CN106754354B - Dissecting aneurysm of aorta wall shear stress, which induces vascular cell release inflammatory factor, influences the micro flow control chip device of pulmonary epithelial cells function - Google Patents
Dissecting aneurysm of aorta wall shear stress, which induces vascular cell release inflammatory factor, influences the micro flow control chip device of pulmonary epithelial cells function Download PDFInfo
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- CN106754354B CN106754354B CN201611026425.5A CN201611026425A CN106754354B CN 106754354 B CN106754354 B CN 106754354B CN 201611026425 A CN201611026425 A CN 201611026425A CN 106754354 B CN106754354 B CN 106754354B
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/16—Microfluidic devices; Capillary tubes
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/34—Internal compartments or partitions
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
Abstract
The invention belongs to cell biomechanics experimental provision technical fields, disclose a kind of micro flow control chip device of dissecting aneurysm of aorta wall shear stress induction vascular cell release inflammatory factor influence pulmonary epithelial cells function.The micro flow control chip device includes hopper, damper, peristaltic pump, air pressure pump control system, vascular endothelial cell VECs/ smooth muscle cell VSMCs Combined culture chamber, elastic cavity A, elastic cavity B, resistance valves A, resistance valves B, pulmonary epithelial cells PECs culture chamber, silica gel pipeline and threeway；Two culture chambers and its peripheral cell, form two relatively independent circulatory systems, and two circulatory systems realize the information communication of VECs/VSMCs and PECs by sharing reservoir.The present invention can be used successfully to oscillatory shear forces and tensile stress induces vascular cell release inflammatory factor and influences the research of pneumonocyte function.
Description
Technical field
The invention belongs to cell biomechanics experimental provision technical fields, and in particular to one kind is for studying dissection of aorta
Tumor shearing force, which induces vascular cell release inflammatory factor, influences the micro flow control chip device of pulmonary epithelial cells function.
Background technique
Dissecting aneurysm of aorta is also referred to as dissection of aorta, be it is common be also most complicated most dangerous cardiovascular disease it
One.When medial retrogression pathological changes or aortic tunica intima breakage cause aortic tunica intima to tear, blood enters master through breach
Arterial wall destroys middle layer and walks to be about to when inner membrance and outer layer are removed referred to as dissection of aorta along aorta.Acute hypoxia mass formed by blood stasis is
The common complication of patients with acute dissection of aorta undergoing generally started to occur in the 2nd~3 day of morbidity, to facing interlayer at any time
For the patient of rupture, this, which is undoubtedly, makes the matter worse, and seriously threatens patient vitals.So far, dissection of aorta causes acute
The mechanism of hypoxemia is unclear.
Patients with acute aortic dissection generally has no primary lung disease, and Lung Injury Complicating is not intrapulmonary factor
It is caused, but induced by lung other factor.Numerous clinical researches show that dissection of aorta patient's body multiple inflammatory factors are aobvious
Write increase, prompt this acute lung injury and hypoxemia may be dissection of aorta cause inflammatory reaction it is secondary.But
Dissection of aorta how to cause inflammatory reaction and this inflammatory reaction be again how subsequent pulmonary damage at present it is still unclear.
Aorta wall is mainly made of the smooth muscle cell of the vascular endothelial cell of inner membrance and middle film.Vascular endothelial cell
(VECs) it is a kind of disperse sexual organ with heterogeneity, can produce cytokine profiles and bioactive substance, has active
Secretion, synthesis, metabolism and immune function.In a sense, the knot of the occurrence and development of a variety of diseases of body and VECs
Structure and changes of function have close relationship.Vascular smooth muscle cells (VSMCs) have stronger plasticity, have and shrink and divide
Secrete function.Under normal physiological condition, VSMCs is in quiescent condition, expresses a series of distinctive contraction of VSMCs of differentiation and maturations
Protein molecular, in contraction phenotype, and when blood vessel is by pathological stimuli, the VSMCs proliferation of differentiation and maturation, transfer ability increase
By force, and cytokine profiles and extracellular matrix are secreted, does not express or low expression shrinks albumen specific to phenotype VSMCs point
Son is become secreting phenotype from contraction phenotype, this process is known as VSMCs Phenotypic change.VSMCs Phenotypic change is in hypertension, hat
It plays a significant role in the proliferating vasculars disease pathology mechanism such as heart trouble.
Under normal circumstances, blood flow can generate mechanical force to vascular wall in Homogeneous Circular artery, these power can divide
For the boundary shear stress parallel with vascular wall along blood flow direction, the compression perpendicular to vascular wall and answering along vascular wall circumferential direction
Power.Wherein wall shear stress and circumferential stress are for maintaining the physiological function of endothelial cell very crucial；And circumferential stress for
Maintain the physiological function of smooth muscle cell mostly important.Studies have shown that the oscillatory shear forces of harmonic(-)mean value and abnormal circumferential direction are answered
Power can induce vascular endothelial cell and smooth muscle cells secrete inflammatory factor, such as interleukin 1, interleukin-6, white thin
Born of the same parents' interleukin -8 and tumor necrosis factor-alpha etc..Due to endangium, middle layer and outer film stripping when dissection of aorta, formed true, false
Two chambers, true and false chamber lead to blood simultaneously.Confirm that true chamber and false chamber oscillation blood flow can all occur and cut using doppler technique detection
Shear force feature.Therefore, endaortic oscillation wall shear stress and the blood vessel circumferential stress of variation may draw when dissection of aorta
The damage for playing blood vessel endothelium and (or) smooth muscle cell releases a variety of inflammatory factors for causing injury of lungs.
Acute lung injury is that the viewpoint of the cell-mediated unsteered inflammatory reaction of inflammation has been recognized, and pathology is raw
Reason basis is damage even necrosis of excessive, the extended alveolar inflammation with alveolar epithelium.These inflammatory cells include multiform
Core neutrophil leucocyte and pulmonary alveolar macrophage etc..Inflammatory cell, which activates and discharges inflammatory mediator, can cause body injury and start inflammation
Disease cascade reaction.In cascade reaction, inflammatory mediator can activate other inflammatory mediators and inflammatory factor, and enhancing and amplification are scorching
Disease reaction, leads to a series of body injuries.It is the important of acute lung injury that polymorphonuclear neutrophils, which are assembled and activated in intrapulmonary,
Feature, it is generated by the tiny blood vessels of lung, after adherency and activation, is migrated in lung tissue and discharging inflammatory mediator and cell factor
(such as proteolytic enzyme and oxygen radical), causes diffusivity injury of lungs.
In summary viewpoint can be with inference: the change of shearing force and blood vessel circumferential stress is drawn in aorta when dissection of aorta
The damage of blood vessel endothelium and (or) smooth muscle cell is played, what is discharged after the latter two damages goes out multiple inflammatory factors, these inflammation
After the factor reaches lung, pulmonary epithelial cells and pulmonary vascular endothelial cell are damaged, and generates acute lung injury.
The disturbing factor as suffered by experiments in vivo is more, and there is an urgent need to one kind to be used for vitro study dissecting aneurysm of aorta
Oscillatory shear forces, which induce vascular cell release inflammatory factor, influences the micro flow control chip device of pulmonary epithelial cells function, device tool
Have can Vitro Simulated in the oscillation wall shear stress of body dissection of aorta and the cell culture chamber of circumferential tensile stress environment, be used for
The Combined culture of vascular endothelial cell (VECs) and smooth muscle cell (VSMCs)；With other cell culture chamber, it to be used for lung
The in vitro culture of chrotoplast (PECs)；Meanwhile the vascular cell release convenient for detection culture causes injury of lungs inflammatory factor and its draws
The case where sending out pulmonary epithelial cells damage, further analyzes relevant Cell. Mol mechanism.
Summary of the invention
The present invention is based on Hemodynamics principle and methods, provide a kind of for studying oscillatory shear forces and tensile stress induction
Vascular cell, which discharges inflammatory factor, influences the micro flow control chip device of pulmonary epithelial cells function, which can be used in analysis actively
Arteries and veins dissecting aneurysm wall shear stress induces vascular endothelial cell or/and smooth muscle cell release inflammatory factor, further influences
Pulmonary epithelial cells biological behaviour and its mechanism.
In order to achieve the above object, technical scheme is as follows:
As shown, the present invention combines Hemodynamics principle and method with microflow control technique, blood is designed and constructed
Endothelial cell VECs/ smooth muscle cell VSMCs Combined culture chamber and 2 cell culture chambers of pulmonary epithelial cells PECs culture chamber and
Its peripheral cell, forms two relatively independent circulatory systems, and two systems realize vascular cell and lung by sharing reservoir
Information communication between epithelial cell.1st cell culture chamber is used for the Combined culture of vascular endothelial cell and smooth muscle cell, the
2 cell culture chambers are used for the culture of pulmonary epithelial cells, by adjusting the 1st forward and backward load of cell culture chamber (compliance and outer
All resistances) size and ratio, realized inside the 1st cell culture chamber different average values and oscillation amplitude oscillatory shear forces and
Circumferential stress environment, thus to cultivate vascular endothelial cell and/or smooth muscle cell application difference in the 1st cell culture chamber
The oscillatory shear stress and tensile stress of feature.The secretion of 1st cell culture chamber inner cell is recycled to reservoir, thus
Connect the pulmonary epithelial cells acted in the 2nd cell culture chamber.
One kind inducing vascular cell release inflammatory factor influence lung epithelial for studying dissecting aneurysm of aorta wall shear stress
The micro flow control chip device of cell function, as shown in the figure.VECs/VSMCs Combined culture chamber and the front and back load cell circulatory system,
PECs culture chamber and its circulatory system constitute micro flow control chip device, and the micro flow control chip device includes hopper, vibration damping
Device, peristaltic pump, air pressure pump control system, three layers of VECs/VSMCs Combined culture chamber, elastic cavity A, elastic cavity B, resistance valves A,
Resistance valves B, the PECs culture chamber of single layer, silica gel pipeline and threeway.
The VECs/VSMCs Combined culture chamber, elastic cavity A, elastic cavity B, resistance valves A, resistance valves B and PECs culture
Chamber is integrated on a micro-fluidic chip, is connected between each element by microchannel.Flow path point on the micro-fluidic chip
At two branches, a branch includes the VECs for being sequentially connected the resistance valves A connect, elastic cavity A, VECs/VSMCs Combined culture chamber
Place chamber, elastic cavity B and resistance valves B, another branch include PECs culture chamber.Two branches merge into a pipeline outflow
Micro-fluidic chip is communicated by silicone tube with hopper, and the circulatory system is constituted；Hopper, peristaltic pump and the damper according to
It is secondary be connected by silicone tube after, damper is connect by silicone tube with micro-fluidic chip.The resistance valves A and resistance valves B is equal
It is connected with pulsometer.
The VECs/VSMCs Combined culture chamber, including upper and lower two rectangular cavities, two rectangular cavities by 500 μ m-thicks bullet
Property porous membrane separates, and the upper and lower faces of elastic cellular film plant smooth muscle cell VSMCs and vascular endothelial cell respectively
VECs；The length of two rectangular cavities is respectively 25mm, 5mm, 0.75mm.Wherein: the upper layer chamber where VECs
It is subjected to the shearing force that flowing generates and the tensile stress effect that pressure generates；Lower chamber is full of static culture solution where VSMCs,
Bear the tensile stress effect that pressure generates.
The peristaltic pump provides driving force for circulating liquid.Pass through air pressure pump control system, quantitatively regulating and controlling air pressure pump pressure
Power controls the open/closed degree, opening/closing time length and closure delay time of resistance valves A and resistance valves B, gives VECs/
VSMCs Combined culture chamber inputs the nearly physiological pulse blood pressure and flow waveform of different amplitudes；Additionally by control resistance valves A's
ON/OFF frequency can change the frequency of nearly physiological pulse blood pressure and flow waveform, and then intracavitary in VECs/VSMCs Combined culture
Obtain the blood pressure and flow waveform of different amplitudes and different frequency, i.e., the circumferential stress and blood flow of different amplitudes and different frequency are cut
Shear force waveform.
When resistance valves A is opened, culture solution enters elastic cavity A and elastic cavity B along pipeline a part, and another part flows through
VECs/VSMCs Combined culture chamber and resistance valves B flow to reservoir, generate forward flow；And when resistance valves A closure, elasticity
Liquid in chamber B can flow to elastic cavity A, Xiang Liudong after occurring in vascular cell culture chamber.To be trained within a cardiac cycle
Oscillation characteristics are presented in the fluid for supporting chamber.It is right by adjusting the compliance of elastic cavity A and B and the ratio between the resistance of resistance valves A and B
Vascular endothelial cell VECs generates the oscillatory shear stress of different average values, backward/forward direction flutter component；Meanwhile it is more to elasticity
The vascular smooth muscle cells of hole film another side culture generate circumferential stress, and stress intensity depends on pressure and elasticity modulus
Size.
Under pseudo steady assumed condition, the shear stress approximate expression for the vascular endothelial cell receiving cultivated in elastic membrane
Are as follows:
Therefore shear stress size τ of the load on endothelial cellwWith cell culture channel total fluid flow Q (t) at just
Than.By the ratio between resistance that the compliance of elastic cavity A and B, resistance valves A and B are set can Human Umbilical Vein Endothelial Cells generate different average values,
Backward/forward direction flutter component oscillatory shear stress.The vascular smooth muscle cells of elastic membrane another side culture are then born pressure and are led
The circumferential tensile stress of cause acts on, and tensile stress size depends on the size of pressure and thin flexible film modulus.
The beneficial effects of the invention are as follows successfully quantitatively can load dynamic vibration by the vascular endothelial cell in vitro Combined culture
Swing shear stress and circumferential stress, to smooth muscle cell load circumferential stress effect, and then quantitative analysis oscillatory shear stress and
The biological behaviour of circumferential stress coordinated regulation vascular endothelial cell and smooth muscle cell and its shadow to pulmonary epithelial cells function
Loud and relevant the Molecular Biology Mechanism.
Detailed description of the invention
Attached drawing 1 is micro flow control chip device schematic diagram of the invention.
Attached drawing 2 is the structural schematic diagram of resistance valves.
Attached drawing 3 is VECs/VSMCs Combined culture cavity configuration schematic diagram.
Attached drawing 4 is pulmonary epithelial cells PECs culture chamber structural schematic diagram.
In figure: 1 hopper；2 peristaltic pumps；3 dampers；
4 vascular endothelial cell VECs/ smooth muscle cell VSMCs Combined culture chambers；
The pulmonary epithelial cells PECs culture chamber of 5 single layer structures；6 elastic cavity A；7 elastic cavity B；
8 resistance valves A；9 resistance valves B；10 pulsometers；11 sheet glass.
Specific embodiment
As shown in the picture, apparatus of the present invention include vascular endothelial cell and smooth muscle cell Combined culture chamber and front and back load
The element circulatory system, pulmonary epithelial cells culture chamber and its circulatory system.The micro flow control chip device includes hopper 1, peristaltic pump
2, damper 3, air pressure pump control system, three-decker vascular endothelial cell VECs/ smooth muscle cell VSMCs Combined culture chamber
4, pulmonary epithelial cells PECs culture chamber 5, the silicone tube of elastic cavity A6, elastic cavity B7, resistance valves A8, resistance valves B9, single layer structure
Road and threeway；Two culture chambers and its peripheral cell, form two relatively independent circulatory systems, and two circulatory systems pass through altogether
The information communication of VECs/VSMCs and PECs is realized with reservoir 1.
In the present embodiment, the place the VECs chamber, elasticity of resistance valves A8, elastic cavity A6, VECs/VSMCs Combined culture chamber 4
Chamber B7 and resistance valves B9 is in turn connected to form a branch；PECs culture chamber 5 forms another branch；Two branches are integrated in one
On a micro-fluidic chip, PDMS- glass structure, the micro-fluidic chip processing technology of the microchannel classics between each element are formed
Production；Resistance valves A8 and resistance valves B9 has two layers of PDMS structure, controls fluid line sectional area by pulsometer and adjusts resistance
Size, damper 3 and elastic cavity A6 and elastic cavity B7 are constituted and are inserted by different tube diameters, the silicone tube for enclosing air column respectively
On microchannel, compliance size is adjusted by adjusting Length of air column.Hopper 1, damper 3, peristaltic pump 2 and pulsometer
The peripheral system of control system composition micro-fluidic chip.Two branches merge into a pipeline outflow micro-fluidic chip, pass through silicon
Sebific duct is communicated with hopper 1.
The present invention can be used successfully to oscillatory shear forces and tensile stress induces vascular cell release inflammatory factor and influences pneumonocyte
The research of function.
Claims (2)
1. a kind of dissecting aneurysm of aorta wall shear stress, which induces vascular cell release inflammatory factor, influences pulmonary epithelial cells function
Micro flow control chip device, which is characterized in that the micro flow control chip device includes hopper (1), peristaltic pump (2), damper
(3), air pressure pump control system, vascular endothelial cell VECs/ smooth muscle cell VSMCs Combined culture chamber (4), pulmonary epithelial cells
PECs culture chamber (5), elastic cavity A (6), elastic cavity B (7), resistance valves A (8), resistance valves B (9), silica gel pipeline and threeway；Two
Culture chamber and its peripheral cell, form two relatively independent circulatory systems, and two circulatory systems are real by sharing reservoir (1)
The information communication of existing VECs/VSMCs and PECs；
The resistance valves A (8), elastic cavity A (6), VECs/VSMCs Combined culture chamber (4) VECs where chamber, elastic cavity B
(7) and resistance valves B (9) is in turn connected to form a branch；PECs culture chamber (5) forms another branch；Two branches are integrated
It on a micro-fluidic chip, is connected between each element by microchannel, it is micro-fluidic that two branches merge into a pipeline outflow
Chip is communicated by silicone tube with hopper (1)；
After the hopper (1), peristaltic pump (2) and damper (3) are sequentially connected, damper (3) is connect with micro-fluidic chip；
The resistance valves A (8) and resistance valves B (9) is connected with pulsometer；
The VECs/VSMCs Combined culture chamber (4) includes upper and lower two rectangular cavities, and two rectangular cavities are by elastic cellular film
It separates, the upper and lower faces of elastic cellular film plant VSMCs and VECs respectively；
The compliance and resistance valves A (8) that the micro flow control chip device passes through adjusting elastic cavity A (6) and elastic cavity B (7)
With the ratio between the resistance of resistance valves B (9), the vibration of different average values, backward/forward direction flutter component is generated to vascular endothelial cell VECs
Swing shear stress；Meanwhile circumferential stress is generated to the vascular smooth muscle cells of elastic cellular film another side culture, stress is big
The small size depending on pressure and elasticity modulus；
By regulating and controlling air pressure pump pressure, open/closed degree, the opening/closing time length of resistance valves A (8) and resistance valves B (9) are controlled
And closure delay time, the nearly physiological pulse blood pressure and flow waves of different amplitudes are inputted for VECs/VSMCs Combined culture chamber (4)
Shape；
By controlling the frequency of ON/OFF frequency shift nearly the physiological pulse blood pressure and flow waveform of resistance valves A (8), in VECs/
The circumferential stress and wall shear stress waveform of different amplitudes and different frequency are obtained in VSMCs Combined culture chamber (4).
2. micro flow control chip device according to claim 1, which is characterized in that the elastic cellular film with a thickness of
500μm。
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| CN201611026425.5A CN106754354B (en) | 2016-11-19 | 2016-11-19 | Dissecting aneurysm of aorta wall shear stress, which induces vascular cell release inflammatory factor, influences the micro flow control chip device of pulmonary epithelial cells function |
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| CN107312713A (en) * | 2017-07-28 | 2017-11-03 | 中科芯瑞(苏州)生物科技有限公司 | A kind of micro-fluidic chip and its application |
| CN108728360B (en) * | 2018-06-09 | 2024-04-09 | 上海天引生物科技有限公司 | Cell co-culture device and method thereof |
| CN108485973A (en) * | 2018-06-20 | 2018-09-04 | 上海长海医院 | A kind of cell co-cultures flat flow chamber and its fluid shear stress stimulating method |
| CN109852548B (en) * | 2019-03-13 | 2022-10-28 | 贵州大学 | Double-circulation tissue engineering blood vessel in-vitro culture system for simulating pulsating blood flow |
| CN111426821B (en) * | 2020-03-24 | 2021-07-06 | 大连理工大学 | Micro-fluidic chip-level extracorporeal circulation system for vascular endothelial cell mechanics biology research |
| CN112980679B (en) * | 2021-02-04 | 2022-10-04 | 大连理工大学 | Endothelial cell in-vitro culture system for optimizing continuous flow artificial heart pulsation working mode |
| CN115350736B (en) * | 2022-08-22 | 2023-09-19 | 华东理工大学 | Detection device and detection method for skin inflammatory factors |
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