CN100369590C - Process for manufacturing wind-up type liver tissue engineering stent - Google Patents
Process for manufacturing wind-up type liver tissue engineering stent Download PDFInfo
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- CN100369590C CN100369590C CNB2005100960420A CN200510096042A CN100369590C CN 100369590 C CN100369590 C CN 100369590C CN B2005100960420 A CNB2005100960420 A CN B2005100960420A CN 200510096042 A CN200510096042 A CN 200510096042A CN 100369590 C CN100369590 C CN 100369590C
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Abstract
The present invention discloses a technology for manufacturing a winding-up type liver tissue engineering bracket. The technology comprises the following steps of firstly, carrying out the bionic design of the blood vessel system and the liver cell hole of a liver tissue engineering bracket according to the inner microstructure of natural liver tissue, using a fast forming micro complex technology to manufacture the silicon rubber mould of the bionic liver tissue engineering bracket, coating biological materials with good bioavailability and biodegradability on the silicon rubber mould, winding up the mould plate of the liver tissue engineering bracket after the mould is removed to realize the spatial structure of the liver tissue engineering bracket, wetting the spatial liver tissue engineering bracket, and then obtaining the spatial liver tissue engineering bracket with high porous property by water absorption, freezing and drying. In external culture, vascular endothelial cells are planted in the blood vessel net system, and liver cells are planted in the holes of the liver cells. With the degradation of the spatial liver tissue engineering bracket, the vascular endothelial cells form blood vessels, and the liver cells breed and reunite to grow into the liver tissue. The present invention establishes a basis for transforming artificial livers into natural livers.
Description
Technical field
The biology that the invention belongs to medical industry is made the field, relates to a kind of manufacturing process of biodegradable wind-up type liver tissue engineering stent.
Background technology
Liver transplantation is the effective means of the heavy hepatopath of treatment, yet is subjected to the restriction of donor scarcity, and the annual liver transplantation quantity in the whole world is 4-6 ten thousand examples only also; Hepatocyte transplantation can be repaired liver function to a certain extent, but since the restriction of factors such as transplanting approach, cell quantity, local microenvironment and immunological rejection fail so far extensively to carry out clinically; The transition means of external artificial liver support system before as liver transplantation can only be finished simple functions such as filtration, purification, is difficult to provide successive treatment, can't replace whole biochemical actions of human liver; Existing liver tissue engineering scaffold adopts traditional pore method preparation, though comprise the hole that is interconnected, the distribution of uncontrollable hole more can't obtain the multistage micro structure of ordered distribution
Summary of the invention
At the limitation of above-mentioned liver disease means, the present invention uses the ultimate principle of organizational project, and bond material science and engineering have proposed a kind of manufacturing process of wind-up type liver tissue engineering stent.This technology can not only realize the ordered distribution of different types of cell at internal stent, can also make support have maximum porosity, it is the mass exchange ability, and along with the degraded of timbering material, internal stent vasoganglion system and hepatic tissue form gradually, realize the conversion of artificial liver to natural liver, i.e. the through engineering approaches manufacturing of hepatic tissue, thus can solve the contradiction of supply and demand in the liver transplantation operation.
Know-why of the present invention is whether according to vasoganglion system regulating liver-QI cell hole being designed on same template, little manufacturing process of liver tissue engineering scaffold is slightly different.For the isolating liver tissue engineering scaffold of vasoganglion system regulating liver-QI cell hole, need to make half penetrating film, be sandwiched in during winding up between the blood vessel vasoganglion system template regulating liver-QI cell hole template, to prevent the counterdiffusion mutually of hepatocyte and vascular endothelial cell.And for the liver tissue engineering scaffold of vasoganglion system regulating liver-QI cell hole on same template, hepatocyte and vascular endothelial cell are planted in respectively in separately the micro structure behind the winding up, can not move mutually, so just there not be the partly manufacture process of penetrating film.
Little manufacturing process is by may further comprise the steps:
(1) internal microstructure of simulating nature liver is carried out the Bionic Design of liver vessel net system, hepatocyte pore space structure and half penetrating film, makes up liver tissue engineering biomimetic scaffolds cad model;
(2) above-mentioned organizational project biomimetic scaffolds cad model is converted into the rapid shaping file format, utilizes rapid shaping technique to prepare the resin die of vasoganglion system regulating liver-QI cell pore space structure;
(3) with half penetrating film of Bionic Design, utilize lithographic printing to make photomask, utilization mask ultraviolet curing process is made half penetrating film resin mould less than 5 μ m holes;
(4), turn over by little replica technology and to be made as silicon rubber mould with the resin die of vasoganglion system regulating liver-QI cell pore space structure;
(5) biomaterial with biocompatibility and biodegradation character is coated on the silicon rubber mould, and realization has little manufacturing of the rami hepatici shut die plate of vasoganglion system regulating liver-QI cell hole after the demoulding;
(6) with the resin die of half penetrating film, utilization micro-embossing technology impresses out the half penetrating film that contains small hole on biomaterial;
(7) for the support of vasoganglion system regulating liver-QI cell pore space structure on same template, directly winding up get final product the liver tissue engineering scaffold of three-dimensional configuration; For the vasoganglion system regulating liver-QI cell pore space structure support on same template not, must press the order of vasoganglion system frame-half penetrating film-hepatocyte pore space structure support and assemble, winding up obtains the liver tissue engineering scaffold of stereochemical structure then;
(8) winding up is obtained in the liver tissue engineering scaffold immersion distilled water it is fully absorbed water the high hole liver tissue engineering scaffold that adopts freeze drying process to obtain then.
The internal microstructure of described simulating nature liver is natural hepatic tissue micro structure to be reduced to comprise the vasoganglion system, the loose structure of hepatocyte hole and half penetrating film, and the hepatocyte hole is a step structure, guarantees maximum cell adhesion surface area; Vasoganglion pipeline yardstick is between 100-200 μ m, and hepatocyte hole yardstick is between 200-300 μ m, and the hole yardstick less than 5 μ m on the half penetrating film only allows nutrient substance, and oxygen and metabolic waste pass through, and hepatocyte and platelet are had iris action.
The manufacture method of the liver tissue engineering scaffold of described stereochemical structure is, liver tissue engineering scaffold template back side coating degradable biological glue with plane, winding up bonding layer by layer then, and guarantee the complete conducting of vasoganglion system and vasoganglion system regulating liver-QI cell hole uniform distribution in the liver tissue engineering scaffold inside of stereochemical structure.
Described freeze drying process be will be fully the liver tissue engineering scaffold of stereochemical structure of suction put into and be lower than-20 ℃ low temperature environment pre-cooling, making the aqueous water quick freezing is tiny ice crystal grain, controls the uniformity of ice crystal grain size by the control chilling temperature; Again the liver tissue engineering scaffold after the pre-freeze is put into the freeze drying equipment drying, with ice crystal grain distillation wherein, can in three-dimensional liver tissue engineering scaffold, stay equally distributed trickle pore space structure with half penetrating function, by reducing the temperature of low temperature environment, can obtain the more micropore hole structure of small scale.
The three-dimensional liver tissue engineering scaffold that the manufacturing process that the present invention proposes obtains, the three-dimensional manufacturing issue of liver tissue engineering scaffold complexity is converted into the simple two-dimensional manufacturing, simple and easy to do, simultaneously, pore combines with being separated will to make pore-forming (vasoganglion system regulating liver-QI cell hole), both can realize the controlled distribution of cell in support, and can guarantee that again support had maximum mass exchange ability (porous structure).Can realize that different types of cell is in the controlled and orderly distribution of internal stent, high pore structure makes support have maximum mass exchange ability, help the picked-up of nutrient substance and the discharge of metabolic waste, can be hepatocellular growth and breeding best external environment condition is provided, accelerate the formation of hepatic tissue.Along with the degraded of timbering material, internal stent vasoganglion system and hepatic tissue form gradually, realize that artificial liver is that the through engineering approaches manufacturing of hepatic tissue lays the foundation to the conversion of natural liver, thereby can solve the contradiction of supply and demand in the liver transplantation operation.Simultaneously, for different hepatopaths, can extract themselves vascular endothelial cell and healthy hepatocyte and three-dimensional hepatic tissue support and carry out In vitro culture after compound, thereby can realize the customization manufacturing of hepatic tissue.
Description of drawings
Fig. 1 is vasoganglion system and the compound rami hepatici shut die of hepatocyte hole plate structure sketch map;
Fig. 2 is a hole isolating construction sketch map;
Fig. 3 is half penetrating membrane structure sketch map;
Fig. 4 is a vasoganglion system structure sketch map;
Fig. 5 is the winding up process schematic representation of three-dimensional liver tissue engineering scaffold;
Fig. 6 is the freeze drying process sketch map that makes the high holeization of rami hepatici frame.
The present invention is described in further detail below in conjunction with drawings and Examples.
The specific embodiment
Referring to accompanying drawing, according to manufacturing process of the present invention, the micro structure of simulating nature hepatic tissue at first comprises the Bionic Design of the liver tissue engineering scaffold template of vasoganglion system regulating liver-QI cell hole micro structure.Convert the cad model of the liver tissue engineering scaffold template that designs to the rapid shaping file format, prepare resin die with rapid shaping technique; The silicon rubber mould that contains hepatocyte hole and vasoganglion micro structure with this resin die by little replica prepared; For half penetrating film, can at first utilize and xerox the mask that the manufacturing of slabstone technology contains trickle pore space structure, by mask ultraviolet curing process or photoetching process molding impressing mould; The biomaterial that will have good biocompatibility and biodegradation character is coated on the mould, and realization has the manufacturing of the biodegradable rami hepatici shut die plate of vasoganglion system regulating liver-QI cell hole after the demoulding.
Fig. 1 is vasoganglion 2 and hepatocyte hole 1 compound liver tissue engineering scaffold template, Fig. 2, Fig. 3 and Fig. 4 are for being respectively the hepatocyte hole, half penetrating film and the isolating liver tissue engineering scaffold template of vasoganglion, before the winding up, must be at microscopically with the hepatocyte pore space structure, half penetrating film and vasoganglion structure are combined with the bio-medical glue bond with good biocompatibility and biodegradability.On a surface of liver tissue engineering scaffold template, apply bio-medical glue, become to include the three-dimensional rami hepatici frame of multiple micro structure then by the mode winding up of Fig. 5.
In Fig. 6, the three-dimensional liver tissue engineering scaffold that winding up is good immerses fully suction in the distilled water, putting into low temperature environment (＜-20 ℃) general aqueous water wherein then freezing is tiny ice crystal grain, by freeze drying process ice crystal grain is wherein distilled, in three-dimensional liver tissue engineering scaffold, stay trickle pore space structure, promptly realize the high holeization of liver tissue engineering scaffold.The yardstick of micropore hole structure can be controlled by the temperature of adjusting low temperature environment.For the compound liver tissue engineering scaffold of vasoganglion regulating liver-QI cell hole, these micropore hole structures can play the effect of half penetrating film.
The rami hepatici frame that utilizes this high hole is when the external structure hepatic tissue, and the hepatocyte that will have the anchor property is planted in the hepatocyte pore space structure, and vascular endothelial cell is planted in the vasoganglion structure; Put into bioreactor then and carry out cell culture.Culture fluid flows through vasoganglion, and nutrient substance and oxygen are transported to the hepatocyte hole for hepatic cell growth and breeding by micropore hole structure, simultaneously the metabolic noxious substance of hepatocyte is taken away, and provides a favourable living environment to hepatocyte.Along with the degraded of Biodegradable material, new vasoganglion and hepatic tissue generate, thereby realize the through engineering approaches manufacturing of artificial liver tissue.
Claims (4)
1. the manufacturing process of a wind-up type liver tissue engineering stent is characterized in that, this technology may further comprise the steps:
(1) internal microstructure of simulating nature liver is carried out the Bionic Design of liver vessel net system, hepatocyte pore space structure and half penetrating film, makes up liver tissue engineering biomimetic scaffolds cad model;
(2) above-mentioned organizational project biomimetic scaffolds cad model is converted into the rapid shaping file format, utilizes rapid shaping technique to prepare the resin die of vasoganglion system regulating liver-QI cell pore space structure;
(3) with half penetrating film of Bionic Design, utilize lithographic printing to make photomask, utilization mask ultraviolet curing process is made half penetrating film resin mould less than 5 μ m holes;
(4) will contain the resin die of vasoganglion system regulating liver-QI cell pore space structure, and turn over by little replica technology and be made as silicon rubber mould;
(5) biomaterial with biocompatibility and biodegradation character is coated on the silicon rubber mould, and realization has the manufacturing of the rami hepatici shut die plate of vasoganglion system regulating liver-QI cell hole after the demoulding;
(6) with the resin die of half penetrating film, utilization micro-embossing technology impresses out the half penetrating film that contains small hole on biomaterial;
(7) for the support of vasoganglion system regulating liver-QI cell pore space structure on same template, directly winding up get final product the liver tissue engineering scaffold of three-dimensional configuration; For the vasoganglion system regulating liver-QI cell pore space structure support on same template not, must press the order of vasoganglion system frame-half penetrating film-hepatocyte pore space structure support and assemble, winding up obtains the liver tissue engineering scaffold of stereochemical structure then;
(8) liver tissue engineering scaffold that winding up is obtained immerses in the distilled water it is fully absorbed water, and adopts freeze drying process can obtain high hole liver tissue engineering scaffold then.
2. technology as claimed in claim 1, it is characterized in that, the internal microstructure of described simulating nature liver is natural hepatic tissue micro structure to be reduced to comprise the vasoganglion system, the loose structure of hepatocyte hole and half penetrating film, the hepatocyte hole is a step structure, guarantees maximum cell adhesion surface area; Vasoganglion pipeline yardstick is between 100-200 μ m, and hepatocyte hole yardstick is between 200-300 μ m, and the hole yardstick less than 5 μ m on the half penetrating film only allows nutrient substance, and oxygen and metabolic waste pass through, and hepatocyte and platelet are had iris action.
3. technology as claimed in claim 1, it is characterized in that, the manufacture method of the liver tissue engineering scaffold of described stereochemical structure is, liver tissue engineering scaffold template back side coating degradable biological glue with plane, winding up bonding layer by layer then, and guarantee the complete conducting of vasoganglion system and vasoganglion system regulating liver-QI cell hole uniform distribution in three-dimensional bracket inside.
4. technology as claimed in claim 1, it is characterized in that, described freeze drying process is the liver tissue engineering scaffold that fully absorbs water to be put into be lower than-20 ℃ low temperature environment pre-cooling, making the aqueous water quick freezing is tiny ice crystal grain, controls the uniformity of ice crystal grain size by the control chilling temperature; Again the liver tissue engineering scaffold after the pre-freeze is put into the freeze drying equipment drying, with ice crystal grain distillation wherein, can in three-dimensional liver tissue engineering scaffold, stay equally distributed trickle pore space structure with half penetrating function, by reducing the temperature of low temperature environment, can obtain the more micropore hole structure of small scale.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2005100960420A CN100369590C (en) | 2005-09-19 | 2005-09-19 | Process for manufacturing wind-up type liver tissue engineering stent |
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| CNB2005100960420A CN100369590C (en) | 2005-09-19 | 2005-09-19 | Process for manufacturing wind-up type liver tissue engineering stent |
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| CN1768718A CN1768718A (en) | 2006-05-10 |
| CN100369590C true CN100369590C (en) | 2008-02-20 |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102188759A (en) * | 2011-03-11 | 2011-09-21 | 西安交通大学 | Liver tissue engineering scaffold for cell composite culture and preparation method thereof |
| CN102488569B (en) * | 2011-11-15 | 2014-07-23 | 西安交通大学 | Layered manufacturing method of three-dimensional microfluidic porous scaffold |
| CN103120807B (en) * | 2013-01-16 | 2015-03-18 | 西北工业大学 | Preparation method of ice-induced microstructure soft tissue |
| CN104027847B (en) * | 2014-06-20 | 2015-08-26 | 西安交通大学 | A kind of manufacture method of artificial soft tissue's body of subsidiary vasoganglion runner |
| CN105666762A (en) * | 2016-03-01 | 2016-06-15 | 贵州师范大学 | Manufacturing method of individual hip joint holder die |
| CN113350574B (en) * | 2021-05-26 | 2022-11-18 | 泸州国之荣耀酒业有限公司 | Method for weaving patterned liver-like small She Weizu |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002053193A2 (en) * | 2001-01-02 | 2002-07-11 | The Charles Stark Draper Laboratory, Inc. | Tissue engineering of three-dimensional vascularized using microfabricated polymer assembly technology |
| US20030044444A1 (en) * | 2001-07-16 | 2003-03-06 | Prasanna Malaviya | Porous extracellular matrix scaffold and method |
| CN1633950A (en) * | 2004-12-24 | 2005-07-06 | 浙江大学 | Bioreactor for artificial liver |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002053193A2 (en) * | 2001-01-02 | 2002-07-11 | The Charles Stark Draper Laboratory, Inc. | Tissue engineering of three-dimensional vascularized using microfabricated polymer assembly technology |
| US20030044444A1 (en) * | 2001-07-16 | 2003-03-06 | Prasanna Malaviya | Porous extracellular matrix scaffold and method |
| CN1633950A (en) * | 2004-12-24 | 2005-07-06 | 浙江大学 | Bioreactor for artificial liver |
Non-Patent Citations (1)
| Title |
|---|
| Materials Selection and Scaffold Construction for Liver TissueEngineering. Qingling Feng.The 5th pacific rim international conference on advanced materials and processing,Vol.5 . 2004 * |
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