3089–3099].But, longer time is remained a need for as outfield using temperature, and required chilling process to a certain degree can damaged
Hinder the function of cell.
Due to TiO2Material has good biocompatibility and distinctive optic response so that it is in bio-medical side
The application in face constantly extends.Hong Yi et al. [Y. Hong et al. Light-induced cell detachment for
Cell sheet technology [J]. Biomaterials (2012) 1-8.] in TiO2Cultivate on nanodot films thin
Born of the same parents, have found that first cell can be from TiO by ultra-vioket radiation2It is desorbed on nanodot films, and extracellular matrix and thin
Born of the same parents' connection albumen can be desorbed together, and the cell for obtaining has good cytoactive.RGD is considered as current
Promote cell adhesion most effective and most widely used peptide sequence.Because various body cells express various integrins, it
Cell adhesion recognition site RGD fragment can be specifically bound, activation signalses path such as MAPK and JNK adjust transcription factor activity,
Instruct cell growth and differentiation.Albumin is the major protein of cell injuring model liquid.It is also egg that material surface adsorbs first
In vain, titanium dioxide has photic hydrophilic, under the exciting of UV 365, anatase TiO2Adsorb albumin on nano dot
Meeting ecto-entad is gradually desorbed, and causes Cell detachment.In order to obtain the higher cell thin of biological activity in vitro faster, if
Count out a kind of material that can accelerate cell in vitro film forming and can quickly be desorbed particularly significant.Therefore in titanium dioxide surface compound
The mixture of layer albumin and biogenic signaling molecule.This composite coating structure, can not only improve the biocompatibility of material also
Adhesion and the desorption of cell can be accelerated, for the development of organizational project, cell biological sensor etc. has important meaning.
The content of the invention
It is an object of the invention to provide it is a kind of with good biocompatibility and preparation method is simple, low cost with can
Titanium dioxide/albumin/biogenic signaling molecule composite coating of photic Cell detachment and preparation method thereof.
The present invention with can photic Cell detachment titanium dioxide/albumin/biogenic signaling molecule composite coating, under
And on have substrate, titanium dioxide nanometer points layer and albumin and biogenic signaling molecule layer successively, wherein, titanium dioxide nanometer points
The size of titanium dioxide nanometer points is 20~300nm in layer, and density is 1.0 × 109~1 × 1011/cm2, albumin and biological letter
Number molecular layer covers titanium dioxide nanometer points full of the gap between titanium dioxide nanometer points, albumin and bio signal point
The mass ratio of son is 1000:1~10:1, the quality sum of albumin and signaling molecule layer is the 0.4- of composite coating gross mass
50%。
The present invention with can photic Cell detachment titanium dioxide/albumin/biogenic signaling molecule composite coating system
Preparation Method, its step is as follows：
1）By titanium source, chelating agent and deionized water in molar ratio 1:0.3:1 ratio is added in dehydrated alcohol, Ran Houjia
Enter polyvinylpyrrolidone, be sufficiently stirred at room temperature, be configured to titanium atom concentration for 0.1-0.5mol/L, polyvinylpyrrolidine
Ketone concentration is the TiO 2 precursor colloidal sol of 10 ~ 100mg/ml；TiO 2 precursor colloidal sol is dropped to into the net substrate of cleaning
Surface is paved with to it, and with the speed spin coating 20-60s of 4000-8000rpm, at 400-600 DEG C is placed on afterwards 0.5- is incubated
2h, obtains titanium dioxide nanometer points layer in substrate；
2) it is 0.5mg/ml ~ 10 mg/ml by biogenic signaling molecule and concentration that concentration is 0.01mg/ml ~ 0.1mg/ml
Albumin albumin in mass ratio：Biogenic signaling molecule=1000:1~10:1 mixing；
3) step is taken by the 0.4-50% that the quality sum of albumin and biogenic signaling molecule layer is composite coating gross mass
2）Mixture, the mixture is dropped to into step 1）Obtained titanium dioxide nanometer points layer up to its be paved with, and with 100-
The speed spin coating of 1000rpm, at being then placed on 37 DEG C 0.5-2h is incubated.
Heretofore described substrate can be quartz glass, silica glass, silicon chip, tantalum piece, pure titanium sheet or Ti-Ni alloy piece.
Heretofore described titanium source can be butyl titanate, titanium tetrachloride, titanous chloride., titanium fluoride or titanium sulfate；
Described chelating agent is acetylacetone,2,4-pentanedione or ethanolamine.
Heretofore described biogenic signaling molecule is：RGD（Arginine-glycine-aspartic acid）、RGDS（Arginine-
Gly-Asp-serine）、GYRGDS（Gly-Tyr-RGDS）.
Heretofore described albumin is bovine serum albumin or human serum albumin.
Preferred albumin and the mass ratio of biogenic signaling molecule are 500 in the present invention:1~100:1.
The titanium dioxide base composite coating of the present invention, due to being received with biogenic signaling molecule modified titanic oxide using albumin
Rice grain, constructs composite coating, and all there is good biocompatibility to be conducive to cell for biogenic signaling molecule and albumin
Initial attachment, propagation and follow-up desorption.The preparation method is simple of the present invention, low cost is easy to utilize, can be widely applied to
The biomedical engineering fields such as Cell culture invitro, organizational project.
Description of the drawings
Fig. 1 be have can photic Cell detachment titanium dioxide/albumin/biogenic signaling molecule composite coating signal
Figure；
Fig. 2 is the SEM figures of titanium dioxide nanometer points thin film；
Fig. 3 is that titanium dioxide/albumin is schemed with the SEM of RGD composite coatings；
Fig. 4 is the composite coating and titanium dioxide nanometer points thin film cultured cells of titanium dioxide/albumin and RGD mixture
Absorbance after 24h；
Fig. 5 is titanium dioxide/albumin with RGD mixture composite coating and titanium dioxide nanometer points thin film with ultraviolet
The desorption rate of UV365 illumination 15min.
Specific embodiment
The present invention is described in detail with reference to embodiment and accompanying drawing, but the present invention is not limited to this.
With can photic Cell detachment titanium dioxide/albumin/biogenic signaling molecule composite coating as shown in figure 1, from
There are substrate 1, titanium dioxide nanometer points layer 2 and albumin and biogenic signaling molecule layer 3 successively on down.Wherein, titanium dioxide
The size of titanium dioxide nanometer points is 20~300nm in nano-dot layer, and density is 1.0 × 109~1 × 1011/cm2, albumin with
Biogenic signaling molecule layer 3 covers titanium dioxide nanometer points full of the gap between titanium dioxide nanometer points, albumin with it is biological
The mass ratio of signaling molecule is 1000:1~10:1, albumin is composite coating gross mass with the quality sum of signaling molecule layer
0.4-50%。
Embodiment 1
1）By butyl titanate, acetylacetone,2,4-pentanedione and deionized water in molar ratio 1:0.3:1 ratio is added to dehydrated alcohol
In, polyvinylpyrrolidone is subsequently adding, it is sufficiently stirred at room temperature, titanium atom concentration is configured to for 0.2 mol/L, polyethylene
Ketopyrrolidine concentration is the TiO 2 precursor colloidal sol of 40mg/ml；TiO 2 precursor colloidal sol is dropped to into the net stone of cleaning
English glass surface is paved with to it, and with the speed spin coating 40s of 8000rpm, 1h is incubated at 500 DEG C are placed on afterwards, in quartz
Titanium dioxide nanometer points layer is obtained on glass (see Fig. 2).The size of the titanium dioxide nanometer points is 30nm~200nm, and density is
3.0×1010/cm2。
2）RGD by the bovine serum albumin of concentration 10mg/ml with concentration for 0.01mg/ml mixes, bovine serum albumin
In vain：The mass ratio of RGD is 100:1；
3) it is the 0.4% of composite coating gross mass to take step 2 by the quality sum of bovine serum albumin and RGD layers）It is mixed
Compound, by the mixture step 1 is dropped to）Obtained titanium dioxide nanometer points layer up to its be paved with, and revolved with the speed of 500rpm
Apply, be incubated 1h at being then placed on 37 DEG C, obtain have can photic Cell detachment titanium dioxide/bovine serum albumin/RGD
Composite coating（See Fig. 3）.
Fiber born of the same parents' In vitro culture is carried out on above-mentioned composite coating, after cultivating one day, using CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), prepared titanium dioxide/Sanguis Bovis seu Bubali
The OD values that pure albumen/RGD composite coatings compare titanium dioxide nanometer points thin film are bigger (see Fig. 4), illustrate titanium dioxide/Sanguis Bovis seu Bubali
Pure albumen/RGD composite coatings have certain facilitation to cell adhesion.And after two days, the acquisition UV 365 of cell
Illumination 15min can be obtained.Obtain unicellular rear titanium dioxide/bovine serum albumin/RGD composite coating CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), desorption rate can be calculated.Titanium dioxide
The desorption rate of titanium/bovine serum albumin/RGD composite coatings and titanium dioxide nanometer points thin film is respectively 98% and 85%.（See figure
5）, illustrate that titanium dioxide/bovine serum albumin is more beneficial for the desorption of cell thin with RGD composite coatings.
Embodiment 2
1）By butyl titanate, acetylacetone,2,4-pentanedione and deionized water in molar ratio 1:0.3:1 ratio is added to dehydrated alcohol
In, polyvinylpyrrolidone is subsequently adding, it is sufficiently stirred at room temperature, titanium atom concentration is configured to for 0.2 mol/L, polyethylene
Ketopyrrolidine concentration is the TiO 2 precursor colloidal sol of 10mg/ml；TiO 2 precursor colloidal sol is dropped to into the net silicon of cleaning
Glass surface is paved with to it, and with the speed spin coating 40s of 4000rpm, 2h is incubated at 400 DEG C are placed on afterwards, in quartzy glass
Titanium dioxide nanometer points layer is obtained on glass.The size of the titanium dioxide nanometer points is 20nm~200nm, and density is 4.0 × 1010/
cm2。
2）RGD by the bovine serum albumin of concentration 10mg/ml with concentration for 0.01mg/ml mixes, bovine serum albumin
In vain：The mass ratio of RGD is 1000:1；
3) it is the 0.8% of composite coating gross mass to take step 2 by the quality sum of bovine serum albumin and RGD layers）It is mixed
Compound, by the mixture step 1 is dropped to）Obtained titanium dioxide nanometer points layer up to its be paved with, and revolved with the speed of 400rpm
Apply, at being then placed on 37 DEG C 1h is incubated.Obtain have can photic Cell detachment titanium dioxide/bovine serum albumin/RGD
Composite coating.
Fiber born of the same parents' In vitro culture is carried out on above-mentioned composite coating, after cultivating one day, using CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), prepared titanium dioxide/Sanguis Bovis seu Bubali
The OD values that pure albumen/RGD composite coatings compare titanium dioxide nanometer points thin film are bigger, illustrate titanium dioxide/bovine serum albumin
In vain/RGD composite coatings have certain facilitation to cell adhesion.And after two days, the acquisition illumination of UV 365 of cell
15min can be obtained.Obtain unicellular rear titanium dioxide/bovine serum albumin/RGD composite coating CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), desorption rate can be calculated.Titanium dioxide
The desorption rate of titanium/bovine serum albumin/RGD composite coatings and titanium dioxide nanometer points thin film is respectively 90% and 85%.Illustrate two
Titanium oxide/bovine serum albumin/RGD composite coatings are more beneficial for the desorption of cell thin.
Embodiment 3
1）Titanous chloride., acetylacetone,2,4-pentanedione and deionized water in molar ratio 1:0.3:1 ratio is added in dehydrated alcohol, so
After add polyvinylpyrrolidone, be sufficiently stirred at room temperature, be configured to titanium atom concentration for 0.2 mol/L, polyvinyl pyrrole
Alkanone concentration is the TiO 2 precursor colloidal sol of 10mg/ml；TiO 2 precursor colloidal sol is dropped to into the net silica glass of cleaning
Surface is paved with to it, and with the speed spin coating 20s of 4500rpm, 0.5h is incubated at 600 DEG C are placed on afterwards, on silica glass
Obtain titanium dioxide nanometer points layer.The size of the titanium dioxide nanometer points is 20nm~300nm, and density is 4.0 × 1010/cm2。
2）RGDS by the bovine serum albumin of concentration 10mg/ml with concentration for 0.01mg/ml mixes, bovine serum albumin
In vain：The mass ratio of RGDS is 50:1；
3) it is the 50% of composite coating gross mass to take step 2 by the quality sum of bovine serum albumin and RGDS layers）It is mixed
Compound, by the mixture step 1 is dropped to）Obtained titanium dioxide nanometer points layer up to its be paved with, and revolved with the speed of 100rpm
Apply, at being then placed on 37 DEG C 1h is incubated.Obtain have can photic Cell detachment titanium dioxide/bovine serum albumin/
RGDS composite coatings.
Fiber born of the same parents' In vitro culture is carried out on above-mentioned composite coating, after cultivating one day, using CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), prepared titanium dioxide/Sanguis Bovis seu Bubali
The OD values that pure albumen/RGDS composite coatings compare titanium dioxide nanometer points thin film are bigger, illustrate titanium dioxide/bovine serum albumin
In vain/RGDS composite coatings have certain facilitation to cell adhesion.And after two days, the acquisition illumination of UV 365 of cell
15min can be obtained.Obtain unicellular rear titanium dioxide/bovine serum albumin/RGDS composite coating CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), desorption rate can be calculated.Titanium dioxide
The desorption rate of titanium/bovine serum albumin/RGDS composite coatings and titanium dioxide nanometer points thin film is respectively 94% and 85%.Illustrate two
Titanium oxide/bovine serum albumin/RGDS composite coatings are more beneficial for the desorption of cell thin.
Embodiment 4
1）By butyl titanate, acetylacetone,2,4-pentanedione and deionized water in molar ratio 1:0.3:1 ratio is added to dehydrated alcohol
In, polyvinylpyrrolidone is subsequently adding, it is sufficiently stirred at room temperature, titanium atom concentration is configured to for 0.2 mol/L, polyethylene
Ketopyrrolidine concentration is the TiO 2 precursor colloidal sol of 100mg/ml；TiO 2 precursor colloidal sol is dropped to into the net silicon of cleaning
Glass surface is paved with to it, and with the speed spin coating 60s of 8000rpm, 2h is incubated at 600 DEG C are placed on afterwards, in silica glass
On obtain titanium dioxide nanometer points layer.The size of the titanium dioxide nanometer points is 30nm~300nm, and density is 9.0 × 1010/
cm2。
2）RGDS by the human serum albumin of concentration 10mg/ml with concentration for 0.01mg/ml mixes, human seralbumin egg
In vain：The mass ratio of RGDS is 50:1；
3) it is the 30% of composite coating gross mass to take step 2 by the quality sum of human serum albumin and RGDS layers）It is mixed
Compound, by the mixture step 1 is dropped to）Obtained titanium dioxide nanometer points layer up to its be paved with, and revolved with the speed of 100rpm
Apply, at being then placed on 37 DEG C 2h is incubated.Obtain have can photic Cell detachment titanium dioxide/bovine serum albumin/
RGDS composite coatings.
Fiber born of the same parents' In vitro culture is carried out on above-mentioned composite coating, after cultivating one day, using CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), prepared titanium dioxide/human blood
The OD values that pure albumen/RGDS composite coatings compare titanium dioxide nanometer points thin film are bigger, illustrate titanium dioxide/human seralbumin egg
In vain/RGDS composite coatings have certain facilitation to cell adhesion.And after two days, the acquisition illumination of UV 365 of cell
15min can be obtained.Obtain unicellular rear titanium dioxide/human serum albumin/RGDS composite coating CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), desorption rate can be calculated.Titanium dioxide
The desorption rate of titanium/bovine serum albumin/RGDS composite coatings and titanium dioxide nanometer points thin film is respectively 93% and 85%.Illustrate two
Titanium oxide/bovine serum albumin/RGDS composite coatings are more beneficial for the desorption of cell thin.
Embodiment 5
1）By titanium tetrachloride, ethanolamine and deionized water in molar ratio 1:0.3:1 ratio is added in dehydrated alcohol, so
After add polyvinylpyrrolidone, be sufficiently stirred at room temperature, be configured to titanium atom concentration for 0.2 mol/L, polyvinyl pyrrole
Alkanone concentration is the TiO 2 precursor colloidal sol of 80mg/ml；TiO 2 precursor colloidal sol is dropped to into the net pure titanium sheet of cleaning
Surface is paved with to it, and with the speed spin coating 45s of 4000rpm, at 600 DEG C are placed on afterwards 1h is incubated, in pure titanium sheet
To titanium dioxide nanometer points layer.The size of the titanium dioxide nanometer points is 30nm~150nm, and density is 1.0 × 1010/cm2。
2）RGDS by the human serum albumin of concentration 10mg/ml with concentration for 0.1mg/ml mixes, human seralbumin egg
In vain：The mass ratio of RGDS is 450:1；
3) it is the 50% of composite coating gross mass to take step 2 by the quality sum of human serum albumin and RGDS layers）It is mixed
Compound, by the mixture step 1 is dropped to）Obtained titanium dioxide nanometer points layer up to its be paved with, and revolved with the speed of 100rpm
Apply, at being then placed on 37 DEG C 2h is incubated.Obtain have can photic Cell detachment titanium dioxide/human serum albumin/
RGDS composite coatings.
Fiber born of the same parents' In vitro culture is carried out on above-mentioned composite coating, after cultivating one day, using CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), prepared titanium dioxide/human blood
The OD values that pure albumen/RGDS composite coatings compare titanium dioxide nanometer points thin film are bigger, illustrate titanium dioxide/human seralbumin egg
In vain/RGDS composite coatings have certain facilitation to cell adhesion.And after two days, the acquisition UV365 illumination 15min of cell
Can obtain.Obtain unicellular rear titanium dioxide/human serum albumin/RGDS composite coating CCK-8 (Cell Counting
Kit-8) method determines OD values (Optical Density, also known as absorbance), can calculate desorption rate.Titanium dioxide/human serum
The desorption rate of albumin/RGDS composite coatings and titanium dioxide nanometer points thin film is respectively 99% and 85%.Illustrate titanium dioxide/cattle
Serum albumin is more beneficial for the desorption of cell thin with the composite coating of RGDS mixture.
Embodiment 6
1）By titanium tetrachloride, ethanolamine and deionized water in molar ratio 1:0.3:1 ratio is added in dehydrated alcohol, so
After add polyvinylpyrrolidone, be sufficiently stirred at room temperature, be configured to titanium atom concentration for 0.2 mol/L, polyvinyl pyrrole
Alkanone concentration is the TiO 2 precursor colloidal sol of 60mg/ml；TiO 2 precursor colloidal sol is dropped to into the net pure titanium sheet of cleaning
Surface is paved with to it, and with the speed spin coating 35s of 7000rpm, at 600 DEG C are placed on afterwards 1h is incubated, in pure titanium sheet
To titanium dioxide nanometer points layer.The size of the titanium dioxide nanometer points is 30nm~100nm, and density is 1.0 × 1011/cm2。
2）GYRGDS by the human serum albumin of concentration 10mg/ml with concentration for 0.05mg/ml mixes, human seralbumin
Albumen：The mass ratio of GYRGDS is 300:1；
3) it is the 45% of composite coating gross mass to take step 2 by the quality sum of human serum albumin and GYRGDS layers）'s
Mixture, by the mixture step 1 is dropped to）Obtained titanium dioxide nanometer points layer up to its be paved with, and with the speed of 100rpm
Spin coating, at being then placed on 37 DEG C 0.5h is incubated.Obtain have can photic Cell detachment titanium dioxide/human seralbumin egg
In vain/GYRGDS composite coatings.
Fiber born of the same parents' In vitro culture is carried out on above-mentioned composite coating, after cultivating one day, using CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), prepared titanium dioxide/human blood
The OD values that pure albumen/GYRGDS composite coatings compare titanium dioxide nanometer points thin film are bigger, illustrate titanium dioxide/human seralbumin
Albumen/GYRGDS composite coatings have certain facilitation to cell adhesion.And after two days, the acquisition UV365 illumination of cell
15min can be obtained.Obtain unicellular rear titanium dioxide/human serum albumin/GYRGDS composite coating CCK-8 (Cell
Counting Kit-8) method measure OD values (Optical Density, also known as absorbance), desorption rate can be calculated.Titanium dioxide
The desorption rate of titanium/human serum albumin/GYRGDS composite coatings and titanium dioxide nanometer points thin film is respectively 99% and 85%.Explanation
Titanium dioxide/bovine serum albumin is more beneficial for the desorption of cell thin with GYRGDS composite coatings.
Claims (6)
1. have can photic Cell detachment titanium dioxide/albumin/biogenic signaling molecule composite coating, it is characterized in that from lower
On have substrate successively（1）, titanium dioxide nanometer points layer（2）And albumin and biogenic signaling molecule layer（3）, wherein, titanium dioxide
The size of titanium dioxide nanometer points is 20~300nm in titanium nano-dot layer, and density is 1.0 × 109~1 × 1011/cm2, albumin
With biogenic signaling molecule layer（3）Full of the gap between titanium dioxide nanometer points, and cover titanium dioxide nanometer points, albumin with
The mass ratio of biogenic signaling molecule is 1000:1~10:1, albumin is the total matter of composite coating with the quality sum of signaling molecule layer
The 0.4-50% of amount, described biogenic signaling molecule is RGD, RGDS or GYRGDS.
2. it is according to claim 1 with can photic Cell detachment titanium dioxide/albumin/biogenic signaling molecule be combined
Coating, it is characterised in that described substrate is quartz glass, silica glass, silicon chip, tantalum piece, pure titanium sheet or Ti-Ni alloy piece.
3. it is according to claim 1 with can photic Cell detachment titanium dioxide/albumin/biogenic signaling molecule be combined
Coating, it is characterised in that albumin is 500 with the mass ratio of biogenic signaling molecule:1~100:1.
4. prepare it is as claimed in claim 1 have can photic Cell detachment titanium dioxide/albumin/biogenic signaling molecule it is multiple
The method for closing coating, it is characterised in that step is as follows：
1）By titanium source, chelating agent and deionized water in molar ratio 1:0.3:1 ratio is added in dehydrated alcohol, is subsequently adding poly-
Vinylpyrrolidone, is sufficiently stirred at room temperature, and it is 0.1-0.5mol/L to be configured to titanium atom concentration, and polyvinylpyrrolidone is dense
Spend the TiO 2 precursor colloidal sol for 10 ~ 100mg/ml；TiO 2 precursor colloidal sol is dropped to into the net substrate surface of cleaning
It is paved with to it, and with the speed spin coating 20-60s of 4000-8000rpm, at 400-600 DEG C is placed on afterwards 0.5-2h is incubated,
Titanium dioxide nanometer points layer is obtained in substrate；
2) it is the white of 0.5mg/ml ~ 10 mg/ml by biogenic signaling molecule and concentration that concentration is 0.01mg/ml ~ 0.1mg/ml
Albumen albumin in mass ratio：Biogenic signaling molecule=1000:1~10:1 mixing；Described biogenic signaling molecule is RGD, RGDS
Or GYRGDS；
3) step 2 is taken by the 0.4-50% that the quality sum of albumin and biogenic signaling molecule layer is composite coating gross mass）'s
Mixture, by the mixture step 1 is dropped to）Obtained titanium dioxide nanometer points layer up to its be paved with, and with 100-1000rpm's
Speed spin coating, at being then placed on 37 DEG C 0.5-2h is incubated.
5. it is according to claim 4 with can photic Cell detachment titanium dioxide/albumin/biogenic signaling molecule be combined
The preparation method of coating, it is characterised in that described titanium source is butyl titanate, titanium tetrachloride, titanous chloride., titanium fluoride or sulfur
Sour titanium；Described chelating agent is acetylacetone,2,4-pentanedione or ethanolamine.
6. it is according to claim 4 with can photic Cell detachment titanium dioxide/albumin/biogenic signaling molecule be combined
The preparation method of coating, it is characterised in that described chelating agent is acetylacetone,2,4-pentanedione or ethanolamine.
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| 组织工程中细胞与材料的粘附作用;秦廷武等;《中国修复重建外科杂志》;19990128;第13卷(第1期);第35页"3.3.2 固定多肽类物质"部分 * |
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