CN104015366B - Biological 3D prints by cross-linked gel status monitoring syringe and method - Google Patents

Abstract

The invention discloses a cross-linked gel state monitoring syringe and a method for bio 3D printing. The syringe includes a needle with a hole made by laser drilling technology, a semi-core transparent shell and a supply mode of two materials participating in the coaxial crosslinking reaction. After one material enters, rotate the needle, and then add another material, which penetrates through the small hole on the needle, and the two contact and cross-linking reaction occurs. According to the degree of reaction, its color change can be observed with a semi-core transparent shell. Excess materials can be collected and recycled. The present invention adopts the original needle on the piston syringe, combines the characteristics of coaxial cross-linking, designs a semi-core transparent shell structure, and realizes the degree and status of cross-linking reaction, the static status of micro-scale cells in the gel and the dynamics of fluid flow The monitoring of images provides good materials for experiments. This device is easy to operate, has good flexibility and good process controllability, and effectively provides the coaxial cross-linking reaction and the biocompatibility of the prepared hollow or solid fibers. The data.

Description

Cross-linked gel state monitoring syringe and method for 3D bioprinting

technical field

The invention relates to a crosslinked gel state monitoring syringe and method for 3D bioprinting, which is used for monitoring the production process of microscale hydrogel scaffold fibers widely used in the field of biomanufacturing, especially for the process of preparing coaxial hollow fibers The cross-linking degree of the material in the medium and the monitoring of the static and fluid dynamic flow images of the cells in the cell-loaded gel can be used in the field of biomanufacturing technology.

Background technique

The cross-linking reaction is the reaction of two or more molecules (usually linear molecules) bonded and cross-linked to form a relatively stable molecule (body-shaped molecule) in a network structure. This reaction converts linear or slightly branched macromolecules into a three-dimensional network structure. At present, in the microfluidic devices studied by many domestic and foreign scholars, it involves making cross-linked solid fibers or hollow fibers through coaxial devices to obtain tissue scaffolds that can be used in the field of biomedicine. In the field of scale, people's research focuses on the transition from the viscous fluid state of materials involved in cross-linking to solid-state scaffolds that can be used in medicine. A single fiber cannot be called a scaffold. It can only be formed after being processed by bio-3D printing technology. Vascular stent needed.

Sodium alginate is the sodium salt of alginic acid, which is degradable and easy to process into a film, and is an environmentally friendly carrier material. In recent decades, sodium alginate has been widely used in food processing, textile processing, biomedicine and environmental protection, and has shown good application prospects. The gelation process of sodium alginate is the cross-linking of the polymer, which is mainly the process of exchanging sodium ions on guluronic acid with divalent cations. Divalent cations help to hold molecules together, and the aggregated nature of the molecules and their bulk and stronger binding of the cations have a synergistic binding effect. Chitosan, sodium alginate and other hydrogels have good biocompatibility and low toxicity, and are widely used in the fields of drug delivery systems and tissue engineering. Because the scaffold produced by the biomanufacturing method will be used in the living body, it is necessary to carry out the work of carrying cells in the biomanufacturing process. In many cases, it is mentioned that in hydrogels such as chitosan or sodium alginate Incorporate the cells, and it is detected that the cells have high activity in it, and the formation of the loaded cells needs to be carried out at room temperature. For example, at present, at room temperature, chitosan is mixed with fibroblasts and sodium tripolyphosphate for cross-linking to form hollow fibers, and a cell survival rate of 92% can be obtained by adjusting it to a suitable pH value. There are also cases where human fibroblasts are loaded into sodium alginate and coaxially cross-linked with calcium chloride to make hollow fibers. The cells are easy to load into the fibers. Not big, so this method can be used in combination with bio-3D printing technology to make nerve or muscle fibers. According to the above examples, dynamic coaxial cross-linking provides a method to form fibers with hollow structures, which combined with bio-3D printing technology provides a new strategy for vascularization.

It is precisely because of the wide range of applications of ionic cross-linking, and because of its good compatibility with biomedicine materials and experimental results, it is currently the focus of research in the field of biomanufacturing, and people's interest in it is increasing day by day. The most important of the research issues is undoubtedly the research on the degree of cross-linking of the two materials involved in cross-linking and the survival rate of cells in the fiber after loading cells. The determination of the degree of cross-linking is of great significance to determine the characteristic parameters such as the diameter of the cross-linked fiber, and whether the cells can survive in the fiber after loading the cells, or the survival rate, this is a problem that we need to care about , the production of hollow or solid fibers is aimed at the field of biomanufacturing. If the fabricated tissue scaffolds are not biocompatible, experiments in animals cannot be performed. Therefore, the determination of these two issues is very urgent for bio-3D printing to fabricate tissue scaffolds. Because the field involved is a micro-scale field, what is involved in ion cross-linking is not what people can see with the naked eye. At present, there are many microfluidic chips on the market, which are used to observe the flow and reaction status of microfluidics. If the produced fiber is cut directly in the axial or radial direction to observe its degree of cross-linking and cell activity, the fiber will be destroyed firstly, and the deformation of the incision will affect the observation of the degree of cross-linking, even It will promote cell death, so that the required real-time data cannot be accurately obtained. However, for the detection of static and fluid dynamic images of coaxial fibers and cells in colloids made of ion cross-linking reactions, because of their tinyness and dynamic conditions, there is no complete observation device to monitor their flow conditions in real time. It is also impossible to know the degree of cross-linking in the experiment, and it is impossible to obtain the desired experimental results according to the actual situation.

Contents of the invention

In order to solve the problems of the prior art, the object of the present invention is to overcome the deficiencies of the prior art, and to provide a cross-linked gel state monitoring syringe for 3D bioprinting and The method is a simple device for image detection of the cross-linking process, the static state of cells in the gel, and the dynamic flow of fluid during the cross-linking process of two substances.

To achieve the above object, the present invention adopts the following technical solutions:

A cross-linked gel state monitoring syringe for 3D bioprinting, comprising a medical needle, an upper cover, a semi-core transparent shell, and a lower cover, characterized in that: the needle, the upper cover, the semi-core transparent shell and The lower cover plate forms a semi-circular cylindrical channel through interference fit; the needle head has a scale of 180 degrees; the upper cover plate is connected to the half-core transparent shell through an upper rubber gasket, and the half-core transparent shell passes through a screw through a lower rubber washer It is connected with the lower cover, and there is also a 180-degree scale on the lower rubber gasket; there is an inlet on the upper part of the semi-core transparent shell, which communicates with the semi-circular cylindrical channel; there is an outlet on the lower cover; the needle tube of the needle faces the semi-circular cylindrical channel Small holes are evenly distributed on one side of the needle tube, so that the inner cavity of the needle tube is connected with the semi-circular cylindrical channel; the inlet on the needle is connected with the semi-circular cylindrical channel through the small holes; one side of the needle barrel is provided with radial holes for material penetration. Small hole, under the pressure of the equipped piston syringe, the material penetrates into the semi-circular cylindrical channel of another material through the small hole on the needle. Before loading the material, the side of the needle without the small hole needs to face halfway. The outer side of the transparent shell of the core, so as to complete the storage of the material in the semi-circular cylindrical channel. If the small hole is aligned with the inner side of the semi-core transparent shell at this time, the material will penetrate into the needle through the small hole. Affect the follow-up experiments; when the semi-circular cylindrical channel is filled with materials, rotate the needle 180 degrees. At this time, the 180-degree scale on the needle and the upper rubber gasket coincides, so that the side with the hole on the needle faces the half-core transparent shell. Inside, another material is loaded through the needle inlet, and the material is added with the equipped medical piston syringe, and the material cross-links with the material in the semi-circular cylindrical channel through the small hole on the needle. The half-core transparent shell was used to observe the degree of cross-linking and the static and dynamic flow of cells.

The shape of the needle is the same as that of the medical needle on the market. Through laser drilling technology, the metal part of the needle tube on the needle is punched to form evenly distributed small holes. The more small holes the better; The 180 degree scale is used for rotation indication.

The structure of the semi-core transparent shell; the interior adopts a semi-circular ring, which is used to position the needle and leave a semi-circular cylindrical channel. The needle can be rotated in the semi-core transparent shell to observe the cross-linking of the material, the cell Static and dynamic flow conditions; the semi-core transparent shell is made of resin material and made by SLA light curing technology.

The upper and lower rubber gaskets not only play a positioning role, but also play a sealing role to prevent the material from seeping out. There is also a 180-degree scale line corresponding to the needle on the upper rubber gasket to indicate that during the process of loading materials The needle spins. The material added before rotating the needle needs to ensure a certain viscosity so that the material can easily penetrate through the small holes on the needle after rotating the needle, while the material added after rotating the needle does not need too much viscosity , so as not to clog when penetrating through the small holes into the storage chamber.

A method for monitoring the state of cross-linked gel for 3D bioprinting, which is operated by using the syringe for monitoring the state of cross-linked gel for 3D bioprinting, and the specific operation steps are as follows:

a. Before adding materials, first align the scale on the needle tube with the scale on the lower rubber gasket, so that the half of the needle tube with holes faces the outside of the semi-core transparent shell, and the half without holes faces inward;

b. Add a material through the inlet on the half-core transparent shell;

c. After the above materials are added, rotate the needle at 180 ^° , so that the half of the needle tube with the hole is facing the inner side of the semi-core transparent shell, and the half without the hole is facing the outside;

d. After the needle is rotated, another material is added through the piston syringe equipped on the needle, and this material reacts with the material added through the inlet of the semi-core transparent shell through the penetration of the small hole on the needle, so that it can pass through the semi-core transparent The housing monitors the degree of reaction of the two materials as well as the static and hydrodynamic flow of the cells.

Compared with the existing microfluidic chip technology, the present invention has the following obvious outstanding substantive features and significant advantages:

1. On the basis of the experiment of coaxial cross-linking to make hollow fibers, the present invention proposes to observe the degree of cross-linking as the goal, and by means of 3D printing technology, to produce a transparent half-core transparent shell, which is suitable for a variety of Types of coaxial crosslinking reactions monitored.

2. The present invention can monitor cell static and fluid dynamic flow.

3. The components of the present invention are simple in structure, easy to manufacture and assemble, and do not need complicated processing. easy to use.

4. The present invention can be used to monitor the reaction degree of coaxial crosslinking in real time. Because there are many kinds of ionic crosslinking reactions, on this basis, the device can also be applied to the observation of crosslinking reactions of the same nature.

5. The present invention collects redundant materials, achieves material recycling, avoids waste of materials, and is of great help in saving costs.

Description of drawings

Fig. 1 is a schematic structural diagram of a microscale half-core-shell gel state monitoring device for 3D bioprinting according to Embodiment 1 of the present invention.

Fig. 2 is a view from direction A in Fig. 1 .

Fig. 3 is a sectional view taken along the line B-B in Fig. 1 .

Fig. 4 is a schematic diagram of a medical needle (1).

Fig. 5 is a schematic diagram of a half-core transparent shell (5).

FIG. 6 is an N-direction view of FIG. 5 .

detailed description

Preferred embodiments of the present invention are described in detail as follows in conjunction with accompanying drawings:

Embodiment one:

Referring to Figures 1 to 6, the cross-linked gel state monitoring syringe for 3D bioprinting includes a medical needle, an upper cover, a half-core transparent shell, and a lower cover, and is characterized in that: the needle and the upper cover, The half-core transparent shell and the lower cover form a semi-circular cylindrical channel through interference fit; there is a 180-degree scale on the needle head; the upper cover is connected to the half-core transparent shell through an upper rubber gasket, and the half-core transparent shell is passed through A lower rubber gasket is connected to the lower cover plate by screws, and there is also a 180-degree scale on the lower rubber gasket; there is an inlet on the upper part of the semi-core transparent shell, which communicates with the semi-circular cylindrical channel; there is an outlet on the lower cover plate; the needle tube of the needle Small holes are evenly distributed on the side facing the semi-circular cylindrical channel, so that the inner cavity of the needle tube is connected with the semi-circular cylindrical channel; the inlet on the needle is connected with the semi-circular cylindrical channel through small holes; Due to the radial small hole for material penetration, the needle is under the pressure of the equipped piston syringe, and the material penetrates into the semi-circular cylindrical channel of another material through the small hole on the needle. One side of the hole needs to face the outside of the half-core transparent shell, so as to complete the storage of materials in the semi-circular cylindrical channel. If the side of the small hole is aligned with the inside of the half-core transparent shell, the material will pass through the The hole penetrates into the needle, which affects the follow-up experiments; when the semi-circular cylindrical channel is filled with materials, the needle is rotated 180 degrees, and at this time the needle and the 180-degree scale on the upper rubber gasket coincide, so that the side with the hole on the needle faces The inner side of the semi-core transparent shell is then filled with another material through the needle inlet, and the equipped medical piston syringe is used to add the material, and the material cross-links with the material in the semi-circular cylindrical channel through the small hole on the needle, At this point, the degree of cross-linking and the static and dynamic flow of cells can be observed through the semi-core transparent shell.

Embodiment two:

This embodiment is basically the same as Embodiment 1, and the special features are as follows:

The shape of the needle is the same as that of the medical needle on the market. Through laser drilling technology, the metal part of the needle tube on the needle is punched to form evenly distributed small holes. The more small holes the better; The 180 degree scale is used for rotation indication.

The structure of the semi-core transparent shell; the interior adopts a semi-circular ring, which is used to position the needle and leave a semi-circular cylindrical channel. The needle can be rotated in the semi-core transparent shell to observe the cross-linking of the material, the cell Static and dynamic flow conditions; the semi-core transparent shell is made of resin material and made by SLA light curing technology. The upper and lower rubber gaskets not only play a positioning role, but also play a sealing role to prevent the material from seeping out. There is also a 180-degree scale line corresponding to the needle on the upper rubber gasket to indicate that during the process of loading materials The needle spins.

Embodiment three:

In this embodiment, referring to Figures 1 to 5, the components of the cross-linked gel state monitoring syringe for 3D bioprinting consist of a syringe needle 1 processed by laser drilling, an upper rubber gasket 2, a screw 3, and an upper cover plate 4 , a half-core transparent shell 5, a lower rubber gasket 6 and a lower cover plate 7.

In this embodiment, referring to Figures 1 to 5, the length of the metal part of the needle is 14.5mm, the diameter of the needle is 0.8mm, the specification of the screw is M1.2, the size of the transparent plate of the half-core shell is 10mm×5mm×14mm, and the spacer The thickness is 0.5mm, and the needle is a medical needle with a diameter of 1.5mm. The diameter of the inlet 9 on the half-core transparent shell is 2mm, and the diameter of the outlet 10 on the lower cover 7 is 1.5mm. As shown in Figure 1, the needle with a hole 1. The rubber gasket 2 is used for axial positioning. The upper cover plate 3 and the semi-core transparent shell are connected and fixed by M1.2 screws. The screws here will not hinder the reaction and observation of the material. It can be clearly seen from Figure 3 that the radial positioning of the needle is supported by the structure of the semi-core transparent shell itself (an arc with the same diameter as the needle), and the rubber gasket at the lower end of the needle and the needle can be used Interference fit, which can not only position the needle, but also maintain its sealing performance well.

In this embodiment, referring to Fig. 1 to Fig. 5, the assembly sequence is semi-core transparent shell-upper cover-screw-up and down rubber gasket-syringe needle-lower cover-screw.

In this embodiment, referring to Figures 1 to 5, taking the cross-linking of chitosan and glutaraldehyde as an example, the specific implementation steps are introduced:

Step 1: Before adding chitosan and glutaraldehyde, rotate the needle with a hole to the designated position, that is, half of the hole is facing the half-core transparent shell M direction (as shown in Figure 5), which prevents chitosan injection Part of the material will be filled into the air through the small hole on the needle, and then cross-linked when the core solution is injected to block the small hole;

Step 2: When the needle with a hole rotates to the specified position, add chitosan through the inlet 9, and use a 10mL syringe needle tube on the inlet 9. When injecting the material, the needle tube compresses the needle, and the rubber gasket 2 is used for axial positioning, and the chitosan Sugar enters in the semi-circular cylindrical channel 13 as shown in Figure 3, at this moment, under the constant situation of the position of the perforated needle, the chitosan added in the channel 13 is in a closed state until the channel 13 is filled complete;

Step 3: After the chitosan is added, rotate the needle with a hole back, that is, the half of the hole is facing the N direction of the semi-core transparent shell (as shown in Figure 5), and the position shown in Figure 3, at this time through the inlet 8 , add another material, glutaraldehyde, with the help of a piston syringe, which also uses a 10mL needle tube, and the added glutaraldehyde passes through the small hole 12 due to the pressure of the syringe, that is, the small hole punched by laser drilling technology, Infiltrate in the chitosan that adds in step 2, both cross-linking reaction takes place;

Step 4: After adding the two materials, the degree of reaction and the real-time situation of the two can be observed through the transparent semi-core transparent shell 5, because the cross-linking of glutaraldehyde and chitosan will make chitosan appear yellow, The degree of yellowing of chitosan can be observed to realize real-time monitoring of its cross-linking degree. Considering that the added glutaraldehyde does not completely permeate into the channel 13 through the small hole 12, so the excess glutaraldehyde is collected through the outlet 10, so as not to waste materials;

Embodiment four:

In this embodiment, referring to Figures 1 to 5, the components of the cross-linked gel state monitoring syringe for 3D bioprinting consist of a syringe needle 1 processed by laser drilling, a rubber gasket 2, a screw 3, an upper cover plate 4, The semi-core transparent casing 5, the rubber gasket 6 and the lower cover plate 7 are composed.

In this embodiment, see Figures 1 to 5, the length of the metal part of the needle is 14.5mm, the diameter of the needle is 0.8mm, the screw specification is M1.2, the size of the transparent cover is 10mm×5mm×14mm, and the thickness of the gasket is 0.5mm mm, as shown in Figure 1, the needle with a hole 1 is axially positioned through the rubber washer 2, the upper cover 3 and the semi-core transparent shell 5 are connected and fixed by M1.2 screws, and the screws here will not hinder Material reaction and observation. It can be clearly seen from Figure 3 that the radial positioning of the needle is supported by the structure of the semi-core transparent shell itself (an arc with the same diameter as the needle), and the rubber gasket at the lower end of the needle and the needle can be used Interference fit, which can not only position the needle, but also maintain its sealing performance well.

In this embodiment, referring to Fig. 1 to Fig. 6, the assembly sequence is the semi-core transparent shell-upper cover-screw-up and down rubber washers-syringe needle-lower cover-screw.

In this example, referring to Figures 1 to 6, taking the cross-linking of sodium alginate (hereinafter directly referred to as sodium alginate) and calcium chloride mixed with mouse fibroblasts as an example, the cross-linking degree and The cell survival rate is the goal, and the specific implementation steps are introduced (because this case needs to be carried out in a sterile environment, the experimental equipment must be sterilized before the experiment, and the experiment is carried out in an ultra-clean bench):

Step 1: Before adding sodium alginate and calcium chloride, rotate the needle with a hole to the designated position, that is, the half of the hole is facing the M direction of the semi-core transparent shell (as shown in Figure 5), which prevents the injection of sodium alginate Part of the material will be filled into the air through the small hole on the needle, and then cross-linked when the core solution is injected to block the small hole;

Step 2: When the needle with a hole rotates to the specified position, add sodium alginate through the inlet 9, and use a 10mL syringe needle tube on the inlet 9. When injecting the material, the needle tube compresses the needle, and the sodium alginate enters the In the semi-circular cylindrical channel 13, at this time, under the condition that the position of the perforated needle remains unchanged, the sodium alginate added to the channel 13 is in a closed state until the channel 13 is completely filled;

Step 3: After the sodium alginate is added, rotate the needle with a hole back, that is, the half of the hole is facing the N direction of the half-core transparent shell (as shown in Figure 5), and the position shown in Figure 3, at this time through the inlet 8 , add another material, calcium chloride, with the help of a piston syringe, which also uses a 10mL needle tube, and the added calcium chloride passes through the small hole 12 due to the pressure of the syringe, that is, the small hole punched by laser drilling technology, Infiltrate into the sodium alginate that adds in step 2, both cross-linking reaction occurs;

Step 4: After adding the two materials, the reaction degree and real-time situation of the two can be observed through the transparent semi-core transparent shell 5, and the cross-linking degree and cell survival rate can be realized (can be carried out after the cross-linking is completed). Infection experiment to observe its survival rate) two aspects of monitoring. Considering that the added calcium chloride will not completely permeate into the channel 13 through the small hole 12, so excess calcium chloride is collected through the outlet 10, so as not to waste materials.

Claims (5)

1. biological 3D prints with a cross-linked gel status monitoring syringe, comprises that medical needle (1), upper cover plate (4), half core are saturatingBright housing (5), lower cover (7), is characterized in that: described syringe needle (1) and upper cover plate (4), half core transparent shell (5) and lower cover(7) form semicircle annular tubular passage (13) by interference fit; On syringe needle (1), there is the scale of 180 degree; Upper cover plate (4) is through oneIndividual upper rubber washer (2) is connected with half core transparent shell (5), and half core transparent shell (5) passes through spiral shell through a lower rubber washer (6)Nail is connected with lower cover (7), also has the scale of 180 degree on lower rubber washer (6); Half core transparent shell (5) top has one to enterMouth (9), is communicated with semicircle annular tubular passage (13); On lower cover (7), there is an outlet (10); Needle tubing on syringe needle (1) towardsIn one side of semicircle annular tubular passage (13), uniform aperture (12), makes syringe intracavity be communicated with semicircle annular tubular passage(13); Import (8) on syringe needle (1) is communicated with semicircle annular tubular passage (13) by aperture (12); Described syringe needle (1) syringeIn one side, be useful on the radial hole (12) of material infiltration, syringe needle (1) makes under the pressure-acting of the piston syringe being equipped withMaterial is penetrated into by the aperture (12) on syringe needle (1) in the semicircle annular tubular passage (13) that another material is housed,Before packing material into, the one side that syringe needle (1) is provided with aperture (12) need to, towards half core transparent shell (5) outside, so just can completeAdding the storage of material in semicircle annular tubular passage (13) after material, if it is saturating now aperture (12) one side to be aimed to half coreThe inner side of bright housing (5), material can be penetrated in syringe needle by aperture, affects subsequent experimental; When semicircle annular tubular leads toRoad (13) is filled after material, by syringe needle (1) Rotate 180 degree, and the now degree of 180 on syringe needle (1) and upper rubber washer (2) scale weightClose, make one of syringe needle (1) upper porose (12) face the inner side of half core transparent shell (5), then fill by syringe needle (1) import (8)Enter another material, adopt the medical piston syringe being equipped with to add material, this material is by the aperture on syringe needle (1)(12) material and in semicircle annular tubular passage (13) occurs crosslinked, now, and just can be by half core transparent shell (5) observationIts crosslinking degree and cell static state and dynamic flow situation.

2. biological 3D according to claim 1 prints with cross-linked gel status monitoring syringe, it is characterized in that by swashingLight punching technology, on the needle tubing on syringe needle (1), metal part is punched, and forms uniform aperture (12), aperture (12) numberMeasure The more the better; On syringe needle 180 degree scale is used for rotating signal.

3. biological 3D according to claim 1 prints with cross-linked gel status monitoring syringe, it is characterized in that: half core is saturatingThe structure of bright housing (5), its inner semicircle annular that adopts, is used for locating syringe needle (1) and reserves semicircle annular tubular passage (13),Syringe needle (1) can be rotated in half core transparent shell (5), the crosslinked situation of observable material, cell static state and dynamicMobility status; Half core transparent shell (5) adopts resin material, makes by SLA photocuring technology.

4. biological 3D according to claim 1 prints with cross-linked gel status monitoring syringe, it is characterized in that: upper and lower rubberSealing function, in playing positioning action, is also played in rubber gasket (2) and (6), prevents that material from infiltrating, at upper rubber blanketOn circle (2), also have and the upper 180 corresponding degree graduation marks of syringe needle (1), to be shown in the syringe needle rotation packing in material process.

5. biological 3D prints and uses a cross-linked gel state monitoring method, adopts biological 3D according to claim 1 to printOperate with cross-linked gel status monitoring syringe, it is characterized in that concrete operation step is as follows:

1), before adding material, first by the scale of the lower rubber washer (6) of scale aligning on syringe needle (1) needle tubing, so that pinHead (1) needle tubing half with holes is towards the outside of half core transparent shell, and half not with holes inwards;

2), add a kind of material by the import (9) on half core transparent shell (5);

3), above-mentioned material add complete after, 180^oRotation syringe needle, makes syringe needle (1) needle tubing half with holes towards half core transparent shell(5) inner side, half not with holes is towards outside;

4), after syringe needle (1) rotation, add another kind of material by the piston syringe being equipped with on syringe needle, this material passes throughThe infiltration of the aperture (12) on syringe needle (1) is reacted with the material adding by half core transparent shell (5) upper inlet (9), thisSample just can be by half core transparent shell (5) extent of reaction to two kinds of materials and cell is static and hydrodynamic is mobile carries outMonitoring.