CN201418901Y - Porous Brick Filled Stent Reactor for Artificial Liver - Google Patents

Abstract

The utility model relates to medical equipment and aims to provide a porous brick-type filled bracket type reactor for an artificial liver. The reactor includes a reactor shell; the interior of the reactor shell is divided into three compartments by two dense grid-covered isolation plates: an oxygenation compartment, a filling bracket compartment, and an immune barrier compartment from bottom to top. The utility model adopts a unique porous brick-type fixed bed as the internal body of the reactor, divides the interior of the reactor into miniature independent units through the slender hole columns in the fixed bed, and divides the inner cavity of the reactor into parts, reducing the The dead space, dead space and liquid flow resistance make the liquid flow inside the reactor uniform. Solve the problems of lack of immune isolation and insufficient oxygen supply in scaffold-type reactors, and make up for the defects of hollow fiber-type reactors, such as insufficient cell growth space, poor cell adhesion, lack of three-dimensional growth environment, and easy to block semipermeable membranes. It has the functions of oxygenation, immune barrier, and three-dimensional culture, with powerful comprehensive performance, simple manufacturing process, low cost, and easy to scale up.

Description

Porous Brick Filled Stent Reactor for Artificial Liver

technical field

The utility model relates to a medical device, which is a core device for the treatment of a biological or hybrid artificial liver-a porous brick-type filling bracket type reactor for artificial liver.

Background technique

As one of the effective treatment methods for patients with severe hepatitis and liver failure, artificial liver support therapy has been recognized all over the world. It includes three categories of non-biological, biological and hybrid artificial livers, of which the latter two are the main international development directions. Bioreactor (i.e., reactor) is the core device of biological and hybrid artificial liver treatment, which not only provides a place for material exchange between exogenous liver cells and patient blood or plasma, but also provides a suitable growth environment for liver cells. The application prospect is broad. An ideal artificial liver bioreactor should meet the following basic requirements: 1. Sufficient space for the growth of liver cells, supporting a patient with liver failure requires about 10% of the normal liver mass, which is equivalent to about 15 billion liver cells. 2. Good biocompatibility can ensure the activity and function of liver cells. 3. Two-way transmission of small and medium substances, immune barrier of macromolecular substances.

So far, there are four main types of bioreactors: hollow fiber type, flat monolayer culture type, perfusion bed type/stent type and cell encapsulation/suspension type. At present, the most clinical application is the hollow fiber reactor, which has the functions of immune isolation and oxygenation, and the cells are less sheared, but has the following disadvantages: 1. The traditional hollow fiber reactor has insufficient space for cell growth and uneven distribution. 2. It can only provide two-dimensional culture, but not three-dimensional culture; 3. The semi-permeable membrane barrier or collagen layer will affect the material exchange efficiency; 4. Cells grow on the surface of the fiber and it is easy to block the fiber hole. The plate monolayer culture type has a small body surface area ratio and is difficult to scale up, and the cell-encapsulated/suspension type has poor stability of the suspended cells and the material exchange of the encapsulated cells is affected. There are not many research reports in recent years. Another type of reactor that is also widely used in animal experiments and cell culture is the perfusion bed/stent reactor. It has the following advantages: 1. The direct contact between liver cells and plasma or blood is beneficial to material exchange; 2. The introduction of bio-scaffold materials increases the space for cell growth; 3. Bio-scaffolds can provide hepatocytes with substrates and three-dimensional Three-dimensional growth environment; as a type of adherent growth cells, hepatocytes must attach to survive and maintain their functions, and the three-dimensional growth environment is conducive to the formation of hepatocyte spheroids, which is conducive to the development and long-term maintenance of cell functions. But it has the following disadvantages: 1. There is a potential safety hazard of lack of immune isolation; 2. The oxygen supply of cells is insufficient; 3. The center and edge of the entire reactor support are prone to produce dead space and dead space; 4. Through a large number of supports The resistance is large, and the required perfusion intensity is high, which causes the cells to be subjected to a large shear force and is difficult to amplify.

How to integrate the advantages of different types of reactors, maximize their strengths and circumvent their weaknesses, and overcome their shortcomings has become a recent research hotspot at home and abroad. In recent years, the research on bio-scaffold materials has made great progress. Emerging nanomaterials, natural materials such as alginic acid-chitosan with a three-dimensional network-like interconnected structure inside, and high-molecular polymer materials modified by galactose and heparin have emerged in large numbers. And widely used in cell culture. The great progress of biological scaffold materials has created a great opportunity for the development of new reactors. The existing reactors with good comprehensive performance in the world often have complex structures, high manufacturing process requirements, and high costs. The development of a new type of bioreactor with good comprehensive performance, simple structure and manufacturing process, and relatively low cost has special and important practical significance for the vast number of developing countries and third world countries.

Utility model content

The purpose of the utility model is to overcome the deficiencies in the prior art, and provide a porous brick for artificial liver with functions of oxygenation and immune isolation, uniform and sufficient perfusion, improvement of dead space and dead space, low production cost and strong practicability Filled scaffold type reactor.

The utility model is achieved through the following technical solutions:

Provide a kind of artificial liver with porous brick-type filled bracket type reactor, including the reactor shell; The interior of the reactor shell is divided into three compartments by two dense grid isolation plates: the oxygenation compartment from bottom to top, Filling the bracket cabin, immune barrier cabin, between the oxygenation cabin and the filled bracket cabin, and between the filled bracket cabin and the immune barrier cabin, there is an isolation plate with a grid;

Air-permeable and impermeable hollow fibers are horizontally arranged in the oxygenation cabin, and a liquid inlet is provided at the bottom of the cabin; the inside of the hollow fiber is a gas passage, and its two ends are respectively connected to the gas inlet and the gas outlet;

The filled scaffold cabin is a fixed bed of transparent plexiglass densely covered with slender holes, and the fixed bed holes are filled with three-dimensional porous biological scaffold beads; the bottom surface and the top surface of the fixed bed are close to the isolation plate, and the isolation plate The side length of the upper grid is smaller than the diameter of the stent ball to prevent the stent ball from leaking out;

The immune barrier compartment is composed of three parts: the shell, the hollow fiber and the sealing bayonet cover; the side wall of the immune barrier compartment is provided with a liquid outlet and is connected with the space outside the hollow fiber; The isolation plates are connected; the hollow fibers are vertically arranged and have the function of water permeability and immune barrier.

As an improvement, the oxygenation compartment and the filling bracket compartment are connected to each other as a whole, and the immune barrier compartment is an independent component.

As an improvement, the diameter of the stent balls is 2mm, and the diameter of the fixed bed hole column is 3mm, which only allows the three-dimensional porous bioscaffold balls to be placed in a single row.

As an improvement, the filled scaffold balls are any of the following: alginate-chitosan scaffold pellets, polyethylene resin scaffold pellets, polyurethane foam scaffold pellets or polycaprolactone scaffold pellets, The inside of the stent balls has a three-dimensional network-like interconnected structure.

As an improvement, the bottom surface of the filling bracket compartment is fixedly connected to the isolation plate, and the isolation plate on the top surface is connected to the sealed bayonet cover of the immune barrier compartment.

As an improvement, the shell of the reactor is a transparent cuboid shell.

As an improvement, the hollow fibers of the oxygenation chamber are air-permeable and impermeable polypropylene hollow fibers, or polysulfone hollow fibers coated with silicone rubber; It is a 50-100kD polysulfone hollow fiber or polyethersulfone hollow fiber.

The beneficial effects of the utility model:

The interior of the traditional perfusion bed/stent reactor is usually a whole connected by liquid flow, and there are problems such as easy formation of dead space, dead space, uneven liquid flow, and large liquid flow resistance. The utility model's filled bracket type reactor adopts a unique porous brick fixed bed as the internal body of the reactor. The inner cavity is divided into zero parts, thereby reducing the dead space and dead space, reducing the liquid flow resistance, and making the liquid flow inside the reactor uniform. At the same time, the utility model utilizes the ingenious combination of filling brackets and hollow fibers, which not only solves the problems of lack of immune isolation and insufficient oxygen supply in the bracket type reactor, but also makes up for the lack of cell growth space and poor cell adhesion of the hollow fiber type reactor. The lack of a three-dimensional growth environment makes it easy to block the defects of the semi-permeable membrane. The utility model has the functions of oxygenation, immune barrier, three-dimensional culture and the like, and has strong comprehensive performance, simple manufacturing process, low cost and easy enlargement.

Description of drawings

Fig. 1 is a schematic diagram of the vertical cross-sectional structure of a porous brick-type filled scaffold type reactor for artificial liver;

Fig. 2 is the transverse cross-sectional structure schematic diagram of the oxygenation chamber of the porous brick type filled support type reactor for artificial liver;

3 is a schematic diagram of a transverse cross-sectional structure of a filled scaffold compartment of a porous brick type filled scaffold type reactor for artificial liver;

Figure 4 is a schematic diagram of the transverse cross-sectional structure of the immune barrier compartment of the porous brick-type filled scaffold-type reactor for the artificial liver;

Fig. 5 is a schematic diagram of the application of the bioartificial liver support system.

Reference signs: porous brick fixed bed 1, support ball 2, hollow fiber 3, hollow fiber 4, isolation plate 5, sealing bayonet cover 6, reactor shell 7, liquid inlet 8, liquid outlet 9, gas inlet 10 , a gas outlet 11 , a patient or an experimental animal 12 , a plasma separator 13 , and an extracorporeal circulation pool 14 .

Detailed ways

The artificial liver porous brick type filled bracket type reactor of the present embodiment comprises a cuboid transparent plexiglass reactor shell 7, and the reactor shell 7 is divided into three compartments by two dense grid-covered partition plates 5, from the bottom to the bottom. The above are the oxygenation cabin, the filling bracket cabin, and the immune barrier cabin. The oxygenation cabin and the filling bracket cabin are integrated, and the immune barrier cabin is an independent component, which is connected with the aforementioned whole through a sealed bayonet cover 6 .

Air-permeable and impermeable hollow fibers 3 are arranged horizontally in the oxygenation chamber, and a liquid inlet 8 is arranged at the bottom of the chamber. The inside of the hollow fiber 3 is a gas channel, and the two ends are respectively connected with a gas inlet 10 and a gas outlet 11 . The hollow fiber 3 of the oxygenation chamber is a breathable and impermeable polypropylene hollow fiber, or a polysulfone hollow fiber coated with silicone rubber.

The main body of the filled scaffold cabin is a unique porous brick fixed bed 1, that is, a transparent plexiglass fixed bed densely covered with slender pore columns, and the porous brick fixed bed 1 pore columns are filled with three-dimensional porous biological scaffold pellets 2. The diameter of the stent ball 2 is 2mm. The diameter of the hole column of the porous brick fixed bed 1 is slightly larger than the diameter of the support balls 2, which is 3mm, and only a single row of support balls 2 is allowed to be inserted. The material of the bracket ball 2 can be a natural material, such as an alginic acid-chitosan bracket, or a polymer compound, such as polyethylene resin, polyurethane foam, polycaprolactone, and the inside of the bracket ball 2 is a three-dimensional network Connected structure, liver cells grow three-dimensionally on the outside and inside.

The bottom surface and the top surface of the porous brick fixed bed 1 are all close to the separation plate 5, and the side length of the grid on the separation plate 5 is less than the diameter of the support ball 2, so as to prevent the support bead 2 from leaking. The isolation plate 5 on the bottom surface is fixed, and the isolation plate 5 on the top surface is connected with the sealing bayonet cover 6, which can be opened to load or pour out the support bead 2.

The immune barrier compartment is composed of a shell, a hollow fiber 4 with an immune barrier effect, and a sealed bayonet cover 6. The hollow fiber 4 is vertically arranged and has the functions of water permeability and immune barrier. The lower end opening of the inner cavity of the hollow fiber 4 communicates with the filling bracket compartment, and the upper end is closed. A liquid outlet 9 is arranged on the side wall of the cabin, and is connected with the space outside the hollow fiber 4 . The hollow fiber 4 of the immune barrier chamber is a polysulfone hollow fiber or a polyethersulfone hollow fiber with a pore size of 0.2 μm and a molecular cut-off of 50-100 kD.

The usage is as follows:

After the blood of the patient or experimental animal 12 is drawn out of the body, it is firstly separated into cell components and plasma components by the plasma separator 13 . The plasma components enter the extracorporeal circulation pool 14, and then enter the oxygenation cabin from bottom to top through the liquid inlet 8 at the bottom of the oxygenation cabin of the reactor. The mixed gas of oxygen and carbon dioxide enters the inner cavity of the hollow fiber 3 of the oxygenation chamber through the gas inlet 10 and flows out through the gas outlet 11 on the opposite side. Plasma passes through the outer cavity of the hollow fiber in the oxygenation cabin to achieve the effect of oxygenation. Then enter the filling bracket compartment through the grid of the isolation plate 5, evenly pass through the filled porous bracket pellet 2, and after fully contacting the hepatocytes on the pellet (hepatocytes are pre-inoculated and cultivated), pass through the top of the filling bracket compartment The grid of the isolation plate 5 enters the inner cavity of the hollow fiber 4 of the immune barrier compartment. The macromolecular immune substances in the plasma are blocked, and the small and medium molecular substances overflow from the fiber hole of the hollow fiber 4 to the outer cavity, and then flow out of the reactor through the liquid outlet 9 and return to the circulation pool 14 . After many cycles, it will be infused back into the body together with the cellular components in the blood.

Hepatocytes can be selected from primary human, porcine hepatocytes or hepatocyte lines, etc. The inoculation and culture steps are briefly described as follows:

1. Open the sealed bayonet cover 6 of the reactor, pour the porous biological support pellet 2 into the elongated hole column of the porous brick fixed bed 1, cover the sealed bayonet cover 6, and connect the immune barrier cabin and the oxygenation cabin , Filling the bracket cabin to form a whole.

2. The reactor, including the porous biological scaffold pellet 2 inside it, is sterilized together.

3. Prepare the hepatocyte suspension with appropriate concentration with the culture medium.

4. Inject the liver cell suspension into the reactor from the liquid inlet 8 at the bottom of the oxygenation chamber until the liquid component in the cell suspension overflows from the liquid outlet 9 on the side wall of the immune barrier chamber. The hepatic cells adhere to the porous bioscaffold ball 2, and are firmly attached after about 24 hours.

5. Fresh culture medium is poured into the liquid outlet 9 on the side wall of the immune barrier chamber, flows out from the liquid inlet 8 after filling the scaffold chamber and the oxygenation chamber, and washes away unattached hepatocytes.

6. The reactor and the extracorporeal circulation pool 14 are connected to form a hepatocyte culture circulation path as shown in the dotted line box in Fig. 5, and fresh culture medium flows into and out of the reactor from the liquid inlet 8 and the liquid outlet 9 respectively, forming a hepatocyte culture cycle. Cells provide nutrients and remove metabolites.

7. After culturing for a suitable time, they can be connected and combined according to Figure 5 to form a biological artificial liver support system.

The scale of the porous brick type filled bracket type reactor for artificial liver in the utility model can be enlarged or reduced according to actual application.

Finally, it should be noted that what is disclosed above are only specific embodiments of the present invention. All deformations that a person skilled in the art can derive or associate directly from the content disclosed in the utility model shall be considered as the protection scope of the utility model.

Claims (7)

1, a kind of perforated brick type filling support type reactor used in artificial liver, comprise shell of reactor, it is characterized in that described shell of reactor inside is divided into three cabins by the division board of two densely covered grids: intercept the cabin for oxygenate cabin, filling bracket cabin, immunity from bottom to top;

Lateral arrangement has the doughnut of permeable watertight in the described oxygenate cabin, and bilge portion establishes the liquid inlet; Doughnut inside is the gas passage, and its two ends connect gas access and gas outlet respectively;

Described filling bracket cabin is a fixed bed that is thick with the transparent organic glass of elongated hole post, fills three-dimensional porous crack biological support bead in the post of fixed bed hole; The all adjacent division board of the bottom surface of fixed bed and end face, the length of side of grid spills to prevent the support bead less than the little bulb diameter of support on the division board;

Described immunity intercepts the cabin and is made up of shell, doughnut and sealed card flap three parts; The sidewall in immunity obstruct cabin is established liquid outlet and is linked to each other with the space of doughnut outside, and the sealed card flap of the bilge links to each other with the division board at top, filling bracket cabin; Doughnut is the doughnut of vertically arranging and having permeable and immune iris action, and its inner chamber lower ending opening is communicated with upper end closed with the filling bracket cabin.

2, perforated brick type filling support type reactor used in artificial liver according to claim 1 is characterized in that, described oxygenate cabin and filling bracket cabin interconnect as a whole, and it is individual member that immunity intercepts the cabin.

3, perforated brick type filling support type reactor used in artificial liver according to claim 1 is characterized in that, the diameter of described support bead is 2mm, and the diameter of fixed bed hole post is 3mm, only allows single the inserting of three-dimensional porous crack biological support bead.

4, perforated brick type filling support type reactor used in artificial liver according to claim 1, it is characterized in that, the support bead inside of described filling is three-dimensional netted connectivity structure, is following any one: alginic acid-chitosan stent bead, polyvinyl resin support bead, polyurethane foam support bead or polycaprolactone support bead.

5, perforated brick type filling support type reactor used in artificial liver according to claim 1 is characterized in that, the bottom surface in described filling bracket cabin is connected with division board is fixed, and the division board of its end face links to each other with the sealed card flap that immunity intercepts the cabin.

6, perforated brick type filling support type reactor used in artificial liver according to claim 1 is characterized in that, the shell of described reactor is transparent cuboid shell.

According to described any one perforated brick type filling support type reactor used in artificial liver of claim 1 to 6, it is characterized in that 7, the doughnut in described oxygenate cabin is the polypropylene hollow fiber of permeable watertight, perhaps the polysulfone hollow fibre of silicon-coating rubber; The doughnut that described immunity intercepts the cabin is that aperture 0.2 μ m, molecular retention amount are polysulfone hollow fibre or the polyether sulfone doughnut of 50～100kD.

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