CN110522946A - A kind of bone renovating material microballoon and preparation method thereof carrying rhBMP-2 - Google Patents
A kind of bone renovating material microballoon and preparation method thereof carrying rhBMP-2 Download PDFInfo
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Abstract
The invention belongs to field of biomedical materials and regenerative medicine field containing activated protein.A kind of bone renovating material, including rhBMP-2(recombinate bone morphogenesis protein-2) and poly (glycolide-lactide) (PLGA) microballoon, rhBMP-2 be wrapped in poly (glycolide-lactide) microballoon, the sea-mussel mucin modification of the surface of microballoon.
Description
Technical field
The invention belongs to field of biomedical materials and regenerative medicine field containing activated protein, and specifically, the application mentions
A kind of bone renovating material microballoon and preparation method thereof for carrying rhBMP-2 is supplied.
Background technique
The treatment of orthopaedics illness usually requires bone collection, as the bridge of new bone growth, such as bone tumour, bone nonunion, cervical vertebra
Disk is prominent, spine osteoporosis is fractured and numerous spine disorders for needing fusion.China has ten hundreds of every year
Patient cause bone defect because of lesions such as wound, infection, tumours, separately have hundreds of thousands of patients underwent spinal fusion surgeries, the two is both needed to
Bone collection is wanted to treat.There is newly-increased operation wound in bone collection, source is restricted, immunological rejection, transmission and expense are high
The problems such as expensive.With the development of material science, new development trend is become with biomaterial substitution bone graft.Ideal bone
Repair materials are not only wanted actively induce new bone formation, quick repairing bone defect, it is necessary to have good biocompatibility and
Degradability, and must be safe and non-toxic.
Bone morphogenesis protein-2 (bone morphogenetic protein-2, BMP-2) is the pass for promoting ostosis
The key factor belongs to TGF- α family member, and BMP-2 is the major signaling molecule that cell differentiation metallogenic material deposits osteoblast, rises
To the effect of induced osteogenesis cell differentiation.It is expressed in limbs growth, endochondral ossification, fracture, in bone growth and development and again
Raw repair plays an important role, and has a good application prospect in clinical treatment bone fracture.It is repaired additionally, due to the common bone for carrying rhBMP
Multiple material is often concentrated release rhBMP-2 using early period in product, not can control the release of rhBMP-2 constantly, micro,
Cause the rhBMP-2 content of local organization in a period of time to be enriched with, with blood flow or tissue fluid flow to tissue regions to be repaired with
Its outer hetero-organization, may cause unnecessary ectopic osteogenesis.In addition, the half-life short of rhBMP-2 in vivo, local use is very
It is just metabolized and absorbs fastly, be difficult to play a role, even if large dosage, which is administered continuously to also tend to that target area cannot be supplied to, persistently to be had
The concentration of effect can not provide good therapeutic effect for the bone for needing to grow and restore for a long time, and be easy to draw
Play low patient's skeletonization quality, inflammatory reaction and other potential risks.The prompt of above reason how to control its is lasting, stable,
Slowly discharge, not only haved the function that effectively to promote bone uptake simultaneously but also can to avoid with to prevent the generation of adverse reaction be to make good use of
A key technology of rhBMP-2.
Poly (glycolide-lactide) (hereinafter referred to as PLGA) microballoon is due to being to answer with preferable biocompatibility and biodegradability
With with study one of widest drug carrier material, U.S. FDA, which has been approved by it, can be used as the original of Implantable Medical Device
Material.
Sea-mussel mucin (mussel adhesive protein, MAP) is the hatching egg that mussel foot gland secrets out of
White compound, superpower viscosity are the key that mussel can be firmly attached on reef and not be afraid of sea beat.Sea-mussel mucin
The advantages that range is wide, and intensity is high, good waterproof performance, bioaffinity and good degradability with bonding, it is considered to be good biology
Adhesive.Mussel adh esive protein is loaded into upper BMP-2, promotees bone uptake functional treatment using its strong bonding characteristic and BMP-2
The fixed difficult situation such as diseases, the especially comminuted fracture such as fracture is a kind of promising medicament.
Summary of the invention
The purpose of the present invention is to provide one kind have preferable biocompatibility and biodegradability, have be sustained,
The bone renovating material of target controlling and releasing.Another object of the present invention is to provide the preparation methods of above-mentioned bone renovating material.
A kind of bone renovating material, including rhBMP-2 (recombination bone morphogenesis protein-2) and poly (glycolide-lactide) (PLGA) it is micro-
Ball, rhBMP-2 are wrapped in poly (glycolide-lactide) microballoon, and the surface of microballoon is modified with sea-mussel mucin.
A kind of preparation method of bone renovating material, includes the following steps:
(1) the PLGA microballoon preparation of rhBMP-2 is loaded；(2) PLGA for sea-mussel mucin being aggregated to load rhBMP-2 is micro-
On ball.
Further, step (1): being dissolved in rhBMP-2 freeze-dried powder in hac buffer, is then added containing PLGA's
Acetonitrile, 3000rpm revolving speed emulsify 30 seconds, are added in the mixed emulsion of atoleine and Tween-80, with the revolving speed of 300rpm or so
Stirring 2-4 hours stands 30 minutes after the completion, then is lyophilized after alternately being washed with petroleum ether and isopropanol, obtains the micro- of drying
Ball.
Further, the PLGA microballoon for loading rhBMP-2 is suspended in aqueous acetic acid by step (2), and mussel is taken to glue egg
It is slowly stirred addition PLGA microsphere suspension liquid thereto under the conditions of white solution tune PH to 6.0,2-8 DEG C and evenly spreads to the viscous egg of mussel
In white solution, it is slowly stirred 3-4 hours, stand 12 hours or more, resulting load rhBMP-2 sea-mussel mucin-PLGA microballoon is used
Petroleum ether and isopropanol are lyophilized after alternately washing, and obtain the PLGA microballoon that dry surface is modified by sea-mussel mucin.
Further, wherein the mass ratio of the sea-mussel mucin and PLGA microballoon is 1:80~120.
There is the PLGA microballoon of rhBMP-2 as primary raw material using, load modified through sea-mussel mucin surface adhesion, has slow
It releases, the effect of target controlling and releasing rhBMP-2, oriented attachment (nerve, blood vessel, cell etc.) the tissue repair factor rush bone uptake, simultaneously
Also have the function of autoadhesion bone cips, is a kind of ideal bone tissue reparation product.
RhBMP-2 is used and is wrapped up with good biocompatibility and biodegradable PLGA microballoon by the present invention, PLGA microballoon
Surface is modified with mussel is viscous, so that PLGA microballoon is utilized the characteristic of the surface-crosslinked absorption of sea-mussel mucin, is used for PLGA
The surface of microballoon is modified.Be adsorbed the PLGA microballoon of sea-mussel mucin molecule formed on surface it is micro- prominent, due between protein molecular
Interaction between the effect and microballoon of power can make microsphere aggregation and form loose porous structure, can promote mind
Growth is walked in absorption through, the growth factor of blood vessel and other cells；Also as the reason of adsorption mucoprotein, reduced
The chance of degradation reaction, the problem of having well solved PLGA burst release and continued controlled release drug.
RhBMP-2PLGA microballoon is carried since sea-mussel mucin is in the oriented attachment of microsphere surface, remains sea-mussel mucin
The double action of bonding and adherency both may be implemented oriented attachment for osseous tissue renovating material orientation and be bonded in the end for needing to repair
At mouthful, material and effective component is avoided, to elsewhere, to cause ectopic osteogenesis with blood flow and tissue infiltration；It simultaneously can be by knochenbruch
Nerve, blood vessel, mesenchymal cell of port surrounding etc. are adsorbed onto knochenbruch surface, advantageously promote the reparation of knochenbruch.
Detailed description of the invention
Fig. 1: cell electron microscope；Wherein, A is to carry rhBMP-2PLGA microballoon group；B, which is that sea-mussel mucin is modified, carries rhBMP-
2PLGA microballoon group；
Fig. 2: ectopic osteogenesis tests X-ray in Mice Body；Wherein A, B, C are test group；D is control group；E is blank control
Group.
Specific embodiment
Embodiment 1: bone renovating material microballoon preparation
Natural sea-mussel mucin extract product: BD Bioscience production is marked as Mytilus galloprovincialis product, containing Mefp-2,
Mefp-2,Mefp-3；
BMP-2 is voluntarily produced by enterprise, and specific production process is referring to applicant CN201910030649.0 Chinese patent
In application production method (expression in escherichia coli, cation exchange chromatography, detection escherichia coli host residual protein <
0.005%), inhibition zone method detects antibiotic residue < 0.1ppm；
Poly (glycolide-lactide) is produced by DURECT company, the U.S..
Preparation method:
(1) preparation of the PLGA microballoon of rhBMP-2 is loaded
RhBMP-2 freeze-dried powder, it is molten to be dissolved in acetate buffer by the PLGA microballoon that load rhBMP-2 is prepared using W/O/W method
In liquid, the acetonitrile containing PLGA is then added, 3000rpm revolving speed emulsifies 30 seconds, and the mixed emulsion of atoleine and Tween-80 is added
In, it is stirred 2-4 hours with the revolving speed of 300rpm or so, after the completion, stands 30 minutes, then alternately washed with petroleum ether and isopropanol
After be lyophilized, obtain the PLGA microballoon of dry load rhBMP-2.
(2) microsphere surface is carried out with sea-mussel mucin to be modified；
The microballoon of above-mentioned preparation is suspended in aqueous acetic acid, solution tune PH to 6.0,2-8 DEG C item of sea-mussel mucin is taken
Be slowly stirred under part thereto be added PLGA microsphere suspension liquid evenly spread in sea-mussel mucin solution (sea-mussel mucin:
PLGA microballoon=1:100), it is slowly stirred 3-4 hours, stand 12 hours or more, resulting load rhBMP-2 sea-mussel mucin-
PLGA microballoon microballoon petroleum ether and isopropanol are lyophilized after alternately washing, and it is micro- to obtain dry new one kind with sea-mussel mucin
Ball.
The PLGA microballoon for the load rhBMP-2 that the present embodiment obtains will using the characteristic of the surface-crosslinked absorption of sea-mussel mucin
Its surface for being used for PLGA microballoon is modified, be adsorbed sea-mussel mucin molecule PLGA microballoon formed on surface it is micro- prominent, due to
Interaction between protein molecular between the effect and microballoon of power can make microsphere aggregation and form loose porous structure,
It can promote the absorption of nerve, the growth factor of blood vessel and other cells, walk growth；Also as adsorption mucoprotein
Reason reduces the chance of degradation reaction, has good biodegradability, overcomes existing PLGA drug bearing microsphere burst release
Problem discharges rhBMP step by step and the bone tissue reparation time is almost the same.
RhBMP-2PLGA microballoon is carried since sea-mussel mucin is in the oriented attachment of microsphere surface, remains sea-mussel mucin
The double action of bonding and adherency both may be implemented oriented attachment for osseous tissue renovating material orientation and be bonded in the end for needing to repair
At mouthful, material and effective component is avoided, to elsewhere, to cause ectopic osteogenesis with blood flow and tissue infiltration；It simultaneously can be by knochenbruch
Nerve, blood vessel, mesenchymal cell of port surrounding etc. are adsorbed onto knochenbruch surface, advantageously promote the reparation of knochenbruch
Test 1: the adherent test of cell promotees bone uptake to verify new microsphere material for nerve, blood vessel, mesenchymal cell etc.
The absorption property of the factor；
Method: the load rhBMP- of the hole 1.0ml/ 1 step containing embodiment (1) acquisition is separately added into 26 well culture plates
The sea-mussel mucin surface for hanging muddy liquid and obtaining containing step (2) of 2PLGA microballoon, which is modified, carries the suspended of rhBMP-2PLGA microballoon
Liquid, 55 DEG C of drying make microballoon be attached to culture plate bottom, add in culture plate spare after water cleans 3 times.Cell culture uses
Mouse skin fibroblast removes the cell in logarithmic growth phase, is separately added into 266 porocyte culture plates got ready
In, the growing state of cell is observed after 37 DEG C of culture 2h.
The result is shown in Figure 1: after culture 2 hours, Figure 1A carries rhBMP-2PLGA microballoon group adherence rate as 35%, Fig. 2 B warp
The modified adherence rate for carrying rhBMP-2PLGA microballoon group of sea-mussel mucin is 85%.
Test 2: the extracorporeal releasing experiment of the PLGA microballoon bone renovating material of rhBMP-2 is carried
Test group: the PLGA microballoon bone renovating material that rhBMP-2 is carried obtained by embodiment is divided into 3 groups of test groups；
Control group: the rhBMP-2 bone renovating material of Jiuyuan Gene Engineering Co., Ltd., Hangzhou's production, product name bone lure
It leads, is made of rhBMP-2 and carrier material, carrier material includes hydroxyapatite, soybean lecithin and pharmagel.
Method: 3 groups of test groups and 1 group of control group are placed in bag filter, and bag filter is placed in 20ml containing 0.2% sodium azide
, in the PBS buffer solution of pH7.0.Be statically placed at 37 DEG C, in 12h, for 24 hours, 120h, 168h, 336h, 504h, 672h, 840h point
Not Qu Yang 1ml (then supplement is containing 0.2% sodium azide, the PBS buffer solution 1ml of pH7.0), according to the explanation of kit with
ELISA method measures rH-BMP2 concentration, the accumulative release percentage of Conversion Calculation.As a result as shown in the table.
Table 1, the accumulative release percentage of rH-BMP2
RH-BMP2 in control material in 168h complete substantially by release, and carry the bone renovating material microballoon of rhBMP-2
Pharmaceutical release time extends to 5 weeks or so, substantially corresponding with the early growth period in Bone Defect Repari.Test 3: practical Bone Defect Repari experiment
One, it designs:
Grouping comparison, multi-angle assess observation experiment.
Two, material:
Mouse strain is ICR or ICR hero mouse, age of mouse 30 days or so；
Test group: the PLGA microballoon bone renovating material for carrying rhBMP-2 is provided by Zhejiang Rui Gu Biotechnology Co., Ltd；
Control group: the rhBMP-2 bone renovating material of Jiuyuan Gene Engineering Co., Ltd., Hangzhou's production, product name bone lure
It leads, is made of rhBMP-2 and carrier material, carrier material includes hydroxyapatite, soybean lecithin and pharmagel.
Three, method:
10 normal ICR or KM male mices are randomly divided into 5 groups, every group 2；3 groups are wherein tested, compares 1 group, blank
Compare 1 group.
After mouse is anaesthetized with 6% yellow Jackets, skin is cut in unhairing disinfection in hind leg thigh, separates spatium intermusculare, implantation is not
Same carrier material, and certain antibiotic is added to prevent from infecting, then layer-by-layer suture muscle and skin, sterilize wound, normal to raise
It supports.
Four, it observes:
After the completion of heeling-in experiment after 4 weeks, mouse is subjected to radiological examination (X-ray), checks whether mouse hind leg bone occurs
Tissue.After 4 weeks progress radiological examinations, mouse is put to death, takes out the bone tissue in heeling-in area, weighs the wet of bone contained by the inside
Weight, and carry out the assessment of HE stained tissue.
Five, the standards of grading of X-ray observation knitting
1.1962 Chai Benfu union measurement standard:
2.Lane-Sandhu X-ray and histological score standard:
Six, result
The mouse of experimental group and control group scoring such as table 1, shown in table 2.
Table is measured in 1 1962 Chai Benfu union of table
| Number | Broken ends of fractured bone edge | Periosteal reaction | Yield of callus | Poroma density | Poroma edge |
| Test group 1 | ++++ | ++++ | ++++ | ++++ | ++++ |
| Test group 2 | +++ | +++ | ++++ | ++++ | +++ |
| Test group 3 | ++++ | +++ | ++++ | ++++ | ++++ |
| Control group | ++ | ++ | ++ | ++ | ++ |
| Blank control | + | + | + | + | + |
Table 2.Lane-Sandhu X-ray and histological score table
| Number | Bon e formation | Bone connection | Bone moulding |
| Test group 1 | 4 | 4 | 4 |
| Test group 2 | 4 | 3 | 4 |
| Test group 3 | 4 | 3 | 4 |
| Control group | 2 | 3 | 2 |
| Blank control | 1 | 2 | 1 |
It is shown by ectopic osteogenesis experiment X-ray in Fig. 2 Mice Body, with the PLGA microballoon prepared by the present invention for carrying rhBMP-2
Bone renovating material is tested for Mouse Muscle bag skeletonization, is seen within 1 month after injection new osteogenic, it was demonstrated that the present invention carries rhBMP-
2 PLGA microballoon bone renovating material has good bone formation performance.
Claims (5)
1. a kind of bone renovating material microballoon for carrying rhBMP-2, it is characterised in that recombinate bone morphogenetic protein-including rhBMP-2(
2) it is wrapped in poly (glycolide-lactide) microballoon with poly (glycolide-lactide) (PLGA) microballoon, rhBMP-2, the surface sea-mussel mucin of microballoon
It is modified.
2. a kind of bone renovating material microballoon for carrying rhBMP-2 according to claim 1, it is characterised in that the mussel glues egg
White and PLGA microballoon mass ratio is 1:80-120.
3. a kind of preparation method of bone renovating material microballoon for carrying rhBMP-2 according to claim 1, it is characterised in that packet
Include following steps:
(1) the PLGA microballoon preparation of rhBMP-2 is loaded；(2) sea-mussel mucin is aggregated to the PLGA microballoon of load rhBMP-2
On.
4. a kind of preparation method of bone renovating material microballoon for carrying rhBMP-2 according to claim 3, it is characterised in that step
Suddenly (1): rhBMP-2 freeze-dried powder being dissolved in hac buffer, the acetonitrile containing PLGA, 3000rpm revolving speed cream is then added
Change 30 seconds, be added in the mixed emulsion of atoleine and Tween-80, stirred 2-4 hours with the revolving speed of 300rpm or so, is completed
Afterwards, 30 minutes are stood, then is lyophilized after alternately being washed with petroleum ether and isopropanol, obtain dry microballoon.
5. a kind of preparation method of bone renovating material microballoon for carrying rhBMP-2 according to claim 3, it is characterised in that step
Suddenly the PLGA microballoon for loading rhBMP-2 is suspended in aqueous acetic acid by (2), takes sea-mussel mucin solution tune PH to 6.0,2-8
It is slowly stirred addition PLGA microsphere suspension liquid thereto under the conditions of DEG C to evenly spread in sea-mussel mucin solution, is slowly stirred 3-
4 hours, stand 12 hours or more, resulting load rhBMP-2 sea-mussel mucin-PLGA microballoon petroleum ether and isopropanol are alternately washed
It is lyophilized after washing, obtains the PLGA microballoon that dry surface is modified by sea-mussel mucin.
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| CN112891636A (en) * | 2021-01-30 | 2021-06-04 | 复旦大学 | Composite biological material with temperature-sensitive and slow-release functions and preparation method and application thereof |
| CN114209890A (en) * | 2021-11-19 | 2022-03-22 | 浙江瑞谷生物科技有限公司 | Bone repair material with strong bone affinity and preparation method thereof |
| CN114404663A (en) * | 2022-01-07 | 2022-04-29 | 浙江瑞谷生物科技有限公司 | Bone repair material with inflammation relieving characteristic and preparation method thereof |
| CN114404664A (en) * | 2022-02-17 | 2022-04-29 | 浙江瑞谷生物科技有限公司 | Bone repair scaffold material with long-acting slow release function and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120253470A1 (en) * | 2011-03-30 | 2012-10-04 | President And Fellows Of Harvard College | Compositions for bone tissue repair and uses thereof |
| CN102755669A (en) * | 2012-07-16 | 2012-10-31 | 姚琦 | Preparation method and application of fibrin glue composite recombinant human bone morphogenetic protein-2 (rhBMP-2) microsphere |
| CN107569714A (en) * | 2017-08-18 | 2018-01-12 | 中国人民解放军第四军医大学 | A kind of preparation method of functionalization fracture adhesive |
| CN108144127A (en) * | 2018-01-25 | 2018-06-12 | 南京医科大学附属口腔医院 | Fibrin gel/poly lactic-co-glycolic acid microsphere support and its preparation method and application |
-
2019
- 2019-09-20 CN CN201910896495.3A patent/CN110522946B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120253470A1 (en) * | 2011-03-30 | 2012-10-04 | President And Fellows Of Harvard College | Compositions for bone tissue repair and uses thereof |
| CN102755669A (en) * | 2012-07-16 | 2012-10-31 | 姚琦 | Preparation method and application of fibrin glue composite recombinant human bone morphogenetic protein-2 (rhBMP-2) microsphere |
| CN107569714A (en) * | 2017-08-18 | 2018-01-12 | 中国人民解放军第四军医大学 | A kind of preparation method of functionalization fracture adhesive |
| CN108144127A (en) * | 2018-01-25 | 2018-06-12 | 南京医科大学附属口腔医院 | Fibrin gel/poly lactic-co-glycolic acid microsphere support and its preparation method and application |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112891636A (en) * | 2021-01-30 | 2021-06-04 | 复旦大学 | Composite biological material with temperature-sensitive and slow-release functions and preparation method and application thereof |
| CN112891636B (en) * | 2021-01-30 | 2022-04-01 | 复旦大学 | Composite biological material with temperature-sensitive and slow-release functions and preparation method and application thereof |
| CN114209890A (en) * | 2021-11-19 | 2022-03-22 | 浙江瑞谷生物科技有限公司 | Bone repair material with strong bone affinity and preparation method thereof |
| CN114404663A (en) * | 2022-01-07 | 2022-04-29 | 浙江瑞谷生物科技有限公司 | Bone repair material with inflammation relieving characteristic and preparation method thereof |
| CN114404664A (en) * | 2022-02-17 | 2022-04-29 | 浙江瑞谷生物科技有限公司 | Bone repair scaffold material with long-acting slow release function and preparation method and application thereof |
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