CN102836461A - Human hair keratin shape righting material with adjustable and controllable mechanical property as well as preparation method thereof - Google Patents
Human hair keratin shape righting material with adjustable and controllable mechanical property as well as preparation method thereof Download PDFInfo
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Abstract
The invention provides a human hair keratin shape righting material with adjustable and controllable mechanical property and a preparation method of the human hair keratin shape righting material. The human hair keratin shape righting material is prepared by a pure keratin solution obtained by treating human hairs or animal hairs by a reduction method; the obtained human hair keratin shape righting material is good in elasticity controllability and excellent in mechanical property; a controllable range of elastic elongation rate is 50-110%, the elastic recovery rate is 98-99%, the breakage strength is 0.5-0.7GPa, and the initial modulus is 0.1-0.75GPa; and requirements of elasticity and mechanical property for shape righting and repairing of loose connective tissues and different muscle body tissues such as blood vessels, ligaments and cartilage tissues can be met. The human hair keratin shape righting material is prepared by pure human hair keratin, can induce collagenous fiber regeneration, is good in biodegradability, can be fully degraded and absorbed by muscle bodies, does not cause tissue adhesion in the shape righting and repairing processes, and has a wide application prospect in the field of biological medicines.
Description
Technical field
It is regulatable from body keratin orthotic material and preparation method thereof to the present invention relates to a kind of mechanical property, belongs to technical field of biological materials.
Background technology
The important subject that the excellent biocompatibility development of new organism orthotic material that utilizes natural biologic material is bio-medical engineering and organizational project replacement therapy field.The human body of human body or animal constitutes by having flexible in a large number and being rich in elastic connective tissue, and these connective tissues are brought into play different functions in different organ or tissues, guarantee the normal operation of human body.Collagen fiber are a kind of important fibers that constitute connective tissue, extensively are present in internal organs, skin, blood vessel, sclera, ligament and the tendon.In order to satisfy the demand of human body different tissues function, also there is difference in the structure of collagen fiber in different tissues, and be different with the difference of its place tissue like the fibril diameter of collagen fiber, thick about 20 times of the fibril in the comparable cornea of the fibril of tendon.Fibril is thicker in the tissue of high load forces, and the size of its load forces becomes positive correlation with the fibril thickness.Therefore, develop the good orthotic material of a kind of adaptability, on the one hand, make it have excellent biological compatibility, can induce collagen to live again; On the other hand, also need consider the elasticity of material and the controllability of mechanical property, make it can satisfy the needs that the human body different tissues is orthopedic and repair.
Already relevant for the report of three-dimensional building process of collagen fiber growth in vitro polymerization and characteristic; This type of work is the basis of developing orthopedic and tissue substitute material; But draw materials from body collagen and to be restricted; Can only be with the collagen tissue of taking from animal body as raw material, therefore, the phenomenon that necroses easily and rejection.And the hair keratin like protein material that is taken from from body is that above-mentioned donor contradiction provides good breach, and the Crinis Carbonisatus that can get the patient prepares the orthotic material reparation that is used to control oneself as raw material.Keratin also has and is self-assembled into fine characteristic, can prepare the orthotic material with excellent mechanical performances.In addition; The keratin macromole is rich in cystine (7%-8%); Cystine make in the keratin macromole and macromole between form cross-bond, thereby given natural keratin material better elastic performance and mechanical property, through taking suitable regulate and control method; Kerateine can prepare and has the regulatable orthotic material of mechanical property, is applicable to the orthopedic repairing and treating of different tissues.
Human the research of from natural material, extracting keratin and being translated into powder, film, coating etc. is started from the seventies in 20th century.After getting into the eighties in 20th century, keratin as the development of emerging biomaterial rapidly, at present, existing many related patent U.S. Patent No. document announcements the method for preparing of keratin orthotic material.
Chinese patent (03819328.0); This invention provides a kind of keratin orthotic material that is used for the bone therapeutic treatment; Described keratin material just has sulfonation and is rich in high-molecular weight intermediate filament protein, processes through compacting solid keratin powder, can make porous; Also can be fine and close keratin material, can be used for the internal fixing device of displacement, increase and the fracture of bone.But the rigidity of powder compaction thing is stronger, and human body mostly tissue all be material with certain flexibility, therefore, the application of this type of orthotic material is limited.
Chinese patent (200780005567.6); This invention provides a kind of charged keratin stage division; Prepared keratin can be used as the coating of biomedical implants; Can effectively reduce bonding to base material of cell and tissue, can be used for developing orthotic materials such as blood vessel graft, conduit, cardioverter, also can be used for the repairing and treating of thromboembolism.But often there is non-degradable property in the matrix of this type of medical implant, need accomplish orthopedic with reply the back operation of treatment back and remove, increased misery to the patient, therefore, should further develop the orthopaedic implant of biodegradation type.
Chinese patent (99107708.3), this patent provides a kind of good biocompatibility, can inducing self-body tendon form manufacture method that can the orthopedic repair materials of absorbed artificial tendon made of human hair keratin in human body.This artificial tendon made of human hair keratin relaxes woollen yarn knitting by a certain percentage by two or more human hair keratin in various degree, that controlled biochemistry is handled and forms.Can be used for the orthopedic reconstructive surgery of medical science uses.This artificial tendon has chemically treated Crinis Carbonisatus fiber woollen yarn knitting to form, and has kept the intensity of Crinis Carbonisatus fiber, can satisfy the mechanical property requirements of the high-strength flatness of tendon, and give from the good biological characteristics of body Crinis Carbonisatus through biochemical treatment.Having overcome the deficiency of " allogeneic tendon ", " carbon fiber tendon ", is the model of exploitation from the orthopedic embedded material of body keratin.
The present invention provides a kind of pure keratin that is made by reducing process, and to be self-assembled into the elasticity that fibre constitutes regulatable from body keratin orthotic material through external, do not contain cross-linking agent or plasticizer.This keratin orthotic material has good biomechanical property; Can induce collagen fiber to live again, have good biodegradability properties, can be by the complete degraded and absorbed of human body; The tissue adhesion is not taken place in the orthopedic repair process, can be widely used in the orthopedic repairing and treating of organ and tissue.
Summary of the invention
It is regulatable from body keratin orthotic material and preparation method thereof to the purpose of this invention is to provide a kind of mechanical property.
In order to achieve the above object, to provide a kind of mechanical property regulatable from body keratin orthotic material for technical scheme of the present invention.It is characterized in that, make by the pure keratin solution that reducing process obtains to have controllability from the elastic performance of body keratin orthotic material, and have good biomechanical property from body keratin orthotic material.The elastic extension controlled range is 40%-190%, and elastic recovery rate is 98%-99%, and fracture strength is 0.5-0.7GPa, and initial modulus is 0.05-0.75GPa.
Described from body keratin orthotic material be preferably Crinis Carbonisatus from body keratin donor from body, also can be the hair of animal self.
Described reducing process agents useful for same is the compositions of Reducing agent, protein denaturant.
Described Reducing agent is sodium sulfite, sodium sulfite, potassium sulfite, Potassium acid sulfite or sodium metabisulfite, and the mass percent concentration of Reducing agent is 7%-12%.
Described protein denaturant is preferably carbamide, thiourea or lithium bromide, and the molar concentration of protein denaturant is 6-10M.
The method for preparing of described pure keratin solution is:
The first step: will be taken from after the hair of the Crinis Carbonisatus of body or animal carries out shredding, remove impurity, cleaning and dried successively; Cut into the segment of fiber of 3-6mm; Be that 2.5-10g/250ml is immersed in the solution that contains Reducing agent and protein denaturant and stirs 2-6h at 50-90 ℃ with the solid-to-liquid ratio, agitator speed is 100-400r/min;
Second step: the mixture that will stir the back gained removes by filter unreacted natural keratin fiber with the 80-120 eye mesh screen, and centrifugal treating is removed the fine impurities in the filtrating, centrifugal speed 3000-7000rpm, and centrifugation time 0.2-1h then gets supernatant.
The 3rd step: under the room temperature condition, quick desalination is removed the small-molecule substance in the solution under oxygen-free environment, then concentrates, and obtains pure keratin solution, and concentration is 2%-5%.
Described method for preparing from body keratin orthotic material is: keratin solution being watered in the mould that casts from different shape, is to solidify 5-24h under 40-60 ℃ the condition in temperature.
Described mould can be to be used for the certain thickness sheet die of having of connective tissue reparation; Also can be the micro-tube mould close with the blood vessel yardstick; Can also be the mould of the elongated shape with certain draw ratio of simulation ligament, tendon tissue, or be used to prepare the block or complex shape mould that cartilaginous tissue is repaired.
The regulatable elastic performance regulate and control method from body keratin orthotic material of described mechanical property is: with concentration is that the 0.02-0.04% aqueous hydrogen peroxide solution slowly splashed in the pure keratin solution in 1: 100 by mass percentage; With macromolecular crosslinking rate and crosslinking degree in the adjustment keratin solution, regulation and control are from the elasticity of body keratin orthotic material.
The invention provides the regulatable method for preparing of a kind of elasticity from body keratin orthotic material; Keratic donor is patient self Crinis Carbonisatus or animal self hair; And do not use cross-linking agent in the system process; Can effectively reduce the bad phenomenon such as rejection of implant, the misery of having avoided graft to draw materials the patient is brought from body.Adopt the method for quick desalination under the oxygen-free environment to avoid the long drawback of CD desalination period; And the active group of farthest having saved the protein macromolecule cross-linking reaction from damage; Keratin molecule oxidation and the recrystallization problem of having avoided long-time dialysis procedure to bring; The macromolecular active group of keratin is remained to greatest extent, and in the material preparation process, effectively utilize the macromolecular active group of keratin, through regulating the cross-linking reaction speed of keratin macromole active group; Improve the elasticity and the toughness of keratin orthotic material from the aspect of keratin macromolecular structure, prepare the orthotic material that various elasticity and mechanical property satisfy instructions for use.Under the prerequisite that guarantees embedded material biocompatibility and safety in utilization, greatly expanded the range of application of keratin embedded material.
The specific embodiment
Specify the present invention through embodiment below.
Embodiment 1
The Crinis Carbonisatus of cleaning is cut into the long segment of fiber of 3-5mm; With the solid-to-liquid ratio is that 3g/100ml is immersed in the solution of 250ml Potassium acid sulfite and carbamide; Wherein, said solution is to obtain through sodium metabisulfite is added in the 7M aqueous solution of urea, and the mass concentration of Potassium acid sulfite is 8%; Under 60 ℃, mixture is stirred 5h, agitator speed 300r/min.After 80 eye mesh screens filter, get supernatant behind the 4000rpm centrifugalize 0.4h, adopt under the nitrogen protection condition filter assemblies quick desalination to remove to concentrate behind the low-molecular-weight salt material in the solution that to obtain mass percent concentration be 3% keratin solution.
With concentration is that 0.01% aqueous hydrogen peroxide solution slowly splashed in the pure keratin solution in 1: 100 by mass percentage, then, keratin solution watered casts from the thick sheet die of 1mm, in temperature is to solidify 17h under 50 ℃ the condition.Obtaining elastic extension is 186.5%, and elastic recovery rate is 98.1%, and fracture strength is 0.51GPa, and initial modulus is the orthopedic repair materials of keratin loose connective tissue of 0.06GPa.
Embodiment 2
The Crinis Carbonisatus of cleaning is cut into the long segment of fiber of 3-5mm; With the solid-to-liquid ratio is that 1g/100ml is immersed in the solution of 250ml sodium metabisulfite and carbamide; Wherein, said solution is to obtain through sodium metabisulfite is added in the 6M aqueous solution of urea, and the mass concentration of sodium metabisulfite is 7%; Under 90 ℃, mixture is stirred 2h, agitator speed 200r/min.After 120 eye mesh screens filter, get supernatant behind the 7000rpm centrifugalize 0.2h, adopt under the nitrogen protection condition filter assemblies quick desalination to remove to concentrate behind the low-molecular-weight salt material in the solution that to obtain mass percent concentration be 2% keratin solution.
With concentration is that 0.02% aqueous hydrogen peroxide solution slowly splashed in the pure keratin solution in 1: 100 by mass percentage, then, keratin solution is watered in the micro-tube mould that casts from the simulated blood vessel yardstick, in temperature is to solidify 24h under 40 ℃ the condition.Obtaining elastic extension is 102.3%, and elastic recovery rate is 98.3%, and fracture strength is 0.52GPa, and initial modulus is the orthopedic renovating bracket material of keratin blood vessel of 0.13GPa.
Embodiment 3
The Mus hair of cleaning is cut into the long segment of fiber of 3-5mm; With the solid-to-liquid ratio is that 4g/100ml is immersed in the solution of 250ml potassium sulfite and thiourea; Wherein, said solution is to obtain through potassium sulfite is added in the 10M thiourea solution, and the mass concentration of potassium sulfite is 12%; Under 50 ℃, mixture is stirred 4h, agitator speed 100r/min.After 100 eye mesh screens filter, get supernatant behind the 3000rpm centrifugalize 1h, adopt under the nitrogen protection condition filter assemblies quick desalination to remove to concentrate behind the low-molecular-weight salt material in the solution that to obtain mass percent concentration be 5% keratin solution.
With concentration is that 0.04% aqueous hydrogen peroxide solution slowly splashed in the pure keratin solution in 1: 100 by mass percentage, then, keratin solution watered casts from the elongate mould with certain draw ratio, in temperature is to solidify 5h under 60 ℃ the condition.Obtaining elastic extension is 43.6%, and elastic recovery rate is 98.7%, and fracture strength is 0.66GPa, and initial modulus is keratin ligament or the orthopedic repair materials of tendon of 0.71GPa.
Embodiment 4
The rabbit hair of cleaning is cut into the long segment of fiber of 3-5mm; With the solid-to-liquid ratio is that 2g/100ml is immersed in the solution of 250ml sodium sulfite and lithium bromide; Wherein, said solution is to obtain through sodium sulfite is added in the 8M lithium bromide water solution, and the mass concentration of sodium sulfite is 10%; Under 70 ℃, mixture is stirred 6h, agitator speed 400r/min.After 80 eye mesh screens filter, get supernatant behind the 5000rpm centrifugalize 0.5h, adopt under the nitrogen protection condition filter assemblies quick desalination to remove to concentrate behind the low-molecular-weight salt material in the solution that to obtain mass percent concentration be 4% keratin solution.
With concentration is that 0.03% aqueous hydrogen peroxide solution slowly splashed in the pure keratin solution in 1: 100 by mass percentage, then, keratin solution is watered in the mould that casts from rabbit nasal cartilages shape, in temperature is to solidify 20h under 50 ℃ the condition.Obtaining elastic extension is 102.3%, and elastic recovery rate is 98.3%, and fracture strength is 0.52GPa, and initial modulus is the keratin orthotic material that is used for the nasal cartilages tissue repair of 0.13GPa.
The relation of high-elastic biomembrane elasticity of table 1 aqueous hydrogen peroxide solution concentration and keratin and mechanical property
Can find out from table 1; Increase with the hydrogenperoxide steam generator addition, the oxidized degree of the macromolecular mercapto groups of the keratin that reducing process makes is high, and the crosslinking degree again of keratin molecule increases; The elastic extension of the keratin orthotic material of gained reduces; Elastic recovery rate raises, and fracture strength increases, and modulus increases.The mechanical property of the keratin orthotic material that crosslinking degree is high is excellent than fibroin; And the elongation at break of elastic extension and wool fiber is close; Therefore, can satisfy the mechanical property requirements of the orthopedic repair materials of ligament, tendon and cartilaginous tissue to substitution material.The keratin orthotic material that crosslinking degree is low shows higher elastic performance, and extension at break and also decline thereupon of modulus, shows excellent flexible characteristic, can be used as the auxiliary performance requirement of repairing orthotic material of loose connective tissue and blood vessel.
Claims (7)
1. high-elastic regenerating ceratin fibres and preparation method thereof; It is characterized in that; Make by the pure keratin solution that reducing process obtains from body keratin orthotic material, have controllability from the elastic performance of body keratin orthotic material, and have good biomechanical property.The elastic extension controlled range is 40%-190%, and elastic recovery rate is 98%-99%, and fracture strength is 0.5-0.7GPa, and initial modulus is 0.05-0.75GPa.
2. a kind of elasticity as claimed in claim 1 is regulatable to be is characterized in that from body keratin orthotic material, is preferably the Crinis Carbonisatus from body from body keratin donor, also can be the hair of animal self.
3. a kind of elasticity as claimed in claim 1 is regulatable to is characterized in that from body keratin orthotic material and preparation method thereof described reducing process agents useful for same is the compositions of Reducing agent, protein denaturant.Described Reducing agent is sodium sulfite, sodium sulfite, potassium sulfite, Potassium acid sulfite or sodium metabisulfite, and the mass percent concentration of Reducing agent is 7%-12%.Described protein denaturant is preferably carbamide, thiourea or lithium bromide, and the molar concentration of protein denaturant is 6-10M.
4. a kind of elasticity as claimed in claim 1 is regulatable to is characterized in that from body keratin orthotic material and preparation method thereof the concrete steps of described pure keratin solution method for preparing are:
1. will be taken from after the hair of the Crinis Carbonisatus of body or animal carries out shredding, remove impurity, cleaning and dried successively; Cut into the segment of fiber of 3-6mm; Be that 2.5-10g/250ml is immersed in the solution that contains Reducing agent and protein denaturant and stirs 2-6h at 50-90 ℃ with the solid-to-liquid ratio, agitator speed is 100-400r/min;
The mixture that 2. will stir the back gained removes by filter unreacted natural keratin fiber with the 80-120 eye mesh screen, and centrifugal treating is removed the fine impurities in the filtrating, centrifugal speed 3000-7000rpm, and centrifugation time 0.2-1h then gets supernatant.
3. under the room temperature condition, quick desalination is removed the small-molecule substance in the solution under oxygen-free environment, then concentrates, and obtains pure keratin solution, and concentration is 2%-5%.
5. a kind of elasticity as claimed in claim 1 is regulatable from body keratin orthotic material and preparation method thereof; It is characterized in that; Described method for preparing is for to water keratin solution in the mould that casts from different shape, in temperature is to solidify 5-24h under 40-60 ℃ the condition.
6. method for preparing as claimed in claim 5; It is characterized in that; Described mould can be to be used for the certain thickness sheet die of having of connective tissue reparation; Also can be the micro-tube mould close with the blood vessel yardstick, can also be the mould of the elongated shape with certain draw ratio of simulation ligament, tendon tissue, or be used to prepare the block or complex shape mould that cartilaginous tissue is repaired.
7. a kind of elasticity as claimed in claim 1 is regulatable from body keratin orthotic material and preparation method thereof; It is characterized in that; The regulatable elastic performance regulate and control method from body keratin orthotic material of described elasticity is: with concentration is that the 0.02-0.04% aqueous hydrogen peroxide solution slowly splashed in the pure keratin solution in 1: 100 by mass percentage; With macromolecular crosslinking rate and crosslinking degree in the adjustment keratin solution, the elasticity of the high-elastic film of regulation and control keratin.
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Cited By (5)
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| CN104941005A (en) * | 2015-05-18 | 2015-09-30 | 天津工业大学 | Preparation method of hydroxyapatite-enhanced keratin composite aggregate |
| CN109481726A (en) * | 2018-11-26 | 2019-03-19 | 广州新诚生物科技有限公司 | A kind of biodegradable hemostasis bone wax and preparation method thereof |
| CN109641079A (en) * | 2016-08-31 | 2019-04-16 | 生物管株式会社 | The forming method of connective tissue body |
| CN113304322A (en) * | 2021-06-11 | 2021-08-27 | 北京中瑞联合生物科技有限公司 | Keratin material and cell compound for cartilage repair and preparation method thereof |
| CN115970056A (en) * | 2023-01-31 | 2023-04-18 | 陕西师范大学 | 3D printing material for bone repair, bone repair material, and preparation method and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104941005A (en) * | 2015-05-18 | 2015-09-30 | 天津工业大学 | Preparation method of hydroxyapatite-enhanced keratin composite aggregate |
| CN104941005B (en) * | 2015-05-18 | 2018-02-02 | 天津工业大学 | A kind of preparation method of hydroxyapatite enhancing keratin compound bone |
| CN109641079A (en) * | 2016-08-31 | 2019-04-16 | 生物管株式会社 | The forming method of connective tissue body |
| CN109481726A (en) * | 2018-11-26 | 2019-03-19 | 广州新诚生物科技有限公司 | A kind of biodegradable hemostasis bone wax and preparation method thereof |
| CN113304322A (en) * | 2021-06-11 | 2021-08-27 | 北京中瑞联合生物科技有限公司 | Keratin material and cell compound for cartilage repair and preparation method thereof |
| CN115970056A (en) * | 2023-01-31 | 2023-04-18 | 陕西师范大学 | 3D printing material for bone repair, bone repair material, and preparation method and application thereof |
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Application publication date: 20121226 |