CN104491929A - Preparation method of cell-containing nanofiber bracket - Google Patents
Preparation method of cell-containing nanofiber bracket Download PDFInfo
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- CN104491929A CN104491929A CN201410790675.0A CN201410790675A CN104491929A CN 104491929 A CN104491929 A CN 104491929A CN 201410790675 A CN201410790675 A CN 201410790675A CN 104491929 A CN104491929 A CN 104491929A
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Abstract
The invention relates to a preparation method of a cell-containing nanofiber bracket. The method is capable of realizing the integrated processing of degradable biological materials and cells and the cell-containing nanofiber bracket is obtained by direct high-voltage electronic injection. The preparation comprises the following steps: (1) preparing a water-soluble degradable high-molecular polymer into a homogeneous solution by deionized water, and adding beta-TCP powder, mixing and grinding by a vibrating ball mill; (2) sterilizing the material by high-temperature steam; (3) adding cell suspension into the sterilized material and stirring uniformly; and (4) extracting the cell-containing material in a micro pump by an injector, starting a high-voltage power supply and the micro pump, and setting corresponding technological parameters to carry out electronic injection so as to obtain the cell-containing nanofiber bracket. The preparation method is simple and feasible, and has important practical significance for the quick repair of bone defects.
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 is that a kind of energy of rising in recent years is quick, the method for simple preparation nano fiber scaffold, and the timbering material porosity utilizing this technology to prepare is high, and hole link is good, can be good at the microenvironment building Growth of Cells.Therefore nano fiber scaffold has a wide range of applications at medical domain.The common methods of current structure nano fibrous tissue engineering nano fiber scaffold is: first prepare nano fiber scaffold, then at rack surface repopulating cell.But the organizational project nano fiber scaffold obtained by the method exists following shortcoming: 1) cell seeding density is little, make the neoblastic speed of growth slow.2) tissue generally comprises various kinds of cell, and the distribution of different types of cell fixed point is difficult to realize.3) cell inwardly moves growth by the surface of support, and this one direction growth environment balanced with the three-dimensional of natural fabric cell falls far short, and have impact on the interaction of cell-ECM, cell-scaffold, and the growth of tissue.4) be 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, finally causes the phenomenon of " sandwich " to occur.
Cell in tissue is all in three-D space structure, accepts signal around.This environment is that the growth of cell provides very favourable condition, and balanced nutrition and the exchange of matter energy, make cell constantly breed and secrete the epimatrix of self.How to build this microenvironment in vitro, the combination and permutation of various kinds of cell will be solved in fact exactly, make cell, somatomedin and epimatrix become an organic coalition, allow their interphase interaction thus obtain certain biological function.Therefore, cell and host material being assembled simultaneously shaping is an effective method.
Summary of the invention
The object of the invention is for above problem, 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, for quick reparation medically provides more advanced technical support.
To achieve these goals, the present invention adopts following technical scheme:
Containing a preparation method for cell nano fibrous framework, nano fiber scaffold is made up of degradable high molecular polymer and cell, utilizes electric jet technology one time to produce, comprises the following steps:
1) 1.5-2.0 gram of water solublity degradable high polymer is taken with electronics Libra, slowly join under agitation in the deionized water of 15-20 gram, 80-100 DEG C of accelerate dissolution is warmed up to after fully swelling, and be incubated 3 hours, after obtained homogeneous macromolecular solution, β-TCP the powder adding 0.10-0.20 gram again mixes, and with shaking ball mill grinding 2 hours;
2) by above-mentioned material at 120 DEG C of high-temperature steam sterilizations;
3) stem cell of the 4th generation people is collected in 50mL centrifuge tube with after the trypsinization of 0.25%, centrifugal 5 min of 1000 r/min, abandon supernatant, cell suspension are moved to the above-mentioned material after sterilization, stir, obtain celliferous macromolecule polymer solution;
4) on the syringe needle of the syringe of electric injection device, load the DC voltage of 6-8KV, the distance between syringe needle and right angle rustless steel dash receiver 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 celliferous macromolecule polymer solution to carry out feed with 150-180ul/min regime flow, 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 following apparent outstanding substantive distinguishing features and remarkable advantage:
Living cells is incorporated into inside material and directly carries out nanofiber spinning by the present invention, disposable shaping has celliferous nano fiber scaffold, cell is made to be evenly distributed in nano fiber scaffold structure, be conducive to cytoskeletal signal transmission, be conducive to sticking of cell, also overcome simultaneously and limit cell migration because supporting structure hole is too little.The method possesses that technique is simple, controllability good and efficiency advantages of higher.
Accompanying drawing explanation
Fig. 1 is the celliferous nano fiber scaffold system schematic of preparation.
Detailed description of the invention
Details are as follows by reference to the accompanying drawings for the preferred embodiments of the present invention:
Be illustrated in figure 1 the celliferous nano fiber scaffold system schematic of preparation.Wherein: computer control system 1 connection control device 2, controller 2 connects high voltage power supply 3 and micro pump 7, right angle rustless steel dash receiver 5 is placed on rustless steel backing plate 4, 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
Containing a preparation method for cell nano fibrous framework, nano fiber scaffold is made up of degradable high molecular polymer and cell, utilizes electric jet technology one time to produce, comprises the following steps:
1) slowly join under agitation in the deionized water of 18.24 grams with the PVA granule that electronics sky takes 1.76 grams, about 90 DEG C accelerate dissolution are warmed up to after fully swelling, and be incubated 3 hours, after obtained transparent 8.8%PVA solution, then 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 collected in 50mL centrifuge tube with after the trypsinization of 0.25%, and centrifugal 5 min of 1000 r/min, abandon supernatant, cell suspension is moved to the above-mentioned material after sterilization and stirs and obtain celliferous PVA solution;
4) celliferous for preparation nano fiber scaffold system is connected, the wide 4cm of right angle rustless steel dash receiver 5, thick 1mm, horizontal edge 6cm, vertical edge 6cm in the present embodiment; Syringe needle is 8cm apart from the vertical distance outside right angle rustless steel dash receiver 5 horizontal sides, and lateral separation is 5cm.
Extract 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.
Get the mid portion of nano fibrous membrane after spinning terminates, put in culture dish and add culture fluid and put into incubator again and cultivate.Finally obtain celliferous nano fiber scaffold.
embodiment 2
This example is substantially identical with example 1, difference is: the PEO granule taking 2 grams with electronics Libra slowly joins in the deionized water of 8 grams under agitation, about 60 DEG C accelerate dissolution are warmed up to after fully swelling, and be incubated 3 hours, after obtained transparent 20%PEO solution, then the β-TCP powder adding 0.15 gram carries out mixing concussion ball mill grinding 2 hours.
embodiment 3
This example is substantially identical with example 1, and difference is: the stem cell that step 3) adopts is mesenchymal stem cells MSCs.
Claims (3)
1., containing a preparation method for cell nano fibrous framework, nano fiber scaffold is made up of degradable high molecular polymer and cell, utilizes electric jet technology one time to produce, it is characterized in that, comprise the following steps:
1) 1.5-2.0 gram of water solublity degradable high polymer is taken with electronics Libra, slowly join under agitation in the deionized water of 15-20 gram, 80-100 DEG C of accelerate dissolution is warmed up to after fully swelling, and be incubated 3 hours, after obtained homogeneous macromolecular solution, β-TCP the powder adding 0.10-0.20 gram again mixes, and with shaking ball mill grinding 2 hours;
2) by above-mentioned material at 120 DEG C of high-temperature steam sterilizations;
3) stem cell of the 4th generation people is collected in 50mL centrifuge tube with after the trypsinization of 0.25%, centrifugal 5 min of 1000 r/min, abandon supernatant, cell suspension are moved to the above-mentioned material after sterilization, stir, obtain celliferous macromolecule polymer solution;
4) on the syringe needle of the syringe of electric injection device, load the DC voltage of 6-8KV, the distance between syringe needle and right angle rustless steel dash receiver 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 celliferous macromolecule polymer solution to carry out feed with 150-180ul/min regime flow, spinning 1-3 hour, finally obtains celliferous nano fiber scaffold.
2. the preparation method containing cell nano fibrous framework according to claim 1, is characterized in that: the water solublity degradable high polymer of described step 1) is polyvinyl alcohol or polyethylene glycol oxide.
3. the preparation method containing cell nano fibrous framework according to claim 1, is characterized in that: the stem cell of described step 3) is fat stem cell or mesenchymal stem cells MSCs.
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| CN201410790675.0A CN104491929B (en) | 2014-12-19 | 2014-12-19 | A kind of preparation method containing cell nano fibrous framework |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104874022A (en) * | 2015-05-14 | 2015-09-02 | 上海大学 | Method for preparing nanofiber scaffold by employing composite cell electrical injection technology |
| CN107281548A (en) * | 2016-04-03 | 2017-10-24 | 井冈山大学 | The celliferous nerve trachea preparation method of Y types |
| CN108525020A (en) * | 2017-03-03 | 2018-09-14 | 井冈山大学 | Celliferous more structure-biological membrane preparation methods |
| CN109420199A (en) * | 2017-09-04 | 2019-03-05 | 井冈山大学 | The cell directional bionical Nerve Scaffold preparation method arranged in parallel with microcellular structure |
| CN111910264A (en) * | 2019-05-10 | 2020-11-10 | 井冈山大学 | Preparation method of bionic nerve nanofiber bundle |
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| 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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2014
- 2014-12-19 CN CN201410790675.0A patent/CN104491929B/en not_active Expired - Fee Related
Patent 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 |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104874022A (en) * | 2015-05-14 | 2015-09-02 | 上海大学 | Method for preparing nanofiber scaffold by employing composite cell electrical injection technology |
| CN107281548A (en) * | 2016-04-03 | 2017-10-24 | 井冈山大学 | The celliferous nerve trachea preparation method of Y types |
| CN107281548B (en) * | 2016-04-03 | 2020-08-07 | 井冈山大学 | Preparation method of Y-shaped nerve conduit containing cells |
| CN108525020A (en) * | 2017-03-03 | 2018-09-14 | 井冈山大学 | Celliferous more structure-biological membrane preparation methods |
| CN108525020B (en) * | 2017-03-03 | 2021-12-24 | 井冈山大学 | Preparation method of multi-structure biological membrane containing cells |
| CN109420199A (en) * | 2017-09-04 | 2019-03-05 | 井冈山大学 | The cell directional bionical Nerve Scaffold preparation method arranged in parallel with microcellular structure |
| 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 |
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| CN104491929B (en) | 2016-08-24 |
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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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