CN1631455A - Vascularization promoting tissue engineering bone and its in vitro construction method - Google Patents
Vascularization promoting tissue engineering bone and its in vitro construction method Download PDFInfo
- Publication number
- CN1631455A CN1631455A CN 200510002378 CN200510002378A CN1631455A CN 1631455 A CN1631455 A CN 1631455A CN 200510002378 CN200510002378 CN 200510002378 CN 200510002378 A CN200510002378 A CN 200510002378A CN 1631455 A CN1631455 A CN 1631455A
- Authority
- CN
- China
- Prior art keywords
- collagen gel
- growth factor
- tissue
- engineered bone
- somatomedin
- 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.)
- Granted
Links
Landscapes
- Materials For Medical Uses (AREA)
Abstract
The invention discloses a tissue engineering bone, wherein the tissue engineering bone constructed in vitro is provided with a plurality of cavities, the cavities are embedded with collagen gel, the collagen gel comprises 50-200 ug/ml of blood vessel endocytosis growth factor, 10-100ug/ml of alkaline desmocyte growth factor, and 400-2000 U/ml of heparinum.
Description
Technical field
The biological substitution thing that the present invention relates to a kind of osseous tissue is a tissue engineered bone, is tissue engineered bone and external structure method that vascularization promoting is handled specifically.
Background technology
Since damaged clinical very common because of the bone due to wound or the osteopathia at orthopaedics, all there is open defect and have treatment means now, make that the progress of application organizes engineering technology repairing bone defect is rapid in recent years.Since Vacanti in 1993 etc. utilize cattle osteoblast and polyglycolic acid to make up tissue-engineered bone, research to experimental projects such as repairing in the seed cell in the bone tissue engineer, timbering material, somatomedin, condition of culture, the body has achieved initial success, Quarto, Yang Zhiming etc. try out this technology in clinical, obtain initial success.The structure basic skills of tissue engineered bone is: on the compound timbering material that is inoculated into as extracellular matrix of the seed cell-osteoblast of In vitro culture, through in the body or In vitro culture be built into tissue engineered bone, implant and carry out the damaged reparation of bone.
The good reparation that bone is damaged must rely on sufficient blood and supply, new vessels is not only the passage of oxygen, nutrient substance and metabolic waste transportation, and be the cell that participates in bone reparation regulation and control, the important path that signaling molecule passes through, i.e. vascularization and osteanagenesis are two links the most basic in the knitting process.The tissue-engineered bone of external structure, must set up sufficient blood after implanting equally rapidly supplies, for the seed cell functional activity provides prescribing adequate nutrition, only in this way could guarantee the activity of tissue-engineered bone and promote the bone defect repair, this seems particularly important when finishing than the DAGU defect repair.
The technical measures of at present short artificial bone vascularization mainly contain 6 kinds.1. wrap up compound osteoblastic biodegradable stent material with vascular bundle, neovascularity is grown in the twig mode in the material; 2. artificial bone is implanted and had in the muscular tissue of axle center blood vessel the tissue-engineered bone flesh bone lobe of pre-structure vascular pedicle; 3. during implanting tissue through engineering approaches bone, the fascial flap of zone of preference vessel pedicle parcel graft nearby; These are to use the vascularization that the blood vessel condition that has existed in the body promotes artificial bone, and existing increases wound, is subjected to shortcoming such as local vascular condition restriction, thereby have limited its clinical application.4. vascular endothelial cell is unified into osteocyte and timbering material is compound; Existence is from the body vascular endothelial cell is difficult for obtaining, amplification in vitro is limited in one's ability deficiency.5. use the somatomedin of short angiogenesis,, adopt the growth factor slow-release technology that evenly is adsorbed in the timbering material, perhaps promote emiocytosis by the gene transfection technology as vascular endothelial cell growth factor (VEGF); Also there is shortcoming in these class methods, and as because somatomedin is the sequential effect of network type to the effect of angiogenesis, and monofactor low concentration action effect is limited, and high concentration VEGF effect has and brings out angiomatous danger.6. attempt the use in conjunction of said method.So, above-mentioned six kinds of vascularization effects that still can not obtain gratifying artificial bone.
Summary of the invention
Deficiency at existing tissue-engineered bone vascularization promoting technology, the object of the present invention is to provide a kind of tissue engineered bone of new vascularization promoting, have lasting, the gentle slow release of limitation after this tissue engineered bone is transplanted and put angiogenic growth factor, start fast when realizing angiogenesis and osteanagenesis, thus the advantage of obvious accelerated bone healing.
In order to achieve the above object, the present invention adopts following technical scheme: on the systematism engineering bone of external structure a plurality of holes are arranged, be embedded with collagen gel in hole, contain the somatomedin of short angiogenesis in collagen gel.
The somatomedin of described short angiogenesis can adopt vascular endothelial cell growth factor, and the weight content in collagen gel is 50～200 μ g/ml.
The basic fibroblast growth factor that can also contain 10～100 μ g/ml in the described collagen gel, form twin long factor combined effect with described vascular endothelial cell growth factor, than adopting single vascular endothelial cell growth factor can better promote the generation of vascular tissue.
Heparin (the standard unit that U is medically commonly used that in collagen gel, can also contain 400～2000U/ml, 125U is equivalent to the dosage of 1mg), as slow releasing agent control growing factor rate of release, as required by adopting different heparin consumptions to design the generating rate of vascular tissue easily.
The optimum content of described vascular endothelial cell growth factor in collagen gel is 100 μ g/ml.
The optimum content of described basic fibroblast growth factor in collagen gel is 50 μ g/ml.
The optimum content of described heparin in collagen gel is 500U/ml.
The present invention also provides the external structure method of the tissue engineered bone of this vascularization promoting, comprising:
(1) external preparation is used for the timbering material of tissue construction: preparation is used for the timbering material that tissue engineered bone makes up, and a plurality of holes are being reserved at the position of composite growth factor;
(2) external structure of tissue engineered bone: the respective seed cell and the timbering material of external a large amount of amplifications is compound, then in external static state or dynamically cultivate certain hour, obtain tissue-engineered bone;
(3) slow release of somatomedin is handled: will urge that the somatomedin of angiogenesis is compound goes in the collagen gel;
(4) embedding of somatomedin: the polylith gel that will contain somatomedin embeds in the reservation hole of tissue-engineered bone of external structure.
In the said method, the somatomedin of described short angiogenesis can be vascular endothelial cell growth factor, and its content in collagen gel is 50～200 μ g/ml; Or the vascular endothelial cell growth factor and the composite twin long factor of basic fibroblast growth factor, wherein vascular endothelial cell growth factor content in collagen gel is 50～200 μ g/ml, and the content of basic fibroblast growth factor in collagen gel is 10～100 μ g/ml.
The heparin that in described collagen gel, can also contain 400～2000U/ml.
In the said method, the optimum content of described vascular endothelial cell growth factor in collagen gel is 100 μ g/ml.
In the said method, the optimum content of described basic fibroblast growth factor in collagen gel is 50 μ g/ml.
In the said method, the optimum content of described heparin in collagen gel is 500U/ml.
The tissue engineered bone of vascularization promoting of the present invention has following beneficial effect owing to be compounded with the somatomedin of short angiogenesis in some regional area holes of systematism engineering bone:
(1) somatomedin compound zone in tissue engineered bone is multiple-point local mode, has avoided the adverse effect of even compound tense to osteogenic response in the tissue engineered bone.
(2) when basic fibroblast growth factor is introduced, make itself and vascular endothelial cell growth factor use in conjunction, coordinate to promote angiogenesis, avoided monofactor dosage to cross curative effect deficiency when low, and dosage more defective of complication when too high.
(3) collagen gel is promptly as the carrier of somatomedin, simultaneously as slow-release material, local release of somatomedin and action time have been prolonged, when the heparin that also is compounded with in the collagen gel as slow releasing agent, variation by the heparin consumption can design the local time that discharges and act on of somatomedin more easily, and vascular tissue is better grown.
(4) construction method of the tissue engineered bone of application vascularization of the present invention makes tissue engineered bone in external structure, combines with the local complex method of somatomedin, by the local concentration and the recombination site of control production factor, can effectively control the formation of vascularization.
(5) compare with prior art, little to patient's response to traume when transplanting by blood vessel tissue's engineering bone of this technique construction, do not damage partial other important blood vessels.
(6) tissue engineered bone along with finishing of tissue repair reconstruction, after local vascular rolls up in the early stage, can reduce gradually in the later stage, thereby coordinate mutually with the local organization function after transplanting.
The specific embodiment
Be described further below in conjunction with the tissue engineered bone and the external structure method thereof of specific embodiment, to help understanding content of the present invention vascularization promoting of the present invention.
Embodiment 1
(1) extracorporeal treatment is used for the timbering material that osseous tissue makes up: preparation or purchase skeletonization timbering material (deproteinization bone derived material, hydroxyapatite, tricalcium phosphate etc.), holed in the position of composite growth factor and reserving a plurality of holes, pore diameter is 2～8mm;
(2) external structure of tissue engineered bone: utilize existing tissue engineering technique that the respective seed cell and the timbering material of external a large amount of amplifications is compound, utilize prior art in external static state or dynamically cultivate certain hour then, obtain tissue-engineered bone;
(3) slow release of somatomedin is handled: vascular endothelial cell growth factor, basic fibroblast growth factor and heparin are pressed mediation, compoundly then go in the collagen gel, make that the content of somatomedin and heparin is VEGF100 μ g/ml, bFGF50 μ g/ml, heparin 500U/ml in the collagen gel after compound;
(4) embedding of somatomedin: the polylith gel that will contain somatomedin embeds in the reservation hole of tissue-engineered bone of external structure.
Embodiment 2
(1) external preparation is used for the timbering material of tissue construction: preparation is used for the timbering material that tissue engineered bone makes up, and is being holed in the position of composite growth factor and is reserving a plurality of holes, and pore diameter is 2～8mm;
(2) external structure of tissue engineered bone: the respective seed cell and the timbering material of external a large amount of amplifications is compound, then in external static state or dynamically cultivate certain hour, obtain tissue-engineered bone;
(3) slow release of somatomedin is handled: vascular endothelial cell growth factor, basic fibroblast growth factor and heparin are pressed mediation, compoundly then go in the collagen gel, make that the content of somatomedin and heparin is VEGF200 μ g/ml, bFGF10 μ g/ml, heparin 400U/ml in the collagen gel;
(4) embedding of somatomedin: the polylith gel that will contain somatomedin embeds in the reservation hole of tissue-engineered bone of external structure.
Embodiment 3
(1) external preparation is used for the timbering material of tissue construction: preparation is used for the timbering material that tissue engineered bone makes up, and is being holed in the position of composite growth factor and is reserving a plurality of holes, and pore diameter is 2～8mm;
(2) external structure of tissue engineered bone: the respective seed cell and the timbering material of external a large amount of amplifications is compound, then in external static state or dynamically cultivate certain hour, obtain tissue-engineered bone;
(3) slow release of somatomedin is handled: vascular endothelial cell growth factor, basic fibroblast growth factor and heparin are pressed mediation, compoundly then go in the collagen gel, make that the content of somatomedin and heparin is VEGF200 μ g/ml, bFGF100 μ g/ml, heparin 2000U/ml in the collagen gel;
(4) embedding of somatomedin: the polylith gel that will contain somatomedin embeds in the reservation hole of tissue-engineered bone of external structure.
Embodiment 4
(1) external preparation is used for the timbering material of tissue construction: preparation is used for the timbering material that tissue engineered bone makes up, and is being holed in the position of composite growth factor and is reserving a plurality of holes, and pore diameter is 2～8mm;
(2) external structure of tissue engineered bone: the respective seed cell and the timbering material of external a large amount of amplifications is compound, then in external static state or dynamically cultivate certain hour, obtain tissue-engineered bone;
(3) slow release of somatomedin is handled: vascular endothelial cell growth factor, basic fibroblast growth factor and heparin are pressed mediation, compoundly then go in the collagen gel, make that the content of somatomedin and heparin is VEGF50 μ g/ml, bFGF30 μ g/ml, heparin 600U/ml in the collagen gel;
(4) embedding of somatomedin: the polylith gel that will contain somatomedin embeds in the reservation hole of tissue-engineered bone of external structure.
Tissue engineered bone among the embodiment 1～4 is implanted into rabbit ulna stage casing or sheep middle tibia bone defect area, take the photograph sheet and transplant regional histopathologic analysis, vascular perfusion experiment etc. through the X line in postoperative 3 days, 2 weeks, 4 weeks, 12 weeks, 18 weeks, find: 2 weeks of postoperative, interior a large amount of cell aggregation with vascular endothelial cell phenotype was in the collagen gel zone, and produce a large amount of netted blood vessels, postoperative 4～12 all blood vessel diameters increase, but quantity reduces gradually, vascularity that 12～18 weeks of postoperative should the zone and the contiguous no significant difference of organizing; The tissue engineered bone of compound angiogenic material and single tissue engineered bone be the bone repairing effect relatively, as seen the former after surgery during 2 weeks skeletonization movable just strong than the latter, 4 weeks of postoperative begin its callus and the bone repairing effect all is better than transplanting single tissue engineered bone.
Embodiment 5
(1) extracorporeal treatment is used for the timbering material that osseous tissue makes up: preparation or purchase skeletonization timbering material (deproteinization bone derived material, hydroxyapatite, tricalcium phosphate etc.), holed in the position of composite growth factor and reserving a plurality of holes, pore diameter is 2～8mm;
(2) external structure of tissue engineered bone: utilize existing tissue engineering technique that the respective seed cell and the timbering material of external a large amount of amplifications is compound, utilize prior art in external static state or dynamically cultivate certain hour then, obtain tissue-engineered bone;
(3) slow release of somatomedin is handled: go in the collagen gel vascular endothelial cell growth factor is compound, making the content of somatomedin in the collagen gel after compound is 200 μ g/m;
(4) embedding of somatomedin: the polylith gel that will contain somatomedin embeds in the reservation hole of tissue-engineered bone of external structure.
Embodiment 6
(1) extracorporeal treatment is used for the timbering material that osseous tissue makes up: preparation or purchase skeletonization timbering material (deproteinization bone derived material, hydroxyapatite, tricalcium phosphate etc.), holed in the position of composite growth factor and reserving a plurality of holes, pore diameter is 2～8mm;
(2) external structure of tissue engineered bone: utilize existing tissue engineering technique that the respective seed cell and the timbering material of external a large amount of amplifications is compound, utilize prior art in external static state or dynamically cultivate certain hour then, obtain tissue-engineered bone;
(3) slow release of somatomedin is handled: vascular endothelial cell growth factor, basic fibroblast growth factor are in harmonious proportion, compoundly then go in the collagen gel, making the content of somatomedin in the collagen gel after compound is VEGF100 μ g/ml, bFGF50 μ g/ml;
(4) embedding of somatomedin: the polylith gel that will contain somatomedin embeds in the reservation hole of tissue-engineered bone of external structure.
With embodiment 5, tissue engineered bone in 6 is implanted into rabbit ulna stage casing bone defect area, process postoperative 3 days, 2 weeks, 4 weeks, 12 weeks, the X line in 18 weeks is taken the photograph sheet, with the histopathologic analysis of transplanting the zone, vascular perfusion experiment etc., find: speed that its short intravascular generates and microvessel density all are lower than implements 1～4 effect, but ratio not single organization's engineering bone of compound short angiogenesis growth factor demonstrates stronger short angiogenesis function, after the concentration of a large amount of raising vascular endothelial cell growth factor, then the downgrade of local vascular generation can produce hemangioma sometimes, side effect such as tissue edema.
Claims (10)
1, a kind of tissue engineered bone of vascularization promoting is characterized in that: on the systematism engineering bone of external structure a plurality of holes are arranged, be embedded with collagen gel at hole, contain the somatomedin of short angiogenesis in collagen gel.
2, the tissue engineered bone of vascularization promoting as claimed in claim 1 is characterized in that: the somatomedin of the short angiogenesis that contains in the described collagen gel is a vascular endothelial cell growth factor, and its content in collagen gel is 50～200 μ g/ml.
3, the tissue engineered bone of vascularization promoting as claimed in claim 2 is characterized in that: the somatomedin at described short angiogenesis also comprises basic fibroblast growth factor, and its content in collagen gel is 10～100 μ g/ml.
4, the tissue engineered bone of vascularization promoting as claimed in claim 3 is characterized in that: the heparin that also contains 400～2000U/ml in the described collagen gel.
5, as the tissue engineered bone of claim 2 or 3 or 4 described vascularization promotings, it is characterized in that: the content of described vascular endothelial cell growth factor in collagen gel is 100 μ g/ml.
6, as the tissue engineered bone of claim 3 or 4 described vascularization promotings, it is characterized in that: the content of described basic fibroblast growth factor in collagen gel is 50 μ g/ml.
7, the tissue engineered bone of vascularization promoting as claimed in claim 4 is characterized in that: the content of described heparin in collagen gel is 500U/ml.
8, the external structure method of the tissue engineered bone of the vascularization promoting described in a kind of claim 1 comprises:
(1) external preparation is used for the timbering material of tissue construction: preparation is used for the timbering material that tissue engineered bone makes up, and a plurality of holes are being reserved at the position of composite growth factor;
(2) external structure of tissue engineered bone: the respective seed cell and the timbering material of external a large amount of amplifications is compound, then in external static state or dynamically cultivate certain hour, obtain tissue-engineered bone;
(3) slow release of somatomedin is handled: will urge that the somatomedin of angiogenesis is compound goes in the collagen gel;
(4) embedding of somatomedin: the polylith gel that will contain somatomedin embeds in the reservation hole of tissue-engineered bone of external structure.
9, the external structure method of the tissue engineered bone of vascularization promoting as claimed in claim 8 is characterized in that: the somatomedin of described short angiogenesis is a vascular endothelial cell growth factor, and its content in collagen gel is 50～200 μ g/ml; Or the vascular endothelial cell growth factor and the composite twin long factor of basic fibroblast growth factor, wherein vascular endothelial cell growth factor content in collagen gel is 50～200 μ g/ml, and the content of basic fibroblast growth factor in collagen gel is 10～100 μ g/ml.
10, the external structure method of the tissue engineered bone of vascularization promoting as claimed in claim 9 is characterized in that: the heparin that also contains 400～2000U/ml in described collagen gel.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510002378 CN1284606C (en) | 2005-01-19 | 2005-01-19 | Vascularization promoting tissue engineering bone and its in vitro construction method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510002378 CN1284606C (en) | 2005-01-19 | 2005-01-19 | Vascularization promoting tissue engineering bone and its in vitro construction method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1631455A true CN1631455A (en) | 2005-06-29 |
| CN1284606C CN1284606C (en) | 2006-11-15 |
Family
ID=34852954
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200510002378 Expired - Fee Related CN1284606C (en) | 2005-01-19 | 2005-01-19 | Vascularization promoting tissue engineering bone and its in vitro construction method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1284606C (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102895704A (en) * | 2012-07-24 | 2013-01-30 | 华东理工大学 | Biological activity factor composition for promoting ossification and vascularization |
| CN105561395A (en) * | 2016-01-05 | 2016-05-11 | 广州医科大学附属第三医院 | Construction and application of coaxial type vascularized tissue-engineered bone |
| CN107349472A (en) * | 2017-06-30 | 2017-11-17 | 浙江德康医疗器械有限公司 | A kind of preparation method of the gradient porous titanium alloy of repetition of promotion bone fusion |
-
2005
- 2005-01-19 CN CN 200510002378 patent/CN1284606C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102895704A (en) * | 2012-07-24 | 2013-01-30 | 华东理工大学 | Biological activity factor composition for promoting ossification and vascularization |
| CN102895704B (en) * | 2012-07-24 | 2016-05-11 | 华东理工大学 | A kind of bioactie agent composition that promotes skeletonization and vascularization |
| CN105561395A (en) * | 2016-01-05 | 2016-05-11 | 广州医科大学附属第三医院 | Construction and application of coaxial type vascularized tissue-engineered bone |
| CN105561395B (en) * | 2016-01-05 | 2018-08-21 | 广州医科大学附属第三医院 | A kind of structure of close coupled type study of vascularized tissue engineering bone and its application |
| CN107349472A (en) * | 2017-06-30 | 2017-11-17 | 浙江德康医疗器械有限公司 | A kind of preparation method of the gradient porous titanium alloy of repetition of promotion bone fusion |
| CN107349472B (en) * | 2017-06-30 | 2020-06-30 | 浙江德康医疗器械有限公司 | Preparation method of repeated gradient porous titanium alloy for promoting bone fusion |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1284606C (en) | 2006-11-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105641753B (en) | A kind of 3D printing biodegradable stent of the achievable blood vessel transfer of compound rhBMP-2 | |
| CN108310467B (en) | Assembled cell-derived extracellular matrix membrane composite bone repair material and preparation method and application thereof | |
| CN102824657B (en) | Bone restoration material and preparation method thereof | |
| CN101568637B (en) | Substrate for culture of cardiovascular tissue | |
| CN104780850B (en) | For by flexible member, particularly natural or synthetic ligament or tendon to be fixed to the equipment on bone | |
| CN1684589A (en) | Vascularization enhanced graft constructs | |
| MX2012008215A (en) | Bioengineered tissue constructs and methods for producing and using thereof. | |
| CN101147810B (en) | Cell-biodegradable material compound and its preparation method and application | |
| CN1713861A (en) | Vascularization enhanced graft constructs | |
| CN1668732A (en) | Treatments with autologous fibroblast | |
| Griffon et al. | Evaluation of vacuum and dynamic cell seeding of polyglycolic acid and chitosan scaffolds for cartilage engineering | |
| EP2035545A2 (en) | Bio-membrane for tissue regeneration | |
| Olender et al. | Revitalization of biostatic tissue allografts: new perspectives in tissue transplantology | |
| CN1284606C (en) | Vascularization promoting tissue engineering bone and its in vitro construction method | |
| Zhang et al. | Biomaterials for periodontal tissue regeneration | |
| Tong et al. | Synthesis of the new-type vascular endothelial growth factor–silk fibroin–chitosan three-dimensional scaffolds for bone tissue engineering and in vitro evaluation | |
| Weng et al. | Engineering of axially vascularized bone tissue using natural coral scaffold and osteogenic bone marrow mesenchymal stem cell sheets | |
| Dong et al. | Polymer scaffolds-enhanced bone regeneration in osteonecrosis therapy | |
| CN101195044A (en) | Tissue engineered fine particle tissue and method for preparing the same | |
| CN112076350B (en) | Biomimetic mineralized hydrogel with nano-micron composite structure and high mineral density as well as preparation method and application thereof | |
| CN1580255A (en) | Tissue engineered peripheral nerve graft | |
| Liu et al. | Recent advancement in vascularized tissue-engineered bone based on materials design and modification | |
| CN103372233A (en) | Preparation method and product of tissue-engineered cartilage graft | |
| CN205569387U (en) | Degradable support of load rhBMP -2 slow release system's blood vessel realized transfer | |
| CN107335096A (en) | A kind of oral cavity sticking patch preparation method of compound oral mucosa epithelial cell |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |