CN102302802A - Medical bone composite scaffold and preparation method thereof - Google Patents
Medical bone composite scaffold and preparation method thereof Download PDFInfo
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- CN102302802A CN102302802A CN201110273701A CN201110273701A CN102302802A CN 102302802 A CN102302802 A CN 102302802A CN 201110273701 A CN201110273701 A CN 201110273701A CN 201110273701 A CN201110273701 A CN 201110273701A CN 102302802 A CN102302802 A CN 102302802A
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Abstract
The invention relates to a scaffold and a preparation method thereof, particularly to a medical bone composite scaffold and a preparation method thereof, belonging to the technical field of medical appliances. The preparation method comprises the following steps of: adding 2 parts by weight of medical calcium sulfate to 23.5 parts by weight of deionized water to obtain a suspension; sufficiently dissolving 1 part by weight of type I collagen in 23.5 parts by weight of 0.5 mol/L acetic acid solution; and slowly dropping the medical calcium sulfate suspension and 50 parts by weight of 0.05 mol/L NaOH solution to the collagen acetic acid solution at the same time, stirring so that flocculent precipitates emerge in the mixed liquid, maintaining for 30 minutes, carrying out soaking and other processes, and finally drying to obtain the scaffold. After being implanted into an organism, the product has no reject reaction and allergic reaction, also has good biocompatibility and meets the requirements of the medical biological materials for application to organisms. The scaffold can be used as an ideal bone substitute and has broad prospects in clinical application.
Description
Technical field
The present invention relates to a kind of support and preparation method thereof, particularly a kind of medical skeleton compound rest and preparation method thereof.Belong to technical field of medical.
Background technology
Owing to factors such as wound, infection, tumor make bone lose some sclerotin, form bigger gap, it is damaged to be called bone.Damaged all being difficult to of most bones heals, and forms bone does not connect at last.Because defect gap is big, osteoblast is difficult to get over the gap and normal agglutination can not take place, only by the fibrous tissue filling.And damaged treatment is a difficulty and the challenging problem in orthopaedics field for bone, clinically mainly through reaching certain therapeutic purposes from body, allosome tissue's transplanting and biological substitution material.Present the most frequently used autograft is because unfavorable factors such as the source is not enough, wound is big, confession district complication have limited and used clinically; And allosome tissue transplant exist allosome rejection, source limited, have a factor such as risk of disease transmission; So, seek and not damage self, target that the person pursues that the substitute that can reach the functional effect of expection bone defect repair again becomes the clinical position.Organizational project learns a skill provides new thinking and method for solving this difficult problem.
The basic skills of organizational project is: be adsorbed in after the amplification of the high concentration histiocyte of In vitro culture good biocompatibility, can be by on the tissue engineering bracket material of human body degraded and absorbed; The function of this material is for cell provides vivosphere, makes cell obtain enough nutrient substance, carries out gas exchange; And cell is grown by the three-dimensional rack of prefabricated form; With the sick position of decreasing of the complex implanting to human body of cell and biomaterial, in biological support degraded and absorbed process, the cell of plantation continues the propagation breeding then; Form new respective organization with original specific function and form and organ, repair wound and the purpose of rebuilding function to reach.
It is an important branch of organizational project that bone tissue engineer is learned, and is expected at first be applied clinical.Formed at present comparatively perfect theory and technology route gradually about seed cell, cell carrier support and tissue construction.But undeniable is that these adjusted range bone tissue engineers finally still have certain distance in Clinical Application.Therefore the focus of present stage research still concentrates on suitable seed cell, material and the construct in vitro mode sought, and seeks three's best of breed, and the natural process that simulation human body bone is repaired is reappeared bone structure and function to greatest extent.At present, the focus of bone tissue engineer research concentrates on seed cell, timbering material, bioactie agent three aspects.Wherein the effect of timbering material in organizational project provides the framework of a kind of porous material as tissue regeneration; Cultured cell in vitro is planted on it; And return and to implant, guiding the continuous growth of required tissue, frame materials fades away simultaneously; Finally in former framework region, form and bear complete lived tissue again, realize permanent reparation.
Ideal timbering material need meet following condition (1) material surface structure and character helps cell absorption, propagation and differentiation; (2) degrade by controllable speed; (3) excellent biological compatibility; (4) be prone to process three-dimensional porous shape, and be prone to be processed into irregular geometric shape; (5) have certain mechanical strength and can support physical stress; (6) can keep the differentiation of pair cell, can not make cell produce variation.The method for preparing of osseous tissue engineering stephanoporate stent material and process are at home to be mature on the whole at present; The timbering material that adopts mainly contains synthetic material and natural biological derived material and composite; Wherein with gather acetic acid (polyglycolic Acid, PGA), polylactic acid (polylactic Acid, PLA), tricalcium phosphate (tricalcium phosphate; TCP), (hydroxy lapatite, HA) grade is comparatively commonly used for hydroxyapatite.Through deeply it is found that of research, above-mentioned different materials its biological degradability and ossification in further research process all have weak point; Though PGA, PLA have better biocompatibility, degradability and absorbability; But there are shortcomings such as expensive, poor plasticity; And the acidic metabolite after the degraded can reduce polymer pH value on every side; Influence the growth of cell and tissue, also can cause the immunoreation of fibrosis and generation surrounding tissue.Be difficult to after HA exist to implant absorb alternative, be stranded in and hinder the reconstruction of osseous tissue and shortcoming such as reparation fully in the body for a long time. we might as well widen thinking and seek some new material-medical grade calcium sulfate (Medical-grade Calcium Sulfate) so; It has excellent biological compatibility, degradability and ossification, makes its raw material as the preparation support will obtain good effect and using value.
Summary of the invention
The object of the invention: invent a kind of medical skeleton compound rest and preparation method thereof.
Through multiple discovering, calcium sulfate is a kind of bone guided property material, and mainly as the implant in space, it can recover the form profile of bone, stops soft tissue to be grown into.It provides the substrate of bone guided property for blood vessel and osteoblastic growing into.
The method for preparing of a kind of medical skeleton compound rest of the present invention, its preparation process is following:
The medical calcium sulfate (the farsighted bio tech ltd that grinds in Shanghai) of 2 weight portions is put into the deionized water of 23.5 weight portions, form muddy suspension;
Again the type i collagen of 1 weight portion fully is dissolved in the acetum of 0.5Mol/L of 23.5 weight portions; Type i collagen: collagen is a kind of natural fibrin; White is opaque, unbranched; Mainly be present in the tissues such as osteoderm tendon of animal; Be structural protein important in the connective tissue, play the support organ, the protection body function
collagen protein be the maximum protein of mammal in-vivo content account for human body or other animal body protein total content 25% to 35%; Existing 27 kinds of the collagen protein of confirming in 2004; Modal collagen protein has I, II, III type, and wherein the type i collagen protein content is maximum, use the most extensive, also maximum to its research of carrying out.The type i collagen that the present invention adopts is purchased the farsighted bio tech ltd that grinds from Shanghai.
Slowly splash into the muddy suspension of medical calcium sulfate in the collagen acetum and form " intermediate liquid ", NaOH (the Tianjin chemical reagent three factories) solution of the 0.05mol/L of 50 weight portions is slowly splashed in " intermediate liquid " and stirs, keep 30min; The gel-shaped precipitate appears in container bottom; Remove the supernatant, keep the gel-shaped precipitate, it is in charge of; Pour centrifuge tube into, put into the xenogenesis deproteinization spongy bone for preparing in advance in the lump; Xenogenesis Grafting Cancellous Bone Bolt piece is adopted 1: 1 chloroform-methanol loss of thick fluid fat 24h, 50 ℃ of distilled water concussion flushings, 20%H2O2 floods 24h, 3 times repeatedly.Bone piece after handling is placed distilled water, under room temperature, soak dialysis 24h, after the oven dry, 60Co gamma-rays 2.5 * 105Gry, behind the irradiation 30min, taking-up places 4 ℃ of refrigerators subsequent use, the centrifugal supernatant that goes.Wherein, The method for preparing of described xenogenesis deproteinization spongy bone is following: get the pig DF spongiosa part of growing up; Do not contain cartilage and cortical bone composition in institute's intercepting bone piece, after-70 ℃ of lyophilizations, process 3.0cm * 0.5cm * 0.5cm size; Its major axis is consistent with the orientation of getting Grafting Cancellous Bone Bolt girder, and the apparent holes gap density of each bone piece of drawing materials is basic identical.
The lyophilization of gained sample obtains type i collagen/medical calcium sulfate/xenogenesis deproteinization spongy bone compound rest; Cryodesiccated condition: temperature :-80 ℃; Pressure: 0.13Pa: time: 24 hours.
The gained material block put in people's 0.25% glutaraldehyde solution (Tianjin chemical reagent three factories) carry out chemical crosslinking 24h;
After the crosslinked completion material block is placed 4 ℃ of 1 weeks of deionized water rinsing, change water every day 2 times, remove remaining not combination glutaraldehyde solution, promptly obtain product of the present invention after the drying.
Beneficial effect
Support top layer and hole wall scribble the medical calcium sulfate of uniformity; No medical calcium sulfate is piled up in the hole, has the pore structure that is interconnected, and porosity is high; The aperture is about 110 ± 12um; Porosity is about (72.1 ± 3.5) %, and the bionic three-dimensional space structure is beneficial to the Oesteoblast growth environment that provides good, and comprcssive strength is that 43 ± 3.2MPa implanting to human body bone defect can satisfy biomechanical strength.
Confirm the irregular high density hole of timbering material grid structure through experiment in vivo and vitro, be similar to the spongy bone structure, have good bone conduction, bone inducibility, help the biodegradation with material self of growing into of osseous tissue; The no rejection that implants does not have irritated reaction, also has excellent biological compatibility, meets the requirement that biomaterial for medical purpose is applied to organism.Support can be used as ideal bone substitute and in clinical practice, can hold out broad prospects.
When medical calcium sulfate is absorbed, new bone is moulding and recover anatomic characteristic and architectural characteristic.Its most important advantage is that its natural infiltration rate is suitable with new bone formation speed.Along with the absorption of medical calcium sulfate implant, new bone recovers to dissect character and construction features gradually.
The specific embodiment
Embodiment 1
The method for preparing of a kind of medical skeleton compound rest of the present invention, its preparation process is following:
The medical calcium sulfate of 2 weight portions is put into the deionized water of 23.5 weight portions, form muddy suspension;
Again in the acetum with the former 0.5Mol/L that fully is dissolved in 23.5 weight portions of the I type glue (the farsighted bio tech ltd that grinds in Shanghai) of 1 weight portion; The type i collagen that adopts is purchased the good and bio tech ltd from Shanghai.
Slowly splash into the muddy suspension of medical calcium sulfate in the collagen acetum and form " intermediate liquid ", the NaOH solution (Tianjin chemical reagent three factories) of the 0.05mol/L of 50 weight portions is slowly splashed in " intermediate liquid " and stirs, keep 30min; The gel-shaped precipitate appears in container bottom; Remove the supernatant, keep the gel-shaped precipitate, it is in charge of; Pour centrifuge tube into, put into the xenogenesis deproteinization spongy bone for preparing in advance in the lump; With the chloroform-methanol loss of thick fluid fat 24h of xenogenesis Grafting Cancellous Bone Bolt piece employing 1:1,50 ℃ of distilled water concussion flushings, 20%H
2O
2Dipping 24h, 3 times repeatedly.Bone piece after handling is placed distilled water, under room temperature, soak dialysis 24h, after the oven dry, 60Co gamma-rays 2.5 * 105Gry, behind the irradiation 30min, taking-up places 4 ℃ of refrigerators subsequent use, the centrifugal supernatant that goes.Wherein, The method for preparing of described xenogenesis deproteinization spongy bone is following: get the pig DF spongiosa part of growing up; Do not contain cartilage and cortical bone composition in institute's intercepting bone piece, after-70 ℃ of lyophilizations, process 3.0cm * 0.5cm * 0.5cm size; Its major axis is consistent with the orientation of getting Grafting Cancellous Bone Bolt girder, and the apparent holes gap density of each bone piece of drawing materials is basic identical.
The lyophilization of gained sample obtains type i collagen/medical calcium sulfate/xenogenesis deproteinization spongy bone compound rest; Cryodesiccated condition: temperature :-80 ℃; Pressure: 0.13Pa: time: 24 hours.
The gained material block put in people's 0.25% glutaraldehyde solution (cleer sea, Beijing ocean Science and Technology Ltd.) carry out chemical crosslinking 24h;
After the crosslinked completion material block is placed 4 ℃ of 1 weeks of deionized water rinsing, change water every day 2 times, remove remaining not combination glutaraldehyde solution, promptly obtain product of the present invention after the drying.
Embodiment 2
The method for preparing of a kind of medical skeleton compound rest of the present invention, its preparation process is following:
The medical calcium sulfate of 2 weight portions is put into the deionized water of 23.5 weight portions, form muddy suspension;
Again the type i collagen of 1 weight portion fully is dissolved in the acetum of 0.5Mol/L of 23.5 weight portions; Type i collagen: collagen is a kind of natural fibrin; White is opaque, unbranched; Mainly be present in the tissues such as osteoderm tendon of animal; Be structural protein important in the connective tissue, play the support organ, the protection body function
collagen protein be the maximum protein of mammal in-vivo content account for human body or other animal body protein total content 25% to 35%; Existing 27 kinds of the collagen protein of confirming in 2004; Modal collagen protein has I, II, III type, and wherein the type i collagen protein content is maximum, use the most extensive, also maximum to its research of carrying out.The type i collagen that the present invention adopts is purchased the farsighted bio tech ltd that grinds from Shanghai.
Slowly splash into the muddy suspension of medical calcium sulfate in the collagen acetum and form " intermediate liquid ", the NaOH solution of the 0.05mol/L of 50 weight portions is slowly splashed in " intermediate liquid " and stirs, keep 30min; The gel-shaped precipitate appears in container bottom; Remove the supernatant, keep the gel-shaped precipitate, it is in charge of; Pour centrifuge tube into, put into the xenogenesis deproteinization spongy bone for preparing in advance in the lump; Xenogenesis Grafting Cancellous Bone Bolt piece is adopted 1: 1 chloroform-methanol loss of thick fluid fat 24h, 50 ℃ of distilled water concussion flushings, 20%H2O2 floods 24h, 3 times repeatedly.Bone piece after handling is placed distilled water, under room temperature, soak dialysis 24h, after the oven dry, 60Co gamma-rays 2.5 * 105Gry, behind the irradiation 30min, taking-up places 4 ℃ of refrigerators subsequent use, the centrifugal supernatant that goes.Wherein, The method for preparing of described xenogenesis deproteinization spongy bone is following: get the pig DF spongiosa part of growing up; Do not contain cartilage and cortical bone composition in institute's intercepting bone piece, after-70 ℃ of lyophilizations, process 3.0cm * 0.5cm * 0.5cm size; Its major axis is consistent with the orientation of getting Grafting Cancellous Bone Bolt girder, and the apparent holes gap density of each bone piece of drawing materials is basic identical.
The lyophilization of gained sample obtains type i collagen/medical calcium sulfate/xenogenesis deproteinization spongy bone compound rest; Cryodesiccated condition: temperature :-80 ℃; Pressure: 0.13Pa: time: 24 hours.
The gained material block put in people's 0.25% glutaraldehyde solution (Tianjin chemical reagent three factories) carry out chemical crosslinking 24h;
After the crosslinked completion material block is placed 4 ℃ of 1 weeks of deionized water rinsing, change water every day 2 times, remove remaining not combination glutaraldehyde solution, promptly obtain product of the present invention after the drying.
After scanning, Electronic Speculum SEM shape appearance figure can be measured, and gained support of the present invention aperture is distributed between 120～280 μ m, interconnect between the hole, and the material internal micropore enriches, is evenly distributed, and presents loose coralliform structure.This similar is in the spongy bone framework of biological living, and it helps the biodegradation of growing into of osseous tissue and material self.When medical calcium sulfate is absorbed, new bone is moulding and recover anatomic characteristic and architectural characteristic.Its most important advantage is that its natural infiltration rate is suitable with new bone formation speed.Along with the absorption of medical calcium sulfate implant, new bone recovers to dissect character and construction features gradually.
Claims (4)
1. the method for preparing of a medical skeleton compound rest, it is characterized in that: preparation process is following:
1) medical calcium sulfate of 2 weight portions is put into the deionized water of 23.5 weight portions, formed muddy suspension;
2) again the type i collagen of 1 weight portion fully is dissolved in the acetum of 0.5Mol/L of 23.5 weight portions;
3) slowly splash into the muddy suspension of medical calcium sulfate in the collagen acetum and form " intermediate liquid ", the NaOH solution of the 0.05mol/L of 50 weight portions is slowly splashed in " intermediate liquid " and stirs, keep 30min; The gel-shaped precipitate appears in container bottom; Remove the supernatant, keep the gel-shaped precipitate, it is in charge of; Pour centrifuge tube into, put into the xenogenesis deproteinization spongy bone for preparing in advance in the lump; Xenogenesis Grafting Cancellous Bone Bolt piece is adopted 1: 1 chloroform-methanol loss of thick fluid fat 24h, 50 ℃ of distilled water concussion flushings, 20%H2O2 floods 24h, 3 times repeatedly.Bone piece after handling is placed distilled water, under room temperature, soak dialysis 24h, after the oven dry, 60Co gamma-rays 2.5 * 105Gry, behind the irradiation 30min, taking-up places 4 ℃ of refrigerators subsequent use, the centrifugal supernatant that goes.
4) gained sample lyophilization obtains type i collagen/medical calcium sulfate/xenogenesis deproteinization spongy bone compound rest;
5) the gained material block is put in people's 0.25% glutaraldehyde solution carried out chemical crosslinking 24h;
6) after the crosslinked completion material block is placed 4 ℃ of 1 weeks of deionized water rinsing, change water every day 2 times, remove remaining not combination glutaraldehyde solution, promptly obtain product of the present invention after the drying.
2. method for preparing as claimed in claim 1; It is characterized in that: type i collagen said step 2): collagen is a kind of natural fibrin; White is opaque, unbranched; Mainly be present in the tissues such as osteoderm tendon of animal; Be structural protein important in the connective tissue, play the support organ, the protection body function
collagen protein be the maximum protein of mammal in-vivo content account for human body or other animal body protein total content 25% to 35%; Existing 27 kinds of the collagen protein of confirming in 2004; Modal collagen protein has I, II, III type, and wherein the type i collagen protein content is maximum, use the most extensive, also maximum to its research of carrying out.The type i collagen that the present invention adopts is purchased the farsighted bio tech ltd that grinds from Shanghai.
3. method for preparing as claimed in claim 1; It is characterized in that: the method for preparing of xenogenesis deproteinization spongy bone is following in the said step 3): get the pig DF spongiosa part of growing up; Do not contain cartilage and cortical bone composition in institute's intercepting bone piece, after-70 ℃ of lyophilizations, process 3.0cm * 0.5cm * 0.5cm size; Its major axis is consistent with the orientation of getting Grafting Cancellous Bone Bolt girder, and the apparent holes gap density of each bone piece of drawing materials is basic identical.
4. method for preparing as claimed in claim 1 is characterized in that: cryodesiccated condition in the said step 4): temperature :-80 ℃; Pressure: 0.13Pa: time: 24 hours.
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| CN201110273701A CN102302802A (en) | 2011-09-16 | 2011-09-16 | Medical bone composite scaffold and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102626526A (en) * | 2012-04-20 | 2012-08-08 | 无锡圆容生物医药股份有限公司 | Novel active absorbable bone cement material |
-
2011
- 2011-09-16 CN CN201110273701A patent/CN102302802A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 永井 裕•藤本大三郎: "《胶原蛋白实验方法》", 31 October 1992, 上海中医学院出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102626526A (en) * | 2012-04-20 | 2012-08-08 | 无锡圆容生物医药股份有限公司 | Novel active absorbable bone cement material |
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