CN103554543B - A kind of preparation method of konjac glucomannan fiber scaffold material - Google Patents
A kind of preparation method of konjac glucomannan fiber scaffold material Download PDFInfo
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- CN103554543B CN103554543B CN201310526509.5A CN201310526509A CN103554543B CN 103554543 B CN103554543 B CN 103554543B CN 201310526509 A CN201310526509 A CN 201310526509A CN 103554543 B CN103554543 B CN 103554543B
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Abstract
The present invention discloses a kind of preparation method of konjac glucomannan fiber scaffold material, belongs to field of biomedical materials; The component of this konjac glucomannan fiber scaffold material is mainly Rhizoma amorphophalli glucomannan, wherein the mass ratio of Rhizoma amorphophalli glucomannan and water is 1:120 ~ 1:70, the volume ratio of sulfuric acid and water is 1:2000 ~ 1:200, in aqua calcis, the massfraction of calcium hydroxide is 0.05% ~ 2%, in crosslinked fluid, the volume fraction of calcium hydroxide aqueous solution and dehydrated alcohol is 1:2 ~ 1:0.5, the present invention is cross-linked by sulfuric acid modified and calcium hydroxide aqueous solution, can calcium sulfate be generated and discharge chondroitin sulfate, making timbering material can better be applied to joint cartilage reparation.
Description
Technical field
The present invention relates to a kind of preparation method of konjac glucomannan fiber scaffold material, this fiber scaffold material can be applied to joint cartilage reparation, belongs to field of biomedical materials.
Background technology
High-energy violence wound, intraarticular infect and other diseases can cause severe bone, also cause articular cartilage damage simultaneously, joint cartilage, once defect and destruction can cause affected joints to occur the dysfunctions such as swelling, hydrops, pain and interlocking, has a strong impact on patient articular's function.In addition, cartilaginous tissue self-healing ability is very limited, once damage will leave over permanent pathology, treatment also suitable difficulty, mainly because: without blood vessel in cartilaginous tissue, and chondrocyte's shortage migrates ability, can not gather rapidly caused by wound site; In joint cartilage, fibre content is few, and only containing a small amount of collegen filament, the main chemical compositions of matrix is protein-polysaccharide; Joint cartilage is different from the tissue of vascularization, supply owing to there is no blood, and chondrocyte is embedded in the extracellular matrix of stiff, damage location cannot be moved to participate in repairing, and lack the necessary parent cell of cytodifferentiation and the necessary ecotopia of cell migration, even so just make minimum cartilage defect also be difficult to nature reparation, the articular cartilage damage therefore caused by wound and osteoarthritis is clinical common diseases, and its clinical treatment is still a huge difficult problem so far.
Rhizoma amorphophalli glucomannan (Konjac glucomannan, KGM) be a kind of extraction from konjac tuber to potential of hydrogen sensitive polysaccharides, can by the gel generation time of the potential of hydrogen control KGM of regulation system and Gel baits, the mechanical property of timbering material is controlled with this, biological degradability, even more noteworthy, KGM has good biocompatibility, it is in hemostasis, antisepsis and anti-inflammation, promote that the aspects such as wound healing also have good effect, large quantity research shows, KGM can be beneficial to cell adhesion and promote cell proliferation and differentiation, and the microtexture of one of composition of itself and cartilage matrix hyaluronate sodium, physico-chemical property is quite similar, therefore a kind of novel bio-medical material can be it can be used as to apply, main application scenario comprises the lapping of surgical wound, styptic sponge, medical degraded auxiliary material and bioengineered scaffolds material etc.KGM also comprises its source comparatively extensively as the application advantage of bio-medical material, with low cost, reducing material cost.
Because KGM solubleness is lower, the feature of the bad and less stable of mobility, therefore has some limitations in application, and investigator often adopts physically modified and the important method of modifying of chemical modification these two kinds to carry out modification to KGM for this reason.Wherein, two classes are mainly contained to the physically modified of KGM: one is adopt sound wave, light wave, thermal field, electric field or magnetic field to KGM process, to change molecular structure or the accumulation shape of KGM; Two is adopt other macromolecular materials and KGM to carry out blended, utilizes the Interaction of substituents between two kinds of differing moleculars, to change the structure of material, and then changes its physicochemical property.Chemical modification for KGM is generally that utilization is introduced or takes off some groups on its molecular chain, the molecular structure of KGM is changed, to optimize the physico-chemical property of KGM; Owing to containing a large amount of hydroxyls and kharophen in KGM molecular chain, the chemical modification methods such as esterification, deacetylation and graft copolymerization therefore can be adopted KGM modification, and then develop the KGM derivative of multiple excellent property.
At present, useful acetic acid, nitric acid, phenylformic acid, xanthan gum and methyl fat, gallic acid, deer horn phycocolloid, starch, soybean protein educt etc. carry out modification as properties-correcting agent to KGM, but modified KGM all has degradation property poor, the shortcomings such as mechanical property is not enough, and be used as properties-correcting agent with strong acid sulfuric acid and modification carried out to KGM and the research of preparing fiber scaffold material is not also found at present.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of konjac glucomannan fiber scaffold material, specifically comprise the steps:
(1) use the distilled water diluting vitriol oil, then add glycerol, and stir and obtain mixed solution A, wherein the volume ratio of the vitriol oil and water is 1:2000 ~ 1:200, and the volume ratio of glycerol and water is 1:1000 ~ 1:100;
(2) added in mixed solution A by Rhizoma amorphophalli glucomannan, and stir and make it dissolve completely to obtain mixed glue solution B, the mass ratio adding water in Rhizoma amorphophalli glucomannan and step (1) is 1:120 ~ 1:70;
(3), after mixed glue solution B being carried out microwave treatment 1 ~ 10min, stir at normal temperatures and make it cooling, filter through 200 ~ 400 orders, under being then placed in the negative pressure of 400 ~ 500 mmhg, deaeration obtains modification composite glue solution C, seals for subsequent use;
(4) glycerol joined in dehydrated alcohol, and stir and obtain mixed solution D, wherein the volume ratio of glycerol and dehydrated alcohol is 1:40 ~ 1:10;
(5) at normal temperatures by modification composite glue solution C curtain coating on mould, and mould is placed in mixed solution D, after carrying out the solidifying of 4 ~ 24h, obtains uncrosslinked fibrous framework F;
(6) after at normal temperatures uncrosslinked fibrous framework F being placed in the crosslinked 4 ~ 10h of mixing solutions of calcium hydroxide aqueous solution and dehydrated alcohol, F taking-up distilled water wash 3 ~ 6 times, then dry 8 ~ 12h at 30 ~ 60 DEG C, namely obtains konjac glucomannan fiber scaffold material.
Mould of the present invention is made up of the gauze above 200 ~ 400 object filter sieve and filter sieve.
The mass percent concentration of calcium hydroxide aqueous solution of the present invention is 0.05% ~ 2%.
The vitriol oil of the present invention is the vitriol oil of commercially available 98%.
The principle of the sulfuric acid modified konjac glucomannan fiber scaffold material that the present invention relates to and preparation method institute foundation is: utilize strong acid sulfuric acid to cut off the long molecular chain of Rhizoma amorphophalli glucomannan, use the mixing solutions of calcium hydroxide aqueous solution and dehydrated alcohol to be cross-linked sulfuric acid modified konjac glucomannan fiber scaffold material can generate calcium sulfate and discharge chondroitin sulfate, calcium sulfate and chondroitin sulfate are all carry out the material required for Bone Defect Repari and Human osteoblast, they respectively to skeleton and cartilage useful, sulfuric acid modified konjac glucomannan fiber scaffold material is so just made to become possibility for cartilage repair material.
The invention has the beneficial effects as follows:
(1) the present invention utilizes strong acid sulfuric acid carry out modification to Rhizoma amorphophalli glucomannan and prepare fiber scaffold material, utilize the mixing solutions of dehydrated alcohol and calcium hydroxide aqueous solution to be cross-linked support, the composition chondroitin sulfate of the calcium sulfate and cartilage that make it generate to have osteoconductive is attached to rack surface;
(2) the present invention controls the degradation rate of fibrous framework by the crosslinking method and crosslinking time regulating Rhizoma amorphophalli glucomannan;
(3) the present invention is cross-linked by sulfuric acid modified and calcium hydroxide aqueous solution, can calcium sulfate be generated and discharge chondroitin sulfate, calcium sulfate and chondroitin sulfate are all carry out the material required for Bone Defect Repari and Human osteoblast, they respectively to skeleton and cartilage useful, make timbering material to repair cartilage defect;
(4) microtexture of one of the composition hyaluronate sodium of raw materials used Rhizoma amorphophalli glucomannan and cartilage, physico-chemical property are quite similar, it is more satisfactory tissue engineering material, and there is good gelation, retentiveness, bridging property, mechanical property and biocompatibility, wide material sources, low price;
(5) sulfuric acid modified konjac glucomannan fiber scaffold material of the present invention has controlled degradation rate, with low cost and good mechanical property, and can provide the calcium sulfate needed for human body cartilage-derived growth and chondroitin sulfate when degrading.
Accompanying drawing explanation
Fig. 1 is the preparation flow figure of fiber scaffold material;
The SEM photo of the konjac glucomannan fiber scaffold material Marrow Mesenchymal Stem Cells In Vitro of Fig. 2 obtained by embodiment 1;
The SEM photo of the konjac glucomannan fiber scaffold material Marrow Mesenchymal Stem Cells In Vitro of Fig. 3 obtained by embodiment 2.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but scope is not limited to described content.
Embodiment 1
As shown in Figure 1, the preparation method of konjac glucomannan fiber scaffold material described in the present embodiment, specifically comprises the steps:
(1) use the distilled water diluting vitriol oil, then add glycerol, and stir and obtain mixed solution A, wherein the volume ratio of the vitriol oil and water is 1:200, and the volume ratio of glycerol and water is 1:100;
(2) added in mixed solution A by Rhizoma amorphophalli glucomannan, and stir and make it dissolve completely to obtain mixed glue solution B, the mass ratio adding water in Rhizoma amorphophalli glucomannan and step (1) is 1:70;
(3), after mixed glue solution B being carried out microwave treatment 10min, stir at normal temperatures and make it cooling, filter through 400 orders, under being then placed in the negative pressure of 500 mmhg, deaeration obtains modification composite glue solution C, seals for subsequent use;
(4) glycerol joined in dehydrated alcohol, and stir and obtain mixed solution D, wherein the volume ratio of glycerol and dehydrated alcohol is 1:10;
(5) at normal temperatures by modification composite glue solution C curtain coating in curtain coating on homemade gauze mould, this gauze mould is made up of the gauze above 200 order filter sieve and filter sieve, and mould is placed in mixed solution D, obtain uncrosslinked fibrous framework F after carrying out the solidifying of 24h;
(6) at normal temperatures uncrosslinked fibrous framework F being placed in mass percent concentration is after the crosslinked 10h of mixing solutions of 2% calcium hydroxide aqueous solution and dehydrated alcohol, F taking-up distilled water wash 6 times, then dry 12h at 60 DEG C, namely obtains konjac glucomannan fiber scaffold material.
The tensile strength of the present embodiment gained konjac glucomannan fiber scaffold material is 7MPa, elongation at break is 2.5%, trimestral degradation in vitro is about 50%, and the SEM photo of the konjac glucomannan fiber scaffold material Marrow Mesenchymal Stem Cells In Vitro that the present embodiment prepares as shown in Figure 2.
Embodiment 2
As shown in Figure 1, the preparation method of konjac glucomannan fiber scaffold material described in the present embodiment, specifically comprises the steps:
(1) use the distilled water diluting vitriol oil, then add glycerol, and stir and obtain mixed solution A, wherein the volume ratio of the vitriol oil and water is 1:1000, and the volume ratio of glycerol and water is 1:500;
(2) added in mixed solution A by Rhizoma amorphophalli glucomannan, and stir and make it dissolve completely to obtain mixed glue solution B, the mass ratio adding water in Rhizoma amorphophalli glucomannan and step (1) is 1:90;
(3), after mixed glue solution B being carried out microwave treatment 5min, stir at normal temperatures and make it cooling, filter through 400 orders, under being then placed in the negative pressure of 450 mmhg, deaeration obtains modification composite glue solution C, seals for subsequent use;
(4) glycerol joined in dehydrated alcohol, and stir and obtain mixed solution D, wherein the volume ratio of glycerol and dehydrated alcohol is 1:30;
(5) at normal temperatures by modification composite glue solution C curtain coating in curtain coating on homemade gauze mould, this gauze mould is made up of the gauze above 200 order filter sieve and filter sieve, and mould is placed in mixed solution D, obtain uncrosslinked fibrous framework F after carrying out the solidifying of 15h;
(6) at normal temperatures uncrosslinked fibrous framework F being placed in mass percent concentration is after the crosslinked 6h of mixing solutions of 1% calcium hydroxide aqueous solution and dehydrated alcohol, F taking-up distilled water wash 4 times, then dry 10h at 45 DEG C, namely obtains konjac glucomannan fiber scaffold material.
The tensile strength of the present embodiment gained konjac glucomannan fiber scaffold material is about 9.5MPa, elongation at break is about 4.5%, trimestral degradation in vitro is about 65%, and the SEM photo of the konjac glucomannan fiber scaffold material Marrow Mesenchymal Stem Cells In Vitro that the present embodiment prepares as shown in Figure 3.
Embodiment 3
As shown in Figure 1, the preparation method of konjac glucomannan fiber scaffold material described in the present embodiment, specifically comprises the steps:
(1) use the distilled water diluting vitriol oil, then add glycerol, and stir and obtain mixed solution A, wherein the volume ratio of the vitriol oil and water is 1:2000, and the volume ratio of glycerol and water is 1:1000;
(2) added in mixed solution A by Rhizoma amorphophalli glucomannan, and stir and make it dissolve completely to obtain mixed glue solution B, the mass ratio adding water in Rhizoma amorphophalli glucomannan and step (1) is 1:120;
(3), after mixed glue solution B being carried out microwave treatment 1min, stir at normal temperatures and make it cooling, filter through 200 orders, under being then placed in the negative pressure of 400 mmhg, deaeration obtains modification composite glue solution C, seals for subsequent use;
(4) glycerol joined in dehydrated alcohol, and stir and obtain mixed solution D, wherein the volume ratio of glycerol and dehydrated alcohol is 1:40;
(5) at normal temperatures by modification composite glue solution C curtain coating in curtain coating on homemade gauze mould, this gauze mould is made up of the gauze above 200 order filter sieve and filter sieve, and mould is placed in mixed solution D, obtain uncrosslinked fibrous framework F after carrying out the solidifying of 4h;
(6) at normal temperatures uncrosslinked fibrous framework F being placed in mass percent concentration is after the crosslinked 4h of mixing solutions of 0.05% calcium hydroxide aqueous solution and dehydrated alcohol, uncrosslinked fibrous framework F taking-up distilled water wash 3 times, then dry 8h at 30 DEG C, namely obtains konjac glucomannan fiber scaffold material.
The tensile strength of the present embodiment gained konjac glucomannan fiber scaffold material is about 12.5MPa, and elongation at break is about 4%, and trimestral degradation in vitro is about 70%, and mesenchymal stem cells MSCs adhesiving effect is better.
Embodiment 4
As shown in Figure 1, the preparation method of konjac glucomannan fiber scaffold material described in the present embodiment, specifically comprises the steps:
(1) use the distilled water diluting vitriol oil, then add glycerol, and stir and obtain mixed solution A, wherein the volume ratio of the vitriol oil and water is 1:800, and the volume ratio of glycerol and water is 1:270;
(2) added in mixed solution A by Rhizoma amorphophalli glucomannan, and stir and make it dissolve completely to obtain mixed glue solution B, the mass ratio adding water in Rhizoma amorphophalli glucomannan and step (1) is 1:90;
(3), after mixed glue solution B being carried out microwave treatment 2min, stir at normal temperatures and make it cooling, filter through 200 orders, under being then placed in the negative pressure of 500 mmhg, deaeration obtains modification composite glue solution C, seals for subsequent use;
(4) glycerol joined in dehydrated alcohol, and stir and obtain mixed solution D, wherein the volume ratio of glycerol and dehydrated alcohol is 1:40;
(5) at normal temperatures by modification composite glue solution C curtain coating in curtain coating on homemade gauze mould, this gauze mould is made up of the gauze above 200 order filter sieve and filter sieve, and mould is placed in mixed solution D, obtain uncrosslinked fibrous framework F after carrying out the solidifying of 15h;
(6) at normal temperatures uncrosslinked fibrous framework F being placed in mass percent concentration is after the crosslinked 6h of mixing solutions of 1% calcium hydroxide aqueous solution and dehydrated alcohol, F taking-up distilled water wash 4 times, then dry 10h at 37 DEG C, namely obtains konjac glucomannan fiber scaffold material.
The tensile strength of the present embodiment gained konjac glucomannan fiber scaffold material is about 8.5MPa, and elongation at break is about 5%, and trimestral degradation in vitro is about 60%, and mesenchymal stem cells MSCs adhesiving effect is better.
Embodiment 5
As shown in Figure 1, the preparation method of konjac glucomannan fiber scaffold material described in the present embodiment, specifically comprises the steps:
(1) use the distilled water diluting vitriol oil, then add glycerol, and stir and obtain mixed solution A, wherein the volume ratio of the vitriol oil and water is 1:500, and the volume ratio of glycerol and water is 1:200;
(2) added in mixed solution A by Rhizoma amorphophalli glucomannan, and stir and make it dissolve completely to obtain mixed glue solution B, the mass ratio adding water in Rhizoma amorphophalli glucomannan and step (1) is 1:80;
(3), after mixed glue solution B being carried out microwave treatment 2min, stir at normal temperatures and make it cooling, filter through 200 orders, under being then placed in the negative pressure of 450 mmhg, deaeration obtains modification composite glue solution C, seals for subsequent use;
(4) glycerol joined in dehydrated alcohol, and stir and obtain mixed solution D, wherein the volume ratio of glycerol and dehydrated alcohol is 1:40;
(5) at normal temperatures by modification composite glue solution C curtain coating in curtain coating on homemade gauze mould, this gauze mould is made up of the gauze above 200 order filter sieve and filter sieve, and mould is placed in mixed solution D, obtain uncrosslinked fibrous framework F after carrying out the solidifying of 6h;
(6) at normal temperatures uncrosslinked fibrous framework F being placed in mass percent concentration is after the crosslinked 8h of mixing solutions of 1% calcium hydroxide aqueous solution and dehydrated alcohol, F taking-up distilled water wash 3 times, then dry 8h at 50 DEG C, namely obtains konjac glucomannan fiber scaffold material.
The tensile strength of the present embodiment gained konjac glucomannan fiber scaffold material is about 6.5MPa, and elongation at break is about 4%, and trimestral degradation in vitro is about 55%, and mesenchymal stem cells MSCs adhesiving effect is better.
Claims (3)
1. a preparation method for konjac glucomannan fiber scaffold material, is characterized in that specifically comprising the steps:
(1) use the distilled water diluting vitriol oil, then add glycerol, and stir and obtain mixed solution A, wherein the volume ratio of the vitriol oil and water is 1:2000 ~ 1:200, and the volume ratio of glycerol and water is 1:1000 ~ 1:100;
(2) added in mixed solution A by Rhizoma amorphophalli glucomannan, and stir and make it dissolve completely to obtain mixed glue solution B, the mass ratio adding water in Rhizoma amorphophalli glucomannan and step (1) is 1:120 ~ 1:70;
(3), after mixed glue solution B being carried out microwave treatment 1 ~ 10min, stir at normal temperatures and make it cooling, filter through 200 ~ 400 orders, under being then placed in the negative pressure of 400 ~ 500 mmhg, deaeration obtains modification composite glue solution C, seals for subsequent use;
(4) glycerol joined in dehydrated alcohol, and stir and obtain mixed solution D, wherein the volume ratio of glycerol and dehydrated alcohol is 1:40 ~ 1:10;
(5) at normal temperatures by modification composite glue solution C curtain coating on mould, and mould is placed in mixed solution D, after carrying out the solidifying of 4 ~ 24h, obtains uncrosslinked fibrous framework F;
(6) after at normal temperatures uncrosslinked fibrous framework F being placed in the crosslinked 4 ~ 10h of mixing solutions of calcium hydroxide aqueous solution and dehydrated alcohol, F taking-up distilled water wash 3 ~ 6 times, then dry 8 ~ 12h at 30 ~ 60 DEG C, namely obtains konjac glucomannan fiber scaffold material.
2. the preparation method of konjac glucomannan fiber scaffold material according to claim 1, is characterized in that described in step (5), mould is made up of the gauze above 200 ~ 400 object filter sieve and filter sieve.
3. the preparation method of konjac glucomannan fiber scaffold material according to claim 1, is characterized in that the mass percent concentration of step (6) calcium hydroxide aqueous solution is 0.05% ~ 2%.
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| CN103007358A (en) * | 2012-12-11 | 2013-04-03 | 昆明理工大学 | Cartilage tissue engineering fiber scaffold material and preparation method thereof |
| CN103044700A (en) * | 2012-12-11 | 2013-04-17 | 昆明理工大学 | Postoperative anti-adhesion membrane material and method for preparing same |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103007358A (en) * | 2012-12-11 | 2013-04-03 | 昆明理工大学 | Cartilage tissue engineering fiber scaffold material and preparation method thereof |
| CN103044700A (en) * | 2012-12-11 | 2013-04-17 | 昆明理工大学 | Postoperative anti-adhesion membrane material and method for preparing same |
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| 雄波 等.浓硫酸法制备魔芋葡甘聚糖硫酸酯(KGMS).《食品工业科技》.2012,(第2期),222-225页. * |
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