CN104491929B - A kind of preparation method containing cell nano fibrous framework - Google Patents
A kind of preparation method containing cell nano fibrous framework Download PDFInfo
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- CN104491929B CN104491929B CN201410790675.0A CN201410790675A CN104491929B CN 104491929 B CN104491929 B CN 104491929B CN 201410790675 A CN201410790675 A CN 201410790675A CN 104491929 B CN104491929 B CN 104491929B
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Abstract
The present invention relates to a kind of preparation method containing cell nano fibrous framework.The method can realize degradable biomaterial and the integrated processing of cell, directly utilizes the celliferous nano fiber scaffold of high pressure EFI.Concretely comprise the following steps: (1) becomes uniform solution with deionized water preparation water solublity degradable high polymer, add β TCP powder and carry out mixing concussion ball mill grinding；(2) by above-mentioned material high-temperature steam sterilization；(3) cell suspension is joined inside the material after above-mentioned sterilization, stir；(4) extract above-mentioned celliferous material with syringe and be placed in micro pump, open high voltage power supply and micro pump, set corresponding technological parameter and carry out EFI, finally obtain celliferous nano fiber scaffold.The whole preparation method of the present invention is simple, quickly repairs Cranial defect etc. and is of great immediate significance.
Description
Technical field
The present invention relates to a kind of preparation method containing cell nano fibrous framework, belong to the preparation of organizational project nano fiber scaffold and reparation field.
Background technology
Method of electrostatic spinning be rising in recent years a kind of energy quickly, the simplicity method of preparing nano fiber scaffold, the timbering material porosity utilizing this technology to prepare is high, and hole link is good, it is possible to well build the microenvironment of cell growth.Therefore nano fiber scaffold has a wide range of applications at medical domain.The common methods building nano fibrous tissue engineering nano fiber scaffold at present is: first prepare nano fiber scaffold, then at rack surface repopulating cell.But the organizational project nano fiber scaffold obtained by the method has the disadvantage in that 1) cell seeding density is little so that and the neoblastic speed of growth is slow.2) tissue generally comprises various kinds of cell, and the fixed point distribution of different types of cell is difficult to.3) cell is grown by the surface inward migration of support, and the growth of this one direction falls far short with the three-dimensional Jun Heng environment of natural fabric cell, have impact on the interaction of cell-ECM, cell-scaffold, and the growth of tissue.4) being not suitable for building large scale support, the cell at heart position makes a difference bad due to the reduction of the penetrating power of liquid in the bracket, causes growth retardation, lacks vascularization, and the phenomenon ultimately resulting in " sandwich " occurs.
Cell in tissue, all in three-D space structure, accepts signal around.This environment is that the growth of cell provides extremely advantageous condition, and the nutrition of equilibrium and the exchange of matter energy make cell constantly breed and secrete the epimatrix of self.The most how to build this microenvironment, the combination and permutation of various kinds of cell to be solved, make cell, somatomedin and epimatrix become an organic coalition, it is allowed to their interphase interaction thus obtain certain biological function.Therefore, cell and host material being assembled shaping simultaneously is an effective method.
Summary of the invention
It is an object of the invention to for problem above, a kind of preparation method containing cell nano fibrous framework is provided, utilize high-tension electricity spray standby, put forth effort on structure and there is biological activity and the equally distributed support of cell, provide the most advanced technical support for quickly reparation medically.
To achieve these goals, the present invention adopts the following technical scheme that
A kind of preparation method containing cell nano fibrous framework, nano fiber scaffold is made up of degradable high molecular polymer and cell, utilizes electric jet technology disposably to prepare, comprises the following steps:
1) 1.5-2.0 gram of water solublity degradable high polymer is weighed with electronics Libra, it is added slowly under agitation in the deionized water of 15-20 gram, it is warmed up to 80-100 DEG C of acceleration dissolving after the most swelling, and it is incubated 3 hours, after preparing homogeneous macromolecular solution, β-TCP the powder adding 0.10-0.20 gram mixes, and with concussion ball mill grinding 2 hours；
2) by above-mentioned material at 120 DEG C of high-temperature steam sterilizations；
3) by the stem cell of the 4th generation people with being collected in 50mL centrifuge tube after the trypsinization of 0.25%, 1000 r/min are centrifuged 5 min, abandon supernatant, cell suspension moves to the above-mentioned material after sterilization, stirs, obtain celliferous macromolecule polymer solution；
4) loading the DC voltage of 6-8KV on the syringe needle of the syringe of electric injection device, the distance between syringe needle and right angle rustless steel reception plate is 7-9cm, and positive pole is carried on the syringe needle of syringe, and negative pole is carried on rustless steel backing plate；Micro pump realizes with 150-180ul/min regime flow, celliferous macromolecule polymer solution is carried out feed, spinning 1-3 hour, finally obtains celliferous nano fiber scaffold.
The water solublity degradable high polymer of described step 1) is polyvinyl alcohol or polyethylene glycol oxide.
The stem cell of described step 3) is fat stem cell or mesenchymal stem cells MSCs.
The present invention compared with prior art, has and the most obviously highlights substantive distinguishing features and remarkable advantage:
Living cells is incorporated into inside material and directly carries out nanofiber spinning by the present invention, disposably it is formed with celliferous nano fiber scaffold, cell is made to be evenly distributed in nano fiber scaffold structure, be conducive to the transmission of cytoskeletal signal, be conducive to sticking of cell, also overcome simultaneously and limit cell migration because supporting structure hole is the least.The method possesses that technique is simple, controllability good and efficiency advantages of higher.
Accompanying drawing explanation
Fig. 1 is for prepare celliferous nano fiber scaffold system schematic.
Detailed description of the invention
It is as follows that the preferred embodiments of the present invention combine detailed description:
It is illustrated in figure 1 and prepares celliferous nano fiber scaffold system schematic.Wherein: computer control system 1 connects controller 2, controller 2 connects high voltage power supply 3 and micro pump 7, and right angle rustless steel receives plate 5 and is placed on rustless steel backing plate 4, and syringe 6 connects micro pump 7, the positive pole of high voltage power supply 3 is carried on the syringe needle of syringe 6, and negative pole is carried on rustless steel backing plate 4.
Embodiment
1
A kind of preparation method containing cell nano fibrous framework, nano fiber scaffold is made up of degradable high molecular polymer and cell, utilizes electric jet technology disposably to prepare, comprises the following steps:
1) weigh the PVA granule of 1.76 grams with electronics sky to be added slowly under agitation in the deionized water of 18.24 grams, it is warmed up to about 90 DEG C acceleration dissolvings after the most swelling, and it is incubated 3 hours, after preparing transparent 8.8%PVA solution, the β-TCP powder adding 0.15 gram carries out mixing concussion ball mill grinding 2 hours；
2) above-mentioned material 120 DEG C is carried out high-temperature sterilization；
3) fat stem cell of the 4th generation people is with being collected in 50mL centrifuge tube after the trypsinization of 0.25%, and 1000 r/min are centrifuged 5 min, abandon supernatant, cell suspension moves to the above-mentioned material after sterilization and is uniformly mixing to obtain celliferous PVA solution；
4) connecting preparing celliferous nano fiber scaffold system, in the present embodiment, right angle rustless steel receives the wide 4cm of plate 5, thick 1mm, horizontal edge 6cm, vertical edge 6cm；The vertical distance that syringe needle receives outside plate 5 horizontal sides away from right angle rustless steel is 8cm, and lateral separation is 5cm.
Extracting the celliferous PVA solution of 8mL with the asepsis injector of 10mL, open high voltage power supply 3, load the DC voltage of 6.5KV, the distance between syringe needle and aseptic rustless steel L-square is adjusted to 8cm.
The feeding speed arranging micro pump is 160ul/min, and spinning prepares nano fiber scaffold in 2 hours.
Spinning takes the mid portion of nano fibrous membrane after terminating, put into and add culture fluid in culture dish and place in incubator and cultivate.Finally obtain celliferous nano fiber scaffold.
Embodiment
2
This example is essentially identical with example 1, difference is: the PEO granule weighing 2 grams with electronics Libra is added slowly in the deionized water of 8 grams under agitation, it is warmed up to about 60 DEG C acceleration dissolvings after the most swelling, and it is incubated 3 hours, after preparing transparent 20%PEO solution, the β-TCP powder adding 0.15 gram carries out mixing concussion ball mill grinding 2 hours.
Embodiment
3
This example is essentially identical with example 1, and difference is: the stem cell that step 3) is used is mesenchymal stem cells MSCs.
Claims (3)
1. the preparation method containing cell nano fibrous framework, nano fiber scaffold is made up of the degradable high molecular polymer of water solublity and cell, utilizes electric jet technology disposably to prepare, it is characterised in that to comprise the following steps:
1) the 1.5-2.0 gram of degradable high molecular polymer of water solublity is weighed with electronics Libra, it is added slowly under agitation in the deionized water of 15-20 gram, it is warmed up to 80-100 DEG C of acceleration dissolving after the most swelling, and it is incubated 3 hours, after preparing homogeneous macromolecular solution, β-TCP the powder adding 0.10-0.20 gram mixes, and with concussion ball mill grinding 2 hours；
2) by above-mentioned material at 120 DEG C of high-temperature steam sterilizations；
3) by the stem cell of the 4th generation people with being collected in 50mL centrifuge tube after the trypsinization of 0.25%, 1000 r/min are centrifuged 5 min, abandon supernatant, cell suspension moves to the above-mentioned material after sterilization, stirs, obtain celliferous macromolecule polymer solution；
4) loading the DC voltage of 6-8KV on the syringe needle of the syringe of electric injection device, the distance between syringe needle and right angle rustless steel reception plate is 7-9cm, and positive pole is carried on the syringe needle of syringe, and negative pole is carried on rustless steel backing plate；Micro pump realizes with 150-180ul/min regime flow, celliferous macromolecule polymer solution is carried out feed, spinning 1-3 hour, finally obtains celliferous nano fiber scaffold.
Preparation method containing cell nano fibrous framework the most according to claim 1, it is characterised in that: the water solublity degradable high polymer of described step 1) is polyvinyl alcohol or polyethylene glycol oxide.
Preparation method containing cell nano fibrous framework the most according to claim 1, it is characterised in that: the stem cell of described step 3) is fat stem cell or mesenchymal stem cells MSCs.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104874022A (en) * | 2015-05-14 | 2015-09-02 | 上海大学 | Method for preparing nanofiber scaffold by employing composite cell electrical injection technology |
| CN107281548B (en) * | 2016-04-03 | 2020-08-07 | 井冈山大学 | Preparation method of Y-shaped nerve conduit containing cells |
| CN108525020B (en) * | 2017-03-03 | 2021-12-24 | 井冈山大学 | Preparation method of multi-structure biological membrane containing cells |
| CN109420199B (en) * | 2017-09-04 | 2022-03-08 | 井冈山大学 | Preparation method of bionic nerve scaffold with directional parallel arrangement and microporous structure of cells |
| CN111910264A (en) * | 2019-05-10 | 2020-11-10 | 井冈山大学 | Preparation method of bionic nerve nanofiber bundle |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101410508A (en) * | 2006-01-27 | 2009-04-15 | 加利福尼亚大学董事会 | Biomimetic scaffolds |
| CN102784413A (en) * | 2012-07-12 | 2012-11-21 | 禹华旭 | Preparation method of composite biological nerve conduit |
| CN103705981A (en) * | 2013-10-10 | 2014-04-09 | 上海师范大学 | Quick preparation method of growth-factor-loading high-cell-load tissue engineering scaffold |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101410508A (en) * | 2006-01-27 | 2009-04-15 | 加利福尼亚大学董事会 | Biomimetic scaffolds |
| CN102784413A (en) * | 2012-07-12 | 2012-11-21 | 禹华旭 | Preparation method of composite biological nerve conduit |
| CN103705981A (en) * | 2013-10-10 | 2014-04-09 | 上海师范大学 | Quick preparation method of growth-factor-loading high-cell-load tissue engineering scaffold |
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Effective date of registration: 20200820 Address after: Room J1377, 3 storeys, 8 blocks, 55 Huiyuan Road, Jiading District, Shanghai, 201815 Patentee after: Shanghai Blue Derivatives Technology Co.,Ltd. Address before: 200444 Baoshan District Road, Shanghai, No. 99 Patentee before: Shanghai University |
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Granted publication date: 20160824 Termination date: 20211219 |