CN203252767U - Tissue engineering blood vessel - Google Patents
Tissue engineering blood vessel Download PDFInfo
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- CN203252767U CN203252767U CN2013202401125U CN201320240112U CN203252767U CN 203252767 U CN203252767 U CN 203252767U CN 2013202401125 U CN2013202401125 U CN 2013202401125U CN 201320240112 U CN201320240112 U CN 201320240112U CN 203252767 U CN203252767 U CN 203252767U
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- blood vessel
- support
- gelatin
- polycaprolactone
- engineering blood
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Abstract
The utility model relates to a tissue engineering blood vessel. A hollow tubular support is arranged in the tissue engineering blood vessel, cells are evenly distributed on the pipe wall, a supporting body is arranged in the hollow inside of the support, the support is manufactured by gelatin/polycaprolactone electricity texture films, and the supporting body can be a PVC pipe. The tissue engineering blood vessel has the advantages that the cells are evenly distributed in the support so as to be closer to the living body blood vessel characteristics, the supporting is manufactured by the gelatin/polycaprolactone electricity texture films so as to be good in supporting function and cytocopatibility, the supporting body is arranged in the support to achieve the supporting and guiding function and help shape the blood vessel shape, and the tissue engineering blood vessel is appropriate in blood elasticity and hardness, is clear in self tissue boundary, compact in tissue structure and excellent in all kinds of performance.
Description
Technical field
This utility model relates to a kind of engineered tissue of implant into body, specifically, is a kind of engineering blood vessel.
Background technology
Annual approximately newborn 150,000 the children with congenital heart disease of China, and need a large amount of blood vessels or biological sticking patch substitute in the congenital heart disease operation.Simultaneously, other vascular lesions such as cardiovascular and cerebrovascular disease, many times all need to take vascular replacement or the treatment of putting up a bridge.The blood vessel that is usually used at present replacing or puts up a bridge is the artificial blood vessel who makes with synthetic materials such as nylon, terylene, polytetrafluoroethylene, implant into body shows without growth property and causes easily narrow calcification, there is stronger rejection, anticoagulant property is not good enough, pogoniasis is operative treatment again, bring greatest misery to the patient, cause simultaneously the waste of a large amount of medical resources.And the advantage such as that the engineering blood vessel of function admirable possesses is clear with autologous tissue boundary, have certain elasticity and hardness, organizational structure is fine and close, cell distribution is even can remedy above-mentioned artificial blood vessel's deficiency, and performance is similar to the effect of intravital blood vessel.
At present, gelatin and polycaprolactone are mixed electrospinning become nano fibrous membrane to be widely used in engineered different tissue, such as skin, nerve, bone, blood vessel etc.The people such as Chong [Chong EJ, et al. Acta Biomaterialia 2007,3 (3): 321-330] with the research of gelatin/polycaprolactone blending nano fibrous membrane for skin wound healing, the result shows that prepared gelatin/polycaprolactone fibrous membrane obviously is conducive to adhesion, increment and the differentiation of fibroblast.People [the Laleh Ghasemi-Mobarakeh such as Laleh Ghasemi-Mobarakeh, et al. Biomaterials 2008,29 (34): 4532-4539] gelatin/polycaprolactone electrospinning is become the very high nano fibrous membrane of the depth of parallelism be used for the reparation of nervous tissue, the cultivation results of neural stem cell shows, gelatin/polycaprolactone nano fibrous membrane is a kind of good tissue engineering bracket material, the parallel nanofiber that spins is conducive to increment and the differentiation of neurocyte, and is conducive to the elongation of nerve synapse.People [the Sepideh Heydarkhan-Hagvall such as Sepideh Heydarkhan-Hagvall, et al. Biomaterials 2008,29 (19): 2907-2914] gelatin is become three-dimensional tissue's engineering rack also successfully be used for the cardiovascular organization engineering with the polycaprolactone electrospinning, the result shows that the nano fibrous membrane of blending shows superior mechanical property and the exclusive cellular affinity of natural material that synthetic material has.Its result shows that all gelatin/polycaprolactone is a kind of good timbering material.
When utilizing at present gelatin/polycaprolactone as the timbering material tissue engineering vessel, all be that gelatin mixes the tubular structure that electrospinning becomes to be similar to blood vessel with polycaprolactone usually, then cell directly be planted in the outer surface of support, and further cultivate.Be the simulation intravital blood vessel, it is basically identical that directly electrospinning becomes wall thickness and the intravital blood vessel thickness of timbering material of tubulose, totally thicker, be unsuitable for cell migration and proliferation, again because cell is directly to be planted in the timbering material outer surface, thereby the inside and outside cell distribution of the engineering blood vessel that causes structure is inhomogeneous, and performance is not good.Therefore need the uniform engineering blood vessel of a kind of cell distribution badly, but yet there are no report about this class blood vessel at present.
Summary of the invention
The purpose of this utility model is that a kind of engineering blood vessel is provided.
For achieving the above object, the technical scheme taked of this utility model is:
A kind of engineering blood vessel is provided with support, and described support is hollow tubular, is evenly distributed with cell in the tube wall, and the inside of hollow is provided with supporter.
Described support is straight tube-like body, U-shaped tubular body, C type tubular body or Y-piece shape body.
Described support is made by gelatin/polycaprolactone electrospinning film.
Described supporter is pvc pipe.
This utility model advantage is:
1, cell is to be uniformly distributed in rack inner wall, outer wall and each position, closer to intravital blood vessel;
2, support is made by gelatin/polycaprolactone electrospinning film, possesses good supportive and cell compatibility;
3, be provided with supporter, rise and support and guide effect, help to mould shape of blood vessel;
4, engineering blood vessel cell distribution of the present utility model evenly, elasticity and hardness is suitable, clear with the autologous tissue boundary, organizational structure is fine and close, properties is all excellent.
Description of drawings
Accompanying drawing 1 is engineering blood vessel structural representation of the present utility model.
Accompanying drawing 2 is engineering blood vessel cross-sectional views of the present utility model.
The specific embodiment
Below in conjunction with embodiment and with reference to accompanying drawing this utility model is further described.
Embodiment 1
The Reference numeral and the ingredient that relate in the accompanying drawing are as follows:
1. support 2. cells
3. supporter
Please refer to Fig. 1, Fig. 1 is engineering blood vessel structural representation of the present utility model, and described engineering blood vessel is provided with support 1, and described support 1 is made by gelatin/polycaprolactone electrospinning film, is hollow tubular.Please refer to Fig. 2, Fig. 2 is engineering blood vessel cross-sectional view of the present utility model, and growth has cell 2 in the tube wall of support 1, and described cell 2 is equally distributed.Please refer to Fig. 1, the inside of described support 1 hollow is provided with supporter 3 again, and described supporter 3 is pvc pipe.
Need to prove that thickness and the intravital blood vessel of described support 1 are suitable, support 1 is tubulose, can be straight tube-like body, U-shaped tubular body, C type tubular body or Y-piece shape body, so that personalized the selection; Described cell 2 is to be uniformly distributed in the support 1, and namely the density of the cell 2 at support 1 inwall, outer wall, each position is suitable; Described supporter 3 is mainly used in keeping shape of blood vessel, is convenient to support 1 and cell 2 molding, and supporter 3 also can be selected other materials, such as hard silica gel.
Engineering blood vessel cell distribution of the present utility model evenly, elasticity and hardness is suitable, clear with the autologous tissue boundary, organizational structure is fine and close, properties is all very excellent.
Embodiment 2
1 materials and methods
1.1 the preparation of gelatin/polycaprolactone electrospinning film
Take by weighing the 0.5g gelatin and the 0.5g polycaprolactone is dissolved in the trifluoroethanol of 10ml with electronic analytical balance, stir 24h to fully dissolving, obtaining concentration is the 10%(grams per milliliter) gelatin/polycaprolactone spinning liquid, in solution, add 10 μ l acetic acid, stir 10-20min and become clarification by muddiness to solution.Select the syringe of 10ml, 1.2mm the syringe needle of internal diameter, extract gelatin/polycaprolactone spinning liquid, be fixed on the electrostatic spinning apparatus, the adjustment electrostatic pressure is 10kv, accepting distance is 12cm, injection rate is 2ml/h, carries out electrospinning, and the employing aluminium foil is receiving device, spinning 3h obtains gelatin/polycaprolactone composite nanometer fiber membrane.When making up blood vessel, be cut into rectangle sticks wide 2 centimetres, long 12 centimetres, it carried out vacuum lyophilization disinfect rear for subsequent use.
1.2 human smooth muscle cell is cultivated
Get human umbilical artery, form tissue in superclean bench supernatant expel stagnation, divest tunica adventitia of artery, vertically cut blood vessel open, passivity is struck off lining endothelium, removes endotheliocyte, residue tunica media tissue.After the normal saline that contains penicillin and streptomycin washes repeatedly, repeatedly cut into the piece of tissue of 1mm * 1mm size with the ophthalmology curved scissors, and will shear the good fritter of organizing and evenly put at the bottom of bottle piece of tissue spacing 0.5cm.Build bottle cap, the culture bottle that overturns gently injects the culture fluid that an amount of low sugar DMEM (Hyclone, the U.S.) contains 10% hyclone (FBS) (Hyclone, the U.S.) up and in bottle at the bottom of allowing bottle, places to contain 5% CO
237 ℃ of incubators in place 2-4h make piece of tissue dry and attach with the bottle wall after, culture bottle slowly overturn to be kept flat, and piece of tissue is immersed in the culture fluid fully, continues to leave standstill to cultivate 3-5d.Wait there being cell after piece of tissue is swum out of, to change liquid on every side, after 90% fusion, go down to posterity.
1.3 scanning electron microscopic observation
Smooth muscle cell is collected in digestion, and be planted on a slice gelatin/polycaprolactone electrospinning film, take out complex behind the In vitro culture 1d, fix through 2.5% glutaraldehyde, then Gradient elution using ethanol is dry, ion sprays the instrument metal spraying, carries out scanning electron microscopic observation and takes pictures, and organizes in contrast with simple gelatin/polycaprolactone electrospinning membrane material group.
" 1.4 egg roll " method tissue engineering vessel
With wide 2 centimetres, long 12 centimetres of disinfecting the rectangle gelatin/polycaprolactone electrospinning film is tiled on the culture dish for subsequent use, collect the smooth muscle cell of cultivating with 0.25% trypsinization, in the low sugar DMEM of 10% FBS culture fluid, cell concentration is 10 * 10 with its Eddy diffusion
6/ ml, inoculation 1ml cell suspension makes cell suspension be paved with whole electrospinning film on gelatin/polycaprolactone electrospinning film, is positioned over to contain 5% CO
237 ℃ of incubators cultivate, behind 30min, add a little culture fluid in material surface, spend the night behind culture dish until add the 10ml culture fluid behind the 2h.Second day is that PVC catheter " egg roll " method of 6mm is curled into tubulose with cell material composite electrospun film along diameter, and puts into the 50ml centrifuge tube after leaving standstill half an hour, adds low sugar DMEM, and the culture fluid of 10% FBS covers complex, places to contain 5% CO
237 ℃ of incubators cultivate.Change liquid every 2 days half amounts, cultivate the rear nude mice by subcutaneous transplanting of 1 week, draw materials after 8 weeks.
1.5 the evaluation of the engineering blood vessel that makes up
1.5.1 histology
HE dyeing: with haematoxylin dyeing 5min after the engineering blood vessel section dewaxing aquation that makes up, the hydrochloride alcohol differentiation several seconds after the flushing, blue 30min is returned in from the beginning washing, ethanol dehydration behind Yihong dyeing 1min, the transparent rear resinene mounting of dimethylbenzene.
Masson dyeing: with again washing after the 1% hydrochloride alcohol differentiation after the WeigerShi Garapa element 5-10min washing after the engineering blood vessel section dewaxing aquation that makes up, Ponceaux acid fuchsin liquid dyes 5-10min, 1% phosphomolybdic acid aqueous solution is processed 5min after the washing, green liquor is redyed 5min, 1% glacial acetic acid is processed 1min, 95% dehydration of alcohol repeatedly, the anhydrous alcohol dehydration, dimethylbenzene is transparent, the neutral gum mounting.
1.5.2 biomechanics detects
Cut off after the engineering blood vessel that makes up is cut into the wide ring-type of 3mm, with detecting its mechanical property by omnipotent pulling force detector (Instron-3343) behind its thickness of miking, the calculating elastic modulus.Getting the little porcine aorta that internal diameter is similarly 6mm organizes in contrast.
2 results
2.1 the evaluation of gelatin/polycaprolactone electrospinning film
The gelatin that the Static Spinning method prepares/polycaprolactone electrospinning membrane material has certain mechanical strength, and operability is more intense.The scanning electron microscope result of simple gelatin/polycaprolactone electrospinning membrane material shows that nanofiber is even, smooth, continuous, the bionical preferably fibre fineness of n cell epimatrix collagen.With the compound rear In vitro culture 1d of human smooth muscular cells, the visible Smooth Muscle Cells Adhesion of scanning electron microscope is in this material surface, and cell is sprawled well.
The outer tissue engineering vessel of " 2.2 egg roll " body of laws
First the gelatin that disinfects/polycaprolactone material is tiled in the large culture dish, the collection human umbilical artery smooth muscle cells is inoculated in uniformly gelatin/polycaprolactone electrospinning film and spends the night behind the surface, second day will have been inoculated the gelatin of cell/polycaprolactone electrospinning film and be wrapped on the PVC catheter, the final engineering blood vessel that forms tubulose, then In vitro culture is transplanted to nude mice by subcutaneous after 1 week, makes it further ripe.
2.3 the detection of the engineering blood vessel that makes up
Nude mice by subcutaneous is drawn materials after transplanting for 8 weeks, substantially sees: the engineering blood vessel of structure and the boundary of nude mice autologous tissue are clear, good springiness, and hardness is moderate, and inner wall smooth becomes tubulose but keep flat independent support.HE section display organization compact structure, cell distribution is even, and Masson dyeing shows that collagen fiber are abundant in the engineering blood vessel that makes up.
The above only is preferred implementation of the present utility model; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from this utility model principle; can also make some improvement and replenish, these improvement and replenish and also should be considered as protection domain of the present utility model.
Claims (4)
1. an engineering blood vessel is provided with support, it is characterized in that, described support is hollow tubular, is evenly distributed with cell in the tube wall, and the inside of hollow is provided with supporter.
2. engineering blood vessel according to claim 1 is characterized in that, described support is straight tube-like body, U-shaped tubular body, C type tubular body or Y-piece shape body.
3. engineering blood vessel according to claim 1 is characterized in that, described support is made by gelatin/polycaprolactone electrospinning film.
4. engineering blood vessel according to claim 1 is characterized in that, described supporter is pvc pipe.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013202401125U CN203252767U (en) | 2013-05-07 | 2013-05-07 | Tissue engineering blood vessel |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013202401125U CN203252767U (en) | 2013-05-07 | 2013-05-07 | Tissue engineering blood vessel |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN203252767U true CN203252767U (en) | 2013-10-30 |
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| CN2013202401125U Expired - Fee Related CN203252767U (en) | 2013-05-07 | 2013-05-07 | Tissue engineering blood vessel |
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2013
- 2013-05-07 CN CN2013202401125U patent/CN203252767U/en not_active Expired - Fee Related
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131030 Termination date: 20170507 |
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| CF01 | Termination of patent right due to non-payment of annual fee |