CN109865159A - A kind of silk network and its preparation method and application - Google Patents
A kind of silk network and its preparation method and application Download PDFInfo
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- CN109865159A CN109865159A CN201811560872.8A CN201811560872A CN109865159A CN 109865159 A CN109865159 A CN 109865159A CN 201811560872 A CN201811560872 A CN 201811560872A CN 109865159 A CN109865159 A CN 109865159A
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Abstract
The invention discloses a kind of silk networks and its preparation method and application, it is used for organizational project, the silk network mainly woven by silk after remove sericin or remove sericin after silk weave, aperture be 0.1~2mm, pore wall thickness be 0.1~1mm.Silk network of the present invention improves comfort level and biocompatibility after implanting, guide tissue regeneration while realizing degradation, while material is easily obtained on the basis of existing tissue engineering bracket, and product is easy to make, sterilizes and save.
Description
Technical field
The present invention relates to technical field of biological material, more precisely a kind of silk network and preparation method thereof.
Background technique
Tissue engineering bracket material can be divided into natural biologic material and artificial synthesized high molecular material two major classes.Natural biological
The advantages of material be be conducive to it includes natural structure cell attachment or keep differentiation function；And the macromolecule material synthesized
Although material can more accurately control molecular weight, degradation time and hydrophilic and hydrophobic etc., the interaction of itself and cell is not enough managed
Think.At present application more absorbable biomaterial be cell free extracellular matrix, products material have pig collagen tissue,
Abortive calfskin skin and people's Allodermis Matrix matrix etc..This kind of material structure compatibility is good, can grow into for autologous tissue and provide sky
Between.However, it is using still by limited scale and high-cost restriction, it is often more important that, nonadjustable physical characteristic is
The defect of such material.
Currently, surgery comes into the new stage of " regenerative medicine ", clinical medicine with the development of organizational project subject
Developed to from " blocking lesion that can only be simple protects remaining function " " not only blocking lesion, but also can regenerating tissues organ
Structure function ".
In numerous natural biologic materials, from the fibroin albumen of natural mulberry silk, due to its easy processing, immunogenicity
It is low, the advantages that good biocompatibility, mechanical strength is high, and degradation rate is slow, and catabolite is amino acid, in recent years, tissue again
Raw field is widely studied.
Compared with other natural materials, silk has excellent tensile strength and elongation at break, with nylon, poly- to benzene two
The polymer fibers such as formic acid glycol ester are close, can be used for making to the higher tissue engineering bracket of mechanics demand.It is strong in mechanics
While spending high, the fibroin albumen from mulberry silk also has good biocompatibility.There are crystal regions in fibroin albumen
With noncrystalline domain mixed structure, basic amino acid existing for noncrystalline domain, surface has positive charge, can be thin with mammal
The negative electrical charge of cellular surface combines, to promote the adherency of cell.In terms of skin tissue engineering, passed with chitosan and collagen etc.
System is compared using more natural macromolecular material, and fibroin albumen film forming is more preferable, can also be maintained for up to 98% or more it is saturating
Lightness, flexibility and shape keeping property under high humidity state can be also more prominent, are conducive to manufacture some in clinical or experiment
In require the high grade of transparency, to observe the biomedical products for extracting biological information or internal high humidity environment and using.Tendon,
In the regenerated research of ligament, fibroin fiber can promote differentiation of the mescenchymal stem cell to Tenocyte cell, and in 3 weeks not
See that silk fibroin bracket has apparent mechanical strength to change, it was demonstrated that its tendon, ligament injury reparation in there is huge answer
With value.
Summary of the invention
The object of the present invention is to provide a kind of silk networks and its preparation method and application, can solve existing skill
Disadvantages mentioned above in art.
The invention adopts the following technical scheme:
A kind of silk network, is used for organizational project, and the silk network is gone after mainly being woven by silk
Except the silk after sericin or removal sericin weaves, aperture is 0.1~2mm, and pore wall thickness is 0.1~1mm.
The silk network is that the special shape for meeting tissue repair requirement is made.
The shape of the silk network is sheet, tubular, cluster integrated conical form, braid shape.
The mesh form is round or polygon.
A kind of preparation method of silk network, comprising the following steps:
The method of the removal sericin is that the silk mesh sheet of silk or sericin to be removed is immersed in Na2CO3
In aqueous solution, heating water bath to temperature reaches 85~100 DEG C, after 20~40min of continuous heating, removes Na2CO3Aqueous solution is set
It is to be cleaned in 4~40 DEG C of water in temperature；After the above degumming and cleaning process repeat 2~5 times, silk after degumming is placed in temperature
It is dried after flattening in 40~90 DEG C for cleaning in 4~40 DEG C of water to cleaning solution 3~10 times.
Silk and Na2CO3The mass volume ratio of solution is 1:(50~200) (kg/L).
Na2CO3The mass concentration of aqueous solution is 0.1%~0.3%.
In cleaning step with the mass volume ratio of water be 1:(50~200) (kg/L).
In degumming step, it is 1:(10~30 that silk after degumming, which is placed in silk quality volume ratio) (kg/L).
A kind of application of silk network is mainly used for repairing impaired abdominal wall soft tissue, rotator cuff injury, ligament damage
The toughness tissues such as wound, tendon and meniscus.
The invention has the advantages that
Silk network of the present invention improves the comfort level after implanting on the basis of existing tissue engineering bracket
With biocompatibility, guide tissue regeneration while realizing degradation, while material is easily obtained, product should be easy to make, sterilize
With preservation.
Detailed description of the invention
Below with reference to embodiment and attached drawing, the present invention is described in detail, in which:
Fig. 1 is the netted supporting structure schematic diagram of silk.Wherein Fig. 1 a is general appearance figure；Fig. 1 b arrives for scanning electron microscopic observation
Part it is mesh-structured.
Fig. 2 is the SD rat animal model of peritonaeum removal.Wherein a is schematic diagram；B is operation modeling figure；C is implantation silk
Network.
Fig. 3 is different time points materials gross findings after the netted stenter to implant SD rat of silk.Scheme a, a ' is implantation
15 days；B, b ' it is implantation 30 days；C, c ' it is implantation 60 days.
Fig. 4 is HE dyeing observation defective region reparation situation after the netted stenter to implant SD rat of silk.A, a ' it is implantation 15 days；
B, b ' it is implantation 30 days；C, c ' it is implantation 60 days；A~c: × 40；A '~c ': × 100.
Fig. 5 is trichrome stain observation defective region reparation situation after the netted stenter to implant SD rat of silk.A, a ' it is implantation 15
It；B, b ' it is implantation 30 days；C, c ' it is implantation 60 days；A~c: × 40；A '~c ': × 100.
Fig. 6 is substantially to scheme after forming abdominal external hernia (a) and cutting off extra hernical sac along hernical sac neck after new zealand rabbit modeling 30 days
(b)。
Fig. 7 substantially schemes when being the netted stenter to implant new zealand rabbit of silk 15 days and 60 days.A is test group 15 days；B is control
Group 15 days；C is test group 60 days；D is control group 60 days.
When Fig. 8 is the netted stenter to implant new zealand rabbit of silk 15 days when test group (a), control group (b) and implantation 60 days
The HE coloration result of test group (c), control group (d).
Specific embodiment
The specific embodiment that present invention is further explained below:
A kind of silk network, is used for organizational project, and the silk network is gone after mainly being woven by silk
Except the silk after sericin or removal sericin weaves, aperture is 0.1~2mm, and pore wall thickness is 0.1~1mm.
The silk network is that the special shape for meeting tissue repair requirement is made.
The shape of the silk network is sheet, tubular, cluster integrated conical form, braid shape.
The mesh form is round or polygon.
A kind of preparation method of silk network, comprising the following steps:
The method of the removal sericin is that the silk mesh sheet of silk or sericin to be removed is immersed in Na2CO3
In aqueous solution, heating water bath to temperature reaches 85~100 DEG C, after 20~40min of continuous heating, removes Na2CO3Aqueous solution is set
It is to be cleaned in 4~40 DEG C of water in temperature；After the above degumming and cleaning process repeat 2~5 times, silk after degumming is placed in temperature
It is dried after flattening in 40~90 DEG C for cleaning in 4~40 DEG C of water to cleaning solution 3~10 times.
Silk and Na2CO3The mass volume ratio of solution is 1:(50~200) (kg/L).
Na2CO3The mass concentration of aqueous solution is 0.1%~0.3%.
In cleaning step with the mass volume ratio of water be 1:(50~200) (kg/L).
In degumming step, it is 1:(10~30 that silk after degumming, which is placed in silk quality volume ratio) (kg/L).
A kind of application of silk network is mainly used for repairing impaired abdominal wall soft tissue, rotator cuff injury, ligament damage
The toughness tissues such as wound, tendon and meniscus.
The invention has the following advantages that
(1) silk network of the present invention has the excellent bionical mechanical characteristic of human body soft tissue.Different from most at present
The synthetic material of high rigidity, low ductility that number product uses, the present invention is innovative to use mulberry silk for raw material, production
Bracket has the mechanical strength to match with tissue and elasticity, can provide bionical mechanics at implantation initial stage and support；Together
When, mulberry silk texture material is soft, and the foreign body sensation that implants, sense of discomfort are small, and usage comfort can be improved.
(2) silk network of the present invention there is suitable cell, tissue and blood vessel to grow into mesh-structured.Silk of the present invention
The design of special mesh diameter that network has is conducive to autologous patient cell and grows into and organize and reconstructing blood vessel.
(3) catabolite (amino acid) of silk network of the present invention can promote tissue synthesis and regeneration.Silkworm of the present invention
Mesh-like bracket is degradable in vivo, and the amino acid for generation of degrading has no toxic side effect, and can be used as the original of extracellular matrix synthesis
Material, while material degradation, the extracellular matrixs such as collagen that injury region is secreted by autogenous cell substitution restores soft tissue
Structure and function, no patch phenomenon.
Embodiment 1:
Silk braiding is reticulated into plain film, being immersed in mass concentration is 0.2%Na2CO3In aqueous solution, wherein silk with
Na2CO3The mass volume ratio of solution is 1:100 (kg/L), and heating water bath to temperature reaches 87 DEG C, after continuous heating 30min, is removed
Remove Na2CO3Aqueous solution, being placed in mass volume ratio is 1:100 (kg/L), and temperature is to clean in 20 DEG C of water；The above degumming and cleaning
After process is repeated 3 times, it is 1:20 (kg/L) that silk after degumming, which is placed in silk quality volume ratio, and temperature is in 20 DEG C of water
Cleaning is to cleaning solution 3 times, 60 DEG C of dryings after mesh sheet is flattened, and through exterior and interior packing, ethylene oxide sterilizing is to get netted to silk
Bracket, as illustrated in figs. 1A and ib.
Embodiment 2:
It is 0.2%Na that silk, which is immersed in mass concentration,2CO3In aqueous solution, wherein silk and Na2CO3The mass body of solution
For product than being 1:100 (kg/L), heating water bath to temperature reaches 87 DEG C, after continuous heating 30min, removes Na2CO3Aqueous solution is set
It is 1:100 (kg/L) in mass volume ratio, temperature is to clean in 20 DEG C of water；It, will after the above degumming and cleaning process are repeated 3 times
It is 1:20 (kg/L) that silk, which is placed in silk quality volume ratio, after degumming, is cleaned in the water that temperature is 20 DEG C to cleaning solution 3 times,
60 DEG C of dryings after silk is flattened, through machinery braiding and shaping and drying after, exterior and interior packing, ethylene oxide sterilizing to get arrive silk
Network.
Embodiment 3: the application of silk network abdominal-wall defect model in SD rat body.
SD rat is anaesthetized using 4% chloraldurate by 7.5mL/kg dosage.After anesthesia, rat abdomen shaving preserved skin,
Rat is fixed on operating table with dorsal position, aseptically, cuts skin along ventrimeson, lower layer's stomach wall fascia is complete
Full exposure cuts off subcutaneous tissue, fascia, muscle and peritonaeum in right rectus aabdominis position, makes the defect of 1.5cm × 2.5cm.It will
Silk network prepared by embodiment 1 is cut into 2.0cm × 3.0cm, is implanted into defect, and using between 4-0 silk thread holostrome
Disconnected no-station pole canopy suture, skin suture, as shown in Figure 2.
In experiment, tested through gross examination of skeletal muscle, adhesion scoring, mechanical test, histological observation etc., it was demonstrated that silk network
The peritonaeum of the can effectively replace that implants defect, no hernia recurrence, as shown in Figure 3；It analyzes from Histological results as it can be seen that silk net
The mesh of shape bracket can promote cell and grow into, until defect area is covered by cambium completely, can when experiment terminates (60 days)
Observe a large amount of blood capillary proliferations, the collagen ordered arrangement newly formed, defect area mechanical property relatively implantation initial stage increases,
Inflammatory cell quantity gradually decreases, as shown in Figure 4, Figure 5.
Embodiment 4: application of the silk network in new zealand rabbit abdominal hernia repairing model.
New zealand rabbit by 4mL/kg dosage inject 10% chloraldurate, accumulated dose half auricular vein inject half abdominal cavity into
Row anesthesia.Abdominal operation region carries out shaving, and experimental animal lies on the back on operating table after the completion, is wiped with Iodophor entire
Abdomen area.Notch from from xiphoid-process to limbs distal end 6cm~9cm cuts skin along ventrimeson, by skin and fascia superficialis with
Muscle layer separates, and cuts about 5cm long along hunter's line, and both ends transverse incision or so is respectively 1cm, forms the holostrome of 5cm × 2cm
Defect (excision includes fascia, muscle and peritonaeum).Then with 4-0 absorbable suture by notching edge reflexed, then with 4-0 silk thread
Interrupted suture skin.
After modeling 30 days, modeling success, it can be seen that the formation of obvious abdominal external hernia, as shown in Figure 6.It is close to one-stage operation to cut
Skin is cut at mouth scar, skin, hernical sac is separated, cuts off extra hernical sac, cicatricial tissue.By trial target, (prepared by embodiment 2
Silk network) or reference substance (super general sticking patch) respectively implantation experiment rabbit it is intraperitoneal, material keep it is smooth, with stomach wall weight
Folded 2cm or more, and sutured using 4-0 silk thread holostrome interruption no-station pole canopy, then conventional batch sutures.
It is shown through experimental results such as gross examination of skeletal muscle, adhesion scoring, histological observations, experimental group and control group are in implantation animal
After in vivo can good reparation abdominal-wall defect, infection or hernia recurrence does not occur in experimentation, as shown in Figure 7.From tissue
Analysis of experimental results is learned, as shown in figure 8, a large amount of cambiums and new vessels can be observed in experimental group, material is combined with tissue
It is close and gradually degrade with the extension of Implantation Time, and control group cambium is few, material with organize to be combined it is not close,
Thus it can be confirmed that the biocompatibility of silk network is better than polymer material.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (10)
1. a kind of silk network, is used for organizational project, which is characterized in that the silk network is mainly by silk
Silk after removing sericin or removal sericin after braiding weaves, and aperture is 0.1~2mm, pore wall thickness 0.1
~1mm.
2. silk network as described in claim 1, which is characterized in that the silk network is to be made to meet tissue
Repair desired special shape.
3. silk network as claimed in claim 2, which is characterized in that the shape of the silk network be sheet,
Tubular, cluster integrate conical form, braid shape.
4. silk network as claimed in claim 2, which is characterized in that the mesh form is round or polygon.
5. a kind of preparation method of silk network as described in claim 1, which comprises the following steps:
The method of the removal sericin is that the silk mesh sheet of silk or sericin to be removed is immersed in Na2CO3It is water-soluble
In liquid, heating water bath to temperature reaches 85~100 DEG C, after 20~40min of continuous heating, removes Na2CO3Aqueous solution is placed in temperature
To be cleaned in 4~40 DEG C of water；After the above degumming and cleaning process repeat 2~5 times, it is 4~40 that silk after degumming, which is placed in temperature,
DEG C water in cleaning to cleaning solution 3~10 times, be dried after flattening in 40~90 DEG C.
6. preparation method according to claim 5, which is characterized in that silk and Na2CO3The mass volume ratio of solution is 1:
(50~200) (kg/L).
7. preparation method according to claim 5, which is characterized in that Na2CO3The mass concentration of aqueous solution be 0.1%~
0.3%.
8. preparation method according to claim 5, which is characterized in that in cleaning step with the mass volume ratio of water be 1:
(50~200) (kg/L).
9. preparation method according to claim 5, which is characterized in that in degumming step, silk after degumming is placed in and silkworm
Yarn quality volume ratio is 1:(10~30) (kg/L).
10. a kind of application of the silk network as described in Claims 1-4 any one, which is characterized in that be mainly used for
Repair impaired abdominal wall soft tissue, rotator cuff injury, the toughness tissue such as ligament injury, tendon and meniscus.
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Cited By (2)
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| CN110947032A (en) * | 2019-12-16 | 2020-04-03 | 浙江大学 | Silk/PET mixed-woven bracket and preparation method and application thereof |
| CN114555108A (en) * | 2019-09-30 | 2022-05-27 | 丝芭博株式会社 | Muscle tissue regeneration agent |
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