CN103751847A - Preparation method of tissue-regeneration-promoting controlled-release multiple-growth-factor self-assembled coating - Google Patents
Preparation method of tissue-regeneration-promoting controlled-release multiple-growth-factor self-assembled coating Download PDFInfo
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Abstract
The invention belongs to the biological regenerative medicine field, and particularly relates to a preparation method of a tissue-regeneration-promoting controlled-release multiple-growth-factor self-assembled coating. The preparation method is as follows: using an organic polymer membrane and a three-dimensional scaffold as templates, and loading bone morphogenetic protein-2 (BMP2), dentin non-collagenous proteins (DNCPs), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and other bioactive factors with tissue regeneration promotion effects by a supramolecular layer by layer self-assembly technique to form a certain layers of supramolecular biological film coating structure simultaneously embedding with different growth factors on the template surface. The tissue-regeneration-promoting functional coating prepared by the preparation method has good biological compatibility and stability, can improve the surface properties of materials, and can induce tissue regeneration by adjusting the release of the loaded growth factors.
Description
Technical field
The invention belongs to bio-regeneration medical domain, be specifically related to the preparation method of the multiple somatomedin self-assembled coating of a kind of short tissue regeneration controlled release.
Background technology
Tissue and organ dysfunction or damaged serious harm human health are to cause one of human diseases and main causes of death.Data shows according to statistics, and the U.S. is every year because various tissues and organ dysfunction or the damaged operation causing are up to up to a million times [1].China need carry out dermatoplastic patient in 1,000,000 because of burn every year, and the patient that need carry out artificial joint replacement because of various bones, cartilage injury is up to 1,000,000.Clinical conventional replacement therapy method comprises allograft, autograft and synthetic tissue substitute, the part but these methods come with some shortcomings.As one of most active research in bio-medical engineering, organizational project is showing huge application prospect [2] aspect reparation, reconstruction and the regeneration of tissue or organ.Organizational project is according to the principle of cytobiology and engineering, the seed tissue cell of In vitro culture is adsorbed on the biomaterial that good biocompatibility and degradable absorb and forms cell-biomaterial composites, the bioactie agent that use in conjunction is different, be implanted into subsequently body disease damage position, along with biomaterial is degraded and seed cell breaks up to specific cells gradually, formation has new tissue or the organ of corresponding form, function, to reach the object [3-5] of repairing and improving.At present, the research in this field has related to Various Tissues or the organs such as tooth, cartilage, bone, tendon, blood vessel, trachea, pancreas, liver.The three elements that organizational project builds comprise seed cell, timbering material and bioactie agent.Wherein, well behaved timbering material and bioactie agent play decisive role in the adhesion of seed cell, propagation, atomization.Timbering material, as one of key element, should possess good mechanical strength, biological activity and supply with the three-D space structure that cell is grown into.Bioactie agent, as somatomedin, mainly obtains by the autocrine of inoculating cell or the approach of the interpolation exogenous growth factor and class growth-factor medication.Because somatomedin existence and stability is poor, the half-life is short and the effect of easy body endoenzyme and the features such as inactivation [6], mostly by means of suitable carrier or slow-released system to protect its biological activity and its continuous action of performance, promote cell proliferation and tissue repair [7].Existing large quantity research [6-13] has been reported the growth of somatomedin to stem cell by In vitro cell experiment, the regulating and controlling effect of propagation and differentiation.Stem cell becomes the somatic cell that becomes with difference in functionality and characteristic from seed cell directed differentiation, somatomedin is vital.And how somatomedin is loaded on timbering material, and realize the orderly controlled release of its timesharing, be key one ring [14] of growth factor solution performance actual application value.Elcin etc. [15] find that the formation speed of local new vessels net is obviously accelerated after adopting alginate beads and VEGF (Vascular Endothelial Growth Factor, VEGF) to build in slow-released system implantation Mice Body.Another group research [16] by wrapping up VEGF in PLGA/ sodium alginate nanoparticle, make VEGF in vitro sustained release over 21 days, and compared to blank group and VEGF single injection group, in human umbilical vein endothelial cell breeding, demonstrate clear superiority in vitro.Zhang etc. [17] study discovery, and the compound rest of being combined with TGF-1L by chitosan/collagen scaffold is compared with simple support, and people's periodontal ligament cell propagation significantly.The research of a nearest relevant osteochondral defect shows, the hyaluronic acid of load fibroblast growth factor (basic fibroblast growth Factor, bFGF) and insulin/
collagen type/fibrin scaffold which can promote the migration of mescenchymal stem cell and be divided into chondrocyte, is conducive to the reparative regeneration [18] of osteochondral defect tissue.More than research is all pointed out by carrier or slow-released system bioactie agent is carried among timbering material, has both protected its biological activity, is conducive to again the efficient performance that continues of biological agent, is conducive to reparation and the regeneration of tissue or organ.Yet, the combination of timbering material and somatomedin and how to reach desirable controlled-release effect and make it approach cells in vivo living environment and remain from now on one of direction of research.At present, in great majority research, somatomedin mode of loading adopts the combination of nanoparticle or microsphere and timbering material more, has comparatively complexity of processing technology, the problem such as in some large area tissue regeneration, is difficult to sprawl.How to build easily the well loaded of somatomedin and timbering material, the composite that formation has certain biological activity and layer functions is problem demanding prompt solution.The mode that the self-assembled coating of the multiple somatomedin of short tissue regeneration controlled release that we prepare has with somatomedin controlled release promotes tissue regeneration, and segmentation is induced, easy and simple to handle, without special installation, be beneficial to the feature such as sprawl, applied widely.And constructed load has the nanoassemble coating structure of somatomedin (BMP-2) to demonstrate the features such as high stability, high cell compatibility, slow release be stable in our recent work.
In sum, in the report of existing document, not yet relevant for the correlational study report of the self-assembled coating of the multiple somatomedin of short tissue regeneration controlled release described in the present invention.
list of references
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[10]Ng?F,?Boucher?S,?Koh?S,?et?al.?PDGF,?TGF-B,?and?FGF?signaling?is?important?for?differentiation?and?growth?of?mesenchymal?stem?cells:?transcriptional?profiling?can?identify?markers?and?signaling?pathways?important?in?differentiation?of?BMSCs?in?to?adipogenic,?chondrogenic,?and?osteogenic?lineages.?Blood?2008;112(2):295-307.
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[15]Elcin?YM,?Dixit?V,?Gitnick,?et?al.?Extensive?in?vivo?angiogenesis?following?controlled?release?of?human?vascular?endothelial?cell?growth?factor:?implications?for?tissue?engineering?and?wound?healing.?Artif?Organs?2001;25(7):558-561.
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summary of the invention
The object of the invention is to propose first the preparation method of the multiple somatomedin self-assembled coating of a kind of short tissue regeneration controlled release, utilize existing organic polymer diaphragm, three-dimensional racks etc. are template, by supermolecule layer-by-layer, to tissue regeneration be there is to the bioactie agent of facilitation, as BMP-2 (BMP-2), dentin non-collagenous protein (DNCPs), VEGF (VEGF), basic fibroblast growth factor (bFGF) etc., load on template surface, formation has the supermolecule biofilm coating structure of certain number of plies the different somatomedin of embedding simultaneously, there is good stability and biocompatibility.
The technical scheme that the present invention provides is:
Take existing organic polymer diaphragm, three-dimensional rack etc. is template, by supermolecule layer-by-layer, bioactie agent tissue regeneration to facilitation is loaded on to template surface, form the supermolecule biofilm coating structure with certain number of plies the different somatomedin of embedding simultaneously.The short tissue regeneration coating making by the inventive method, has good stability and biocompatibility, then after sterilization packaging, obtains product.
A preparation method for the multiple somatomedin self-assembled coating of short tissue regeneration controlled release, is characterized in that, specifically comprises the steps:
Step 1, prepare mould material:
Adopt host material, comprise as: take a kind of in polylactic acid (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), Poly(D,L-lactide-co-glycolide (PLGA), collagen (collagen), hydroxyapatite (HA), beta-calcium phosphate (β-TCP), sodium alginate (ALG), gelatin (gelatin) or arbitrarily several mixture by electrostatic spinning technique, prepare organic polymer diaphragm as raw material, or take gelatin and modification of chitosan and by photocuring, obtain porous three-dimensional host material as raw material, but be not restricted to this two example.
Select a class somatomedin for tissue regeneration being there is to the bioactive molecule of facilitation, comprise as BMP-2 (BMP-2), dentin non-collagenous protein (DNCPs), VEGF (VEGF), basic fibroblast growth factor (bFGF) etc., but be not restricted to this four example.Somatomedin lysate of the present invention is acid solution, and wherein HCL concentration is 2-6mM.And somatomedin need to be dissolved in polyelectrolyte solution, require somatomedin and polyelectrolyte solution not with contrary electrically.Polyelectrolyte solution of the present invention is: polycyclic aromatic hydrocarbon (Polycyclic Aromatic Hydrocarbon, PAH) and polyacrylic acid (Polyacrylic Acid, PAA), polystyrolsulfon acid (Polystyrene sulfonic acid, PSS), diallyl dimethyl ammoniumchloride (Poly dimethyl diallyl ammonium chloride, PDDA), compound concentration is 0.5-3mg/ml, and the NaCl content in solution is 0.05-0.4M.The cleaning solution adopting (washing liquid) is: deionized water (deionized water), MiniQ ultra-pure water (Millipore), acetic acid-sodium-acetate buffer (Sodium acetate buffer solution), or Tris-hydrochloric acid (Tris-HCl) buffer.
Step 3, use supermolecule layer-by-layer:
Use supermolecule layer-by-layer, it is static layer-by-layer, this know-why is to utilize the weak interaction force (as model Dehua power, electrostatic attraction, hydrogen bond etc.) of existence between macromole, the orderly alternating deposit of realization to biomacromolecule and polyelectrolyte, so that all kinds of polyelectrolyte of high-efficient simple ground assembling, thereby form the ultrathin membrane of nano thickness, maintain biological activity and the space structure of biomacromolecule, can on the device of complex space configuration, operate again.The preparation condition adopting in the present invention is: template time of immersion in every kind of solution is 5-20min, template dip-coating in every kind of solution be sequentially " ...-positive charge solution-washing liquid-negative charge solution-washing liquid-positive charge solution-washing liquid-negative charge solution-... " so circulation.The solution that ground floor soaks, what require solution electrically must be electrically contrary with template surface.
Step 4, makes to be cross-linked and forms supermolecule biofilm coating structure:
After self assembly completes, can use cross-linking agent (cross-linking agent), as carbodiimides (EDC), N-hydroxy-succinamide (NHS), but be not restricted to this two example.Finally at template surface, form the supermolecule biofilm coating structure with certain number of plies the somatomedin of embedding simultaneously.
Advantage of the present invention compared with prior art: the multiple somatomedin self-assembled coating of this short tissue regeneration controlled release structure has good biocompatibility and stability, can regulate and control growth factor-loaded release and have operability on the device of complex space configuration, the original position induction that is beneficial to tissue regeneration activates.
Accompanying drawing explanation
Fig. 1 is embodiment 1 load { [(PAH/BMP-2)/PSS]/[COL/ALG] }
20the PCL support of supermolecular film (B support), the controlled release curve of BMP-2 somatomedin in 21 days.
Fig. 2 is the Electronic Speculum pattern of embodiment 1 empty PCL thin film (A support).
Fig. 3 is the Electronic Speculum pattern of B support in embodiment 1.
Fig. 4 is the Electronic Speculum pattern that embodiment 1 periodontal ligament stem cell is inoculated 4 days on A support.
Fig. 5 is the Electronic Speculum pattern that embodiment 1 periodontal ligament stem cell is inoculated 4 days on B support.
Fig. 6 is embodiment 1 periodontal ligament stem cell while inoculating 7 days on A support, the high power Electronic Speculum pattern that cell distributes at material surface.
Fig. 7 is embodiment 1 periodontal ligament stem cell while inoculating 7 days on B support, the high power Electronic Speculum pattern that cell distributes at material surface.
Fig. 8 is the alkaline phosphatase activities that embodiment 1 periodontal ligament stem cell is grown on A, two kinds of supports of B.
Fig. 9 is the MTT cell proliferation curve that embodiment 1 periodontal ligament stem cell is grown on A, two kinds of supports of B.
Figure 10 is embodiment 1{[(PAH/BMP-2)/PSS]/[COL/ALG] }
20supermolecular film is structured in the schematic diagram on blank PCL electrostatic spinning support.
The specific embodiment
embodiment 1
Below in conjunction with drawings and Examples, further introduce technical solution of the present invention.
Step 1, employing PCL is raw material, is dissolved in chloroform, makes mass ratio and be 8% solution.This solution is joined in the electrostatic spinning syringe of controlling by syringe pump, and high tension generator voltage is set as 15KV, and collecting distance is 16cm, and syringe flow velocity is 0.5ml/min, on square aluminium foil, collects electrostatic spinning film.After natural drying, then after using ethanol and deionized water wash repeatedly, nitrogen drying is stand-by.
Step 3, immerses above-mentioned stand-by thin film after (PAH/BMP-2) solution 10min, with deionized water wash, then immerses PSS solution 10min, with deionized water wash, forms the somatomedin structure coating of [(PAH/BMP-2)/PSS]; In like manner making barrier layer is: [COL/ALG].[structure coating+barrier layer], through 20 circulations, forms { [(PAH/BMP-2)/PSS]/[COL/ALG] }
20supermolecular film structure, controlled release BMP-2.This step is as shown in Figure 10: be illustrated as the Build Order that an one functional layer adds one deck barrier layer, utilize the static of the positive and negative charge power that attracts each other, 5 kinds of compositions are adsorbed on to PCL rack surface.
Step 4, by above-mentioned electrostatic spinning film after coating is processed, immerses the solution crosslinking 30min that contains cross-linking agent EDC/NHS, washing, and-20 ° of C are freezing stand-by.
The accompanying drawing that the present embodiment provides has compared the difference on pattern, performance, cell compatibility of two kinds of timbering materials, is respectively blank PCL electrostatic spinning support (A support) and load { [(PAH/BMP-2)/PSS]/[COL/ALG] }
20the PCL support of coating (B support).
Fig. 1 has illustrated { [(PAH/BMP-2)/PSS]/[COL/ALG] }
20the common B support forming of coating+PCL thin film can sustained release BMP-2 in 21 days.
According to step shown in embodiment 1, by { [(PAH/BMP-2)/PSS]/[COL/ALG] }
20supermolecular film structural load on PCL electrostatic spinning support, form B support.By area, be 1CM
2b support material be immersed in the PBS buffer of 2ml.Therefrom take out 0.5ml buffer continuous 21 day every day, and supplement simultaneously and inject 0.5ml buffer.In detecting 21 days by ELISA method, the release concentration of BMP-2, draws concentration release profiles.
Fig. 2 and Fig. 3: the Electronic Speculum pattern that has compared A support and B support.As shown in the figure, { [(PAH/BMP-2)/PSS]/[COL/ALG] }
20coating makes PCL film surface by fine thread, roughness, becomes smooth.
Fig. 4: the pattern of periodontal ligament stem cell on electric Microscopic observation A support, cell surface is smooth; Now cell was material surface inoculation 4 days.
Fig. 5: the pattern of periodontal ligament stem cell on electric Microscopic observation B support, cell surface has a large amount of mineralising secretory vacuoles, and forms mineralising crystal, and mineralising crystal is as shown in white arrow.Now cell was material surface inoculation 4 days.
The explanation load { [(PAH/BMP-2)/PSS]/[COL/ALG] } of above Fig. 1-5
20the PCL support of coating can discharge BMP-2, makes material have biological activity, and induces periodontal ligament stem cell to possess mineralization ability.
Fig. 6-9: compared blank PCL electrostatic spinning support (A support) and load { [(PAH/BMP-2)/PSS]/[COL/ALG] }
20the PCL support of coating (B support) cell proliferation dry to periodontal membrane, the impact of alkaline phosphatase activities.
Fig. 6: electric Microscopic observation periodontal ligament stem cell is seeded in after A support the pattern of the 7th day.Illustrate that cell is slower in the propagation of the 11st day cell.
Fig. 7: electric Microscopic observation periodontal ligament stem cell is seeded in after B support the pattern of the 7th day.Illustrate that cell is very fast in the propagation of the 11st day cell.
Fig. 6 and 7 explanations { [(PAH/BMP-2)/PSS]/[COL/ALG] }
20coating has improved cytoactive and the growth rate of periodontal ligament stem cell on PCL support.
Fig. 8: the alkali phosphatase that periodontal ligament stem cell is grown on A, two kinds of supports of B is lived and contrasted.By alkaline phosphatase activities test kit method, compared periodontal ligament stem cell after being inoculated on A, two kinds of supports of B, the 36th hour, the alkaline phosphatase activities of the 4th, 7,11 days, was presented at above-mentioned four time points { [(PAH/BMP-2)/PSS]/[COL/ALG] }
20the be significantly increased alkaline phosphatase activities of periodontal ligament stem cell of coating, illustrates that this coating has induction periodontal ligament stem cell to the function with the cell differentiation of mineralization ability.
Fig. 9: the cell proliferation curve that periodontal ligament stem cell is grown on A, two kinds of supports of B.By MTT agent box method, compared periodontal ligament stem cell after being inoculated on A, two kinds of supports of B, the 36th hour, the cell proliferation situation of the 4th, 7,9,11 days, was presented at the 7th, 9,11 days, { [(PAH/BMP-2)/PSS]/[COL/ALG] }
20coating significantly improves the cell proliferation speed of periodontal ligament stem cell, illustrates that this coating has improved the cell compatibility of PCL spinning thin film.
Figure 10: { [(PAH/BMP-2)/PSS]/[COL/ALG] } is described
20supermolecular film is structured in principle and the process on blank PCL electrostatic spinning support.
Step 1, adopting PLGA and collagen protein (collagen) is raw material (mass ratio is 6:1), is dissolved in hexafluoro propanol, makes mass percent and be 7% solution.This solution is joined in the electrostatic spinning syringe of controlling by syringe pump, and high tension generator voltage is set as 13KV, and collecting distance is 14cm, and syringe flow velocity is 0.02ml/min, collects electrostatic spinning film on aluminium foil.After collection, inserted vacuum drying oven (temperature is 35 ° of C) inner drying, by the EDC solution of the thin film immersion 5% of dry gained, under room temperature, be cross-linked 1 hour subsequently, after using ethanol and deionized water wash repeatedly, nitrogen drying is stand-by.
Step 3, immerses above-mentioned stand-by thin film after DNCPs solution 10min, with deionized water wash, circulates 2 times.Immerse respectively again PAH and each 10min of PAA solution, with deionized water wash, form [(DNCPs)
2-(PAH/PAA)
1] structure coating (subscript represents the number of plies that self assembly is layer by layer constructed); The structure that in like manner forms intermediate layer is: [(PAH/PAA-DNCPs)
2-(PAH/PAA)
3]; Surface structure is: [(PAH/PAA)
2-(DNCPs)
1].[structure coating+intermediate layer+top layer] can be as required, through 15 more somatomedin of repeated loading.
Step 4, by above-mentioned electrostatic spinning film after coating is processed, immerses in the solution that contains collagen protein after 10min, through washing, then immerses the solution that contains cross-linking agent genipin the collagen protein of coating is cross-linked to 20min, then washs stand-by.
embodiment 3
Step 1, adopting gelatin (Gelatin) and modification of chitosan (chitosan) is raw material (mass ratio is 2:3), be dissolved in N-N-dimethyl acetylamide (DMA), and adding therein 1.0% (wt) 2-dimethylamino-2-benzyl-1-[4-(4-morpholinyl) phenyl]-1-butanone is as trigger for optical solidification, at 365nm, 10 w/cm
2under ultraviolet light, irradiation 20min, obtains gel rubber material.Gained gel is washed under deionized water effect, remove DMA wherein, obtain porous three-dimensional matrix scaffold.After collection, inserted vacuum drying oven (temperature is 35 ° of C) inner drying, subsequently the timbering material of dry gained being immersed to concentration is in 2mg/ml polymine (PEI) solution, soaks after 10min, and with deionized water wash, nitrogen drying is stand-by.
Step 3, immerses above-mentioned timbering material after bFGF solution 20min, uses deionized water wash.Immerse respectively again after PAH and PAA solution 20min, with deionized water wash, circulate 3 times.Form [(bFGF)
1-(PAH/PAA)
3] structure coating; The structure that in like manner forms intermediate layer is: [(PAH/PAA-bFGF)
3-(PAH/PAA)
2]; Surface structure is: [(PAH/PAA)
1-(bFGF)
2].[structure coating+intermediate layer+top layer] can be as required, through 25 more somatomedin of repeated loading.
Step 4, by above-mentioned timbering material deionized water wash after coating is processed, stream of nitrogen gas dried for standby.
embodiment 4
Step 1, adopting gelatin (Gelatin) and modification of chitosan (chitosan) is raw material (mass ratio is 1:2), be dissolved in dimethyl sulfoxine (DMSO), and add therein 0.5% (wt) 1-[4-(2-hydroxy ethoxy)-phenylene]-2-hydroxyl-2 ', 2 '-dimethyl ethyl ketone is as trigger for optical solidification, at 200nm, 5 w/cm
2under ultraviolet light, irradiation is 12 minutes, obtains gel rubber material.Gained gel is washed under deionized water effect, remove DMSO wherein, obtain porous three-dimensional matrix scaffold.After collection, inserted vacuum drying oven (temperature is 35 ° of C) inner drying, subsequently the timbering material of dry gained being immersed to concentration is in 1.5mg/ml polymine (PEI) solution, soaks after 10min, and with deionized water wash, nitrogen drying is stand-by.
Step 3, immerses above-mentioned timbering material after VEGF solution 15min, uses deionized water wash.Immerse respectively again after PAH and PAA solution 15min, with deionized water wash, circulate 2 times.Form [(VEGF)
1-(PAH/PAA)
2] structure coating; The structure that in like manner forms intermediate layer is: [(PAH/PAA-VEGF)
1-(PAH/PAA)
3]; Surface structure is: [(PAH/PAA)
3-(VEGF)
2].[structure coating+intermediate layer+top layer] can be as required, through 30 more somatomedin of repeated loading.
Step 4, by above-mentioned timbering material deionized water wash after coating is processed, stream of nitrogen gas dried for standby.
Claims (1)
1. a preparation method for the multiple somatomedin self-assembled coating of short tissue regeneration controlled release, is characterized in that, specifically comprises the steps:
Step 1, prepare mould material:
Adopt host material comprise as: take in polylactic acid (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), Poly(D,L-lactide-co-glycolide (PLGA), collagen (collagen), hydroxyapatite (HA), beta-calcium phosphate (β-TCP), sodium alginate (ALG), gelatin (gelatin) a kind of or arbitrarily several mixture by electrostatic spinning technique, prepare organic polymer diaphragm as raw material, or take gelatin and modification of chitosan and by photocuring, obtain porous three-dimensional host material as raw material;
Step 2, adopts somatomedin to promote the regeneration to tissue:
Select a class somatomedin for tissue regeneration being there is to the bioactive molecule of facilitation, be specially BMP-2 (BMP-2), dentin non-collagenous protein (DNCPs), VEGF (VEGF), basic fibroblast growth factor (bFGF)
The somatomedin lysate adopting is acid solution, and wherein HCL concentration is 2-6mM, and somatomedin need to be dissolved in polyelectrolyte solution, requires somatomedin and polyelectrolyte solution not with contrary electrical,
The polyelectrolyte solution adopting is: polycyclic aromatic hydrocarbon (Polycyclic Aromatic Hydrocarbon, PAH) and polyacrylic acid (Polyacrylic Acid, PAA), polystyrolsulfon acid (Polystyrene sulfonic acid, PSS), diallyl dimethyl ammoniumchloride (Poly dimethyl diallyl ammonium chloride, PDDA), compound concentration is 0.5-3mg/ml, and the NaCl content in solution is 0.05-0.4M
The cleaning solution adopting is: deionized water (deionized water), MiniQ ultra-pure water (Millipore), acetic acid-sodium-acetate buffer (Sodium acetate buffer solution), or Tris-hydrochloric acid (Tris-HCl) buffer;
Step 3, use supermolecule self assembly layer by layer:
Use supermolecule layer-by-layer, utilize between macromole and exist weak interaction force to realize the orderly alternating deposit to biomacromolecule and polyelectrolyte, form the ultrathin membrane of nano thickness,
The preparation condition adopting is: template time of immersion in every kind of solution is 5-20min, template dip-coating in every kind of solution be sequentially " ...-positive charge solution-washing liquid-negative charge solution-washing liquid-positive charge solution-washing liquid-negative charge solution-... " so circulation, the solution that ground floor soaks, what require solution electrically must be electrically contrary with template surface;
Step 4, makes to be cross-linked and forms supermolecule biofilm coating structure:
After self assembly completes, can use cross-linking agent (cross-linking agent), finally at template surface, form the supermolecule biofilm coating structure with certain number of plies the somatomedin of embedding simultaneously.
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN113710292A (en) * | 2019-02-13 | 2021-11-26 | 法国国家健康与医学研究院 | Biomaterial comprising a scaffold containing mineral compounds and its use as bone substitute |
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| CN113183594A (en) * | 2021-04-07 | 2021-07-30 | 广东工业大学 | Preparation method of electrostatic spinning functional nanofiber film for epidermis repair |
| CN115227872A (en) * | 2021-04-25 | 2022-10-25 | 上海大学 | Collagen fiber-based drug sustained-release system and construction method and application thereof |
| CN113769167A (en) * | 2021-09-14 | 2021-12-10 | 深圳先进技术研究院 | Preparation method of orthopedic implant material capable of sequentially releasing bioactive factors, product and application thereof |
| CN113769167B (en) * | 2021-09-14 | 2023-07-25 | 深圳先进技术研究院 | Preparation method of orthopedic implant material capable of sequentially releasing bioactive factors, product and application thereof |
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