CN102552987B - 4 DEG C are in a liquid state 37 DEG C in solid-state biomaterial and preparation method with purposes - Google Patents
4 DEG C are in a liquid state 37 DEG C in solid-state biomaterial and preparation method with purposes Download PDFInfo
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Abstract
The invention discloses a kind of 4 DEG C and be in a liquid state 37 DEG C in solid-state biomaterial and preparation method with purposes, product through edible and medical fungi enzymolysis, cell culture, organize the steps such as enzymolysis to obtain.Can be used as nasolacrimal duct, tumor vessel or deferential transient or permanent occlusion, half blocking material.Manufacture method of the present invention is easy, raw material is easy to get, easy to operate, obtained product is in a liquid state when four degrees celsius, be solid-state when 37 degree, can be used for having tube chamber (as nasolacrimal duct, tumor vessel, deferent duct etc.) in the transient of therapeutic purposes or permanent occlusion, half occluder, can be used as medicine, gene, the slow release platform of vaccine and targetable drug carriers.
Description
Technical field
The present invention relates to a kind of biological medicine material.
Background technology
Existing medical material is a lot, but also do not have a kind of can in the acceptable temperature range of human body liquid and solid-state between the material of change, therefore the use of a lot of occasion is restricted.
Summary of the invention
A kind of 4 DEG C are the object of the present invention is to provide to be in a liquid state 37 DEG C in solid-state biomaterial and preparation method with purposes.
Technical solution of the present invention is:
4 DEG C are in a liquid state 37 DEG C in a solid-state biomaterial, and it is characterized in that: be in a liquid state 4 DEG C time, be solid-state 37 DEG C time; Its preparation method is:
(1) edible and medical fungi is carried out enzymolysis processing under the cryogenic conditions of 4-29 DEG C, obtain low temperature edible and medical fungi extracting solution after filtration; Preserve under being placed in 4 DEG C of aseptic conditions;
(2) cell culture: by one or several in fibrocyte, epithelial cell, osteoblast, mixing or single culture; During cell culture, in regular growth culture fluid, add the above-mentioned edible and medical fungi extracting solution of 1-40% percentage by weight, cultivate through 1-12 week, supernatant Collection and conservation is for subsequent use in 4 DEG C of gnotobasiss;
(3) enzyme digestion reaction: fetch and come from bathypelagic fish or mammiferous corium or skeletal tissue; The supernatant above-mentioned tissue and step (2) obtained adds according to 1-12: 1 weight ratio in the enzymolysis solution of 4 DEG C and reacts; Filter under 4 DEG C of gnotobasiss after reaction, filtering accuracy scope: 0.10 μm-0.22 μm; Filtered solution PH is adjusted to 7.4, obtains product, is stored in 4 DEG C of gnotobasiss.
4 DEG C are in a liquid state 37 DEG C in a preparation method for solid-state biomaterial, it is characterized in that: comprise the following steps:
(1) edible and medical fungi is carried out enzymolysis processing under the cryogenic conditions of 4-29 DEG C, obtain low temperature edible and medical fungi extracting solution after filtration; Preserve under being placed in 4 DEG C of aseptic conditions;
(2) cell culture: by one or several in fibrocyte, epithelial cell, osteoblast, mixing or single culture; During cell culture, in regular growth culture fluid, add the above-mentioned edible and medical fungi extracting solution of 1-40% percentage by weight, cultivate through 1-12 week, supernatant Collection and conservation is for subsequent use in 4 DEG C of gnotobasiss;
(3) enzyme digestion reaction: fetch and come from bathypelagic fish or mammiferous corium or skeletal tissue; The supernatant above-mentioned tissue and step (2) obtained adds according to 1-12: 1 weight ratio in the enzymolysis solution of 4 DEG C and reacts; Filter under 4 DEG C of gnotobasiss after reaction, filtering accuracy scope: 0.10 μm-0.22 μm; Filtered solution PH is adjusted to 7.4, obtains product, is stored in 4 DEG C of gnotobasiss.
Described edible and medical fungi is one or several in chanterelle, GANBAJUN, bolete, Termitomyces albuminosus, Morchella esculenta (L.) Pers, Tricholoma lobayense Heim, Pnoliota adiposa(Fr.) Quel., Armillariella mellea, blue or green mushroom, Hericium erinaceus (Bull. Ex Fr.) Pers., red mushroom, Grifola frondosa, Agrocybe aegerita (Brig.) Sing., Ganoderma, Clitocybe maxima, agrocyb eaegerita, pholiota nameko, Mushroom Corals, Oudemansiella Radicata, Pleurotus citrinopileatus, Hypsizygus marmoreus, asafoetida mushroom, Pleurotus eryngii, Pleurotus abalonus, but is not limited only to this.
During the enzymolysis processing of step (1), the enzyme of use is pectase, one or several in cellulase, hemicellulase, α-amylase, glucoamylase, beta amylase; Feed liquid weight ratio: 1: 1-19; The enzyme digestion reaction time: 1-190 hour; PH value: 3-9; Enzyme concentration: be with edible and medical fungi weight ratio: enzyme 0.01-5: edible and medical fungi 100; Filtering accuracy in step (1) is 0.10 μm-0.22 μm.
Enzymatic hydrolysis condition in step (3) is: pH value: 3-5.4; Enzyme digestion reaction temperature: 4-9 degree Celsius; Feed liquid weight ratio: 1: 1-19; The enzyme digestion reaction time: 72-480 hour; The kind of enzyme: one or several in metalloproteases, thiol protease, serine carboxypeptidase, cathepsin, endopeptidase, ficin, pepsin, but be not limited only to this; Enzyme concentration: with raw material weight ratio be: enzyme 0.01-5: raw material 100.
A kind of 4 DEG C of 37 DEG C of application in solid-state biomaterial in preparation in transient or permanent occlusion, half occluder that are in a liquid state.
As nasolacrimal duct, tumor vessel or deferential transient or permanent occlusion, half blocking material.
Manufacture method of the present invention is easy, raw material is easy to get, easy to operate, obtained product is in a liquid state when four degrees celsius, be solid-state when 37 degree, can be used for having tube chamber (as nasolacrimal duct, tumor vessel, deferent duct etc.) in the transient of therapeutic purposes or permanent occlusion, half occluder, can be used as medicine, gene, the slow release platform of vaccine and targetable drug carriers.
Below in conjunction with embodiment, the invention will be further described.
Detailed description of the invention
4 DEG C are in a liquid state 37 DEG C in a preparation method for solid-state biomaterial, comprise the following steps:
(1) edible and medical fungi is carried out enzymolysis processing under the cryogenic conditions of 4-29 DEG C (example 4 DEG C, 15 DEG C, 29 DEG C), obtain low temperature edible and medical fungi extracting solution after filtration; Preserve under being placed in 4 DEG C of aseptic conditions;
(2) cell culture: by one or several in fibrocyte, epithelial cell, osteoblast, mixing or single culture; During cell culture, the above-mentioned edible and medical fungi extracting solution of 1-40% (example 1%, 20%, 40%) percentage by weight is added in regular growth culture fluid, cultivate through 1-12 week (example 1 week, 7 weeks, 12 weeks), supernatant Collection and conservation is for subsequent use in 4 DEG C of gnotobasiss;
(3) enzyme digestion reaction: fetch and come from bathypelagic fish or mammiferous corium or skeletal tissue; The supernatant above-mentioned tissue and step (2) obtained adds in the enzymolysis solution of 4 DEG C according to 1-12: 1 (example 1: 1,6: 1,12: 1) weight ratio and reacts; Filter under 4 DEG C of gnotobasiss after reaction, filtering accuracy scope: 0.10 μm-0.22 μm (example 0.10 μm, 0.18 μm, 0.22 μm); Filtered solution PH is adjusted to 7.4, obtains product, is stored in 4 DEG C of gnotobasiss.
Described edible and medical fungi is one or several in chanterelle, GANBAJUN, bolete, Termitomyces albuminosus, Morchella esculenta (L.) Pers, Tricholoma lobayense Heim, Pnoliota adiposa(Fr.) Quel., Armillariella mellea, blue or green mushroom, Hericium erinaceus (Bull. Ex Fr.) Pers., red mushroom, Grifola frondosa, Agrocybe aegerita (Brig.) Sing., Ganoderma, Clitocybe maxima, agrocyb eaegerita, pholiota nameko, Mushroom Corals, Oudemansiella Radicata, Pleurotus citrinopileatus, Hypsizygus marmoreus, asafoetida mushroom, Pleurotus eryngii, Pleurotus abalonus, but is not limited only to this.
During the enzymolysis processing of step (1), the enzyme of use is pectase, one or several in cellulase, hemicellulase, α-amylase, glucoamylase, beta amylase; Feed liquid weight ratio: 1: 1-19 (example 1: 1,1: 10,1: 19); Enzyme digestion reaction time: 1-190 hour (example 1 hour, 100 hours, 190 hours); PH value: 3-9 (example 3,6,9); Enzyme concentration: be with edible and medical fungi weight ratio: enzyme 0.01-5: edible and medical fungi 100 (example 0.01: 100,3: 100,5: 100); Filtering accuracy in step (1) be 0.10 μm-0.22 μm (example 0.10 μm, 0.18 μm, 0.22 μm).
Enzymatic hydrolysis condition in step (3) is: pH value: 3-5.4 (example 3,4,5); Enzyme digestion reaction temperature: 4-9 degree Celsius (example 4 DEG C, 6 DEG C, 9 DEG C); Feed liquid weight ratio: 1: 1-19 (example 1: 1,1: 10,1: 19); Enzyme digestion reaction time: 72-480 hour (example 72 hours, 200 hours, 480 hours); The kind of enzyme: one or several in metalloproteases, thiol protease, serine carboxypeptidase, cathepsin, endopeptidase, ficin, pepsin, but be not limited only to this; Enzyme concentration: with raw material (namely organizing and supernatant) weight ratio be: enzyme 0.01-5: raw material 100 (example 0.01: 100,3: 100,5: 100).
Above-mentioned 4 DEG C of 37 DEG C of application in solid-state biomaterial in preparation in transient or permanent occlusion, half occluder that are in a liquid state are as nasolacrimal duct, tumor vessel or deferential transient or permanent occlusion, half blocking material.
Claims (7)
1. 4 DEG C are in a liquid state 37 DEG C in a solid-state biomaterial, and it is characterized in that: be in a liquid state 4 DEG C time, be solid-state 37 DEG C time; Its preparation method is:
(1) edible and medical fungi is carried out enzymolysis processing under the cryogenic conditions of 4-29 DEG C, obtain low temperature edible and medical fungi extracting solution after filtration; Preserve under being placed in 4 DEG C of aseptic conditions;
(2) cell culture: by one or several in fibrocyte, epithelial cell, osteoblast, mixing or single culture; During cell culture, in regular growth culture fluid, add the above-mentioned edible and medical fungi extracting solution of 1-40% percentage by weight, cultivate through 1-12 week, supernatant Collection and conservation is for subsequent use in 4 DEG C of gnotobasiss;
(3) enzyme digestion reaction: fetch and come from bathypelagic fish or mammiferous corium or skeletal tissue; The supernatant above-mentioned tissue and step (2) obtained adds according to 1-12: 1 weight ratio in the enzymolysis solution of 4 DEG C and reacts; Filter under 4 DEG C of gnotobasiss after reaction, filtering accuracy scope: 0.10 μm-0.22 μm; Filtered solution PH is adjusted to 7.4, obtains product, is stored in 4 DEG C of gnotobasiss.
2. 4 DEG C according to claim 1 are in a liquid state 37 DEG C in a preparation method for solid-state biomaterial, it is characterized in that: comprise the following steps:
(1) edible and medical fungi is carried out enzymolysis processing under the cryogenic conditions of 4-29 DEG C, obtain low temperature edible and medical fungi extracting solution after filtration; Preserve under being placed in 4 DEG C of aseptic conditions;
(2) cell culture: by one or several in fibrocyte, epithelial cell, osteoblast, mixing or single culture; During cell culture, in regular growth culture fluid, add the above-mentioned edible and medical fungi extracting solution of 1-40% percentage by weight, cultivate through 1-12 week, supernatant Collection and conservation is for subsequent use in 4 DEG C of gnotobasiss;
(3) enzyme digestion reaction: fetch and come from bathypelagic fish or mammiferous corium or skeletal tissue; The supernatant above-mentioned tissue and step (2) obtained adds according to 1-12: 1 weight ratio in the enzymolysis solution of 4 DEG C and reacts; Filter under 4 DEG C of gnotobasiss after reaction, filtering accuracy scope: 0.10 μm-0.22 μm; Filtered solution PH is adjusted to 7.4, obtains product, is stored in 4 DEG C of gnotobasiss.
3. 4 DEG C according to claim 2 are in a liquid state 37 DEG C in the preparation method of solid-state biomaterial, it is characterized in that: described edible and medical fungi is one or several in chanterelle, GANBAJUN, bolete, Termitomyces albuminosus, Morchella esculenta (L.) Pers, Tricholoma lobayense Heim, Pnoliota adiposa(Fr.) Quel., Armillariella mellea, blue or green mushroom, Hericium erinaceus (Bull. Ex Fr.) Pers., red mushroom, Grifola frondosa, Agrocybe aegerita (Brig.) Sing., Ganoderma, Clitocybe maxima, agrocyb eaegerita, pholiota nameko, Mushroom Corals, Oudemansiella Radicata, Pleurotus citrinopileatus, Hypsizygus marmoreus, asafoetida mushroom, Pleurotus eryngii, Pleurotus abalonus.
4. 4 DEG C of 37 DEG C of preparation methoies in solid-state biomaterial that are in a liquid state according to claim 2, it is characterized in that: during the enzymolysis processing of step (1), the enzyme of use is pectase, one or several in cellulase, hemicellulase, α-amylase, glucoamylase, beta amylase; Feed liquid weight ratio: 1: 1-19; The enzyme digestion reaction time: 1-190 hour; PH value: 3-9; Enzyme concentration: be with edible and medical fungi weight ratio: enzyme 0.01-5: edible and medical fungi 100; Filtering accuracy in step (1) is 0.10 μm-0.22 μm.
5. 4 DEG C according to claim 2,3 or 4 are in a liquid state 37 DEG C in the preparation method of solid-state biomaterial, it is characterized in that: the enzymatic hydrolysis condition in step (3) is: pH value: 3-5.4; Enzyme digestion reaction temperature: 4-9 degree Celsius; Feed liquid weight ratio: 1: 1-19; The enzyme digestion reaction time: 72-480 hour; The kind of enzyme: one or several in metalloproteases, thiol protease, serine carboxypeptidase, cathepsin, endopeptidase, ficin, pepsin; Enzyme concentration: with raw material weight ratio be: enzyme 0.01-5: raw material 100.
6. 4 DEG C of 37 DEG C of application in solid-state biomaterial in preparation in transient or permanent occlusion, half occluder that are in a liquid state according to claim 1.
7. 4 DEG C of 37 DEG C of application in solid-state biomaterial in preparation in transient or permanent occlusion, half occluder that are in a liquid state according to claim 6, is characterized in that: be as nasolacrimal duct, tumor vessel or deferential transient or permanent occlusion, half blocking material.
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| CN1709915A (en) * | 2005-06-23 | 2005-12-21 | 南京大学医学院附属鼓楼医院 | Polysaccharide material for promoting cell tactophily and preparation thereof |
| CN101921821A (en) * | 2010-08-06 | 2010-12-22 | 郁小兵 | Method for extracting high-quality collagen at low temperature |
| CN101947245A (en) * | 2010-08-27 | 2011-01-19 | 郁小兵 | Method for extracting effective components of edible and medical fungi in ordinary state |
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| JPS6028936A (en) * | 1983-07-27 | 1985-02-14 | Koken:Kk | Atherocollagen aqueous solution and its preparation |
| US5648208A (en) * | 1991-08-01 | 1997-07-15 | Coletica | Use of a collagen as solid binding substrate for a ligand capable of reacting specifically with an element to be detected in a biological medium, reactant and implementation |
| CN101869263A (en) * | 2010-05-27 | 2010-10-27 | 郁小兵 | Edible fungi dietary fiber and production method thereof |
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| CN1709915A (en) * | 2005-06-23 | 2005-12-21 | 南京大学医学院附属鼓楼医院 | Polysaccharide material for promoting cell tactophily and preparation thereof |
| CN101921821A (en) * | 2010-08-06 | 2010-12-22 | 郁小兵 | Method for extracting high-quality collagen at low temperature |
| CN101947245A (en) * | 2010-08-27 | 2011-01-19 | 郁小兵 | Method for extracting effective components of edible and medical fungi in ordinary state |
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