CN110404109A - Soft tissue filler and preparation method thereof - Google Patents
Soft tissue filler and preparation method thereof Download PDFInfo
- Publication number
- CN110404109A CN110404109A CN201910754249.4A CN201910754249A CN110404109A CN 110404109 A CN110404109 A CN 110404109A CN 201910754249 A CN201910754249 A CN 201910754249A CN 110404109 A CN110404109 A CN 110404109A
- Authority
- CN
- China
- Prior art keywords
- collagen
- weight
- composite microballoon
- collagen composite
- soft tissue
- 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.)
- Pending
Links
Classifications
-
- 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
- A61L27/20—Polysaccharides
-
- 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
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
-
- 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
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/24—Collagen
-
- 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
-
- 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
-
- 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
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/06—Flowable or injectable implant compositions
-
- 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
- A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
Abstract
The present invention relates to a kind of soft tissue filler and preparation method thereof, which includes collagen composite microballoon and dispersion liquid, and the collagen composite microballoon is scattered in the dispersion liquid, and the partial size of the collagen composite microballoon is 20 μm~200 μm.The soft tissue filler has good excellent flowability, and convenient for injection and moulding, the retention time is longer in body, is a kind of safe and efficient and good biocompatibility biomaterial.
Description
Technical field
The present invention relates to technical field of biological material, more particularly to a kind of soft tissue filler and preparation method thereof.
Background technique
The thickness of dermal tissue and the number of its fibr tissue and matrix are in close relations, and with the compactness of skin, full
Degree, relaxation are closely related with wrinkling.Under body naturally-aged and extraneous various stimulations, collagen in skin corium,
The speed that elastin laminin is lost is accelerated, and supplies not as good as consume, shows as that corium is thinning, and subcutaneous fat and fat pad wither in histology
Contracting, aesthetic obviousization for showing as bone mark, blood vessel and wrinkle.
Ideal filler is lasting when introducing below skin or skin, soft, and fine needle easy to use is implanted into patient's body
Interior and need low injection extrusion force, very little or none discomfort for patient will not generate adverse side effect.Height is handed over
Connection and/or the collagen of high concentration are all highly viscous dermal fillers, and often the duration is longer in vivo, but are injected by needle
Difficulty it is larger, and usually require more small dimension needle.
Patent application AU2016201976A1 provides a kind of soft tissue filling material, and main component is hyaluronic acid.
But hyaluronic acid degradation speed in body is fast, and body cannot be stimulated to form regeneration, therefore needs periodically constantly to be injected, with
Guarantee functional form, the aesthetic requirement etc. of tissue.And the soft tissue filling material that this method provides is form of hydrogels, flowing
Property is poor, causes larger difficulty to injection.
Summary of the invention
Based on this, it is necessary to which it is long to provide one kind retention time in body, and excellent flowability, convenient for the soft tissue of injection
Filler.
A kind of soft tissue filler, including collagen composite microballoon and dispersion liquid, the collagen composite microballoon are scattered in described
In dispersion liquid, the partial size of the collagen composite microballoon is 20 μm~200 μm.
In one of the embodiments, on the basis of the soft tissue filler of 100 parts by weight, the collagen composite is micro-
The content of ball is 10~40 parts by weight, and the content of the dispersion liquid is 60~90 parts by weight.
The collagen composite microballoon includes collagen and chondroitin sulfate in one of the embodiments,.
The weight ratio of the collagen and chondroitin sulfate is 100:(5~20 in one of the embodiments).
The collagen is selected from least one of Type I collagen and III Collagen Type VI in one of the embodiments,.
The dispersion liquid is the solution containing elastin laminin in one of the embodiments,.
The concentration of the elastin laminin is 0.1 weight of weight %~5 % in one of the embodiments,.
The elastin laminin is selected from animal derived elastin laminin and genetic engineering elastin laminin in one of the embodiments,
At least one of.
The viscosity-adjusting agent that the dispersion liquid is also 1 weight of weight %~15 % containing concentration in one of the embodiments,
The anesthetic that agent and concentration are 0.1 weight of weight %~0.3 %.
In one of the embodiments, the viscosity modifier in carboxymethyl cellulose and hyaluronic acid at least one
Kind.
The anesthetic is lidocaine in one of the embodiments,.
A method of soft tissue filler is prepared, this method comprises:
Use microsyringe that the solution containing collagen is injected chondroitin sulfate solution with the speed of 2 μ of μ L/s~40 L/s
In, the solution containing collagen composite microballoon is obtained, solid material and drying is collected, obtains collagen composite microballoon；
The collagen composite microballoon is mixed with crosslinking agent, carries out cross-linking reaction, the collagen composite microballoon after being crosslinked；
Collagen microballoon after the crosslinking is mixed with dispersion liquid, obtains soft tissue filler.
The preparation step of the solution containing collagen includes: that collagen is dissolved in first is molten in one of the embodiments,
In agent, and 4h~8h is stirred under the revolving speed of 400rpm~1000rpm.
The pH of the solution containing collagen is 2~5 in one of the embodiments, and the concentration of the collagen is 0.5 weight
The weight of %~3.5 % is measured, first solvent is acetum, hydrochloric acid solution, citric acid solution, formic solutions and sulfuric acid solution
At least one of.
The chondroitin sulfate solution includes chondroitin sulfate and the second solvent, the sulphur in one of the embodiments,
The concentration of aching and limp ossein is 0.5 weight of weight %~5 %.
Second solvent includes water and organic solvent, the body of the water and organic solvent in one of the embodiments,
Product is than being 1:(0.25~1).
The organic solvent is selected from least one of ethyl alcohol and acetone in one of the embodiments,.
The weight ratio of the solution containing collagen and chondroitin sulfate solution is 1:(50 in one of the embodiments,
~300).
It, will be described micro- containing collagen composite after obtaining the solution containing collagen composite microballoon in one of the embodiments,
The solution of ball stirs 2h~6h under 100rpm~300rpm revolving speed, is then collected the step of solid material and drying again.
In one of the embodiments, before mixing the collagen composite microballoon with crosslinking agent, first the collagen is answered
It closes microballoon to be sieved, the collagen composite microballoon that collection cut size is 20 μm~200 μm.
The weight ratio of the collagen composite microballoon and crosslinking agent is 1:(100~1000 in one of the embodiments).
The crosslinking agent includes glutaraldehyde, carbodiimides, oxidized sodium alginate and Jing Ni in one of the embodiments,
At least one of put down.
In one of the embodiments, the condition of the cross-linking reaction include: temperature be 2 DEG C~15 DEG C, the time be 12h~
48h。
The dispersion liquid is the solution containing elastin laminin, the concentration of the elastin laminin in one of the embodiments,
For 0.1 weight of weight %~5 %.
Viscosity modifier of the dispersion liquid also containing 1 weight of weight %~15 % and dense in one of the embodiments,
Degree is the anesthetic of 0.1 weight of weight %~0.3 %.
This method further includes that the soft tissue filler is carried out sterile filling and electron beam in one of the embodiments,
The step of processing sterilization treatment.
In above-mentioned soft tissue filler, the partial size of collagen composite microballoon is smaller, can be dispersed in dispersion liquid, has good
Good mobility convenient for injection and moulding, and is degraded slowly, and the retention time is longer in body, and subcutaneous injection can maintain effect
Up to 12~24 months.Collagen composite microballoon can stimulate body constantly to generate new autologous collagen in degradation process, be conducive to
The reparation of body tissue and reconstruction are a kind of safe and efficient and good biocompatibility biomaterials.
Detailed description of the invention
Fig. 1 is the flow chart of the method for preparing soft tissue filler of an embodiment.
Specific embodiment
To facilitate the understanding of the present invention, below by specific embodiment to invention is more fully described.It is specific real
The mode of applying gives preferred embodiment of the invention.But the invention can be realized in many different forms, and unlimited
In embodiment described herein.On the contrary, purpose of providing these embodiments is makes understanding to the disclosure
It is more thorough and comprehensive.
Unless otherwise defined, all technical and scientific terms used herein and belong to technical field of the invention
The normally understood meaning of technical staff is identical.Term as used herein in the specification of the present invention is intended merely to description tool
The purpose of the embodiment of body, it is not intended that in the limitation present invention.
The soft tissue filler of one embodiment, including collagen composite microballoon and dispersion liquid, collagen composite microballoon are scattered in
In dispersion liquid, the partial size of collagen composite microballoon is 20 μm~200 μm.
The excellent flowability of collagen composite microballoon with above-mentioned particle size range, being dispersed in is that can infuse in dispersion liquid
The soft tissue filler penetrated.The soft tissue filler is in the intracorporal stability of machine and good biocompatibility, and the retention time is long, collagen
Body can be stimulated constantly to generate new autologous collagen in degradation process, will not filling position formed cavity, because without
Continuous injection is carried out in order to maintain institutional framework and performance.
In one embodiment, on the basis of the soft tissue filler of 100 parts by weight, the content of collagen composite microballoon is 10
~40 parts by weight, the content of dispersion liquid are 60~90 parts by weight.Soft tissue filler in above-mentioned content range has more excellent
Mobility, it is preferable in the intracorporal maintenance effect of machine.
Further, collagen composite microballoon includes collagen and chondroitin sulfate.Further, collagen and chondroitin sulfate
Weight ratio be 100:(5~20).The composition and content of collagen composite microballoon allow to Soft-tissue operation within the above range
Agent has suitable viscosity and higher elasticity, preferable in the intracorporal maintenance effect of machine.
Wherein, collagen be selected from least one of Type I collagen and III Collagen Type VI, can for animal derived collagen (such as ox,
Sheep, pig, humanized's collagen etc.) and genetic engineering collagen, the tropocollagen molecule of mentioned kind is with complete triple-helix structure, flowing
Property is excellent.The tissue-derived position of collagen can be the collagen contents such as heel string, skin, bone, small intestinal submucosa, peritonaeum, bladder
More rich tissue.
Chondroitin sulfate can improve the flexibility of tropocollagen molecule, may advantageously facilitate tropocollagen molecule and form nanometer microfibre knot
Structure further increases the biocompatibility of soft tissue filler.
Dispersion liquid is the solution containing elastin laminin.Further, the concentration of elastin laminin is 0.1 weight of weight %~5
Measure %.Elastin laminin is hydrophilic, lipophilic, has elasticity, toughness and Thermo-sensitive.Elastin laminin not only decide skin elasticity and
Flexibility, also having to skin physiology ageing process caused by the chemical factors such as light prevents aging and the regenerated work of promotion
With.When dispersion liquid contains elastin laminin, Shi Kecheng solution state is injected in room temperature, convenient for injection, and is then agglomerated after being injected in vivo
For colloidal state, there is certain viscoplasticity and cohesiveness, will not be defeated and dispersed in vivo, extend maintenance effect in vivo.In addition, bullet
Property albumen be conducive to supplement the elastin laminin being lost in vivo, and while degrading in body, can equally stimulate body to regenerate, and be formed new
Elastin laminin, not only make body tissue structure restore elasticity, be more advantageous to reparation and the reconstruction of body tissue.
Elastin laminin is selected from animal derived elastin laminin (such as ox, sheep, pig, humanized's elastin laminin) and genetic engineering bullet
At least one of property albumen.Tissue-derived position can be rich in the tissue of elastomer for ligament, blood vessel wall etc..
Further, dispersion liquid also contains viscosity modifier and anesthetic.Viscosity modifier is for adjusting Soft-tissue operation
The viscosity of agent is conducive to the mobility and biocompatibility that improve soft tissue filler.Further, viscosity modifier is selected from
At least one of carboxymethyl cellulose and hyaluronic acid, the concentration of viscosity modifier can be 1 weight of weight %~15 %.Fiber crops
Liquor-saturated dose, for implementing anaesthetic effect, the step of anesthetic can save preparatory injecting anesthetic is added in dispersion liquid, alleviates note
Penetrate bring sense of discomfort.Further, anesthetic can for lidocaine etc., the concentration of anesthetic can for 0.1 weight %~
0.3 weight %.
Above-mentioned soft tissue filler is easy to inject and moulding, and the duration is long in vivo, and degradation process is slow, subcutaneous injection
The form of injection site can be made to maintain effect up to 12~24 months.In addition, collagen composite microballoon can be used as a variety of drugs and biology
The slow-released carrier of active factors so that above-mentioned soft tissue filler treatment and in terms of application it is more extensive.
The method for preparing above-mentioned soft tissue filler of one embodiment, with reference to Fig. 1, this method comprises:
S1: collagen is dissolved in the first solvent, and 4h~8h is stirred under the revolving speed of 400rpm~1000rpm, is contained
There is the solution of collagen.
First solvent is the substance that can dissolve collagen, and in the obtained solution containing collagen, the concentration of collagen is 0.5
The weight of weight %~3.5 %.Further, the pH of the solution containing collagen be 2~5, pH within the above range when be conducive to promote
Into the dissolution of collagen, in order to obtain with the solution of above-mentioned pH, the first solvent can be acetum, hydrochloric acid solution, citric acid
At least one of solution, formic solutions and sulfuric acid solution.
S2: use microsyringe that the solution containing collagen is injected chondroitin sulfate with the speed of 2 μ of μ L/s~40 L/s
In solution, the solution containing collagen composite microballoon is obtained, solid material and drying is collected, obtains collagen composite microballoon.
By will the solution containing collagen with specific injection speed inject chondroitin sulfate solution in, tropocollagen molecule can be made
It is compound with chondroitin sulfate, formed it is more submissive and can self assembly nanometer micro-fiber structure, obtained collagen composite microballoon
In the intracorporal anti-degradation function enhancing of machine, histocompatbility is more preferable.The process is easy to operate, without adding additional adjuvant,
Free from admixture residual and security risk, meet bionics theory.
In order to stablize injection speed within the above range, generally can be used with the matched sample introduction of automatic fine syringe pump
Device, for example, by using HARVARD high pressure syringe pump PHD 22/2000Hpsi.
In one embodiment, chondroitin sulfate solution includes chondroitin sulfate and the second solvent, chondroitin sulfate it is dense
Degree can be 0.5 weight of weight %~5 %.Further, the second solvent includes water and organic solvent, the body of water and organic solvent
Product ratio can be 1:(0.25~1).Further, water is injection water, organic solvent in ethyl alcohol and acetone at least one
Kind.Above-mentioned second solvent is conducive to improve the dissolubility of chondroitin sulfate, promotes the formation of collagen composite microballoon.
When further, using the solution and chondroitin sulfate solution containing collagen in above-mentioned concentration range, control contains
The weight ratio of the solution and chondroitin sulfate solution that have collagen is 1:(50~300) so that prepared collagen composite microballoon
In collagen and chondroitin sulfate weight ratio in 100:(5~20).
In one embodiment, after injection finishes to obtain the solution containing collagen composite microballoon, this is contained into collagen and is answered
The solution for closing microballoon stirs 2h~6h under 100rpm~300rpm revolving speed, is then collected the step of solid material and drying again
Suddenly.In this way, be conducive to improve collagen composite microballoon stability, prevent from occurring in subsequent operating procedure it is defeated and dispersed, while into
One step extends maintenance effect in vivo.
It is then possible to collect solid material by the way of centrifugation, the condition of centrifugation for example can be with are as follows: revolving speed is
6000rpm~10000rpm, time are 10min~30min.In one embodiment, dry to be freeze-dried, freeze-drying
Condition includes: pre-cooling 2h~5h at -50 DEG C to -25 DEG C, is then freeze-dried for 24 hours~36h.
S3: collagen composite microballoon is sieved, the collagen composite microballoon that collection cut size is 20 μm~200 μm.
Pass through screening, it can be ensured that the particle size range of used collagen composite microballoon is at 20 μm~200 μm, such glue
Former complex microsphere has good mobility, convenient for injection and moulding.
S4: collagen composite microballoon is mixed with crosslinking agent, carries out cross-linking reaction, the collagen composite microballoon after being crosslinked.
In one embodiment, the weight ratio of collagen composite microballoon and crosslinking agent is 1:(100~1000).Dosage is above-mentioned
When in range, the collagen composite microballoon after capable of making crosslinking has the suitable degree of cross linking and viscosity.
Further, crosslinking agent includes at least one in glutaraldehyde, carbodiimides, oxidized sodium alginate and Geniposide
Kind.Crosslinking agent can be the form of the aqueous solution containing above-mentioned substance, and wherein the concentration of aqueous solution can be 0.2 weight %~5
Weight %.
The condition of cross-linking reaction includes: that temperature is 2 DEG C~15 DEG C, and the time is 12h~48h.Further, cross-linking reaction can
To carry out under stirring conditions, the revolving speed of stirring can be 200rpm~500rpm.After the completion of cross-linking reaction, when there is extra friendship
When joining agent residual, subsequent operation is carried out again after remaining crosslinking agent being separated, organic substance is avoided to remain.Isolated mode
It such as can be to be centrifuged, be filtered, washed.
S5: the collagen microballoon after crosslinking is mixed with dispersion liquid, obtains soft tissue filler.
Dispersion liquid includes elastin laminin, third solvent and optional viscosity modifier, optional anesthetic.Wherein,
Three solvents can be injection water.In order to obtain with the soft tissue filler of good fluidity and suitable viscosity, implement at one
Example in, can under agitation first disperse the collagen microballoon after crosslinking in third solvent, add elastin laminin and
Optional viscosity modifier, optional anesthetic, obtain soft tissue filler, wherein the condition stirred may include: that revolving speed is
200rpm~500rpm, time are 8h~15h.Wherein, used elastin laminin is generally the form of freeze-dried powder.
S6: soft tissue filler is subjected to sterile filling and electron beam process sterilization treatment.
As needed, the soft tissue filler of above-mentioned preparation can be perfused to different model by aseptic filling technology and is advised
In the syringe of lattice, electron beam process sterilization treatment is carried out after packaging again, to ensure the safety in utilization of product.
The present invention is further illustrated by the following examples, but is not used in the limitation present invention.
In embodiment, the partial size of collagen composite microballoon is surveyed using Malvern MASTERSIZER2000 laser particle analyzer
Examination, by collagen composite microballoon to be detected after the concentration dispersion of 20mg/100mL.
Collagen employed in embodiment is ox heel string Type I collagen, according to the method system of embodiment 1 in CN103966294B
It is standby to obtain.Elastin laminin freeze-dried powder cuts fritter the preparation method comprises the following steps: taking ox paxwax tissue for ox paxwax elastin laminin, cleans
Homogenate blends, and with grease removal such as organic solvent-acetones, then is swollen in 40 DEG C of soaking and stirrings with 1M sodium hydroxide, is extracted with sodium chloride
Afterwards, the collagenase solution that 5-10U/mL is added removes wherein remaining collagen component, then is extracted twice with sodium chloride, then dialyse
Purifying freeze-drying.Chondroitin sulfate, carboxymethyl cellulose and lidocaine are commercial products.
Embodiment 1
(1) collagen is dissolved in dilute acetic acid solution, stirs 6h under 600rpm revolving speed, until uniformly, obtaining containing collagen
Solution, pH=3.5, the concentration of collagen is 2 weight %.
(2) solution by above-mentioned containing collagen be packed into the matched microsyringe of automatic fine syringe pump (brand:
HARVARD model: PHD 22/2000Hpsi) in, microsyringe is shaken to guarantee that no air is mixed into and forms cavity.Later
Micro-sampling pin is assembled, micro-injection pump is opened and sample introduction syringe needle is submerged into 1.5 after seeing has drop to drip from syringe needle
In water/ethanol solution (water: ethyl alcohol volume 70:30) of weight % chondroitin sulfate, solution and chondroitin sulfate containing collagen
The weight ratio of solution is 1:100, and sample injector injection speed is 10 μ L/s, continues slow magnetic agitation 6h (stirring after being added dropwise
Revolving speed 300rpm), centrifugation 20min collects solid material under the conditions of revolving speed is 8000rpm, then 5h is pre-chilled at -25 DEG C, then
For 24 hours, screening obtains the collagen composite microballoon that partial size is 20 μm~200 μm, wherein the weight of collagen and chondroitin sulfate for freeze-drying
Amount is than being 100:15.3.
(3) above-mentioned collagen composite microballoon is added in the crosslinking agent glutaraldehyde solution that concentration is 2 weight %, collagen composite
The weight ratio of microballoon and crosslinking agent is 1:100, and (speed of agitator 200rpm) is stirred at 15 DEG C and carries out cross-linking reaction 48h, centrifugation
Collagen composite microballoon after collecting crosslinking, then cleaned with injection water, until cross-linking agent-free residual in cleaning solution.
(4) the collagen composite microballoon after crosslinking is distributed in injection water, slow magnetic agitation (speed of agitator 300rpm),
It is slow added into elastin laminin freeze-dried powder, is stirred until homogeneous, carboxymethyl cellulose and lidocaine are added, is allowed to sufficiently molten
Solution, the concentration of elastin laminin are 2 weight %, and the concentration of carboxymethyl cellulose is weight 5%, and the concentration of lidocaine is 0.2 weight
% is measured, obtains the soft tissue filler that collagen composite microballoon is dispersed in dispersion liquid, wherein the content of collagen composite microballoon
For 10 weight %, the content of dispersion liquid is 90 weight %.
(5) above-mentioned soft tissue filler is perfused into the syringe of different model specification using aseptically filling technology, is wrapped
It installs and carries out electron beam process sterilization treatment after finishing.
Embodiment 2
The soft tissue filler preparation process of the present embodiment is roughly the same with embodiment 1, and difference is, in step (2), contains
The weight ratio of the solution and chondroitin sulfate solution that have collagen is 1:30, and collagen and sulfuric acid are soft in obtained collagen composite microballoon
The weight ratio of ossein is 100:2.2.
Embodiment 3
The soft tissue filler preparation process of the present embodiment is roughly the same with embodiment 1, and difference is, step (4): will hand over
Collagen composite microballoon after connection is distributed in injection water, and slow magnetic agitation (speed of agitator 300rpm) is slow added into elasticity
Protein freeze-dried powder is stirred until homogeneous, and adds carboxymethyl cellulose and lidocaine, is allowed to sufficiently dissolve, elastin laminin it is dense
Degree is 6 weight %, and the concentration of carboxymethyl cellulose is weight 5%, and the concentration of lidocaine is 0.2 weight %, and it is multiple to obtain collagen
It closes microballoon and is dispersed in the soft tissue filler in dispersion liquid, wherein the content of collagen composite microballoon is 5 weight %, dispersion
The content of liquid is 95 weight %.
Test case 1
The viscous modulus and elasticity modulus of soft tissue filler prepared by testing example 1~3, are as a result listed in table 1.Viscosity
Modulus and elasticity modulus are tested using Anton Paar rotational rheometer (MCR301), and testing conditions: being warming up to 25 DEG C of room temperature for sample,
Constant temperature 30min again, 0.1Hz is frequency scanned under 0.1% strain.
Table 1
| Embodiment | Viscous modulus, Pa | Elasticity modulus, Pa |
| Embodiment 1 | 25.8 | 336.2 |
| Embodiment 2 | 95.3 | 179.5 |
| Embodiment 3 | 74.9 | 204.6 |
Seen from table 1, soft tissue filler of the invention has suitable viscosity and higher elasticity.
Test case 2
The cytotoxicity of soft tissue filler prepared by testing example 1~3, is as a result listed in table 2.
Test reference standard: GB/T16886.5-2017.
Sample preparation reference standard: GB/T16886.12-2017.
Extract medium: MEM complete medium.
Blank control: it is extracted under extraction medium, with sample similarity condition.
Negative control: high density polyethylene (HDPE) is extracted, with 0.22um filter membrane in the ratio of 0.2g/mL with material same procedure
Filtration sterilization.
Positive control: 5%DMSO (Sigma Products, article No. D4540) is prepared with extraction medium.
Cell strain: L929 l cell, the cell line recommended by ISO 10993-5.Cell culture sets 5%
37 degree of CO2 incubator cultures are spare to increased logarithmic phase.
MEM culture medium: Gibco Products, MEM powder culture medium article No. 11700；MEM liquid is without phenol red medium.
Fetal calf serum: Gibco Products, article No. 10270.
MTT solution: Sigma Products.With the MEM culture medium dissolved powders without phenol red and serum, it is made into 5mg/mL
Concentration use.
Cell toxicity test --- MTT colorimetric method: preparing density is 2 × 104A/mL~3 × 104A/mL cell suspension is standby
With；By cell suspension inoculation into 96 orifice plates, every 100 μ L of hole, culture is for 24 hours.The original fluid in 96 orifice plates is discarded, every hole is added
The leaching liquor of 100 μ L continues culture for 24 hours；Leaching liquor is sopped up, fresh MEM is added and is mixed without phenol red medium with MTT solution
Liquid continues to cultivate 4h；Liquid in hole is removed, 150 μ L DMSO shake culture 10min are added, measures absorbance A with microplate reader.It presses
Following formula calculates opposite proliferation rate (RGR).
Wherein, RGR represents opposite proliferation rate, the absorbance of A representative sample group (negative, positive), A0Represent blank control
The absorbance of group.
Table 2
| Embodiment | Cytotoxicity, rank |
| Embodiment 1 | 1 |
| Embodiment 2 | 0 |
| Embodiment 3 | 1 |
As can be seen from Table 2, soft tissue filler of the invention biological safety with higher.
Test case 3
Soft tissue filler prepared by Examples 1 to 3 is subcutaneously injected, test reference standard is GB/T16886.6-
2015.Specific test method: selecting family's rabbit back to carry out injection fillers, and after the unhairing of rabbit anesthesia back, every side is implanted into 5 points, often
A point is separated by 2cm~3cm, injection volume 0.2mL, the degradation situation of observation filler in animal body.
Test result: it is convenient that the soft tissue filler of Examples 1 to 3 is injected, and without obvious inflammatory reaction after subcutaneous filling, fills out
Filling effect can keep 12 months or more, illustrate that the biocompatibility of soft tissue filler of the invention is good, can in body
It maintains for a long time, does not need to carry out continuous injection to maintain institutional framework and performance.
Each technical characteristic of embodiment described above can be combined arbitrarily, for simplicity of description, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, all should be considered as described in this specification.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection of the invention
Range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of soft tissue filler, which is characterized in that including collagen composite microballoon and dispersion liquid, the collagen composite microballoon point
It dissipates in the dispersion liquid, the partial size of the collagen composite microballoon is 20 μm~200 μm.
2. soft tissue filler according to claim 1, which is characterized in that with the Soft-tissue operation of 100 parts by weight
On the basis of agent, the content of the collagen composite microballoon is 10~40 parts by weight, and the content of the dispersion liquid is 60~90 parts by weight.
3. soft tissue filler according to claim 1, which is characterized in that the collagen composite microballoon includes collagen and sulphur
Aching and limp ossein；
Further, the weight ratio of the collagen and chondroitin sulfate is 100:(5~20), and/or, the collagen is selected from I type
At least one of collagen and III Collagen Type VI.
4. soft tissue filler according to claim 1, which is characterized in that the dispersion liquid is to contain the molten of elastin laminin
Liquid；
Further, the elastin laminin is selected from least one of animal derived elastin laminin and genetic engineering elastin laminin；
And/or the concentration of the elastin laminin is 0.1 weight of weight %~5 %.
5. soft tissue filler according to claim 4, which is characterized in that it is 1 weight that the dispersion liquid, which also contains concentration,
The anesthetic that the viscosity modifier and concentration for measuring the weight of %~15 % are 0.1 weight of weight %~0.3 %；
Further, the viscosity modifier is selected from least one of carboxymethyl cellulose and hyaluronic acid；And/or it is described
Anesthetic is lidocaine.
6. a kind of method for preparing soft tissue filler, which is characterized in that this method comprises:
Microsyringe is used to inject the solution containing collagen in chondroitin sulfate solution with the speed of 2 μ of μ L/s~40 L/s,
The solution containing collagen composite microballoon is obtained, solid material and drying is collected, obtains collagen composite microballoon；
The collagen composite microballoon is mixed with crosslinking agent, carries out cross-linking reaction, the collagen composite microballoon after being crosslinked；
Collagen microballoon after the crosslinking is mixed with dispersion liquid, obtains soft tissue filler.
7. according to the method described in claim 6, it is characterized in that, the preparation step of the solution containing collagen include: by
Collagen is dissolved in the first solvent, and 4h~8h is stirred under the revolving speed of 400rpm~1000rpm；
Further, the pH of the solution containing collagen is 2~5, and the concentration of the collagen is 0.5 weight of weight %~3.5
% is measured, first solvent is at least one in acetum, hydrochloric acid solution, citric acid solution, formic solutions and sulfuric acid solution
Kind.
8. according to the method described in claim 6, it is characterized in that, the chondroitin sulfate solution includes chondroitin sulfate and
Two solvents, the concentration of the chondroitin sulfate are 0.5 weight of weight %~5 %；
Further, second solvent includes water and organic solvent, the volume ratio of the water and organic solvent be 1:(0.25~
1), the organic solvent is selected from least one of ethyl alcohol and acetone.
9., will be described according to the method described in claim 6, it is characterized in that, after obtaining the solution containing collagen composite microballoon
Solution containing collagen composite microballoon stirs 2h~6h under 100rpm~300rpm revolving speed, is then collected solid material again
And dry step；And/or
Before mixing the collagen composite microballoon with crosslinking agent, first the collagen composite microballoon is sieved, collection cut size
For 20 μm~200 μm of collagen composite microballoon.
10. according to the method described in claim 6, it is characterized in that, the dispersion liquid be the solution containing elastin laminin, it is described
The concentration of elastin laminin is 0.1 weight of weight %~5 %；Further, it is 1 weight %~15 that the dispersion liquid, which also contains concentration,
The viscosity modifier and concentration of weight % is the anesthetic of 0.1 weight of weight %~0.3 %；And/or
The weight ratio of the collagen composite microballoon and crosslinking agent is 1:(100~1000)；And/or
The crosslinking agent includes at least one of glutaraldehyde, carbodiimides, oxidized sodium alginate and Geniposide；And/or
The condition of the cross-linking reaction includes: that temperature is 2 DEG C~15 DEG C, and the time is 12h~48h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910754249.4A CN110404109A (en) | 2019-08-15 | 2019-08-15 | Soft tissue filler and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910754249.4A CN110404109A (en) | 2019-08-15 | 2019-08-15 | Soft tissue filler and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110404109A true CN110404109A (en) | 2019-11-05 |
Family
ID=68367749
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910754249.4A Pending CN110404109A (en) | 2019-08-15 | 2019-08-15 | Soft tissue filler and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110404109A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111544656A (en) * | 2020-06-16 | 2020-08-18 | 深圳兰度生物材料有限公司 | Dermal filler and preparation method thereof |
| CN111848991A (en) * | 2020-07-21 | 2020-10-30 | 华熙生物科技股份有限公司 | Preparation method of crosslinked hyaluronic acid microspheres |
| CN113058074A (en) * | 2020-01-02 | 2021-07-02 | 华利源(上海)生物医药科技有限公司 | Temperature-sensitive filler composition |
| CN113144288A (en) * | 2021-04-26 | 2021-07-23 | 磐升瑞祥(山东)生物工程有限公司 | Composite multi-component collagen micro-emulsion filler and preparation method thereof |
| CN113521392A (en) * | 2020-10-19 | 2021-10-22 | 红色未来科技(北京)有限公司 | Filler and preparation method of filler |
| CN114225118A (en) * | 2021-12-27 | 2022-03-25 | 深圳齐康医疗器械有限公司 | Injectable artificial dermis for promoting wound healing and preparation method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5658593A (en) * | 1992-01-16 | 1997-08-19 | Coletica | Injectable compositions containing collagen microcapsules |
| CN105126166A (en) * | 2015-09-17 | 2015-12-09 | 北京爱美客生物科技有限公司 | Injection amphiphilic-microsphere-containing hyaluronic acid mixed gel and preparation method thereof |
| CN106176292A (en) * | 2016-07-16 | 2016-12-07 | 江苏华亿美素生物组织工程有限公司 | A kind of biodegradable injection molding process decorative material and preparation method thereof |
| CN107261207A (en) * | 2017-06-28 | 2017-10-20 | 广州新诚生物科技有限公司 | A kind of collagen gel and preparation method thereof |
| CN108653818A (en) * | 2018-05-24 | 2018-10-16 | 上海其胜生物制剂有限公司 | A kind of reversible collagen stimulation filler and preparation method thereof |
-
2019
- 2019-08-15 CN CN201910754249.4A patent/CN110404109A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5658593A (en) * | 1992-01-16 | 1997-08-19 | Coletica | Injectable compositions containing collagen microcapsules |
| CN105126166A (en) * | 2015-09-17 | 2015-12-09 | 北京爱美客生物科技有限公司 | Injection amphiphilic-microsphere-containing hyaluronic acid mixed gel and preparation method thereof |
| CN106176292A (en) * | 2016-07-16 | 2016-12-07 | 江苏华亿美素生物组织工程有限公司 | A kind of biodegradable injection molding process decorative material and preparation method thereof |
| CN107261207A (en) * | 2017-06-28 | 2017-10-20 | 广州新诚生物科技有限公司 | A kind of collagen gel and preparation method thereof |
| CN108653818A (en) * | 2018-05-24 | 2018-10-16 | 上海其胜生物制剂有限公司 | A kind of reversible collagen stimulation filler and preparation method thereof |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113058074A (en) * | 2020-01-02 | 2021-07-02 | 华利源(上海)生物医药科技有限公司 | Temperature-sensitive filler composition |
| CN113058074B (en) * | 2020-01-02 | 2022-04-29 | 华利源(上海)生物医药科技有限公司 | Temperature-sensitive filler composition |
| CN111544656A (en) * | 2020-06-16 | 2020-08-18 | 深圳兰度生物材料有限公司 | Dermal filler and preparation method thereof |
| CN111848991A (en) * | 2020-07-21 | 2020-10-30 | 华熙生物科技股份有限公司 | Preparation method of crosslinked hyaluronic acid microspheres |
| CN111848991B (en) * | 2020-07-21 | 2022-03-29 | 华熙生物科技股份有限公司 | Preparation method of crosslinked hyaluronic acid microspheres |
| CN113521392A (en) * | 2020-10-19 | 2021-10-22 | 红色未来科技(北京)有限公司 | Filler and preparation method of filler |
| CN113521392B (en) * | 2020-10-19 | 2022-07-22 | 红色未来科技(北京)有限公司 | Filler and preparation method of filler |
| CN113144288A (en) * | 2021-04-26 | 2021-07-23 | 磐升瑞祥(山东)生物工程有限公司 | Composite multi-component collagen micro-emulsion filler and preparation method thereof |
| CN114225118A (en) * | 2021-12-27 | 2022-03-25 | 深圳齐康医疗器械有限公司 | Injectable artificial dermis for promoting wound healing and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110404109A (en) | Soft tissue filler and preparation method thereof | |
| DE60117984T2 (en) | COMPOSITION AND METHOD FOR REPAIRING AND REGENERATING CARTIL AND OTHER WOVEN FABRICS | |
| CN102238970B (en) | Method for manufacturing a porous three-dimensional support using powder from animal tissue, and porous three-dimensional support manufactured by same | |
| US20120165935A1 (en) | Compositions and methods for tissue filling and regeneration | |
| CN107224617B (en) | Hydrogel taking spleen extracellular matrix as raw material and preparation method thereof | |
| CN104055795B (en) | A kind of injectable implant and preparation method thereof | |
| EP2851096B1 (en) | Composition for repairing cartilage tissue, method for producing same, and use thereof | |
| CN103656749B (en) | A kind of compound degradable antibacterial artificial dura mater and preparation method thereof | |
| CN110354311A (en) | Extracellular matrix composite transparent matter acid gel and preparation method thereof, application and biomaterial | |
| US20140328939A1 (en) | Method for preparing biocompatible small intestinal mucosa hydrogel capable of controlling in-vivo degradation period | |
| BR112020015616B1 (en) | BIOINK COMPOSITION FOR DERMIS REGENERATION SHEET, METHOD FOR MANUFACTURING CUSTOM DERMIS REGENERATION SHEET USING THE SAME AND CUSTOM DERMIS REGENERATION SHEET MANUFACTURED USING THE MANUFACTURING METHOD | |
| Apelgren et al. | Vascularization of tissue engineered cartilage-Sequential in vivo MRI display functional blood circulation | |
| EP4259223A1 (en) | Particulate materials for tissue mimics | |
| CN107693844A (en) | A kind of composition gels and application | |
| DE102006026591A1 (en) | Isolated nature-identical collagen | |
| CN113877001A (en) | Silk fibroin composite gel for injection | |
| CN109503863B (en) | Injectable hydrogel and preparation method and application thereof | |
| US20200230177A1 (en) | Implants, methods for making implants and methods of treating lipoatrophy defects therewith | |
| CN104307040A (en) | Injectable hydrogel with controlled release capacity and for tissue engineering, and application thereof | |
| US20200282107A1 (en) | Sterile clear concentrated solution of biocompatible collagen, process for the preparation and use thereof | |
| CN103371857A (en) | Composite biological cannula and preparation method and application thereof | |
| CN114451397B (en) | Gel preparation for preserving stem cells, preparation method thereof and pharmaceutical composition containing gel preparation and stem cells | |
| CN115501393A (en) | Hydrogel for repairing nerve defects and preparation method and application thereof | |
| CN104324417A (en) | Tissue engineering neural restoration material constructed by autologous plasma and preparation method thereof | |
| Hagmeijer et al. | Surgical feasibility of a one-stage cell-based arthroscopic procedure for meniscus regeneration: a cadaveric study |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191105 |
|
| RJ01 | Rejection of invention patent application after publication |