CN103357067B - Fibroin-base artificial ligament repair material and preparation method thereof - Google Patents
Fibroin-base artificial ligament repair material and preparation method thereof Download PDFInfo
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- CN103357067B CN103357067B CN201310284212.2A CN201310284212A CN103357067B CN 103357067 B CN103357067 B CN 103357067B CN 201310284212 A CN201310284212 A CN 201310284212A CN 103357067 B CN103357067 B CN 103357067B
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Abstract
The invention discloses a fibroin-base artificial ligament repair material and a preparation method thereof. The ligament repair material comprises stents and laid-in yarns, wherein the stents are produced by carrying out biaxial laid-in warp knitting on natural mulberry silks; the laid-in yarns are regenerated silkworm silk fibroin filament yarns and regenerated silkworm silk nanofiber yarns. The preparation method specifically comprises the following steps of: (1) preparing biaxial laid-in warp knitting stent materials; (2) preparing regenerated silk fibroin filament yarns by utilizing a dry or wet spinning method; (3) preparing and dissloving a regenerated silk fibroin film, and preparing nanometer silk fibroin fiber yarns by utilizing an electrostatic spinning method; (4) taking a pure fibroin warp knitting material as a base material, taking the regenerated silkworm silk fibroin filament yarns and the nanometer silk fibroin fiber yarns as the laid-in yarns, weaving the base material and the laid-in yarns into the warp knitting stent material to obtain the artificial ligament repair material, and sewing two sides of a ligament material into a columnar structure. The preparation method is simple and efficient, and the artificial ligament repair material prepared by utilizing the preparation method has excellent compatibility, degradability and mechanical properties.
Description
Technical field
The present invention relates to a kind of artificial ligament repair materials, be specifically related to a kind of fibroin base artificial ligament repair materials and preparation method thereof, belong to Implantable Medical Device technical field.
Background technology
Along with the rise of tissue engineering, artificial ligament obtains people's attention again as Healing in Anterior Cruciate Ligament Reconstruction material, for ligament is provided by the effective approach that provides.The development of the subjects such as the comprehensive life sciences of tissue engineering artificial ligament and engineering, is used biological fiber and synthetic material as tough belt supporting frame, promotes in vitro and in vivo sticking and growing of cell and tissue, to substitute normal anterior cruciate ligament.
In prior art, the artificial ligament material that belongs to study hotspot and obtained certain progress is permanent type ligament material, as: carbon fibre ligament; Dacron ligament, material is polyethylene terephthalate; Gore-Tex ligament, material is politef; LAD strengthens ligament, and material is polypropylene; LARS polyester ligament etc.But all Shortcomings of above-mentioned ligament material: carbon fibre degeneration is fast, can not provide enough tensile strength, and histocompatibility is poor, and cracking granule causes heavier inflammatory reaction, and compliance is poor; Dacron ligament material is prone to complication, as ligament rapture and synovitis; Although Gore-Tex ligament material intensity is large, rebuild early stage results fair, can continue in time generation degeneration, develop complications; LAD strengthen ligament material implant can cause that reactivity is oozed out behind knee joint inside, joint fibrosis and synovitis, increase infection chance; LARS polyester ligament cannot degradation in vivo, can not guide Growth of Cells to promote the formation of human body self ligament tissue.So first suitable artificial ligament repair materials will possess excellent biocompatibility and degradability.
Fibroin albumen is the natural polymer fibrin extracting from silkworm silk, has good biocompatibility and good physical and chemical performance, becomes in recent years one of focus object of tissue engineering bracket material research.Research shows that silk fibroin material has than other natural fibers and the unique mechanical property of high-performance synthetic fibers; Fiber surface is easy to chemical covalent modification and sticks site and cytokine, can be processed and be obtained variform and change surface property by distinct methods; Can slow degradation be aminoacid in vivo, and catabolite to organism without danger.
The people such as Altman adopt fibroin design organization engineering anterior cruciate ligament, its 3-D space structure is referred to as steel cable type structure, its mechanical property and fatigue durability are all satisfactory (referring to Gregory H. Altman, David L. Kaplan et al. Biomaterials. 23 (2002) 4131 – 4141), but in cell cultivation process, because steel cable type ligament structure is tight, cell can only be attached to the surface of support, cannot enter internal stent; The people such as Haifeng Liu have studied fibroin frame-type ligament material, silk stent material is immersed to the final 3-D of formation space structure after silk fibroin protein solution (referring to Haifeng Liu, Hongbin Fan et al. Biomaterials. 29 (2008) 662 – 674), but the stability of the porous fibroin material forming is restive; The people such as Laurencin are taking PLLA as raw material, weave banded timbering material and form 3-D space structure (referring to Cooper JA, Lu HH et al. Biomaterials. 26 (2005) 1523 – 1532), its ligament is that fibre bundle interlacing becomes band, the intersecting angle at ligament stage casing and two ends difference to some extent, but stage casing yarn line structure has intersection, can some effects cell adhesion.
So, need to develop new artificial ligament repair materials.
Summary of the invention
The object of the invention is to provide a kind of fibroin base artificial ligament repair materials, utilize natural mulberry silk, regeneration mulberry silk and regeneration mulberry silk nano-fibre yams jointly to build, to obtain the material of good biocompatibility, improve cell adhering to and growing in artificial ligament repair materials.
Another object of the present invention is to provide the preparation method of above-mentioned material.
For achieving the above object, the technical solution used in the present invention is, a kind of fibroin base artificial ligament repair materials, comprises support and stuffer weft, and described support adopts twin shaft to form through compiling to inlaid thread by natural mulberry silk; Stuffer weft is Regenerated Bombyx mori Silk Fibroin long filament and regeneration mulberry silk nano-fibre yams, and described stuffer weft inweaves in the warp knitting fabrics structure of support.
In technique scheme, natural mulberry silk consumption accounts for 30~70% of artificial ligament repair materials total amount; Regenerated Bombyx mori Silk Fibroin long filament consumption accounts for 10~30% of artificial ligament repair materials total amount; Regeneration mulberry silk nano-fibre yams consumption accounts for 20~40% of artificial ligament repair materials total amount.
In technique scheme, described artificial ligament repair materials length is 27.5~41cm; The diameter of described Regenerated Bombyx mori Silk Fibroin long filament is 15~80 μ m; The diameter of described regeneration mulberry silk nano-fibre yams is 50~2000nm.
The diameter of artificial ligament repair materials disclosed by the invention can be according to the diameter design of the ligament that will repair, is preferably 0.8~1.5 times of the ligament diameter that will repair.
In preferred technical scheme, described artificial ligament repair materials also comprises reinforcing fibre, and it inweaves in warp knitting fabrics structure as stuffer weft, and consumption accounts for 10~90% of the total consumption of stuffer weft; Described reinforcing fibre is carbon fiber, superhigh molecular weight polyethylene fibers or polyester fiber.
The invention also discloses a kind of method of preparing above-mentioned artificial ligament repair materials, comprise the following steps:
(1) adopt natural mulberry silk braiding twin shaft to inlaid thread through compiling timbering material, after obtain Bombyx mori Silk Fibroin through coming unstuck through compiling timbering material;
(2) natural mulberry silk is prepared regenerated silk fibroin solution after coming unstuck, dissolve, dialysing, and adopts dry method or wet spinning to prepare Regenerated Bombyx mori Silk Fibroin long filament;
(3) natural mulberry silk is prepared regenerated silk fibroin film after coming unstuck, dissolve, dialyse, being dried, and uses hexafluoroisopropanol or formic acid to dissolve fibroin protein film, then adopts method of electrostatic spinning preparation regeneration mulberry silk nano-fibre yams;
(4) taking fibroin through compile timbering material as base material, Regenerated Bombyx mori Silk Fibroin long filament and regeneration mulberry silk nano-fibre yams inweave in warp knitting fabrics structure as stuffer weft, reel and form artificial ligament material, finally ligament material both sides are sewed up and obtained described fibroin base artificial ligament repair materials.
In technique scheme, in mass ratio, natural mulberry silk: Regenerated Bombyx mori Silk Fibroin long filament: regeneration mulberry silk nano-fibre yams is (30~70): (10~30): (20~40).
In preferred technical scheme, in step (4), inweave natural mulberry silk prepared by step (1) using prepared regenerated silk fibroin long filament in carbon fiber, superhigh molecular weight polyethylene fibers or polyester fiber and step (2), (3) and regeneration mulberry silk nano-fibre yams as stuffer weft in compiling timbering material, jointly build artificial ligament repair materials; The consumption of carbon fiber, superhigh molecular weight polyethylene fibers or polyester fiber accounts for 10~90% of the total consumption of stuffer weft.
In the present invention, braiding all belongs to prior art through compiling timbering material, natural silk degumming, dissolving, dialysis, dry spinning, wet spinning, electrostatic spinning, coiling and sewing up, and those skilled in the art can select voluntarily according to product.The preferred condition of the present invention is as follows:
In above-mentioned steps (1), weave to tricot weft insertion machine through its tissue of braiding employing RSM twin shaft of compiling timbering material, organize technique, threading yarn technique as follows:
Organize technique:
GB1:2-3/3-2/1-0/2-3/1-0/0-1/2-3/1-0//
GB2:1-0/0-1//×4;
Threading yarn technique:
GB1: completely wear
GB2: completely wear;
Plane No.: E12
In above-mentioned steps (2), the preparation of Regenerated Bombyx mori Silk Fibroin long filament be by natural mulberry silk through coming unstuck, to form mass fraction after dissolving, dialysis treatment be 4% silk fibroin water solution, being positioned over stirring at low speed in Polyethylene Glycol (PEG) solution, to be concentrated into mass fraction be 22%, 27%, 32% silk fibroin water solution; Be 25 DEG C of left and right in temperature, within the resting period of 0-5d, carry out wet spinning, spun filament cellulose fiber is immersed in dehydrated alcohol, sealing, is placed in 24 h under room temperature, carries out drawing-off in the deionized water more than 60 DEG C, by one times of as-spun fibre drawing-off, heat setting 30 min at 120 DEG C; Again spun filament cellulose fiber is positioned over to mass ratio and is in 1-(3-dimethylaminopropyl)-3-ethyl carbodiimides (EDC)/N-maloyl imines (NHS) of 2: 1, add by a certain percentage ethanol/water (V
c2H5OH/ V
h2O=90/10) mass fraction being mixed with in is 4.5% crosslinked fluid, crosses 24 h, and drawing-off in the deionized water more than 60 DEG C, is placed on heat setting 30 min at 120 DEG C, obtains the Regenerated Bombyx mori Silk Fibroin long filament after EDC/NHS is cross-linked.
In above-mentioned steps (3), method of electrostatic spinning be by natural mulberry silk through coming unstuck, to form mass fraction after dissolving, dialysis treatment be 4~8% silk fibroin water solution, aqueous solution is poured on vinyon flat board, at ambient temperature the dry fibroin membrane that obtains; Regenerated Silk Fibroin Film is dissolved in hexafluoroisopropanol (HFIP) or formic acid, in 25 DEG C of water-baths of constant temperature, vibrating, within one week, to make mass fraction be 10~12% spinning liquid, spinning liquid is poured into and in syringe, carried out electrostatic spinning, No. 9 syringe needles, spinning flow 2mL/h, adjust voltage and curing distance, make field intensity reach 1kV/cm, make regeneration mulberry silk nano-fibre yams.
In above-mentioned steps (4), adopt serger that ligament material both sides are sewed up, the artificial ligament repair materials obtaining is column construction, overall length 27.5~41cm, parallel fiber zone length 1.5~5cm, two ends woollen yarn knitting zone length is respectively 10~15cm, and consolidation zone length of field is 3cm, and diameter is 8~15mm.This ligament repair material, taking the warp knitting fabrics structure of natural mulberry silk weaving as base rack, has ensured the mechanical property of ligament structure; Increase regenerated silk fibroin long filament and regeneration mulberry silk nano-fibre yams in the inside of support simultaneously, its fiber size is more conducive to guide Growth of Cells, thereby accelerate the formation of self ligament tissue, can in the process that self ligament tissue forms, degrade gradually simultaneously, reach the advantageous property of organizational project ligament, also for cambium provides support, and keep certain hour to cambium to there is the biomechanics characteristic of growing.
Because technique scheme is used, the present invention compared with prior art has following advantages:
(1) the present invention is raw materials used is pure natural fibroin albumen, has excellent biocompatibility, nontoxic, be beneficial to cell adhesion, growth and differentiation, and final catabolite is aminoacid, harmless; When the artificial ligament repair materials of preparation has good biocompatibility and degradability thus, there is high mechanical strength;
(2) ligament repair material disclosed in this invention presents 3-D solid structure, has the feature of high porosity, high-specific surface area, can provide sufficient space to be beneficial to cell adhesion growth and nutrient substance enters, the discharge of metabolite;
(3) preparation method disclosed in this invention is simple, and easy operating is applicable to suitability for industrialized production.
Brief description of the drawings
Fig. 1 is through compiling the structure chart of support in embodiment mono-;
Fig. 2 is the structural representation of artificial ligament repair materials in embodiment mono-;
Fig. 3 is the longitudinal scanning Electronic Speculum figure of Regenerated Bombyx mori Silk Fibroin filament fiber in embodiment mono-;
Fig. 4 is the scanning electron microscope (SEM) photograph of mulberry silk nano-fibre yams of regenerating in embodiment mono-.
Detailed description of the invention
Below in conjunction with drawings and Examples, the invention will be further described:
Embodiment mono-:
(1) natural mulberry silk is weaved to tricot weft insertion machine through its tissue of braiding employing RSM twin shaft of compiling timbering material, organizes technique, threading yarn technique as follows:
A. organize technique:
GB1:2-3/3-2/1-0/2-3/1-0/0-1/2-3/1-0//
GB2:1-0/0-1//×4
B. threading yarn technique:
GB1: completely wear;
GB2: completely wear
C. plane No.: E12;
(2) by natural mulberry silk through coming unstuck, to form mass fraction after dissolving, dialysis treatment be 5% silk fibroin water solution, being positioned over stirring at low speed in PEG solution, to be concentrated into mass fraction be 22% silk fibroin water solution; Be 25 DEG C of left and right in temperature, within the resting period of 5d, carry out wet spinning, spun filament cellulose fiber is immersed in dehydrated alcohol, sealing, is placed in 24 h under room temperature, carries out drawing-off in the deionized water more than 60 DEG C, by one times of as-spun fibre drawing-off, heat setting 30 min at 120 DEG C.Spun filament cellulose fiber is positioned over to EDC/NHS(wt/wt=2/1) join by a certain percentage ethanol/water (V
c2H5OH/ V
h2O=90/10) mass fraction being mixed with in is 24 h in 4.5% crosslinked fluid, and drawing-off in the deionized water more than 60 DEG C, is placed on heat setting 30 min at 120 DEG C, obtains the Regenerated Bombyx mori Silk Fibroin long filament of EDC/NHS after crosslinked;
(3) by natural mulberry silk through coming unstuck, to form mass fraction after dissolving, dialysis treatment be 5% silk fibroin water solution, aqueous solution is poured on vinyon flat board, at ambient temperature the dry fibroin membrane that obtains; Regenerated Silk Fibroin Film is dissolved in HFIP, and in 25 DEG C of water-baths of constant temperature, vibrating, within one week, to make mass fraction be 10% spinning liquid.Spinning liquid is poured in syringe and carried out electrostatic spinning, No. 9 syringe needles, spinning flow 2mL/h, adjusts voltage and curing distance, makes field intensity reach 1kV/cm, makes regeneration mulberry silk nano-fibre yams;
(4) inweave natural mulberry silk prepared by step (1) using prepared Regenerated Bombyx mori Silk Fibroin long filament in fortifying fibre carbon fiber and step (2) (3) and regeneration mulberry silk nano-fibre yams as stuffer weft in compiling timbering material, the common artificial ligament repair materials that builds, adopt again serger that ligament material both sides are sewed up and formed column structure, the overall length 29cm of artificial ligament repair materials, parallel fiber zone length 3cm, two ends woollen yarn knitting zone length is respectively 10cm, consolidation zone length of field is 3cm, and diameter is 10mm; The ultimate strength of this artificial ligament repair materials is 1250N.
Accompanying drawing 1 is above-mentioned through compiling the structure chart of support; Accompanying drawing 2 is the structural representation of above-mentioned artificial ligament repair materials, A: parallel fiber district; B: knitting zone; C: consolidation zone; Accompanying drawing 3 is the longitudinal scanning Electronic Speculum figure of above-mentioned Regenerated Bombyx mori Silk Fibroin filament fiber; Accompanying drawing 4 is the scanning electron microscope (SEM) photograph of above-mentioned regeneration mulberry silk nano-fibre yams.
Referring to accompanying drawing 1, Fig. 2, Fig. 3 and Fig. 4, be 29cm by the prepared artificial ligament repair materials of the present embodiment total length, parallel fiber zone length 3cm, two ends woollen yarn knitting zone length is respectively 10cm, and consolidation zone length of field is 3cm, and diameter is 10mm; The average diameter of Regenerated Bombyx mori Silk Fibroin long filament is about 12 μ m; The diameter of regeneration mulberry silk nano-fibre yams is 280nm.
Embodiment bis-:
(1) braiding through compiling timbering material is with embodiment mono-;
(2) by natural mulberry silk through coming unstuck, to form mass fraction after dissolving, dialysis treatment be 5% silk fibroin water solution, being positioned over stirring at low speed in PEG solution, to be concentrated into mass fraction be 27% silk fibroin water solution; Be 25 DEG C of left and right in temperature, within the resting period of 3d, carry out wet spinning, spun filament cellulose fiber is immersed in dehydrated alcohol, sealing, is placed in 24 h under room temperature, carries out drawing-off in the deionized water more than 60 DEG C, by one times of as-spun fibre drawing-off, heat setting 30 min at 120 DEG C.Spun filament cellulose fiber is positioned over to EDC/NHS(wt/wt=2/1) join by a certain percentage ethanol/water (V
c2H5OH/ V
h2O=90/10) mass fraction being mixed with in is 24 h in 4.5% crosslinked fluid, and drawing-off in the deionized water more than 60 DEG C, is placed on heat setting 30 min at 120 DEG C, obtains the Regenerated Bombyx mori Silk Fibroin long filament of EDC/NHS after crosslinked;
(3) by natural mulberry silk through coming unstuck, to form mass fraction after dissolving, dialysis treatment be 7% silk fibroin water solution, aqueous solution is poured on vinyon flat board, at room temperature the dry fibroin membrane that obtains; Regenerated Silk Fibroin Film is dissolved in formic acid, and in 25 DEG C of water-baths of constant temperature, vibrating, within one week, to make mass fraction be 11% spinning liquid.Spinning liquid is poured in syringe and carried out electrostatic spinning, No. 9 syringe needles, spinning flow 2mL/h, adjusts voltage and curing distance, makes field intensity reach 1kV/cm, makes regeneration mulberry silk nano-fiber material;
(4) inweave natural mulberry silk prepared by step (1) using prepared regenerated silk fibroin long filament in fortifying fibre superhigh molecular weight polyethylene fibers and step (2) (3) and regeneration mulberry silk nano-fibre yams as stuffer weft in compiling timbering material, jointly build artificial ligament repair materials, further improve the mechanical property of material.Adopt again serger that ligament material both sides are sewed up and formed column structure, the overall length 27.5cm of artificial ligament repair materials, parallel fiber zone length 1.5cm, two ends woollen yarn knitting zone length is respectively 10cm, and consolidation zone length of field is 3cm, and diameter is 6mm.
Claims (9)
1. a fibroin base artificial ligament repair materials, comprises support and stuffer weft, it is characterized in that: described support adopts twin shaft to form through compiling to inlaid thread by natural mulberry silk; Stuffer weft is Regenerated Bombyx mori Silk Fibroin long filament and regeneration mulberry silk nano-fibre yams, and described stuffer weft inweaves in the warp knitting fabrics structure of support and reels and forms artificial ligament material; The both sides of described artificial ligament material are sewed up and are obtained described fibroin base artificial ligament repair materials.
2. artificial ligament repair materials according to claim 1, is characterized in that: in described artificial ligament repair materials, natural mulberry silk consumption accounts for 30~70% of artificial ligament repair materials total amount; Regenerated Bombyx mori Silk Fibroin long filament consumption accounts for 10~30% of artificial ligament repair materials total amount; Regeneration mulberry silk nano-fibre yams consumption accounts for 20~40% of artificial ligament repair materials total amount.
3. artificial ligament repair materials according to claim 1, is characterized in that: described artificial ligament repair materials length is 27.5~41cm; The diameter of described Regenerated Bombyx mori Silk Fibroin long filament is 15~80 μ m; The diameter of described regeneration mulberry silk nano-fibre yams is 50~2000nm.
4. artificial ligament repair materials according to claim 1, is characterized in that: described artificial ligament repair materials also comprises reinforcing fibre; Described reinforcing fibre is carbon fiber, superhigh molecular weight polyethylene fibers or polyester fiber.
5. artificial ligament repair materials according to claim 4, is characterized in that: in described artificial ligament repair materials, reinforcing fibre consumption accounts for 10~90% of the total consumption of stuffer weft.
6. a method of preparing artificial ligament repair materials claimed in claim 1, is characterized in that, comprises the following steps:
(1) adopt natural mulberry silk braiding twin shaft to inlaid thread through compiling timbering material, after obtain Bombyx mori Silk Fibroin through coming unstuck through compiling timbering material;
(2) natural mulberry silk is prepared regenerated silk fibroin solution after coming unstuck, dissolve, dialysing, and adopts dry method or wet spinning to prepare Regenerated Bombyx mori Silk Fibroin long filament;
(3) natural mulberry silk is prepared regenerated silk fibroin film after coming unstuck, dissolve, dialyse, being dried, and uses hexafluoroisopropanol or formic acid to dissolve fibroin protein film, then adopts method of electrostatic spinning preparation regeneration mulberry silk nano-fibre yams;
(4) taking fibroin through compile timbering material as base material, Regenerated Bombyx mori Silk Fibroin long filament and regeneration mulberry silk nano-fibre yams inweave in warp knitting fabrics structure as stuffer weft, reel and form artificial ligament material, finally ligament material both sides are sewed up and obtained described fibroin base artificial ligament repair materials.
7. preparation method according to claim 6, is characterized in that: in described step (4), stuffer weft also comprises carbon fiber, superhigh molecular weight polyethylene fibers or polyester fiber.
8. preparation method according to claim 6, is characterized in that: in described step (2), the regeneration Bombyx mori Silk Fibroin long filament of preparation also passes through 1-(3-dimethylaminopropyl)-3-ethyl carbodiimides and N-maloyl imines crosslinking Treatment.
9. preparation method according to claim 6, is characterized in that: in mass ratio, and natural mulberry silk: Regenerated Bombyx mori Silk Fibroin long filament: regeneration mulberry silk nano-fibre yams is (30~70): (10~30): (20~40).
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CN106929997A (en) * | 2015-12-31 | 2017-07-07 | 苏州微创脊柱创伤医疗科技有限公司 | A kind of artificial soft tissue's braid and preparation method thereof |
CN106975104A (en) * | 2016-01-19 | 2017-07-25 | 苏州微创脊柱创伤医疗科技有限公司 | A kind of artificial ligament braid and its production and use |
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CN106048765B (en) * | 2016-07-14 | 2018-05-08 | 苏州经贸职业技术学院 | A kind of preparation method of artificial ligament material |
CN106730016A (en) * | 2016-12-09 | 2017-05-31 | 苏州纳贝通环境科技有限公司 | A kind of activity modifying modifying artificial ligament and its preparation technology |
CN110331486A (en) * | 2019-07-25 | 2019-10-15 | 东华大学 | A kind of multilayered structure nano-fibre yams knitting tendon scaffold and its preparation and application |
CN110947032A (en) * | 2019-12-16 | 2020-04-03 | 浙江大学 | Silk/PET mixed-woven bracket and preparation method and application thereof |
CN113663128B (en) * | 2021-09-29 | 2022-05-10 | 东华大学 | High-strength composite coating artificial ligament and preparation method and application thereof |
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