阅读说明：本技术 一种面向药物微球的多浓度微液滴芯片及其制造方法 (Multi-concentration micro-droplet chip for drug microspheres and manufacturing method thereof ) 是由 刘海强 许依海 杨晨 吕明 于 2021-07-19 设计创作，主要内容包括：本发明公开了一种面向药物微球的多浓度微液滴芯片及其制造方法。多浓度微液滴芯片包括下基片和上盖片,在上盖片上设置有注入液体的通孔,在下基片上设置了由圆心向外辐射分布的曲线形浓度梯度流道和微液滴流道。浓度梯度流道可以稳定实现药物的浓度梯度分布,微液滴流道通过矿物油控制药物形成大小可控的微液滴。芯片的制造过程中通过3D打印技术打印上盖片与下基片的阳模,然后通过浇注法与热键合得到多浓度微液滴芯片。该制备方法操作简单,成本低廉,可以快速、大量生产微液滴芯片。本发明可以实现多浓度、粒径可控的药物微球的制备。为研究药物缓释凝胶微球最佳浓度及粒径大小提供了更可靠的方案。(The invention discloses a multi-concentration micro-droplet chip for drug microspheres and a manufacturing method thereof. The multi-concentration micro-droplet chip comprises a lower substrate and an upper cover plate, wherein a through hole for injecting liquid is formed in the upper cover plate, and a curved concentration gradient runner and a micro-droplet runner which are radially distributed from the center of a circle to the outside are formed in the lower substrate. The concentration gradient flow channel can stably realize the concentration gradient distribution of the medicine, and the micro-droplet flow channel forms micro-droplets with controllable sizes through the mineral oil medicine control agent. In the manufacturing process of the chip, male molds of the upper cover plate and the lower substrate are printed by a 3D printing technology, and then the multi-concentration micro-droplet chip is obtained by casting and thermal bonding. The preparation method is simple to operate and low in cost, and micro-droplet chips can be rapidly produced in large quantities. The invention can realize the preparation of the drug microspheres with multiple concentrations and controllable particle sizes. Provides a more reliable scheme for researching the optimal concentration and the particle size of the drug sustained-release gel microspheres.)

1. A multi-concentration micro-droplet chip for drug microspheres is characterized in that: comprises a circular upper cover plate (2) and a lower base plate (1) with the same diameter;

the lower substrate (1) is provided with a concentration gradient flow channel (6), a micro-droplet flow channel (7), a micro-droplet outlet, a medicinal solution inlet (3), a diluted solution inlet (4) and a plurality of mineral oil inlets (5);

the drug solution inlet (3) and the diluted solution inlet (4) are symmetrically arranged at two sides of the circle center of the lower substrate (1); the concentration gradient flow channel (6) is formed by sequentially arranging a concentric circular arc flow channel and a bent flow channel and radiates from the circle center of the lower substrate (1) to the circumference; i +2 bent flow channels are arranged between the ith concentric circular arc flow channel and the (i + 1) th concentric circular arc flow channel from inside to outside; a plurality of output ports of the last concentric arc runner are respectively converged into a plurality of micro-droplet runners (7) through a plurality of bent runners; the drug solution inlet (3) and the diluted solution inlet (4) are connected with a first concentric circular arc flow channel of the concentration gradient flow channel (6); one end of the micro-droplet flow channel (7) is a mineral oil inlet (5), and the other end is a micro-droplet outlet;

the upper cover plate (2) is provided with a plurality of through holes which respectively correspond to the drug solution inlet (3), the diluted solution inlet (4) and the mineral oil inlets (5) on the lower substrate (1).

2. The multi-concentration micro-droplet chip oriented to drug microspheres of claim 1, wherein: each curved flow passage is bent twice.

3. The multi-concentration micro-droplet chip oriented to drug microspheres of claim 1, wherein: the plurality of concentric circular arc runners are arranged at equal intervals.

4. The multi-concentration micro-droplet chip oriented to drug microspheres of claim 1, wherein: the radius of the upper cover plate (2) and the radius of the lower base plate (1) are both 40 mm.

5. The multi-concentration micro-droplet chip oriented to drug microspheres of claim 1, wherein: the length and the width of the cross section of the concentration gradient flow channel and the micro-droplet flow channel are both 0.4 mm.

6. The method for manufacturing a multi-concentration micro-droplet chip oriented to drug microspheres according to claim 1, wherein the method comprises the following steps: the method specifically comprises the following steps:

step 1, manufacturing an upper cover plate male die of a multi-concentration micro-droplet chip by using a 3D printer, putting a printed lower substrate male die into an isopropanol solution, soaking for 8-10 min, and then putting into a 98% ethanol solution, and soaking for 5 min;

step 2, respectively preparing A, B parts of PDMS and a curing agent according to the proportion of 10:1 and 15:1, fully stirring and mixing the materials, and then placing the materials into a defoaming machine for defoaming treatment;

step 3, spraying a paper-based mold and a glass mold by using a release agent; placing the lower substrate male die processed in the step one into a paper-based die, and respectively pouring the paper-based die and a glass die by using a material A and a material B;

step 4, firstly, defoaming the mold after pouring in the step 3, then sequentially putting the mold into a constant-temperature drying box for curing, and after curing, putting the mold into a dust-free environment to cool the mold to room temperature; wherein the curing temperature of the paper-based mold is 40-50 ℃, and the curing time is 4-4.5 hours; the curing temperature of the glass mold is 75-80 ℃, and the curing time is 35 minutes;

step 5, respectively stripping the lower substrate and the upper cover plate from the paper-based mold and the glass mold, cutting the lower substrate and the upper cover plate into circles with the radius of 40mm, and then punching holes in the positions, corresponding to solution inlets in the lower substrate, on the upper cover plate, wherein the hole diameter is 1.5 mm;

step 6, overlapping the upper cover plate and the lower substrate processed in the step 5, aligning the through holes with the solution inlets one by one, and then placing the aligned through holes in a constant-temperature drying box for bonding to obtain a multi-concentration micro-droplet chip; wherein the bonding temperature is 70-80 ℃, and the bonding time is 1 hour.

7. The method for manufacturing a multi-concentration micro-droplet chip oriented to drug microspheres according to claim 6, wherein the method comprises the following steps: in step 2, the time for defoaming A, B was 5 minutes.

Technical Field

The invention belongs to the field of micro-droplet chip manufacturing, and particularly relates to a multi-concentration micro-droplet chip for drug microspheres and a manufacturing method thereof.

Background

Microfluidic chip technology (Microfluidics) is a technology that can integrate operations of biological culture, chemical reaction, and medical diagnosis processes that need to be performed on sample reagents into a chip with a micron-scale flow channel.

In drug delivery systems, drug microspheres are receiving increasing attention due to their good drug-loading properties and controlled release properties. The main influences on the performance of a drug microsphere are the concentration of drug wrapped by the microsphere and the particle size of the microsphere, and in addition, the influences have great influence on the administration mode. The gel microsphere is micron-sized or submicron spherical particles with a high molecular network structure, and due to the special structure, when the gel microsphere is immersed in a proper solution, the gel microsphere can absorb the volume expansion of the solution, and a reagent in the gel can be slowly released through the network. Therefore, the gel microsphere has potential application prospects in the fields of medicine and protein carriers, sensing, photonic crystal construction, catalysis, separation, purification and the like. The drug gel microspheres are widely applied to improve the efficiency of disease treatment by increasing the drug concentration of a specific part and solving the problems of low solubility and low specificity of small molecule drugs and rapid elimination of the small molecule drugs. Strict control and prediction of the size, dispersity and drug loading of gel microsphere particles are critical to the pharmacokinetic and pharmacodynamic efficacy. However, it is not easy to prepare complex gel microspheres at high levels and reproducibly. Although micron-scale gel microspheres can be prepared by using the traditional preparation methods such as a solvent extraction method, a phase separation method, an emulsion crosslinking method and the like, the size of the obtained gel microspheres is not uniform and is difficult to regulate. The micro-droplet with uniform and adjustable size can be prepared by utilizing the traditional micro-droplet chip, but micro-mixing is needed to be carried out on multi-concentration micro-droplets after the micro-droplets are generated, and the insufficient mixing effect can cause a great amount of medicine loss after the cross-linking reaction, so that the medicine effect is not ideal.

The commonly used micro-fluidic chip preparation methods at present mainly include a photoetching method and an etching method, and when the micro-fluidic chip material is a high-molecular organic polymer, the processing method mainly includes the following four methods: hot pressing, molding, LIGF, laser ablation. Although the technology can process the runner chip with higher precision, the process is complex, the equipment is expensive, and the flow is complicated.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a multi-concentration micro-droplet chip facing to drug microspheres and a manufacturing method thereof, the micro-droplet chip for generating a concentration gradient flow channel along the circumference is designed, different solutions are fully mixed through a bent flow channel, the concentration gradient distribution of drugs is stabilized, and then micro-droplets are generated. The chip material adopts PDMS, and the positive membrane of preparation chip is printed through 3D, adopts the injection molding method to pour out the different positions of chip on the positive membrane, and the method is simple, reduces manual operation to can reuse positive membrane preparation a large amount of chips, reduce preparation cost.

A multi-concentration micro-droplet chip facing to drug microspheres comprises a circular upper cover plate and a lower substrate with the same diameter.

The lower substrate is provided with a concentration gradient flow channel, a micro-droplet outlet, a medicinal solution inlet, a diluted solution inlet and a plurality of mineral oil inlets.

The drug solution inlet and the diluting solution inlet are symmetrically arranged at two sides of the circle center of the lower substrate. The concentration gradient flow channel is formed by sequentially arranging a concentric circular arc flow channel and a bent flow channel and radiates from the circle center of the lower substrate to the circumference; and i +2 bent flow channels are arranged between the ith concentric circular arc flow channel from inside to outside and the (i + 1) th concentric circular arc flow channel. And a plurality of output ports of the last concentric arc runner are respectively merged into a plurality of micro-droplet runners through a plurality of bent runners. The drug solution inlet and the dilute solution inlet are connected with a first concentric circular arc flow channel of the concentration gradient flow channel. One end of the micro-droplet flow channel is a mineral oil inlet, and the other end is a micro-droplet outlet.

The upper cover plate is provided with a plurality of through holes which respectively correspond to the drug solution inlet, the diluted solution inlet and the mineral oil inlets on the lower substrate.

Preferably, each meandering channel is bent twice.

Preferably, the plurality of concentric circular arc runners are arranged at equal intervals.

Preferably, the radii of the upper cover sheet and the lower base sheet are both 40 mm.

Preferably, the concentration gradient flow channel and the micro-droplet flow channel have a cross section with a length and a width of 0.4 mm.

A manufacturing method of a multi-concentration micro-droplet chip for drug microspheres specifically comprises the following steps:

step 1, manufacturing a lower substrate male die of the multi-concentration micro-droplet chip by using a 3D printer, putting the printed lower substrate male die into an isopropanol solution, soaking for 8-10 minutes, and then putting the lower substrate male die into a 98% ethanol solution, and soaking for 5 minutes.

Step 2, preparing A, B parts of materials by the PDMS and the curing agent according to the proportion of 10:1 and 15:1 respectively, fully stirring and mixing the materials, and then placing the materials into a defoaming machine for defoaming treatment.

And 3, spraying a paper-based mold and a glass mold by using a release agent. And (4) placing the lower substrate male die processed in the step one into a paper-based die, and respectively pouring the paper-based die and a glass die by using the material A and the material B.

And 4, firstly, defoaming the mold after pouring in the step 3, then sequentially putting the mold into a constant-temperature drying box for curing, and after curing, putting the mold into a dust-free environment to cool the mold to room temperature. Wherein the curing temperature of the paper-based mold is 40-50 ℃, and the curing time is 4-4.5 hours; the curing temperature of the glass mold is 75-80 ℃, and the curing time is 35 minutes.

And 5, respectively stripping the lower substrate and the upper cover plate from the paper-based mold and the glass mold, cutting the lower substrate and the upper cover plate into circles with the radius of 40mm, and then punching holes in the positions, corresponding to solution inlets in the lower substrate, on the upper cover plate, wherein the hole diameter is 1.5 mm.

And 6, overlapping the upper cover plate processed in the step 5 with the lower substrate, aligning the through holes with the solution inlets one by one, and then placing the aligned through holes in a constant-temperature drying box for bonding to obtain the multi-concentration micro-droplet chip. Wherein the bonding temperature is 70-80 ℃, and the bonding time is 1 hour.

Preferably, the time for defoaming A, B for two portions is 5 minutes.

The invention has the following beneficial effects:

1. the concentration gradient flow channel of the multi-concentration micro-droplet chip is formed by arranging a concentric circular arc flow channel and a bent flow channel in sequence, when liquid flows in the micro-fluidic chip, the liquid is in a laminar flow state, the mixing efficiency among fluids can be improved by increasing the length of the flow channel and changing the bending direction, the contact surface of the mixed liquid is increased, and the liquid with stable concentration gradient distribution is obtained.

2. The liquid in the concentration gradient flow passage finally converges into the micro-droplet flow passage, and the added mineral oil generates the drug microspheres with controllable particle diameters, so that the diameter of the micro-droplet flow passage is reduced, and the droplets with smaller diameters can be generated.

3. In the manufacturing process of the multi-concentration micro-droplet chip, PDMS/curing agents with different proportions are respectively poured to obtain a lower substrate and an upper cover plate, and then the upper cover plate and the lower substrate are thermally bonded in a heating mode, so that the bonding strength and the bonding tightness are improved.

Drawings

FIG. 1 is a perspective view of a bottom substrate of a multi-concentration micro-droplet chip;

FIG. 2 is a perspective view of an upper cover plate of a multi-concentration micro-droplet chip;

FIG. 3 is a flow chart of a method for manufacturing a multi-concentration micro-droplet chip.

Detailed Description

The invention is further explained below with reference to the drawings;

a multi-concentration micro-droplet chip facing to drug microspheres comprises a circular upper cover plate 2 and a lower substrate 1, wherein the radii of the circular upper cover plate and the lower substrate are both 40 mm.

As shown in fig. 1, the lower substrate 1 has a thickness of 5mm, and is provided with a concentration gradient channel 6, a micro droplet channel 7, a micro droplet outlet, a drug solution inlet 3, a diluting solution inlet 4, and 5 mineral oil inlets 5.

The drug solution inlet 3 and the diluting solution inlet 4 are symmetrically arranged on two sides of the center of the lower substrate 1. The concentration gradient flow channel 6 is formed by sequentially arranging concentric circular arc flow channels and bent flow channels, radiates from the circle center of the lower substrate 1 to the circumference to form a 3-level gradient flow channel, and finally converges into the micro-droplet flow channel 7. Wherein the distance between the arc runners of each circle center is equal, and each curved runner is bent twice. The drug solution inlet 3 and the diluting solution inlet 4 are connected to a first concentric circular arc flow path of the concentration gradient flow path 6. One end of the micro-droplet flow channel 7 is a mineral oil inlet 5, and the other end is a micro-droplet outlet. The length and the width of the cross section of the concentration gradient flow channel and the micro-droplet flow channel are both 0.4 mm.

As shown in FIG. 2, the upper cover plate 2 has a thickness of 6mm, and is provided with a drug solution through hole 8, a diluting solution through hole 9, and a mineral oil through hole 10, which correspond to the drug solution inlet 3, the diluting solution inlet 4, and the 5 mineral oil inlets 5 of the lower substrate 1, respectively.

As shown in fig. 3, a method for manufacturing a multi-concentration micro-droplet chip facing to a drug microsphere specifically includes the following steps:

example one

Step 1, manufacturing an upper cover plate male die of the multi-concentration micro-droplet chip by using a 3D printer, putting the printed lower substrate male die into an isopropanol solution to be soaked for 8 minutes, and then putting the lower substrate male die into a 98% ethanol solution to be soaked for 5 minutes.

Step 2, preparing A, B parts of materials by the PDMS and the curing agent according to the proportion of 10:1 and 15:1 respectively, fully stirring and mixing the materials, and then placing the materials into a defoaming machine for defoaming treatment.

Step 3, spraying a paper-based mold and a glass mold by using a release agent; and (4) placing the lower substrate male die processed in the step one into a paper-based die, and respectively pouring the paper-based die and a glass die by using the material A and the material B.

And 4, firstly, defoaming the mold after pouring in the step 3, then sequentially putting the mold into a constant-temperature drying box for curing, and then putting the mold into a dust-free environment for cooling to room temperature after curing. Wherein the curing temperature of the paper-based mold is 40 ℃, and the curing time is 4.5 hours; the curing temperature of the glass mold was 75 ℃ and the curing time was 35 minutes.

And 5, respectively stripping the lower substrate and the upper cover plate from the paper-based mold and the glass mold, cutting the lower substrate and the upper cover plate into circles with the radius of 40mm, and then punching holes in the positions, corresponding to solution inlets in the lower substrate, on the upper cover plate, wherein the hole diameter is 1.5 mm.

And 6, overlapping the upper cover plate processed in the step 5 with the lower substrate, aligning the through holes with the solution inlets one by one, and then placing the aligned through holes in a constant-temperature drying box for bonding to obtain the multi-concentration micro-droplet chip. Wherein the bonding temperature is 70 ℃ and the bonding time is 1 hour.

Example two

Step 1, manufacturing an upper cover plate male die of the multi-concentration micro-droplet chip by using a 3D printer, putting the printed lower substrate male die into an isopropanol solution to be soaked for 10 minutes, and then putting the lower substrate male die into a 98% ethanol solution to be soaked for 5 minutes.

Step 2, preparing A, B parts of materials by the PDMS and the curing agent according to the proportion of 10:1 and 15:1 respectively, fully stirring and mixing the materials, and then placing the materials into a defoaming machine for defoaming treatment.

Step 3, spraying a paper-based mold and a glass mold by using a release agent; and (4) placing the lower substrate male die processed in the step one into a paper-based die, and respectively pouring the paper-based die and a glass die by using the material A and the material B.

And 4, firstly, defoaming the male mold after pouring in the step 3, then sequentially putting the male mold into a constant-temperature drying box for curing, and after curing, putting the male mold into a dust-free environment to cool the male mold to room temperature. Wherein the curing temperature of the paper-based mold is 50 ℃, and the curing time is 4 hours; the curing temperature of the glass mold was 80 ℃ and the curing time was 35 minutes.

And 5, respectively stripping the lower substrate and the upper cover plate from the paper-based mold and the glass mold, cutting the lower substrate and the upper cover plate into circles with the radius of 40mm, and then punching holes in the positions, corresponding to solution inlets in the lower substrate, on the upper cover plate, wherein the hole diameter is 1.5 mm.

And 6, overlapping the upper cover plate processed in the step 5 with the lower substrate, aligning the through holes with the solution inlets one by one, and then placing the aligned through holes in a constant-temperature drying box for bonding to obtain the multi-concentration micro-droplet chip. Wherein the bonding temperature is 80 ℃ and the bonding time is 1 hour.

EXAMPLE III

Step 1, manufacturing an upper cover plate male die of the multi-concentration micro-droplet chip by using a 3D printer, putting the printed lower substrate male die into an isopropanol solution to be soaked for 9 minutes, and then putting the lower substrate male die into a 98% ethanol solution to be soaked for 5 minutes.

Step 2, preparing A, B parts of materials by the PDMS and the curing agent according to the proportion of 10:1 and 15:1 respectively, fully stirring and mixing the materials, and then placing the materials into a defoaming machine for defoaming treatment.

Step 3, spraying a paper-based mold and a glass mold by using a release agent; and (4) placing the lower substrate male die processed in the step one into a paper-based die, and respectively pouring the paper-based die and a glass die by using the material A and the material B.

And 4, firstly, defoaming the male mold after pouring in the step 3, then sequentially putting the male mold into a constant-temperature drying box for curing, and after curing, putting the male mold into a dust-free environment to cool the male mold to room temperature. Wherein the curing temperature of the paper-based mold is 45 ℃, and the curing time is 4.5 hours; the curing temperature of the glass mold was 77 ℃ and the curing time was 35 minutes.

And 5, respectively stripping the lower substrate and the upper cover plate from the paper-based mold and the glass mold, cutting the lower substrate and the upper cover plate into circles with the radius of 40mm, and then punching holes in the positions, corresponding to solution inlets in the lower substrate, on the upper cover plate, wherein the hole diameter is 1.5 mm.

And 6, overlapping the upper cover plate processed in the step 5 with the lower substrate, aligning the through holes with the solution inlets one by one, and then placing the aligned through holes in a constant-temperature drying box for bonding to obtain the multi-concentration micro-droplet chip. Wherein the bonding temperature is 75 ℃ and the bonding time is 1 hour.