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3D biological protein and preparation method and applications thereof

A biological protein, 3D technology, applied in the field of 3D biological protein and its preparation, can solve the problems of affecting the experimental effect, porosity and hardness, and large pores of 3D biological protein.

Active Publication Date: 2018-06-15
深圳灵赋拓普生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the traditional biological protein obtained is porous and not strong, so that when some cells are planted on the material, the proteins, growth factors and hormones secreted by the body cannot fully act between cells, and some will penetrate into the cell. The biological protein material itself affects the experimental results
[0004] In summary, the traditional 3D biological protein has large pores, large surface roughness, and low hardness.

Method used

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  • 3D biological protein and preparation method and applications thereof
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  • 3D biological protein and preparation method and applications thereof

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preparation example Construction

[0030] A method for preparing a 3D biological protein in one embodiment includes the following steps S110-S120.

[0031] S110, mixing the fluorescein-labeled albumin and the photosensitizer to obtain a mixed solution.

[0032] In one embodiment, in the fluorescein-labeled albumin, the fluorescein is selected from at least one of FITC (fluorescein isothiocyanate) and PE (phycoerythrin). The albumin is at least one selected from bovine serum albumin (BSA), human serum albumin (HSA), human recombinant serum albumin, sheep serum albumin and rabbit serum albumin.

[0033] In one embodiment, the photosensitizer is rose bengal sodium salt (RB).

[0034] Specifically, the final concentration of fluorescein-labeled albumin in the mixed solution is 50 mg / mL-100 mg / mL. The final concentration of the photosensitizer is 0.05w / v%-0.15w / v%.

[0035] Specifically, w / v% represents the mass-to-volume ratio, that is, the final concentration of the photosensitizer in the mixed solution is 0.5g...

Embodiment 1

[0063] Mix FITC-BSA and RB photosensitizer, and add a certain amount of pure water to prepare a mixed solution. The final concentration of FITC-BSA in the mixed solution is 100mg / mL, and the final concentration of RB photosensitizer is 0.1w / v%.

[0064] See figure 1 , place a glass slide in a Petri dish, and drop 100 μL of the above mixture onto the glass slide through a pipette gun. Then the culture dish is moved into the high oxygen airtight chamber, and the oxygen partial pressure in the high oxygen airtight chamber is set to be 0.3Kpa, the oxygen concentration to be 75%, and the humidity to be 60%. A 40× oily objective lens was used, and a two-photon exciter was used to scan the mixture layer by layer in a high-oxygen airtight chamber to solidify the mixture. The wavelength of the excitation light is set to 700nm, the energy is 20mW, and the area of ​​each laser beam scanning section is 1μm 2 . The laser penetrates the slide into the mixture, and scans upward layer by l...

Embodiment 2

[0066] Mix FITC-BSA and RB photosensitizer, and add a certain amount of pure water to prepare a mixed solution. The final concentration of FITC-BSA in the mixed solution is 75mg / mL, and the final concentration of RB photosensitizer is 0.15w / v%.

[0067] See figure 1 , place a glass slide in a Petri dish, and drop 100 μL of the above mixture onto the glass slide through a pipette gun. Then move the petri dish into the high oxygen airtight chamber, set the oxygen partial pressure in the high oxygen airtight chamber to be 0.35Kpa, the oxygen concentration to be 80%, and the humidity to be 70%. The mixed solution is scanned layer by layer in the oxygen-tight chamber to solidify the mixed solution. The wavelength of the excitation light is set to 750nm, the energy is 10mW, and the area of ​​each laser scanning section is 0.8μm 2 . The laser penetrates the slide into the mixture, and scans upward layer by layer from the surface of the slide (the liquid bottom of the mixture). Aft...

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Abstract

The invention discloses a 3D biological protein and a preparation method and applications thereof. The 3D biological protein is prepared by following steps: mixing fluorescein labeled albumin with a photo sensitizer to form a mixed solution; scanning the mixed solution by exciting light with a wave length of 700-750 nm and energy of 10-30 mW under following conditions: oxygen partial pressure: 0.25-0.35 KPa, and oxygen concentration: 70-80%; and finally solidifying the mixed solution to obtain the 3D biological protein. The experiment results show that the prepared 3D biological protein has the advantages of small pores, smooth surface, and high hardness.

Description

technical field [0001] The invention relates to the field of biological materials, in particular to a 3D biological protein and its preparation method and application. Background technique [0002] The traditional way of culturing stem cells in vitro is to plant stem cells in a petri dish, but this environment is single and only provides a flat two-dimensional structural space for cells. In order to be closer to the three-dimensional space effect of the human body, some degradable polymer biomaterials have appeared, such as preparing some three-dimensional patterns for cell growth through photolithography technology, or electrospinning technology, etc., but these technologies have complex manufacturing processes. The cycle is long, the production process has the disadvantages of adding toxic substances, the structure is single and random, and the polymer material itself is not favored by cells. Therefore, a layer of protein needs to be plated on the surface of the material ...

Claims

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Application Information

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IPC IPC(8): A61L27/34A61L27/22A61L27/50
CPCA61L27/227A61L27/34A61L27/50A61L27/507A61L2400/18A61L2420/02A61L2430/02C08L89/00
Inventor 马名泽
Owner 深圳灵赋拓普生物科技有限公司
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