CN101766836A - Preparation method of nano silver cordspinal cord and peripheral nerve repairing material - Google Patents
Preparation method of nano silver cordspinal cord and peripheral nerve repairing material Download PDFInfo
- Publication number
- CN101766836A CN101766836A CN200910001072A CN200910001072A CN101766836A CN 101766836 A CN101766836 A CN 101766836A CN 200910001072 A CN200910001072 A CN 200910001072A CN 200910001072 A CN200910001072 A CN 200910001072A CN 101766836 A CN101766836 A CN 101766836A
- Authority
- CN
- China
- Prior art keywords
- collagen
- preparation
- nanometer silver
- chitosan
- microtubule
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Materials For Medical Uses (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Prostheses (AREA)
Abstract
The invention relates to a preparation method of a nano silver material for repair of damage of a spinal cord and peripheral nerves, which belongs to the field of repair of damage of human or animal spinal cords and peripheral nerves with a tissue engineering method. The repairing material is prepared by mixing collagens (I, III and IV type), chitosan and nano silver. The speed for lowering a silicone tube filled with mixture of the nano silver, the collagens and the chitosan into cold leaching solution and/or the temperature of the cold leaching solution are/is changed so that the internal structure of the material for repairing the tissue engineering of the spinal cords and the peripheral nerves can be provided with an axial micro tube matrix, and the inner surfaces of micro tubes are provided with positive charges uniformly distributed. The nerve repairing material for tissue engineering can substitute the material for grafting of autologous free nerves, avoid the iatrogenic damage to the supply area, speed up the growth of the regenerated axon of the peripheral nerve, and shorten the nerve repairing time. Therefore, the invention can be widely applied in the surgical repair of clinical spinal cord injury and long-term peripheral nerve defects.
Description
Technical field
The invention belongs to the using-system engineering method and repair human body or animal spinal cord and peripheral nerve injury.Relate generally to a kind of use that nanometer silver, collagen and chitosan make have a preparation technology with the micro-tubular structure support of damage location nervous tissue equidirectional.
Background technology
Extracellular matrix (extracellular matrix) composition such as collagen etc. play an important role in the adhesion of cell and the process of dividing a word with a hyphen at the end of a line.Collagen is the most important water-insoluble fibrin in extracellular, is the skeleton that constitutes extracellular matrix.Collagen forms the fiber of semi-crystal in extracellular matrix, provide tension stress and elasticity to cell, and works in the migration of cell with in growing.Collagen all has existence in various animals.Collagen in the vertebrates in tendon, cartilage and the bone is very abundant, has almost accounted for half of albumen gross weight.
Chitosan is the product of chitin behind deacetylation.The biocompatibility of chitosan is good, and is extremely extensive in the application of aspects such as biomedicine and pharmacy, can be used as burn dressing and Wound-healing agent, the wrapping gauze with treatment with chitosan after, speed of wound healing can improve 75%.With the absorbability sutures that chitosan is made, mechanical strength height, but long term store can be sterilized with conventional method, can dye, and can mix medicament, can be organized degraded and absorbed, exempt the misery that the patient takes out stitches.
The safety of nanometer silver is that international medical community generally acknowledges that human body cell is not had negative effect.U.S.'s poisonous substance and disease registry (Toxicological Profile Information Sheet, ATSDR) and U.S. sanitary and (the United States Department of Health and Human Services of public service portion, HHS) toxicity of silver is investigated, content relates to the absorption, distribution, metabolism, drainage of silver and other to health affected, finds that so far silver is to people's toxic reaction or influence function and the growth and the genotoxic report of immune system, cardiovascular system, reproductive system.Secondly, the nano-Ag particles stable in properties can be easy to add in the timbering material with variable concentrations.Because its surface has positive divalent charge, between the granule because of the Coulomb repulsion effect is difficult for accumulation, so be evenly distributed.Again secondly, the nano-Ag particles size is stable, and diameter can not be by immune system recognition, so can not produce immunological rejection between the 3-30 nanometer.Can slowly discharge with material degradation, be difficult for accumulating in vivo.At last, the antibiotic property of nanometer silver is admitted by medical circle that the timbering material that uses nano-Ag particles to make so can play good antibacterial properties at injury region.Particularly after wound, can effectively reduce the infiltration of inflammatory cell, can play further facilitation the regeneration of nerve.
Summary of the invention
The objective of the invention is the using-system engineering method and prepare a kind of engineered artificial neuron with axial micro-tubular structure to repair spinal cord and peripheroneural damaged.
The technical solution used in the present invention is: use nanometer silver, collagen protein and chitosan to mix the back according to a certain percentage as material and inject silica gel tube, under certain cryogenic conditions, change and to be marked with nanometer silver, collagen and chitosan compound silica gel tube and to fall into speed in the stranguria of cold type liquid and/or stranguria of cold type liquid temp and make spinal cord, peripheral nerve tissue's engineering repair materials on internal structure, have axial microtubule matrix to arrange and microtubule inner surface uniform distribution positive charge characteristic.Use cross-linking agent crosslinked on the support that begins to take shape, can form the tissue engineering bracket that has axial micro-tubular structure and have certain mechanical strength.Concrete making step is as follows:
1) on the make takes by weighing 75 milligrams of 150 milligrams of collagen protein and chitosans, be dissolved in 0.05 mol acetum.In 4 degrees centigrade of isoperibols,, make the collagen suspension with 1000 rev/mins of stirrings 30 minutes;
2) 0.01-0.1 mg/litre concentration nanometer silver solution is pressed 1: 8-1: 12 volume ratios add the collagen suspension.In 4 degrees centigrade of isoperibols,, make nanometer silver, collagen and chitosan suspension equally with 1000 rev/mins of stirrings 30 minutes;
3) with the silica gel tube of nanometer silver, collagen and chitosan suspension injection internal diameter 2-15 millimeter, sealing both ends is by 2 * 10
-5Meter per second speed direct-axis is in immersing condensing agent;
4) suspension and the freezing thing of silica gel tube are cut into respective length according to application, (40 degrees centigrade, 100 millitorrs) lyophilizing is 48 hours in the lyophilization machine;
5) take out exsiccant nanometer silver, collagen and chitosan stent, immerse concentration and be in 5-15 grams per liter genipin (Genipin) solution after crosslinked 48 hours, (40 degrees centigrade, 100 millitorrs) lyophilizing is 24 hours in the lyophilization machine.
The specific embodiment
Embodiment 1: according to technical solution of the present invention, be example with type i collagen albumen, nanometer silver and chitosan, its manufacture method is carried out according to the following steps:
1. take by weighing 75 milligrams of 0 milligram of type i collagen protein 15 and chitosans, be dissolved in 0.05 mol acetum.In 4 degrees centigrade of isoperibols,, make the collagen suspension with 1000 rev/mins of stirrings 30 minutes;
2. 0.01 mg/litre concentration nanometer silver solution is joined the collagen suspension by 1: 8 volume ratio.In 4 degrees centigrade of isoperibols,, make nanometer silver, collagen and chitosan suspension equally with 1000 rev/mins of stirrings 30 minutes;
3. nanometer silver, collagen and chitosan suspension are injected the silica gel tube of internal diameter 2-15 millimeter, sealing both ends is by 2 * 10
-5Meter per second speed direct-axis is in immersing condensing agent;
4. suspension and the freezing thing of silica gel tube are cut into respective length according to application, (40 degrees centigrade, 100 millitorrs) lyophilizing is 48 hours in the lyophilization machine;
5. take out exsiccant nanometer silver, collagen and chitosan stent, immerse concentration and be in the 5-15 grams per liter genipin solution after crosslinked 48 hours, (40 degrees centigrade, 100 millitorrs) lyophilizing is 24 hours in the lyophilization machine.
Prepared spinal cord, the peripheral nerve repairing material of the present invention has following characteristics:
1. the preparation of this spinal cord, peripheral nerve repairing material mainly uses nanometer silver, collagen and chitosan to be primary raw material, and timbering material can be neuranagenesis in vivo bionical temporary structure is provided, but and natural degradation.No immunologic rejection and body accumulation toxicity.
2. the internal stent microtubule has axial matrix structure arranged feature, the microtubule controllable diameter is built in 20 microns～300 microns, the aixs cylinder oriented growth helps regenerating, also make the seed cell of transplanting can be, thereby make the nerve of damage obtain effective for repairing regeneration to greatest extent along microtubule wall axially-aligned, migration.The outer surface of material is a compact texture, can effectively stop growing into of the interior fibrous connective tissue of body.Can keep certain profile for a long time in vivo, still can keep the better space structure after 4 months, this material both can directly be implanted the bridge joint neurologic defect, also can be used as the carrier of seed cell plantation.
3. a large amount of nano-Ag particles of support microtubule inner surface uniform distribution.Because nano-Ag particles is met water power and is made the support microtubule inner surface positive divalent charge that evenly and in a large number distributing in vivo from the characteristics that go out positive bivalence silver ion.The inner surface of uniform distribution positive charge can be directly and the electronegative group of the molecule of neuronal cell surface interact, with cell membrane generation non-specific adsorption, help neural regeneration.
4. use plant extract composition genipin as cross-linking agent, can strengthen the timbering material mechanical strength, and no cytotoxicity.
5. the nano-Ag particles in the timbering material can play good antibacterial properties at injury region.
Following example 2～12.
Embodiment 2: adopt type i collagen albumen, nanometer silver solution (1: 12 volume ratio, 0.01 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Embodiment 3: adopt type i collagen albumen, nanometer silver solution (1: 8 volume ratio, 0.1 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Embodiment 4: adopt type i collagen albumen, nanometer silver solution (1: 12 volume ratio, 0.1 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Embodiment 5: adopt III collagen type, nanometer silver solution (1: 12 volume ratio, 0.01 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Embodiment 6: adopt III collagen type, nanometer silver solution (1: 12 volume ratio, 0.1 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Embodiment 7: adopt III collagen type, nanometer silver solution (1: 8 volume ratio, 0.1 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Embodiment 8: adopt III collagen type, nanometer silver solution (1: 8 volume ratio, 0.01 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Embodiment 9: adopt IV collagen type, nanometer silver solution (1: 12 volume ratio, 0.01 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Embodiment 10: adopt IV collagen type, nanometer silver solution (1: 12 volume ratio, 0.1 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Embodiment 11: adopt IV collagen type, nanometer silver solution (1: 8 volume ratio, 0.1 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Embodiment 12: adopt IV collagen type, nanometer silver solution (1: 8 volume ratio, 0.01 mg/litre concentration) and chitosan to prepare as original material, its implementation process is with embodiment 1.
Nanometer silver, collagen and the chitosan stent of this patent preparation have the axial matrix structure arranged of microtubule feature, and the microtubule controllable diameter is built in 30 μ m～300 μ m, and the outer surface of material is a compact texture, can effectively stop growing into of the interior fibrous connective tissue of body.Can keep certain profile for a long time in human body, still can keep the better space structure after 4 months, this material both can directly be implanted the bridge joint neurologic defect, also can be used as the carrier of seed cell plantation.
The above only is the preferred embodiments of the disclosure; but protection scope of the present invention is not limited thereto; any people who is familiar with this technology can expect similarly changing and replacing in technical scope disclosed in this invention easily, all should be encompassed in protection scope of the present invention.
Claims (6)
1. the preparation method of a spinal cord, peripheral nerve repairing material, this spinal cord, peripheral nerve repairing material use I, III and IV Collagen Type VI and chitosan and nanometer silver are mixed and made into, under certain cryogenic conditions, change and to be marked with nanometer silver, collagen and chitosan compound silica gel tube and to fall into speed in the stranguria of cold type liquid and/or stranguria of cold type liquid temp and make spinal cord, peripheral nerve tissue's engineering repair materials have axial microtubule matrix on internal structure to arrange and microtubule inner surface uniform distribution positive charge characteristic, its preparation method carries out according to the following steps:
1) takes by weighing 75 milligrams of 150 milligrams of collagen protein and chitosans, be dissolved in 0.05 mol acetum, in 4 degrees centigrade of isoperibols,, make the collagen suspension with 1000 rev/mins of stirrings 30 minutes;
2) with 0.01-0.1 mg/litre concentration nanometer silver solution according to 1: 8-1: 12 volume ratios join the collagen suspension, equally in 4 degrees centigrade of isoperibols, stir 30 minutes with 1000 rev/mins, make nanometer silver-collagen-chitin suspension;
3) nanometer silver-collagen-chitin suspension is injected the silica gel tube of 2 millimeters-15 millimeters of internal diameters, sealing both ends is by 2 * 10
-5Meter per second speed direct-axis is in immersing condensing agent;
4) suspension and the freezing thing of silica gel tube are cut into respective length according to application, at-40 degrees centigrade, lyophilizing is 48 hours in the 100 millitorr lyophilization machines;
5) take out exsiccant nanometer silver-collagen-chitin support, immerse concentration and be in the 5-15 grams per liter genipin solution after crosslinked 48 hours, at-40 degrees centigrade, lyophilizing is 24 hours in the 100 millitorr lyophilization machines.
2. preparation method as claimed in claim 1, it is characterized in that, the preparation of this spinal cord, peripheral nerve repairing material mainly uses I, III and IV Collagen Type VI and chitosan and nanometer silver to be primary raw material, nanometer silver-the collagen-chitin support can be neuranagenesis in vivo provides bionical temporary structure, but and natural degradation, no immunologic rejection and body accumulation toxicity.
3. preparation method as claimed in claim 1, it is characterized in that, the internal stent microtubule has axial matrix structure arranged feature, the microtubule controllable diameter is built in 20 microns～300 microns, the aixs cylinder oriented growth helps regenerating, also make the seed cell of transplanting can be along microtubule wall axially-aligned, migration, thereby the nerve that makes damage to greatest extent obtains effective for repairing regeneration, the outer surface of material is a compact texture, can effectively stop growing into of the interior fibrous connective tissue of body, can keep certain profile for a long time in vivo, still can keep the better space structure after 4 months, this material both can directly be implanted the bridge joint neurologic defect, also can be used as the carrier of seed cell plantation.
4. preparation method as claimed in claim 1, it is characterized in that, the a large amount of nano-Ag particles of support microtubule inner surface uniform distribution, because nano-Ag particles is met water power and is made the support microtubule inner surface positive divalent charge that evenly and in a large number distributing in vivo from the characteristics that go out positive bivalence silver ion, the inner surface of uniform distribution positive charge can be directly and the electronegative group of the molecule of neuronal cell surface interact, with cell membrane generation non-specific adsorption, help neural regeneration.
5. preparation method as claimed in claim 1 is characterized in that, uses plant extract composition genipin as cross-linking agent, can strengthen the timbering material mechanical strength, and no cytotoxicity.
6. preparation method as claimed in claim 1 is characterized in that, uses the timbering material of nano-Ag particles preparation to have good antibiotic property at injury region.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910001072A CN101766836B (en) | 2009-01-21 | 2009-01-21 | Preparation method of nano silver cordspinal cord and peripheral nerve repairing material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910001072A CN101766836B (en) | 2009-01-21 | 2009-01-21 | Preparation method of nano silver cordspinal cord and peripheral nerve repairing material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101766836A true CN101766836A (en) | 2010-07-07 |
| CN101766836B CN101766836B (en) | 2012-09-05 |
Family
ID=42500036
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910001072A Expired - Fee Related CN101766836B (en) | 2009-01-21 | 2009-01-21 | Preparation method of nano silver cordspinal cord and peripheral nerve repairing material |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101766836B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103007350A (en) * | 2012-12-14 | 2013-04-03 | 中国人民解放军第三军医大学第二附属医院 | Modified rat accellular spinal cord bracket material and preparation method thereof |
| CN105709278A (en) * | 2015-05-06 | 2016-06-29 | 周晗 | Tissue engineering spinal marrow containing directional axons in various areas |
| CN106110390A (en) * | 2016-06-30 | 2016-11-16 | 丁坦 | A kind of RAPA PLGA support |
| CN111359019A (en) * | 2020-03-16 | 2020-07-03 | 四川大学 | Preparation method of graphene and chitosan composite conductive nerve scaffold with longitudinal pore channels |
| CN111936151A (en) * | 2017-02-22 | 2020-11-13 | 铠耀纳米银Rx(香港)有限公司 | Compositions and methods for neuroprotection using nanoparticulate silver |
| WO2022193599A1 (en) * | 2021-03-19 | 2022-09-22 | 潍坊奥精医学研究有限公司 | Method for preparing spinal cord regeneration and repair material |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1380115A (en) * | 2002-03-25 | 2002-11-20 | 中国人民解放军第四军医大学 | Preparation process of spinal cord and peripheral nerve repairing material |
| CN1256155C (en) * | 2003-12-15 | 2006-05-17 | 中国人民解放军第四军医大学 | Tissue engineering compound material for repairing nerve injury and its preparation |
| CN1303946C (en) * | 2004-06-25 | 2007-03-14 | 清华大学 | Nerve tissue engineering tube type bracket and method for making same |
| CN101096424A (en) * | 2007-06-27 | 2008-01-02 | 东华大学 | Glutin nano fabric film containing nano silver and preparation and application thereof |
-
2009
- 2009-01-21 CN CN200910001072A patent/CN101766836B/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103007350A (en) * | 2012-12-14 | 2013-04-03 | 中国人民解放军第三军医大学第二附属医院 | Modified rat accellular spinal cord bracket material and preparation method thereof |
| CN105709278A (en) * | 2015-05-06 | 2016-06-29 | 周晗 | Tissue engineering spinal marrow containing directional axons in various areas |
| CN105709278B (en) * | 2015-05-06 | 2019-05-21 | 上海蓝十字脑科医院有限公司 | Contain the tissue engineering spinal cord of orientation aixs cylinder in subregion |
| CN106110390A (en) * | 2016-06-30 | 2016-11-16 | 丁坦 | A kind of RAPA PLGA support |
| CN111936151A (en) * | 2017-02-22 | 2020-11-13 | 铠耀纳米银Rx(香港)有限公司 | Compositions and methods for neuroprotection using nanoparticulate silver |
| CN111359019A (en) * | 2020-03-16 | 2020-07-03 | 四川大学 | Preparation method of graphene and chitosan composite conductive nerve scaffold with longitudinal pore channels |
| WO2022193599A1 (en) * | 2021-03-19 | 2022-09-22 | 潍坊奥精医学研究有限公司 | Method for preparing spinal cord regeneration and repair material |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101766836B (en) | 2012-09-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Islam et al. | Chitosan based bioactive materials in tissue engineering applications-A review | |
| Yan et al. | Injectable alginate/hydroxyapatite gel scaffold combined with gelatin microspheres for drug delivery and bone tissue engineering | |
| CN101766836B (en) | Preparation method of nano silver cordspinal cord and peripheral nerve repairing material | |
| CN102526798B (en) | Injectable compound bone cement and preparation method thereof | |
| CN100408112C (en) | Injection aquagel of sodium alginate cross-linking gelatin comprising biphase calcium phosphor granule, method for making same and use thereof | |
| CN104271166B (en) | Biomaterial based on absorbent cellulosic and implant | |
| CN101502678B (en) | Mixed gel for injection and preparation method thereof | |
| CN101557801A (en) | Hydrogels containing low molecular weight alginates and biostructures made therefrom | |
| CN101905038A (en) | Growth factor-loaded collagen group composite material as well as preparation method and application thereof | |
| EP2358758A1 (en) | Bioactive hydrogel | |
| EP0977780B1 (en) | Hetero-polysaccharide conjugates, s-inp polysaccharide gels and methods of making and using the same | |
| CN102813961A (en) | Injection gel containing submicron hyaluronic acid microspheres and preparation method thereof | |
| CN103007358B (en) | Cartilage tissue engineering fiber scaffold material and preparation method thereof | |
| Liu et al. | Biomedical applications of bacterial cellulose based composite hydrogels | |
| Zhang et al. | Chitosan/gelatin-tannic acid decorated porous tape suture with multifunctionality for tendon healing | |
| CN110790951A (en) | In-situ crosslinked gamma-polyglutamic acid hydrogel and preparation method and application thereof | |
| CN102552983B (en) | Year enzyme calcium phosphate bone cement of prevention and therapy infection of bone and preparation method thereof | |
| CN1539514A (en) | Method for preparing multifunctional biological repair material | |
| CN102552985B (en) | Silk fibroin/calcium phosphate bone cement-based porous composite material and preparation method thereof | |
| CN107982575A (en) | Preparation method of LDI-PCLLA-PEG-PCLLA/ hydroxyapatite electrospun fiber membranes and products thereof and application | |
| CN105148322A (en) | Injectable hydrogel and method for preparing same | |
| CN110721348B (en) | Natural silk reinforced hydroxyapatite/chitosan composite membrane and preparation method thereof | |
| CN109316623B (en) | Double-layer porous biodegradable material coated with active molecules and preparation method and application thereof | |
| CN113941033B (en) | Double-drug-loading nanofiber hydrogel composite cartilage repair system and preparation method thereof | |
| Kaur et al. | Biomedical applications of synthetic and natural biodegradable polymers |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120905 Termination date: 20130121 |