阅读说明：本技术 一种适用于化学交联成型的微流控生物纺丝技术 (Microfluidic biological spinning technology suitable for chemical crosslinking forming ) 是由 刘凯 何浩男 张磊 张洪杰 于 2019-10-15 设计创作，主要内容包括：本发明提供了一种适用于化学交联成型的微流控生物纺丝技术,包括：将生物基溶液经导管和注射器通入到同轴双通道微流控芯片的第一微管；将交联剂溶液经导管和注射器通入到同轴双通道微流控芯片的第二微管；第一微管的生物基溶液和第二微管的交联剂溶液混合交联,得到纺丝液；将纺丝液经第一微管出口挤出进入凝固浴中,通过转筒收集器收集得到纤维。本发明提供了选取生物基材料作为纺丝原料,同时结合特定的新型微流控芯片,通过调节上述微流控芯片的参数及纺丝参数,可制备具有需要尺寸和优异力学性能参数的生物纤维材料。本发明所制备的化学交联生物纤维材料的力学性能明显优于纯生物纤维。(The invention provides a microfluidic biological spinning technology suitable for chemical crosslinking forming, which comprises the following steps: introducing the bio-based solution into a first micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector; introducing the cross-linking agent solution into a second micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector; mixing and crosslinking the bio-based solution of the first microtube and the crosslinking agent solution of the second microtube to obtain a spinning solution; and extruding the spinning solution into a coagulating bath through a first micro-pipe outlet, and collecting the spinning solution by a rotary drum collector to obtain the fibers. The invention provides a method for preparing a biological fiber material with required size and excellent mechanical property parameters by selecting a biological base material as a spinning raw material, simultaneously combining a specific novel micro-fluidic chip and adjusting the parameters and spinning parameters of the micro-fluidic chip. The mechanical property of the chemical crosslinking biological fiber material prepared by the invention is obviously superior to that of pure biological fiber.)

1. A microfluidic biological spinning technology suitable for chemical crosslinking molding is characterized by comprising the following steps:

A) introducing the bio-based solution into a first micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector; introducing the cross-linking reagent solution into a second micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector;

B) mixing and crosslinking the bio-based solution of the first microtube and the crosslinking agent solution of the second microtube to obtain a spinning solution;

C) and extruding the spinning solution into a coagulating bath through a first micro-pipe outlet, and collecting the spinning solution by a rotary drum collector to obtain the fibers.

2. The preparation method according to claim 1, wherein the coaxial two-channel microfluidic chip comprises a second microtube and a first microtube which are coaxially arranged, and the first microtube is a square glass microtube; the second microtube is a cylindrical glass microtube; the cylindrical glass micro-tube is arranged in the square glass micro-tube, and the outlet of the square glass micro-tube is in a tapered shape with the tube diameter gradually reduced; the inner diameter of the conical outlet is 0.05-0.50 mm.

3. The preparation method of claim 1, wherein the bio-based solution is prepared by dissolving a bio-based material solid powder in a buffer solution, and the concentration of the bio-based solution is 50-800 mg/mL.

4. The method according to claim 3, wherein the buffer solution is a PBS buffer solution, and the pH of the buffer solution is 8.

5. The method according to claim 1, wherein the cross-linking agent solution has a mass concentration of 0.1 to 10.0%.

6. The method of claim 1, wherein the coagulation bath is 50% to 100% wt methanol/water solution.

7. The method of claim 1, wherein the flow rate of the bio-based solution is 2 to 20 μ L/min.

8. The method according to claim 7, wherein the flow rate of the crosslinking agent solution is 2 to 15. mu.L/min.

9. A microfluidic biological spinning technology suitable for chemical crosslinking molding is characterized by being prepared by the preparation method of any one of claims 1-8.

Technical Field

The invention relates to the technical field of biological fiber preparation, in particular to a chemical crosslinking formed biological fiber material and a preparation method thereof.

Background

Artificial biofiber materials are a class of fibrous materials formed by secondary spinning of biobased materials extracted from plants or animals, which often have low density, high mechanical strength, excellent ductility, degradability and good biocompatibility. However, such fiber materials have not yet achieved the excellent properties of some natural fiber materials, such as spider silk fibers. Therefore, in the process of preparing the bio-fiber, the original bio-fiber needs to be modified. Chemical crosslinking is a common biomaterial modification method, a stable covalent bond is formed by a chemical crosslinking reagent, the common biomaterial chemical crosslinking reagent comprises glutaraldehyde, genipin, hexamethylene dicyanate, EDC/NHS and the like, and the crosslinking reagents can enable a biomaterial to obtain a more stable structure, so that corresponding application is increased. However, at present, the crosslinking effect of these crosslinking reagents is very different, and taking the most common EDC/NHS as an example, the crosslinking speed of the crosslinking reagent is slow, the number of processing procedures is large, and it is difficult to obtain the desired crosslinked product in a short time, so the application of the crosslinking reagent in fiber crosslinking is greatly limited.

The microfluidic spinning technology is a technology for controlling fiber formation at a microscale, and the technology is an emerging technology developed in recent years and mainly depends on a microscale chip. By controlling the relevant parameters of external conditions such as flow rate, concentration, viscosity and the like, the micro-fluidic chip can realize the efficient and rapid reaction of the solution under the micro-scale, and further control the generation of fibers. Due to its great advantages, the technology has been successfully developed into a popular research field integrating biology, chemistry, electronics, materials, mechanical and electrical and other subjects.

Most of the cross-linked products of the traditional biomaterials are gel substances, and are used in the fields of drug slow release or biological adhesives and the like, and the application potential is not further developed. At present, no technology for realizing rapid crosslinking and mass preparation of the biological fiber material, obtaining the fiber material with better performance and wider application range exists.

Disclosure of Invention

In view of the above, the technical problem to be solved by the present invention is to provide a microfluidic biological spinning technology suitable for chemical crosslinking formation, and the chemical crosslinking biological fiber prepared by the present invention has good mechanical properties.

The invention provides a preparation method of a crosslinking agent coupled protein fiber, which comprises the following steps:

A) introducing the bio-based solution into a first micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector; introducing the cross-linking agent solution into a second micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector;

B) mixing and crosslinking the bio-based solution of the first microtube and the crosslinking agent solution of the second microtube to obtain a spinning solution;

C) and extruding the spinning solution into a coagulating bath through a first micro-pipe outlet, and collecting the spinning solution by a rotary drum collector to obtain the fibers.

Preferably, the coaxial two-channel microfluidic chip comprises a second micro-tube and a first micro-tube which are coaxially arranged, and the first micro-tube is a square glass micro-tube; the second microtube is a cylindrical glass microtube; the cylindrical glass micro-tube is arranged in the square glass micro-tube, and the outlet of the square glass micro-tube is in a tapered shape with the tube diameter gradually reduced; the inner diameter of the conical outlet is 0.05-0.0.50 mm.

Preferably, the bio-based solution is prepared by dissolving bio-based material solid powder into a buffer solution, and the concentration of the bio-based solution is 50-800 mg/mL.

Preferably, the buffer solution is a PBS buffer solution, and the pH value of the buffer solution is 8.

Preferably, the mass concentration of the cross-linking agent solution is 0.1-10.0%.

Preferably, the coagulating bath is 50-100 wt% methanol water solution.

Preferably, the flow rate of the bio-based solution is 2-20 mu L/min,

preferably, the flow rate of the cross-linking agent solution is 2-15 mu L/min.

The invention provides a microfluidic biological spinning technology suitable for chemical crosslinking molding, which is prepared by the preparation method in any one of the technical schemes.

Compared with the prior art, the invention provides a microfluidic biological spinning technology suitable for chemical crosslinking forming, which comprises the following steps: A) introducing the bio-based solution into a first micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector; introducing the cross-linking agent solution into a second micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector; B) mixing and crosslinking the bio-based solution of the first microtube and the crosslinking agent solution of the second microtube to obtain a spinning solution; C) and extruding the spinning solution into a coagulating bath through a first micro-pipe outlet, and collecting the spinning solution by a rotary drum collector to obtain the fibers. The invention provides a method for preparing biological fiber with required size and excellent mechanical property parameters by selecting biological materials as spinning raw materials, simultaneously combining a specific novel micro-fluidic chip and adjusting the parameters and spinning parameters of the micro-fluidic chip. The mechanical property of the chemical crosslinking fiber material prepared by the invention is obviously superior to that of pure biological fiber, and the application of the crosslinking biological fiber material in biology, materials and medical fields is developed; the chemical crosslinking biological fiber can be applied to the fields of artificial ligaments, sports bandages, surgical sutures, medical biological scaffolds and the like.

Drawings

FIG. 1 is a schematic view of an overall process of microfluidic spinning according to an embodiment of the present invention;

FIG. 2 is a schematic view of a microfluidic chip according to an embodiment of the present invention;

FIG. 3 is a graph of a fiber tensile test of example 1 of the present invention;

FIG. 4 is a graph of a fiber tensile test of example 2 of the present invention;

FIG. 5 is a graph of a fiber tensile test of example 3 of the present invention;

FIG. 6 is a graph of the fiber tensile test of example 4 of the present invention.

Detailed Description

The invention provides a microfluidic biological spinning technology suitable for chemical crosslinking molding, and a person skilled in the art can appropriately improve process parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the scope of the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.

The invention provides a preparation method of a crosslinking agent coupled protein fiber, which comprises the following steps:

A) introducing the bio-based solution into a first micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector; introducing the cross-linking agent solution into a second micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector;

B) mixing and crosslinking the bio-based solution of the first microtube and the crosslinking agent solution of the second microtube to obtain a spinning solution;

C) and extruding the spinning solution into a coagulating bath through a first micro-pipe outlet, and collecting the spinning solution by a rotary drum collector to obtain the fibers.

The invention provides a microfluidic spinning device which comprises an injection pump, wherein the injection pump is connected with a second micro-tube of a microfluidic chip through an inner-phase injector and a guide tube; the injection pump is connected with the first micro-tube of the micro-fluidic chip through the external phase injector and the guide tube, the coagulating bath arranged below the micro-fluidic chip, the rotary drum collector connected with the coagulating bath and the motor for driving the rotary drum collector.

Fig. 1 of the present invention is a schematic view of an overall microfluidic spinning process in an embodiment of the present invention.

Wherein, 1 is an injection pump; 2 is an internal phase injector; 3 is an external phase injector; 4 is a micro-fluidic chip; 5, coagulating bath; 6 is a rotating drum collector; and 7 is a motor.

The sources and specific arrangements of the above-mentioned components are not limited in the present invention, and those skilled in the art will be familiar with them.

According to the invention, the first microtube is a square glass microtube; the inner diameter of the square glass micro-tube is 1.00mm, the length of the square glass micro-tube is 5.00cm, and the second micro-tube is a cylindrical glass micro-tube; the inner aperture of the cylindrical glass micro-tube is 0.400-0.600 mm, the outer diameter is 0.900mm, and the length is 5.00 cm.

The coaxial two-channel micro-fluidic chip comprises a cylindrical glass micro-tube and a square glass micro-tube which are coaxially arranged, wherein the cylindrical glass micro-tube is arranged in the square glass micro-tube.

The outlet of the square glass micro-tube is in a tapered shape with the tube diameter gradually reduced; the inner diameter of the conical outlet is 0.05-0.50 mm. The special treatment of the invention can increase the shearing force and the extrusion force, and improve the ordered arrangement among molecules and the mechanical property of spinning. Specifically, as shown in fig. 2, 8 is a second microtube, 9 is a first microtube, and 10 is a conical head.

The invention leads the biology-based solution into the first microtube of the coaxial double-channel micro-fluidic chip through the guide tube and the injector. Preferably, the biological material solid powder is placed in a centrifuge tube, a buffer solution is added, and the foam is removed by shaking and centrifugation to obtain the biological base solution.

According to the invention, the biological-based solution is preferably prepared by dissolving solid biological material powder in a buffer solution, and the concentration of the biological-based solution is preferably 50-800 mg/mL. The buffer solution is PBS buffer solution, and the pH value of the buffer solution is 8.

Introducing the cross-linking agent solution into a second micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector;

the mass concentration of the cross-linking agent solution is preferably 0.1-10.0%.

Mixing and crosslinking the bio-based solution of the first microtube and the crosslinking agent solution of the second microtube to obtain a spinning solution;

the preferable concrete is as follows: respectively introducing the bio-based solution and the cross-linking agent solution into a square micro-tube and a cylindrical micro-tube of the micro-fluidic chip through an injector and a guide tube, mixing and cross-linking, and controlling the injector to adjust the extrusion speed of the two-phase solution by an injection pump.

The bio-based solution is introduced into a square glass micro-tube from an external phase through an injection pump, an injector and a guide tube, the cross-linking agent solution is introduced into a cylindrical glass micro-tube from an internal phase, the two-phase solutions start to be mixed after being contacted, and the cross-linking reaction occurs.

The flow rate of the bio-based solution is preferably 2-20 mu L/min, more preferably 3-15 mu L/min, most preferably 4-13 mu L/min, and the flow rate of the cross-linking agent solution is preferably 2-15 mu L/min; more preferably 3-12 muL/min; more preferably 4 to 10. mu.L/min.

Extruding the spinning solution into a coagulation bath through a first micro-tube outlet, introducing the mixed solution into the coagulation bath through a conical outlet, and performing dehydration on the crosslinking bio-based solution and a denaturation reaction on the biological material. The resulting fibers were then collected by a drum collector. The spinning solution was passed to a methanol/water system coagulation tank and collected by a drum collector.

The extrusion speed is preferably 10-30 mu L/min.

Wherein the coagulating bath is 60-100 wt% methanol water solution.

The speed of the rotary drum collector is preferably 2-10 cm/s.

The degree of the cross-linking reaction of the biomaterial of the present invention is determined by the specific cross-linker solution concentration and pumping rate. Further, the bio-fiber size parameter is determined by the inner diameter of the tapered outlet and the extrusion speed. Further, the surface smoothness and mechanical property parameters of the biological fibers can be determined by the stretching action of the drum collector.

The invention expands the application of the microfluidic technology in the field of biomaterial spinning. Cheap and easily-obtained regenerated biological materials are selected as spinning raw materials, a simple novel micro-fluidic chip is designed and prepared, and biological fibers with different sizes and mechanical property parameters can be prepared by adjusting design parameters and experimental parameters of the micro-fluidic chip. The performance of the prepared chemical crosslinking biological fiber material is obviously superior to that of pure biological fiber, and the application of the biological fiber material in biology, materials and medical fields is developed.

The invention provides a chemical crosslinking biological fiber which is prepared by the preparation method of any one of the technical schemes.

The invention provides a microfluidic biological spinning technology suitable for chemical crosslinking forming, which comprises the following steps: A) introducing the bio-based solution into a first micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector; introducing the cross-linking agent solution into a second micro-tube of the coaxial two-channel micro-fluidic chip through a guide tube and an injector; B) mixing and crosslinking the bio-based solution of the first microtube and the crosslinking agent solution of the second microtube to obtain a spinning solution; C) and extruding the spinning solution into a coagulating bath through a first micro-pipe outlet, and collecting the spinning solution by a rotary drum collector to obtain the fibers. The invention provides a method for preparing biological fiber with required size and excellent mechanical property parameters by selecting a regenerated biological material as a spinning raw material, simultaneously combining a specific novel micro-fluidic chip and adjusting the parameters and spinning parameters of the micro-fluidic chip. The mechanical property of the cross-linking agent coupled biological fiber material prepared by the invention is obviously better than that of the pure biological fiber, and the application of the biological fiber material in biology, materials and medical fields is developed; the cross-linked biological fiber can be applied to the fields of artificial ligaments, sports bandages, surgical sutures, medical biological scaffolds and the like.

In order to further illustrate the present invention, the following will describe a chemically crosslinked bio-fiber and a method for preparing the same in detail with reference to the following examples.