CN106822991B - Small-molecular peptides biology bone renovating material and preparation method thereof - Google Patents
Small-molecular peptides biology bone renovating material and preparation method thereof Download PDFInfo
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- CN106822991B CN106822991B CN201710148011.8A CN201710148011A CN106822991B CN 106822991 B CN106822991 B CN 106822991B CN 201710148011 A CN201710148011 A CN 201710148011A CN 106822991 B CN106822991 B CN 106822991B
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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
- A61L27/20—Polysaccharides
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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
- A61L27/02—Inorganic materials
- A61L27/12—Phosphorus-containing materials, e.g. apatite
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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
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
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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
-
- 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
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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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
- A61L27/56—Porous materials, e.g. foams or sponges
-
- 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
- A61L2300/10—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing inorganic materials
- A61L2300/112—Phosphorus-containing compounds, e.g. phosphates, phosphonates
-
- 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
-
- 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
- A61L2300/25—Peptides having up to 20 amino acids in a defined sequence
-
- 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
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
Abstract
The invention belongs to osseous tissue renovating material technical fields, specifically disclose a kind of small-molecular peptides biology bone renovating material and preparation method thereof.The small-molecular peptides biology bone renovating material includes the following component of mass percentage: chitin 33%~58%；Tricalcium phosphate 27%~52%；Small-molecular peptides 8.0%~14%；Crosslinking agent 1%~7%.Small-molecular peptides biology bone renovating material provided by the invention can be used for the substitution of bone, and bone collection operation has and promotes bone defect healing, promotes the effect of Oesteoblast growth.
Description
Technical field
The present invention relates to osseous tissue renovating material technical field more particularly to a kind of small-molecular peptides biology bone renovating material and
Preparation method.
Background technique
Bone tissue damage is a great problem that human health faces.With the development of medical science, bone cement material and people
Work implantation technique is applied to clinic relatively broadly, but there are still some problems so far, for example, the biology of organic matter cement
Poor compatibility, forming process heat release are big, it is possible to the state of an illness of making patients；Inanimate matter bone cement then can form solidification in generating body
Rate is slow, the poor phenomenon of suppleness；Artificial bone graft's therapeutic scheme limits there is source be immunized with that may be present always
Problem.And organizational project is a kind of important potential replacement therapy measure, is expected to finally solve these problems.
The purpose of tissue engineering bracket material is that a three-dimensional rack is provided for cell, conducive to the sticking of cell, be proliferated and
Differentiation provides suitable Environmental Support for cell growth.Being produced in Tissue Engineering Study of bone renovating material occupies very heavy
The position wanted, there is good biocompatibility to have and close that is, without apparent cytotoxicity, inflammatory reaction and immunological rejection for it
Suitable biodegradable absorbability, i.e. the degradation absorption rate adaptable with cell, tissue growth rate, with implant site tissue
The structural strength that mechanical property matches keeps stable structure and integrality in vivo in biomethanics microenvironment, and is implantation
Cell provides suitable microstress environment.In view of bone cement material and artificial graft's technology there are the problem of and organizational project branch
Frame material there are the advantages of, it is necessary to propose a kind of new tissue engineering bracket material.
Summary of the invention
The present invention is for poor biocompatibility existing for bone cement material and artificial graft's technology, forming process heat release is big,
The problems such as solidification rate is slow, suppleness is poor provides a kind of small-molecular peptides biology bone renovating material and its method.
To achieve the above object of the invention, the embodiment of the present invention uses the following technical solution:
A kind of small-molecular peptides biology bone renovating material includes the following component of mass percentage:
And a kind of preparation method of small-molecular peptides biology bone renovating material, it at least includes the following steps:
Small-molecular peptides biology bone renovating material weighs chitin, tricalcium phosphate, small-molecular peptides and crosslinking as described above
Agent；
The weighed chitin and solvent are subjected to mixing treatment, obtain the first solution；
The weighed tricalcium phosphate and small-molecular peptides and first solution are subjected to mixing treatment, obtain mixing molten
Liquid；
The mixed solution is subjected to freeze forming processing and first time freeze-drying process, is then added weighed described
Crosslinking agent carries out cross-linking reaction processing, carries out second of freeze-drying process after cross-linking reaction processing, obtains small-molecular peptides biology
Bone renovating material；Or 3D printing processing is carried out after mixing the crosslinking agent with the mixed solution immediately, obtain small molecule
Peptide biology bone renovating material.
The above embodiment of the present invention using chitin, tricalcium phosphate, small-molecular peptides as primary raw material, in conjunction with crosslinking agent,
Obtained small-molecular peptides biology bone renovating material porosity reaches 85%~94%, and aperture is 200 μm~400 μm, compression strength
It for 1.3KPa~1.6KPa, strains as 8.8mm~10.8mm, compression modulus is 3MPa~12MPa, and with good biofacies
Capacitive, forming process heat release is few in vivo, it is suppressed that in exothermic process a possibility that inflammation, has controlled degradation, sustained release
And have no toxic side effect, it is suitble to the clinical application of bone collection.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
The embodiment of the present invention provides a kind of small-molecular peptides biology bone renovating material, includes the following group of mass percentage
Point:
In a preferred embodiment, the mass percentage of the small-molecular peptides is 10%~14%, and molecular weight ranges are
659-3000, the molecular weight ranges of this preferred small-molecular peptides have bioactivity high, and human body is easy absorption, promote osteoblast raw
Long fast feature.
The crosslinking agent is sodium tripolyphosphate solution；The sodium tripolyphosphate solution concentration is 40g/L~60g/L.It can also
It is substituted with pharmagel, the solution concentration of pharmagel is 0.5g/L~1.0g/L.
Small-molecular peptides can obviously increase the calcium content of bone of ovariectomized female rats, increase bone density, can prevent sclerotin well and dredge
Pine, fracture, and Osteoblast Differentiation can be promoted, promote union；Chitin has good biology as a kind of natural organic matter
Compatibility, controllable degradability, sustained release agent effect have no toxic side effect, inhibit the effect of inflammation, but poor mechanical property, it is difficult to answer
Defect for weight bearing area；Tricalcium phosphate as bioceramic have no antigen, no absorbability, foreign body reaction very little,
Adsorbable biotic factor, stem cell, chemotherapeutics, but without bone inductive effect.Therefore by this triplicity, homogenous material can be overcome
Deficiency, preferably meet the needs of tissue engineering bracket material.
The above embodiment of the present invention creatively uses chitin, tricalcium phosphate, small-molecular peptides as primary raw material, in conjunction with
Crosslinking agent, obtained small-molecular peptides biology bone renovating material porosity reach 85%~94%, and aperture is 200 μm~400 μm, is resisted
Compressive Strength is 1.3KPa~1.5KPa, is strained as 8.8mm~10.8mm, and compression modulus is 3MPa~12MPa, and with good
Biocompatibility, forming process heat release is few in vivo, it is suppressed that in exothermic process a possibility that inflammation, has controllable drop
It solves, be sustained and have no toxic side effect, be suitble to the clinical application of bone collection.
Correspondingly, the present invention is on the basis of the above embodiments, it is further provided small-molecular peptides of the embodiment of the present invention are raw
The preparation method of object bone renovating material.
In embodiment, the preparation method of the small-molecular peptides biology bone renovating material at least includes the following steps:
Small-molecular peptides biology bone renovating material weighs chitin, tricalcium phosphate, small-molecular peptides and crosslinking as described above
Agent；
The weighed chitin and solvent are subjected to mixing treatment, obtain the first solution；
The weighed tricalcium phosphate and small-molecular peptides and first solution are subjected to mixing treatment, obtain mixing molten
Liquid；
The mixed solution is subjected to freeze forming processing and first time freeze-drying process, is then added weighed described
Crosslinking agent carries out cross-linking reaction processing, carries out second of freeze-drying process after cross-linking reaction processing, obtains small-molecular peptides biology
Bone renovating material；Or 3D printing processing is carried out after mixing the crosslinking agent with the mixed solution immediately, obtain small molecule
Peptide biology bone renovating material.
Preferably, the solvent is glacial acetic acid solution or dioxanes.
Preferably, the temperature of above-mentioned freeze forming processing is -15 DEG C~-25 DEG C.
Preferably, the temperature of above-mentioned first time freeze-drying process is -50 DEG C~-60 DEG C, and the time is 20h~for 24 hours.
Preferably, the temperature of second of freeze-drying process is -50 DEG C~-60 DEG C, and the time is 3h~8h.
Preferably, the time of above-mentioned cross-linking reaction processing is 1h~2h.
Preferably, 3D printing processing uses 3D printer.
In order to more preferably reduce the intensity for influencing material due to there are bubble during freeze forming, freeze forming it
Before, further include that obtained mixed solution is stood into a period of time, realizes deaeration processing by standing.
The preparation method for the small-molecular peptides biology bone renovating material that the above embodiment of the present invention provides, raw material sources are wide
It is general, without adding other auxiliary agents, for freeze-drying or 3D printing technique, entire processing technology simple possible, small point of acquisition
Sub- peptide biology bone renovating material performance is stablized, and porosity reaches 85%~94%, and aperture is 200 μm~400 μm, and compression strength is
1.3KPa~1.5KPa strains as 8.8mm~10.8mm, and compression modulus is 3MPa~12MPa, and with good bio-compatible
Property, forming process heat release is few in vivo, it is suppressed that in exothermic process a possibility that inflammation, have controlled degradation, sustained release and
It has no toxic side effect, is suitble to large-scale production.
In the embodiment of the present invention, using chitin and tricalcium phosphate as carrier, the small-molecular peptides that small-molecular peptides preparation is added are raw
Object bone renovating material has the function of promoting union and accelerates bone uptake at bone defect.The small-molecular peptides biological bone is repaired
Multiple material has physicochemical properties similar with bone tissue, and has good biocompatibility, biological degradability and bone
Conducting power, can be used for the substitution of bone, and bone collection operation has the function of promoting bone defect healing.
The small-molecular peptides biology bone renovating material and preparation method thereof that embodiment provides in order to better illustrate the present invention, under
Face is illustrated by embodiment.
Embodiment 1
A kind of small-molecular peptides biology bone renovating material, including following component:
Embodiment 1 provide small-molecular peptides biology bone renovating material the preparation method is as follows:
4.2g chitin is dissolved in the glacial acetic acid solution of 60ml 1%, obtains the first solution after completely dissolution, to first
The tricalcium phosphate powder and 1.2g small-molecular peptides of 4.0g are added in solution, mixes well to obtain gel mixed solution, stands
Carry out processing deaeration；
Obtained gel mixed solution is transferred in 96 orifice plates, preforming processing 8h in -20 DEG C of refrigerator is placed in；So
After go in freeze drier, be freeze-dried at -55 DEG C for 24 hours, take out, be added account for total amount 6% sodium tripolyphosphate solution it is (dense
Degree is 50g/L), cross-linking reaction 1h；
Cross-linking reaction terminates, and is transferred in freeze drier, and 5h is freeze-dried under the conditions of -55 DEG C, obtains small-molecular peptides
Biological bone renovating material；It is detected mean porosities and reaches 90.6%, average pore size is 280 μm, and mean compressive strength is
1.52KPa, mean strain 9.21mm, average compression modulus are 5.31MPa.
Embodiment 2
A kind of small-molecular peptides biology bone renovating material, including following component:
Embodiment 2 provide small-molecular peptides biology bone renovating material the preparation method is as follows:
5.9g chitin is dissolved in the glacial acetic acid solution of 60mL1%, obtains the first solution after completely dissolution, it is molten to first
In liquid be added 2.7g tricalcium phosphate powder and 0.8g small-molecular peptides, mix well to obtain gel mixed solution, stand into
Row processing deaeration；
Obtained gel mixed solution is transferred in 96 orifice plates, preforming processing 8h in -20 DEG C of refrigerator is placed in；So
After go in freeze drier, be freeze-dried at -55 DEG C for 24 hours, take out, be added account for total amount 6% sodium tripolyphosphate solution it is (dense
Degree is 50g/L), cross-linking reaction 1h；
Cross-linking reaction terminates, and is transferred in freeze drier, and 5h is freeze-dried under the conditions of -55 DEG C, obtains small-molecular peptides
Biological bone renovating material.
It is detected mean porosities and reaches 83%, average pore size is 338 μm, and mean compressive strength 1.1KPa is averagely answered
Become 11.7mm, average compression modulus is 7.4MPa.
Embodiment 3
A kind of small-molecular peptides biology bone renovating material, including following component:
Embodiment 3 provide small-molecular peptides biology bone renovating material the preparation method is as follows:
3g chitin is dissolved in the glacial acetic acid solution of 60mL1%, obtains the first solution after completely dissolution, to the first solution
The middle tricalcium phosphate powder and 1.2g small-molecular peptides that 5.2g is added mixes well to obtain gel mixed solution, stands and carry out
Handle deaeration；
Obtained gel mixed solution is transferred in 96 orifice plates, preforming processing 8h in -20 DEG C of refrigerator is placed in；So
After go in freeze drier, be freeze-dried at -55 DEG C for 24 hours, take out, be added account for total amount 6% sodium tripolyphosphate solution it is (dense
Degree is 50g/L), cross-linking reaction 1h；
Cross-linking reaction terminates, and is transferred in freeze drier, and 5h is freeze-dried under the conditions of -55 DEG C, obtains small-molecular peptides
Biological bone renovating material.
It is detected mean porosities and reaches 80%, average pore size is 318 μm, and mean compressive strength 1.04KPa is averagely answered
Become 11.8mm, average compression modulus is 7.8MPa.
Embodiment 4
A kind of small-molecular peptides biology bone renovating material, including following component:
Embodiment 4 provide small-molecular peptides biology bone renovating material the preparation method is as follows:
4.5g chitin is dissolved in the glacial acetic acid solution of 60mL1%, obtains the first solution after completely dissolution, it is molten to first
In liquid be added 3.6g tricalcium phosphate powder and 1.2g small-molecular peptides, mix well to obtain gel mixed solution, stand into
Row processing deaeration；
Obtained gel mixed solution is transferred in 96 orifice plates, preforming processing 8h in -20 DEG C of refrigerator is placed in；So
After go in freeze drier, be freeze-dried at -55 DEG C for 24 hours, take out, be added account for total amount 6% sodium tripolyphosphate solution it is (dense
Degree is 50g/L), cross-linking reaction 1h；
Cross-linking reaction terminates, and is transferred in freeze drier, and 5h is freeze-dried under the conditions of -55 DEG C, obtains small-molecular peptides
Biological bone renovating material.
It is detected mean porosities and reaches 63%, average pore size is 290 μm, and mean compressive strength 1.42KPa is averagely answered
Become 9.5mm, average compression modulus is 10.6MPa.
Embodiment 5
A kind of small-molecular peptides biology bone renovating material, including following component:
Embodiment 5 provide small-molecular peptides biology bone renovating material the preparation method is as follows:
4.2g chitin is dissolved in the glacial acetic acid solution of 60mL1%, obtains the first solution after completely dissolution, it is molten to first
In liquid be added 4.0g tricalcium phosphate powder and 1.2g small-molecular peptides, mix well to obtain gel mixed solution, stand into
Row processing deaeration；The sodium tripolyphosphate solution (concentration 50g/L) for accounting for total amount 6% is added, is uniformly mixed.
Gel mixed solution is carried out by 3D printing using 3D printing equipment, obtains small-molecular peptides biology bone renovating material.
It is detected mean porosities and reaches 93%, average pore size is 285 μm, and mean compressive strength 1.54KPa is averagely answered
Become 9.24mm, average compression modulus is 5.66MPa.
In order to verify the toxicity of small-molecular peptides biology bone renovating material prepared by the embodiment of the present invention 1~5, below by it
Growth experiment verifying to experimental mouse osteocyte.
By osteoblast with 103The density in a/hole is laid on 96 orifice plates (6 plates), after cell is adherent, in each orifice plate
It is separately added into the small-molecular peptides biology bone renovating material leaching liquor that 50g/L is prepared by embodiment 1-5, in different time points (24,
48,72h) 3- (4,5- dimethylthiazole -2) -2,5- diphenyltetrazolium bromide bromide (English abbreviation: MTT) solution is added and is incubated for 4h,
Ferric tri-dodecanesulfonate is added afterwards to stay overnight in the incubator, absorbance value is measured at 570nm wavelength.It is yin with DMEM culture solution
Property control, measure the cell survival rate of compound bone biologic material.Cell survival rate (%)=(each cell mean/control of experimental group
Cell mean) × 100%.
Test result is shown, relative to negative control group, each compound bone biologic material group is showed no in 24,48 and 72h
The proliferation for inhibiting osteoblast illustrates that the bone biologic material acellular poison acts on.Wherein by embodiment 1 and embodiment 5 formula and
Compound bone biologic material prepared by technique can be obviously promoted the proliferation of osteoblast, and be substantially better than by embodiment 2,3 and 4
Bone biologic material prepared by formula rate (P < 0.01), the results are shown in Table 1.
Influence (n=5) of the 1 each group bone biologic material of table to osteoclast survival rate
* indicate that P < 0.05 compared with negative control group, * * indicate P < 0.01 compared with negative control group, Δ indicates and implements
1 group of example is compared P < 0.05.It can be seen that the small-molecular peptides biology bone renovating material that the embodiment of the present invention 1~5 obtains does not have cell toxicant
Property.
It is right for the performance of further description small-molecular peptides biology bone renovating material provided in an embodiment of the present invention
The rat at 3 monthly age of SPF grade carries out biological living experiment.
1. experimental material: 70 weight (218.8 ± 47.30) g of male SD rat, 35 weight (187.3 of female sd inbred rats
± 9.06) g is provided by Guangdong Medical Lab Animal Center, medical experiment animal certificate number: check and affirm word 2008A020 in Guangdong.
2. rearing conditions: special room sub-cage rearing, the next day replace padding, freely take the photograph water, ingest (standard feed nursing), weekly
It weighs primary.
3. experimental design and grouping
3 monthly age cleaning grade rats are chosen, the weight application micro excel of male rat is generated into random number series, and benefit
It is carried out being randomly divided into 7 groups with the random number series of generation, is shown in Table 2.
The grouping of 2 experimental animal of table and dosage
4. experimental method
The foundation of 4.1 rat bone defect models and the filling of biomaterial
All rat pre-operative anxieties 6 hours, surgical instrument is through high pressure steam sterilization.0.3mL/ is pressed with 7% chloraldurate
100g intraperitoneal injection of anesthesia rat, is disappeared with righting response or clamp hind leg ungauged regions are to anaesthetize successfully reference standard.Prone position
It is fixed, the hair of periphery 2cm at the shin bone body surface projection of right side is shaved, it is conventional to spread aseptic operation towel with 75% alcohol disinfecting 3 times.Edge
Make a long longitudinal cut for being about 1cm in front of shin bone long axis direction, cut skins, subcutaneous tissue, passivity point with No. 11 sharp knives
From exposure shin bone, take 2.0 × 250mm Kirschner wire as drill bit, making a diameter at 1-1.5mm below shin bone epiphysis with electric drill is
Drill-through shin bone bilateral cortical bone is subject in the drilling of 3.5mm, drilling, and it is clean then to extract normal saline flushing with 5mL syringe
Hole intraclast, and bone biologic material is filled to bone defect, it sews up the incision after operation and records operating time.Wait anaesthetize
After recovery, rat is put back in former cage.Compounding medicine and surgical procedure keep sterile working.
The scanning of 4.2X light ray
Experiment carries out surrounding altogether, carries out x-ray scanning to all rats weekly and takes pictures, to rat tibia X-ray
The gray scale of picture is handled, and calculates corresponding gray value as bone density, judges that bone renovating material heals at bone defect
Influence.
4.3 serum alkaline phosphatases (ALP) measurement
7,14,28d rat blood serum is taken after administration, utilizes its ALP content of kit measurement.
5. experimental result
5.1 bone densities calculate scanning 7d, 14d, 28d and intervention group are respectively administered compared with simple bone defect group, scan gray scale
Increase, i.e., bone density increases (P < 0.01), illustrates that bone defect position bone tissue growth is all right after intervening.And in the corresponding time
5 groups of point, embodiment 1 and embodiment are compared with giving chitin/tricalcium phosphate group, and scanning gray scale significantly increases, i.e., bone is close
Angle value increases obvious P < 0.05), it is shown in Table 3.
3 rat femur bone density result of table
* indicate that P < 0.05 compared with bone defect group, * * indicate P < 0.01 compared with bone defect group, Δ indicates and bone defect group
+ chitin/tricalcium phosphate group compares P < 0.05.
Chitin/tricalcium phosphate is given after 5.2 serum alkaline phosphatases (ALP) level and simple bone defect group and bone defect
Group is compared, and in 7d, each embodiment group of 14d, 28d can promote the vigor (P < 0.05) of ALP, wherein the promotion effect of embodiment 1 and 5
Fruit more preferably, illustrates the bone biologic material with that can significantly improve osteoblast viability, promoting bone growing the results are shown in Table 4.
The content of ALP in 4 rat blood serum of table
* indicate that P < 0.05 compared with bone defect group, * * indicate P < 0.01 compared with bone defect group, Δ indicates and bone defect group
+ chitin/tricalcium phosphate group compares P < 0.05, and Δ Δ indicates P < 0.01 compared with bone defect group+chitin/tricalcium phosphate group.
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 modification, equivalent replacement or improvement etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (10)
1. a kind of small-molecular peptides biology bone renovating material, it is characterised in that: include the following component of mass percentage:
The small-molecular peptides biology bone renovating material porosity is 85%~94%, and aperture is 200 μm~400 μm, compression strength
For 1.3kPa~1.6kPa, strain as 8.8mm~10.8mm, compression modulus is 3MPa~12MPa.
2. small-molecular peptides biology bone renovating material as described in claim 1, it is characterised in that: the molecular weight of the small-molecular peptides
Range is 659~3000.
3. small-molecular peptides biology bone renovating material as described in claim 1, it is characterised in that: the crosslinking agent is tripolyphosphate
Sodium solution；The sodium tripolyphosphate solution concentration is 40g/L~60g/L.
4. a kind of preparation method of small-molecular peptides biology bone renovating material, it is characterised in that: at least include the following steps:
Chitin, tricalcium phosphate, small is weighed according to the small-molecular peptides biology bone renovating material as described in any one of claims 1 to 3
Molecular peptide and crosslinking agent；
The weighed chitin and solvent are subjected to mixing treatment, obtain the first solution；
The weighed tricalcium phosphate and small-molecular peptides and first solution are subjected to mixing treatment, obtain mixed solution；
The mixture is subjected to freeze forming processing and first time freeze-drying process, the weighed crosslinking is then added
Agent carries out cross-linking reaction processing, carries out second of freeze-drying process after cross-linking reaction processing, obtain small-molecular peptides biological bone and repair
Multiple material；Or 3D printing processing is carried out after mixing the crosslinking agent with the mixed solution immediately, it is raw to obtain small-molecular peptides
Object bone renovating material.
5. the preparation method of small-molecular peptides biology bone renovating material as claimed in claim 4, it is characterised in that: the solvent is
Glacial acetic acid solution or dioxanes.
6. the preparation method of small-molecular peptides biology bone renovating material as claimed in claim 4, it is characterised in that: described to be frozen into
The temperature of type processing is -15 DEG C~-25 DEG C.
7. the preparation method of small-molecular peptides biology bone renovating material as claimed in claim 4, it is characterised in that: the first time
The temperature of freeze-drying process is -50 DEG C~-60 DEG C, and the time is 20h~for 24 hours.
8. the preparation method of small-molecular peptides biology bone renovating material as claimed in claim 4, it is characterised in that: described second
The temperature of freeze-drying process is -50 DEG C~-60 DEG C, and the time is 3h~8h.
9. the preparation method of small-molecular peptides biology bone renovating material as claimed in claim 4, it is characterised in that: the crosslinking is anti-
The time that should be handled is 1h~2h.
10. the preparation method of small-molecular peptides biology bone renovating material as claimed in claim 4, it is characterised in that: the freezing
It further include standing and defoaming processing before molding.
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Family Cites Families (4)
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| TWI267378B (en) * | 2001-06-08 | 2006-12-01 | Wyeth Corp | Calcium phosphate delivery vehicles for osteoinductive proteins |
| CN1403166A (en) * | 2002-09-26 | 2003-03-19 | 东南大学 | Absorbable active tissue matter for repairing hard tissue and its prepn |
| CN104292309B (en) * | 2014-09-29 | 2017-04-12 | 广西中医药大学 | Micro-molecule polypeptide |
| CN105944077A (en) * | 2016-04-29 | 2016-09-21 | 郑州京科强直中西医结合医院 | Application of small molecular peptide and small molecular minimally invasive treatment method |
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