CN106075575A - A kind of composite for knee osteoarthritis repair of cartilage and preparation method thereof - Google Patents
A kind of composite for knee osteoarthritis repair of cartilage and preparation method thereof Download PDFInfo
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- CN106075575A CN106075575A CN201610490909.9A CN201610490909A CN106075575A CN 106075575 A CN106075575 A CN 106075575A CN 201610490909 A CN201610490909 A CN 201610490909A CN 106075575 A CN106075575 A CN 106075575A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/26—Mixtures of macromolecular compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/23—Carbohydrates
- A61L2300/232—Monosaccharides, disaccharides, polysaccharides, lipopolysaccharides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
- A61L2300/602—Type of release, e.g. controlled, sustained, slow
- A61L2300/604—Biodegradation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/06—Materials or treatment for tissue regeneration for cartilage reconstruction, e.g. meniscus
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Abstract
The present invention discloses a kind of composite for knee osteoarthritis repair of cartilage, is made up of fibroin albumen, collagen and hyaluronic acid, wherein fibroin albumen: collagen: the mass ratio of hyaluronic acid is 30:60 ~ 70:2.5 ~ 10.Invention additionally discloses its preparation method, the method process is simple, and raw material is easy to get inexpensively.Gained composite can improve collagen mechanical performance deficiency, too fast shortcoming of degrading, and has the lubrication of hyaluronic acid concurrently simultaneously, promotes the propagation of cell, breaks up and improve the important physiological process in joint.The composite that the present invention obtains has the feature of good biocompatibility, degradability and non-immunogenicity, can effectively facilitate reconstruction and the regeneration of cartilage, be a kind of be preferably applicable in, the medical material of knee osteoarthritis cartilage defect repair in early days.
Description
Technical field
The invention belongs to bio-medical field, be specifically related to a kind of composite for knee osteoarthritis repair of cartilage
And preparation method thereof.
Background technology
Knee osteoarthritis (KOA) is a kind of due to the appearance deformation of patient's knee cartilage, hyperosteogeny and necrosis etc.
The joint disease that Lymph nodes is characterized, even disappears as major pathologic features with articular cartilage degenerative change, at present
The clinical treatment to KOA is broadly divided into operative treatment and non-operative treatment two kinds.Operative treatment main method has: autologous or of the same race
Allograph bone cartilage transplantation, tissue transplantation, joint debridement art, Drilling and knee joint peripheral osteotomy, knee fusion, people
Work total knee arthroplasty (TKA) etc..Although TKA treatment KOA is effective, but is mainly used in patients with terminal, there is also cost simultaneously
The problems such as high, big, the prosthesis wear of operation wound.For the patient that young KOA articular cartilage abrasion in early days is lighter, TKA is not
Optimum selection.In other operation method, compared with Drilling, arthroplasty art, microfrature etc., clinical confirmation is autologous inlays
Formula bone cartilage transplantation has long-term substantially clinical efficacy, but its shortcoming is to sacrifice other cartilages as cost.Therefore, development one can
The material effectively repairing KOA cartilage defect is imperative.
In cartilage tissue engineered natural material, fibroin albumen has good biocompatibility, can maintain soft
Osteocyte normal growth state and performance, for cartilage and bone remoulding and regeneration., distribution the abundantest as animal in-vivo content is
Protein widely, collagen has triple-helix structure, low antigenicity, avirulence, good biocompatibility and the biology of uniqueness
The features such as degradability, have been widely used in tissue engineering bracket material, medicine control slow-released system, gene delivery vector, U.S.
Hold the bio-medical fields such as surgery.
Hyaluronic acid is the bio-molecules being widely present in skin and other tissue, be composition human body cell interstitial,
The main component of the connective tissues such as knuckle synovia, it is filled among cell and collagen fiber space and covers in some epidermis group
Knit, be distributed widely in the soft connective tissue of human body, almost exist with pure form in articular cavity.It can promote cell
Propagation, differentiation, scavenging activated oxygen, promote wound healing, regulation osmotic pressure, lubricate, promote cytothesis and improve joint
Important function.In clinical KOA non-operative treatment, supplemented with exogenous hyaluronic acid is one of common method, has raising synovial fluid
Middle hyaluronic acid contents, the physiological function of synovial fluid under pathologic condition of improving, arthralgia of alleviating, protection cartilage and synovial membrane are from broken
The effects such as evil idea.
The present invention combines the advantage of natural collagen, fibroin albumen and hyaluronic acid, and preparation one can be used for kneecap joint
The composite of scorching repair of cartilage.The support that it can adhere to as interior originality bone marrow derived stem cells, breed and break up, leads to simultaneously
While crossing the hyaluronic acid enhancing repairing effect that material degradation produces, reach to alleviate and the purpose for the treatment of KOA.
Summary of the invention
It is an object of the invention to provide a kind of composite for knee osteoarthritis repair of cartilage and preparation method thereof.
Gained composite, has the feature of good biocompatibility, degradability and non-immunogenicity, can effectively facilitate cartilage
Rebuild and regeneration, be a kind of be preferably applicable in, the medical material of knee osteoarthritis cartilage defect repair in early days.
For achieving the above object, the present invention is by the following technical solutions:
A kind of composite for knee osteoarthritis repair of cartilage is with silk fibroin protein solution, collagen, hyaluronic acid as raw material
Prepare, wherein fibroin albumen: collagen: the mass ratio of hyaluronic acid is 30:60 ~ 70:2.5 ~ 10.
Described collagen is for coming from fish skin, Corii Sus domestica, Corii Bovis seu Bubali or beef tendon.
The described preparation method for the composite of knee osteoarthritis repair of cartilage comprises the steps:
(1) silk fibroin protein solution is prepared；
(2) respectively collagen, hyaluronic acid are dissolved in concentration be 1 mol/L malonic acid is prepared as certain density collagen solution and
Hyaluronic acid solution；Described collagen is 0.4:100 with the mass ratio of malonic acid；Described hyaluronic acid with the mass ratio of malonic acid is
0.5:100；
(3) step (1) gained silk fibroin protein solution is mixed with collagen solution and the hyaluronic acid solution of step (2) gained, stir
Mix uniformly, add cross-linking agent after lyophilization and carry out cross-linking 10h；Described cross-linking agent is by 1-ethyl-3(3-dimethyl aminopropyl) carbon
The ethanol solution changing diimine (EDC), N-maloyl (NHS) and 90% prepares, and wherein the mass ratio of EDC/NHS is 5:2.
(4) repeatedly removing unnecessary cross-linking agent with distilled water or deionized water rinsing after crosslinking, secondary freeze drying is the most handy
Composite in knee osteoarthritis repair of cartilage.
Wherein the preparation method of silk fibroin protein solution is as follows:
1) Bombyx bombycis shredded is put into Na2CO3In solution, place 30min in a water bath, remove colloid on Bombyx bombycis, repeatable operation 3
Secondary post-drying, obtains fibroin；Wherein Na2CO3The mass fraction of solution is 0.5%, and bath temperature is 98 ± 2 DEG C.
2) fibroin described in step 1) is weighed, at 78 DEG C, at CaCl2/H2O/C2H5OH mixed solution dissolves, gauze
Filter, obtain coarse filtration liquid；Wherein CaCl2/H2O/C2H5CaCl in OH mixed solution2/H2O/C2H5Mol ratio 1:8:2 of OH；Fibroin
With CaCl2/H2O/C2H5The mass ratio of OH mixed solution is 1:25.
3) by step 2) gained coarse filtration liquid pours 3 d that dialyse in bag filter into, removes calcium chloride and ethanol；Refilter, obtain
Silk fibroin protein solution；The molecular cut off of bag filter used is 8000-14000.
Composite for knee osteoarthritis repair of cartilage prepared by the present invention is tridimensional network, has good
Hygroscopicity, preparation process is simple, cheap and easy to get.Resulting materials can improve collagen mechanical performance deficiency, too fast shortcoming of degrading,
Hyaluronic acid can play lubrication simultaneously, promotes the propagation of cell, breaks up and improve the important physiological process in joint.This is combined
Material has the feature of good biocompatibility, degradability and non-immunogenicity, can effectively facilitate the reconstruction and again of cartilage
Raw, be a kind of be preferably applicable in, the medical material of knee osteoarthritis cartilage defect repair in early days.
Accompanying drawing explanation
Fig. 1 is the infrared spectrogram of the composite for knee osteoarthritis repair of cartilage, a: before crosslinking, b: crosslinking
After.
Fig. 2 is the scanning electron microscope (SEM) photograph of the composite for knee osteoarthritis repair of cartilage, represents enforcement respectively from A-E
First group to the 5th group gained composite in example.
Detailed description of the invention
Preparation for the composite of knee osteoarthritis repair of cartilage
Embodiment 1
1) Bombyx bombycis shredded is put into 0.5% Na2CO3Solution, places 30 min in 98 ± 2 DEG C of water-baths, remove Bombyx bombycis gluing
Matter, repeatable operation 3 times.After putting into 60 DEG C of oven for drying, obtain fibroin silk.Weigh fibroin silk after above-mentioned degumming, 78 DEG C of ternarys
Solution C aCl2/H2O/C2H5OH solution (mol ratio 1:8:2) dissolves, bath raio 1:25, filtered through gauze.Solution is poured into and retains point
Son measures 3 d that dialyse in the bag filter for 8000-14000, removes calcium chloride and ethanol；Filter, obtain silk fibroin protein solution.
2) weigh 1.3 g beef tendon collagens being dissolved in 323.7 g concentration is in 1 mol/L malonic acid solution, is placed on agitator
Upper stirring and dissolving, prepares the collagen solution that mass fraction is 0.4%.
3) weigh 0.089 g hyaluronic acid being dissolved in 19.9 g concentration is in 1 mol/L malonic acid solution, is placed on agitator
Upper stirring and dissolving, prepares the collagen solution that mass fraction is 0.5%.
4) silk fibroin protein solution is mixed with beef tendon collagen solution, be placed on magnetic stirring apparatus and stir, more slowly add
Enter hyaluronic acid solution, make the fibroin albumen/collagen/hyaluronic acid solution of mix homogeneously, its fibroin albumen (SF)/collagen
(Col) mass ratio of/hyaluronic acid (HA) is 30:67.5:2.5.Lyophilization；Utilize the compound of lyophilizing of cross-linking agents
Material, removes unnecessary cross-linking agent with deionized water rinsing repeatedly, and its cross-linking agent is by 1-ethyl-3(3-dimethyl aminopropyl) carbonization
Diimine, N-maloyl and 90% ethanol solution prepare, and wherein the mass ratio of EDC/NHS is 5:2.Then secondary freezing is done
The dry composite i.e. obtained for knee osteoarthritis repair of cartilage, is designated as first group, composite red before and after its crosslinking
Outer figure such as Fig. 1.As shown in Figure 1: after crosslinking, composite is at 1234 cm-1With 1450 cm-1Place peak absorbance ratio close to
1, show that in material, collagen protein still keeps complete tertiary structure, biologic activity preserves good；Compare with (Fig. 1 a) before crosslinking,
Composite 1630 cm after crosslinking-1(amide Ⅰ C=O stretching vibration peak) and 1234 cm-1The position of (Amide Ⅲ band) place absworption peak
Put and there occurs that change, material there occurs the anti-degradation property of effectively crosslinking, beneficially composite and mechanical performance with intensity
Improve.
Embodiment 2
Changing the mass ratio of fibroin albumen/collagen/hyaluronic acid in step 4) is 30:65:5, other steps and embodiment 1 phase
With, prepare composite C.
Embodiment 3
Changing the mass ratio of fibroin albumen/collagen/hyaluronic acid in step 4) is 30:62.5:7.5, other steps and embodiment 1
Identical, prepare composite D.
Embodiment 4
Changing the mass ratio of fibroin albumen/collagen/hyaluronic acid in step 4) is 30:60:10, other steps and embodiment 1 phase
With, prepare composite E.
Comparative example:
Changing the mass ratio of fibroin albumen/collagen/hyaluronic acid in step 4) is 30:70:0, other steps and embodiment 1 phase
With, prepare composite A.
Above-mentioned gained is for the water absorption rate test of the composite of knee osteoarthritis repair of cartilage:
Weigh 0.01 g composite for knee osteoarthritis repair of cartilage respectively (for W1), it is put in culture dish every hole and adds
Entering 1800 mLPBS buffer solution, place 24h at 37 DEG C, its weight (is called W respectively2), water absorption rate (%)=(W2-W1)/W1
× 100%, each experiment is parallel to be done 3 times, takes its meansigma methods, and result is as follows:
The water absorption rate of table 1. above-described embodiment gained composite.
As can be known from the above table, above-described embodiment and comparative example gained for knee osteoarthritis repair of cartilage composite with
The rising of hyaluronic acid contents, water absorption rate reduces, but overall difference is little.
Above-described embodiment gained is that tridimensional network is (such as Fig. 2 for the composite of knee osteoarthritis repair of cartilage
Shown in), there is good hygroscopicity, the feature of biocompatibility, degradability and non-immunogenicity.
The foregoing is only presently preferred embodiments of the present invention, all impartial changes done according to scope of the present invention patent with
Modify, all should belong to the covering scope of the present invention.
Claims (7)
1. the composite for knee osteoarthritis repair of cartilage, it is characterised in that: with fibroin albumen, collagen, transparent
Matter acid is that raw material prepares, described fibroin albumen: collagen: the mass ratio of hyaluronic acid is 30:60 ~ 70:2.5 ~ 10.
The most according to claim 1 for the composite of knee osteoarthritis repair of cartilage, it is characterised in that: described collagen
For coming from fish skin, Corii Sus domestica, Corii Bovis seu Bubali or beef tendon.
3. a preparation method for the composite for knee osteoarthritis repair of cartilage as claimed in claim 1, it is special
Levy and be: comprise the steps:
(1) silk fibroin protein solution is prepared；
(2) respectively collagen, hyaluronic acid are dissolved in malonic acid and are prepared as certain density collagen solution and hyaluronic acid solution；
(3) step (1) gained silk fibroin protein solution is mixed with collagen solution and the hyaluronic acid solution of step (2) gained, stir
Mix uniformly, add cross-linking agent after lyophilization and cross-link；
(4) repeatedly rinsing with water after crosslinking, secondary freeze drying i.e. obtains the composite for knee osteoarthritis repair of cartilage.
Composite material and preparation method thereof for knee osteoarthritis repair of cartilage the most according to claim 3, its feature exists
In: collagen described in step (2) is 0.4:100 with the mass ratio of malonic acid；Described hyaluronic acid with the mass ratio of malonic acid is
0.5:100。
Composite material and preparation method thereof for knee osteoarthritis repair of cartilage the most according to claim 3, its feature exists
In: cross-linking agent described in step (3) is by 1-ethyl-3(3-dimethyl aminopropyl) carbodiimides, N-maloyl be dissolved in
Volumetric concentration be 90% ethanol solution in prepare, wherein 1-ethyl-3(3-dimethyl aminopropyl) carbodiimides and N-hydroxyl
The mol ratio of succinyl is 5:2.
The preparation method of the composite for knee osteoarthritis repair of cartilage the most according to claim 3, its feature
It is: the preparation method of described silk fibroin protein solution is as follows:
(1) Bombyx bombycis shredded being put into mass fraction is 0.5%Na2CO3Solution, places 30min in 98 ± 2 DEG C of water-baths, removes
Colloid on Bombyx bombycis, 3 post-dryings of repeatable operation, obtain fibroin；
(2) fibroin described in step (1) is weighed, at 78 DEG C, at CaCl2/H2O/C2H5OH mixed solution dissolves, gauze mistake
Filter, obtains coarse filtration liquid；Described CaCl2/H2O/C2H5The middle CaCl of OH mixed solution2/H2O/C2H5The mol ratio of OH is 1:8:2；
(3) step (2) gained coarse filtration liquid is poured 3 d that dialyse in bag filter into, remove calcium chloride and ethanol；Refilter, obtain silk
Fibroin solution；The molecular cut off of described bag filter is 8000-14000.
Composite material and preparation method thereof for knee osteoarthritis repair of cartilage the most according to claim 4, its feature exists
In: described fibroin and CaCl2/H2O/C2H5The mass ratio of OH mixed solution is 1:25.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108310461A (en) * | 2018-02-09 | 2018-07-24 | 武汉纺织大学 | Porous fibroin spinal cord rack with targeted release function and preparation method thereof prepares mold and application |
| CN111701072A (en) * | 2020-06-01 | 2020-09-25 | 天津大学 | Joint injection preparation based on collagen-binding polypeptide modified hyaluronic acid and preparation method and application thereof |
| CN111701073A (en) * | 2020-06-01 | 2020-09-25 | 天津大学 | Joint injection preparation based on collagen mimetic peptide modified hyaluronic acid and preparation method and application thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108310461A (en) * | 2018-02-09 | 2018-07-24 | 武汉纺织大学 | Porous fibroin spinal cord rack with targeted release function and preparation method thereof prepares mold and application |
| CN111701072A (en) * | 2020-06-01 | 2020-09-25 | 天津大学 | Joint injection preparation based on collagen-binding polypeptide modified hyaluronic acid and preparation method and application thereof |
| CN111701073A (en) * | 2020-06-01 | 2020-09-25 | 天津大学 | Joint injection preparation based on collagen mimetic peptide modified hyaluronic acid and preparation method and application thereof |
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Application publication date: 20161109 |