CN-117138731-B - Collagen microsphere reaction device and application method thereof

CN117138731BCN 117138731 BCN117138731 BCN 117138731BCN-117138731-B

Abstract

The invention discloses a collagen microsphere reaction device and an application method thereof, wherein the device is based on the fact that structural members provided with a through hole array are mutually matched to form a serial type focusing through hole array, the problems of high integration difficulty and complex operation of a pipeline microfluidic technology are solved, the serial type focusing through hole array can be used for realizing instant mixing and emulsification balling of a collagen solution and a cross-linking agent, the problem of solution fluidity change caused by pre-mixing of the cross-linking agent and early solidification of the solution is solved, in addition, a plurality of containers synchronously generate monodisperse collagen microspheres in the focusing through hole array by adopting a homologous driving force, standardized batch production of the collagen microspheres can be realized, the problem of large batch-to-batch variability of the traditional method is solved, and the device can conveniently and efficiently realize collagen microsphere preparation and can be widely applied to the technical field of microsphere synthesis.

Inventors

GONG YAO
YOU KAI
GAO BOTAO
ZOU LILI

Assignees

广东省科学院生物与医学工程研究所

Dates

Publication Date: 20260512
Application Date: 20230818

Claims (10)

1. The collagen microsphere reaction device is characterized by comprising a first container, a second container, a third container and a sealing cover plate, wherein the tops of the first container, the second container and the third container are all provided with openings; the bottom of the first container is provided with a first through hole array, and the first container is used for loading a continuous phase solution; the bottom of the second container is provided with a second through hole array, the second container is used for loading a cross-linking agent solution, the second container is nested and installed in the first container, a first annular cavity is arranged between the outer diameter of the second container and the inner diameter of the first container, and a first gap is arranged between the lower surface of the bottom of the second container and the upper surface of the bottom of the first container; The bottom of the third container is provided with a third through hole array, the third container is used for loading discrete phase solution, the third container is nested and installed in the second container, a second annular cavity is formed between the outer diameter of the third container and the inner diameter of the second container, and a second gap is formed between the lower surface of the bottom of the third container and the upper surface of the bottom of the second container; The sealing cover plate covers the tops of the first container, the second container and the third container and is used for isolating the first container, the first annular cavity and the second annular cavity into independent spaces, a first air passage interface, a second air passage interface and a third air passage interface are sequentially arranged above the first container, above the first annular cavity and above the second annular cavity, through holes in the first through hole array, the second through hole array and the third through hole array are coaxially arranged, and the first through hole array, the second through hole array and the third through hole array form a series focusing through hole array based on the first gap and the second gap.
2. The collagen microsphere reaction apparatus according to claim 1, wherein a first positioning blind hole array is further arranged at the bottom of the first container, a first positioning shaft array and a first protruding cylindrical array are further arranged at the bottom of the second container, and the second through hole array is arranged in the first cylindrical array, and the first positioning blind hole array and the first positioning shaft array are aligned with the first through hole array and the second through hole array through interference fit.
3. The collagen microsphere reaction device according to claim 1, wherein a second positioning blind hole array is further arranged at the bottom of the second container, a second positioning shaft array and a protruding second cylindrical array are further arranged at the bottom of the third container, and the third through hole array is arranged in the second cylindrical array, wherein the second positioning blind hole array and the second positioning shaft array are aligned with the second through hole array and the third through hole array through interference fit.
4. The collagen microsphere reaction apparatus according to claim 1, further comprising a driving device, wherein the driving device is connected to the first air passage interface, the second air passage interface, and the third air passage interface, respectively, and the driving device is configured to provide driving pressure to the first container, the second container, and the third container, respectively.
5. The collagen microsphere reaction apparatus according to claim 1, wherein the first array of through holes, the second array of through holes, and the third array of through holes comprise different sizes of through holes to form different sizes of focused through hole arrays.
6. An application method of a collagen microsphere reaction device, a collagen microsphere reaction apparatus according to any one of claims 1 to 5, wherein the application method comprises: Adding the continuous phase solution to the first container through the first gas path interface, adding the cross-linking agent solution to the second container through the second gas path interface, and adding the discrete phase solution to the third container through the third gas path interface; And under the preset reaction temperature, inputting preset pneumatic pressure to the first container, the second container and the third container through the first air passage interface, the second air passage interface and the third air passage interface, so that the discrete phase solution is mixed with the cross-linking agent solution in the second gap through the third through hole array and enters the second through hole array, further, the continuous phase solution and the mixed solution of the cross-linking agent solution and the discrete phase solution in the second through hole are vertically intersected in the circumferential direction of the first gap between the second through hole and the first through hole, the mixed solution is separated into independent microspheres, and the independent microspheres are conveyed to the collecting liquid through the first through hole array to complete cross-linking solidification, so that the collagen microspheres are obtained.
7. A collagen according to claim 6 an application method of a protein microsphere reaction device, the application method is characterized by further comprising the following steps: adding the collagen freeze-dried solid into deionized water or phosphate buffer solution, and dissolving in water bath to obtain recombinant collagen solution as the discrete phase solution.
8. A collagen according to claim 6 an application method of a protein microsphere reaction device, the application method is characterized by further comprising the following steps: Mixing paraffin oil with a surfactant, and preparing the continuous phase solution by shaking and magnetic stirring.
9. A collagen according to claim 6 an application method of a protein microsphere reaction device, the application method is characterized by further comprising the following steps: Mixing 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride solid and N-hydroxysuccinimide solid, adding phosphate buffer solution for complete dissolution, and obtaining the cross-linking agent solution.
10. The method according to claim 6, further comprising a driving device connected to the first air passage interface, the second air passage interface, and the third air passage interface, wherein the inputting of the preset pneumatic pressure to the first container, the second container, and the third container through the first air passage interface, the second air passage interface, and the third air passage interface, respectively, comprises: The first air passage interface, the second air passage interface and the third air passage interface which are connected through the driving device respectively input different preset pneumatic pressures to the first container, the second container and the third container.

Description

Collagen microsphere reaction device and application method thereof Technical Field The invention relates to the technical field of microsphere synthesis, in particular to a collagen microsphere reaction device and an application method thereof. Background The existing collagen microsphere preparation method comprises a solvent evaporation method, a seed swelling method, a polymerization method, a sintering method, a spray drying method, a phase separation method, a microfluidic method and the like. The method has the advantages that the method is low in monodispersity, complex in preparation procedure and difficult to accurately control microsphere particle size, the method is remarkable in preparation of monodispersed microspheres, commercially available microfluidic technologies mainly comprise injection chip microfluidic methods and pipeline microfluidic methods, the injection chip microfluidic methods are high in technical threshold and depend on expensive equipment and dies, the injection chip and other equipment are high in integration difficulty, the whole microsphere production efficiency is low, the pipeline microfluidic methods are low-cost solutions, but have the problems that coaxiality assembly errors among pipelines are large, multichannel assembly operation is complex, and differences among batches of generated microspheres are large. Disclosure of Invention In view of the above, the embodiment of the invention provides a collagen microsphere reaction device and an application method thereof, which can conveniently and efficiently prepare collagen microspheres. In one aspect, the embodiment of the invention provides a collagen microsphere reaction device, which comprises a first container, a second container, a third container and a sealing cover plate, wherein the tops of the first container, the second container and the third container are all provided with openings; The bottom of the first container is provided with a first through hole array, and the first container is used for loading the continuous phase solution; The bottom of the second container is provided with a second through hole array, the second container is used for loading a cross-linking agent solution, the second container is nested and arranged in the first container, a first annular cavity is arranged between the outer diameter of the second container and the inner diameter of the first container, and a first gap is arranged between the lower surface of the bottom of the second container and the upper surface of the bottom of the first container; the bottom of the third container is provided with a third through hole array, the third container is used for loading discrete phase solution, the third container is nested and arranged in the second container, a second annular cavity is formed between the outer diameter of the third container and the inner diameter of the second container, and a second gap is formed between the lower surface of the bottom of the third container and the upper surface of the bottom of the second container; The sealing cover plate is covered on the tops of the first container, the second container and the third container and is used for isolating the first container, the first annular cavity and the second annular cavity into independent spaces, a first air passage interface, a second air passage interface and a third air passage interface are sequentially arranged above the first container, above the first annular cavity and above the second annular cavity, through holes in the first through hole array, the second through hole array and the third through hole array are coaxially arranged, and the first through hole array, the second through hole array and the third through hole array form a serial focusing through hole array based on the first gap and the second gap. Optionally, the bottom of the first container is also provided with a first positioning blind hole array, the bottom of the second container is also provided with a first positioning shaft array and a first protruding cylindrical array, and the second through hole array is arranged in the first cylindrical array, wherein the first positioning blind hole array and the first positioning shaft array are aligned with the first through hole array and the second through hole array through interference fit. Optionally, a second positioning blind hole array is further arranged at the bottom of the second container, a second positioning shaft array and a convex second cylindrical array are further arranged at the bottom of the third container, and a third through hole array is arranged in the second cylindrical array, wherein the second positioning blind hole array and the second positioning shaft array are aligned with the second through hole array and the third through hole array through interference fit. Optionally, the device further comprises a driving device, wherein the driving device is respectively connected with the first air passa