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CN-121971703-A - Fiber-reinforced 3D printing biological ink and preparation method thereof

CN121971703ACN 121971703 ACN121971703 ACN 121971703ACN-121971703-A

Abstract

The application relates to a fiber reinforced 3D printing biological ink and a preparation method thereof, wherein the preparation method comprises the following raw materials, by mass, 1.0-20.0 parts of gelatin, 0.1-10.0 parts of sodium alginate, 0.01-10 parts of pretreated bacterial cellulose dispersion liquid and 60-100 parts of water, the preparation method comprises the following steps of adding water into bacterial cellulose gel to break, circulating for 2-4 times by a colloid mill at a rotating speed of 5000-15000rpm, a gap of 30-100 mu m, and carrying out vacuum defoaming for 10-20min to obtain the pretreated bacterial cellulose dispersion liquid.

Inventors

  • LIU SIYUN
  • YU KUI
  • Liang Liting
  • LI ZHAO
  • HUANG SHA
  • WANG YAOWEN
  • FENG YONGLIANG
  • FU XIAOBING

Assignees

  • 苏州汇涵医用科技发展有限公司
  • 中国人民解放军总医院

Dates

Publication Date
20260505
Application Date
20260204

Claims (10)

  1. 1. The fiber reinforced 3D printing biological ink is characterized by comprising, by mass, 1.0-20.0 parts of gelatin, 0.1-10.0 parts of sodium alginate, 0.01-10 parts of pretreated bacterial cellulose dispersion liquid and 60-100 parts of water, wherein the pretreated bacterial cellulose dispersion liquid comprises the following preparation steps of adding water into bacterial cellulose gel to break, circulating for 2-4 times by a colloid mill, rotating at 5000-15000rpm, and conducting vacuum defoaming for 10-20min to obtain the pretreated bacterial cellulose dispersion liquid.
  2. 2. The 3D printing bio-ink of claim 1 wherein said bacterial cellulose fiber bundles in said pretreated bacterial cellulose dispersion are 20-80 μm in length and 50-200nm in diameter.
  3. 3. The 3D printing bio-ink of claim 1, wherein the pretreated bacterial cellulose dispersion liquid comprises a double-modified pretreated bacterial cellulose dispersion liquid, the double-modified pretreated bacterial cellulose dispersion liquid comprises the following preparation steps of adding epoxypropyl trimethyl ammonium chloride and sodium hydroxide into the pretreated bacterial cellulose dispersion liquid, stirring for reaction, then purifying and washing to obtain a quaternized bacterial cellulose dispersion liquid, adding triethylamine into the quaternized bacterial cellulose dispersion liquid, dropwise adding methacryloyl chloride under an ice bath, and performing secondary purification to obtain the double-modified pretreated bacterial cellulose dispersion liquid.
  4. 4. The fiber reinforced 3D printing bio-ink according to claim 1, wherein the sodium alginate comprises methacryloyl modified sodium alginate, and the preparation method comprises the steps of dispersing sodium alginate in water, stirring uniformly, adjusting pH value, cooling, dropwise adding methacryloyl chloride, adjusting pH value, dialyzing, and freeze-drying to obtain the methacryloyl modified sodium alginate.
  5. 5. The 3D printing bio-ink of claim 4 wherein the gelatin comprises a methacryloyl modified gelatin, comprising the steps of dispersing gelatin in phosphate buffer, adding an acid binding agent, adjusting pH, cooling, dropwise adding methacrylic anhydride, adding deionized water, dialyzing, and freeze-drying to obtain the methacryloyl modified gelatin.
  6. 6. The 3D printing bio-ink of claim 4 wherein the molar ratio of said methacryloyl chloride to sodium alginate is (0.3-1): 1.
  7. 7. A fiber-reinforced 3D printing bio-ink according to claim 5 wherein the molar ratio of methacrylic anhydride to gelatin is (0.5-2): 1.
  8. 8. The fiber-reinforced 3D printing bio-ink according to claim 5, wherein the mass ratio of the methacryloyl modified gelatin to the methacryloyl modified sodium alginate is (2-5): 1.
  9. 9. A fiber-reinforced 3D printing bio-ink according to claim 3 wherein the mass ratio of said double modified pretreated bacterial cellulose dispersion to gelatin is (0.005-0.3): 1.
  10. 10. A preparation method of the fiber-reinforced 3D printing bio-ink according to any one of claims 1 to 9 is characterized by comprising the following preparation steps of mixing a suspension with a gelatin solution at 40 to 50 ℃ and a sodium alginate solution, stirring at 600 to 1000rpm for 3 to 8min, defoaming for 5min at 30 to 50 ℃ and- (0.06 to 0.10) MPa for two times, and preserving at 0 to 8 ℃ in a dark place to obtain the fiber-reinforced 3D printing bio-ink.

Description

Fiber-reinforced 3D printing biological ink and preparation method thereof Technical Field The application relates to a biological ink, in particular to a fiber reinforced 3D printing biological ink and a preparation method thereof. Background The bio-ink is a special material for 3D bio-printing, and is prepared by removing cells from tissues or organs, and the main components include hydrogel and biomolecules. The core technology is that the water-based gel is used as a carrier, and tissue structures are formed through the differentiation of mesenchymal stem cells, so that the use limit of the traditional collagen-based ink is broken through. The combination of Sodium Alginate (SA) and Gelatin (Gelatin) is one of classical double-network hydrogel systems in the field of biological printing at present. According to the combination, sodium alginate and gelatin are dispersed in a calcium chloride solution according to a certain proportion to form a network hydrogel, the network hydrogel is used as a common 3D biological printing ink, the gelatin provides a temperature-sensitive reversible gel network, the temperature-sensitive reversible gel network is convenient to form at room temperature, and the sodium alginate is crosslinked through Ca < 2+ > ions to form an irreversible network, so that a printed product is endowed with a certain shape retaining capability. The raw materials are common and easy to obtain, the crosslinking condition is mild, the biocompatibility is good, the printability is strong, and the skin fibroblasts can be loaded and the rapid in-situ forming can be realized. However, the traditional sodium alginate and gelatin system has the problems of weak mechanical strength, quick degradation and poor structural fidelity, and the simple additional reinforcing component can partially reinforce, but has difficult dispersion, easy stress concentration, insufficient biological activity, and the pure bacterial cellulose bracket has better mechanical property and slow degradation, but has poor dispersibility in the sodium alginate/gelatin system, so that uneven ink can be caused, and a spray nozzle can be blocked. Therefore, there is a need in the industry for a bio-ink solution that maintains good 3D printability while being fully scalable in terms of mechanical properties, stability and multi-level tissue suitability. Disclosure of Invention In order to improve the strength and stability of a sodium alginate and gelatin system bio-ink product, the application provides a fiber reinforced 3D printing bio-ink and a preparation method thereof. In the first aspect, the application provides a fiber-reinforced 3D printing biological ink, which comprises, by mass, 1.0-20.0 parts of gelatin, 0.1-10.0 parts of sodium alginate, 0.01-10 parts of pretreated bacterial cellulose dispersion liquid and 60-100 parts of water, wherein the pretreated bacterial cellulose dispersion liquid comprises the following preparation steps of adding water into bacterial cellulose gel to break, circulating for 2-4 times by a colloid mill at a rotating speed of 5000-15000rpm, a gap of 30-100 mu m, and performing vacuum defoaming for 10-20min to obtain the pretreated bacterial cellulose dispersion liquid. Preferably, vacuum defoamation is carried out for 10-20min at 30 ℃ and minus 0.09 MPa. Preferably, the length of the bacterial cellulose fiber bundles in the pretreated bacterial cellulose dispersion is 20-80 μm and the diameter is 50-200nm. The process obtains fiber bundles with specific size through mild depolymerization of a colloid mill, depolymerizes rather than breaks, has good enhancement effect, and has the size suitable for printing and being used as biological ink. Preferably, the pretreated bacterial cellulose comprises double-modified pretreated bacterial cellulose, and the preparation method comprises the following steps of adding epoxypropyl trimethyl ammonium chloride and sodium hydroxide into a pretreated bacterial cellulose dispersion liquid, stirring for reaction, purifying and washing to obtain a quaternized bacterial cellulose dispersion liquid, adding triethylamine into the quaternized bacterial cellulose dispersion liquid, dropwise adding methacryloyl chloride under ice bath, and purifying the dispersion liquid for the second time to obtain the double-modified pretreated bacterial cellulose. In the first step of quaternization modification, epoxypropyl trimethyl ammonium chloride is subjected to ring opening reaction with hydroxyl on the surface of bacterial cellulose under the catalysis of sodium hydroxide, quaternary ammonium cations are grafted on the surface of the fiber, triethylamine is used as an acid binding agent in the second step of methacryloylation modification, residual hydroxyl on the surface of the bacterial cellulose is subjected to esterification reaction with methacryloyl chloride under the ice bath condition, and carbon-carbon double bonds are introduced. This double modification conver