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CN-121987847-A - Polymer frame ink, method for collaborative 3D printing of polymer frame and cell-loaded gel and structure

CN121987847ACN 121987847 ACN121987847 ACN 121987847ACN-121987847-A

Abstract

The invention provides a method and a structure for collaborative 3D printing of polymer frame ink, a polymer frame and a cell-loaded gel. The polymer frame ink contains a high polymer, an organic solvent and an organic modified synthetic sheet silicate, wherein the organic solvent comprises a first solvent and a second solvent, the first solvent is ethanol aqueous solution and/or absolute ethanol, and the second solvent is methyl acetate and/or isoflurane. The polymer frame ink provided by the invention can be quickly solidified by directly volatilizing an organic solvent, higher fidelity is obtained by organically modifying and synthesizing sheet silicate, the effect of quick forming is achieved while solidification is achieved, different pore sizes in polymer filaments can be obtained by controlling the temperature difference between the polymer frame ink and the environment, and finally, the synergistic printing of the polymer frame and the cell-carrying gel can be realized under a mild condition, and the activity of cells is ensured.

Inventors

  • XIONG ZHUO
  • TIAN YUEMING
  • FANG YONGCONG
  • ZHANG TING
  • ZHANG ZHENRUI
  • LIU ZIBO

Assignees

  • 清华大学

Dates

Publication Date
20260508
Application Date
20251230

Claims (13)

  1. 1.A polymer frame ink, wherein the polymer frame ink contains a high molecular polymer, an organic solvent and an organic modified synthetic sheet silicate; the organic solvent comprises a first solvent and a second solvent, wherein the first solvent is ethanol water solution and/or absolute ethanol, and the second solvent is methyl acetate and/or isoflurane; The concentration of the high molecular polymer in the organic solvent is 0.1-5 g/mL; The concentration of the organic modified synthetic sheet silicate in the organic solvent is 0.001-0.02 g/mL.
  2. 2. The polymer frame ink according to claim 1, wherein the high molecular polymer comprises one or a combination of two or more of polycaprolactone, racemic polylactic acid, polyurethane, polylactic acid-glycolic acid copolymer.
  3. 3. The polymer frame ink of claim 1, wherein the viscosity of the polymer frame ink is 50-1000 mPa s, preferably 300-600 mPa s.
  4. 4. The polymer frame ink of claim 1, wherein the volume ratio of the first solvent to the second solvent is 1 (9-40).
  5. 5. The polymer frame ink of claim 1, wherein the polymer frame ink further comprises a water-soluble porogen having a concentration in the organic solvent of 0.01-0.5 g/mL; preferably, the water-soluble pore-forming agent comprises one or a combination of more than two of glucose, sodium chloride and sucrose.
  6. 6. A method of co-3D printing of a polymer frame and a cell-loaded gel, wherein the method comprises: Printing polymer frame ink onto a bottom plate to form polymer filaments which are distributed at intervals to obtain a polymer frame layer; Printing cell-carrying gel ink to gaps among the polymer filaments, and solidifying and crosslinking the cell-carrying gel ink to form a polymer framework layer containing cells; Repeating the first step and the second step, and repeatedly printing the cell-containing polymer frame layer by layer on the cell-containing polymer frame layer to obtain a cell-containing structure; wherein the polymer frame ink is the polymer frame ink of any one of claims 1-5; In the printing process from the first step to the third step, the ambient temperature is 5-37 ℃, the temperature of the bottom plate is 5-37 ℃, the temperature of the polymer frame ink is 5-37 ℃, and the temperature of the cell-carrying gel ink is 15-37 ℃.
  7. 7. The method of collaborative 3D printing according to claim 6, wherein the cell-loaded gel ink contains cells, gel material and photoinitiator, the density of cells in the cell-loaded gel ink being from 0.5 x 10 6 to 10 x 10 6 cells/mL; Preferably, the cells comprise one or a combination of more than two of bone marrow mesenchymal stem cells, induced pluripotent stem cells, periosteum stem cells, angiogenic cells, bone-related tumor cells.
  8. 8. The method for collaborative 3D printing according to claim 6, wherein the printing process parameters meet the conditions of a printing speed of 0.1-6 mm/s, an extrusion flow of 0.012-20 μL/s, and an inside diameter of 0.09-2 mm.
  9. 9. The method for collaborative 3D printing of a polymer frame and a cell-loaded gel according to claim 6, wherein in step three, when the layer-by-layer printing of the polymer frame layer containing cells is repeated, the arrangement direction of the polymer filaments of the adjacent two layers is rotated by 0-90 °; Preferably, the arrangement directions of the polymer filaments between two adjacent layers are mutually perpendicular.
  10. 10. A structure printed from the polymer frame of any one of claims 6-9 and a cell-loaded gel in conjunction with a 3D printing process.
  11. 11. The structure of claim 10, wherein the polymer frame layer has a wire diameter of 100-2000 μιη.
  12. 12. The structure of claim 10, wherein the polymeric frame layer has a pore structure having a pore size of 0.1-200 μιη.
  13. 13. The structure of claim 10, wherein the structure has a side dimension of 500 μm to 500 mm; Preferably, the structure comprises a tissue structure, organoid, organ structure.

Description

Polymer frame ink, method for collaborative 3D printing of polymer frame and cell-loaded gel and structure Technical Field The invention belongs to the technical fields of tissue engineering and biological manufacturing, and particularly relates to a method and a structure for collaborative 3D printing of polymer frame ink, a polymer frame and cell-carrying gel. Background To cope with the increasing demands of tissue regeneration and repair reconstruction, tissue structures need to have material/cell heterogeneity, good biocompatibility, degradability and sufficient mechanical strength. Biological 3D printing techniques, which can provide specific spatial geometries, controlled microstructures, and precise localization of multiple cell types, are important techniques for constructing heterogeneous tissue structures. At present, the common biological 3D printing technology mainly comprises an extrusion type, a photo-curing type and an ink-jet type, wherein the extrusion type technology can accurately arrange a polymer frame and a cell-carrying hydrogel material, has the capacity of synchronously forming multiple materials, and is widely applied to constructing heterogeneous tissue structures. Currently, the framing material commonly used for printing heterogeneous tissue structures is based on high molecular weight polymers such as Polycaprolactone (PCL), polylactic acid (PLA), polylactic-co-glycolic acid (PLGA), and polyurethane (TPU). They typically have a relatively high melting point (greater than 100 ℃) and are based on Fused Deposition Modeling (FDM) printing. However, higher forming temperatures can cause thermal damage to cells/biological factors when co-printed with cell-containing hydrogels. Meanwhile, continuous gradient construction of a multi-tissue interface (such as bone-cartilage and tendon-muscle) requires collaborative printing of multiple polymers, but the difference of phase transition temperatures of different materials (such as 60 ℃ C, PLA for 190 ℃ for PCL) can cause thermal stress concentration during coextrusion, so that interface bonding strength is reduced, and the integrated forming capability of a complex tissue body is severely restricted. In addition, the proliferation, migration, etc. of cells within the tissue structure are still restricted. In order to solve the above problems, there is a prior art in which polymer ink is dissolved in an organic solvent and extrusion printing is performed in a room temperature environment. Yong Huang team （Sole-Gras, M., Ren, B., Ryder, B.J. et al. Vapor-induced phase-separation-enabled versatile direct ink writing. Nat Commun 15, 3058 (2024).） developed a non-solvent induced phase separation 3D printing technique whereby polymer fibers can be cured to shape in a room temperature environment and in a water vapor atmosphere by displacing the solvent with a non-solvent. Daniel Therriault team （Bodkhe S, Turcot G, Gosselin FP, et al. One-Step Solvent Evaporation-Assisted 3D Printing of Piezoelectric PVDF Nanocomposite Structures. ACS Appl Mater Interfaces. 2017 9 (24), 20833-20842. Guo SZ, Heuzey MC, Therriault D. Properties of polylactide inks for solvent-cast printing of three-dimensional freeform microstructures. Langmuir 2014 30 (4), 1142-1150.） developed evaporation-induced phase separation 3D printing techniques whereby polymer fibers can be cured into shape in a room temperature environment by solvent evaporation. Anabel Renteria team （Rodriguez, A., Gonzalez, S., Cruz, A. et al. Solvent evaporation-assisted direct ink writing using polar solvent for flexible polyvinyl fluoride piezoelectric composites in enhancement of piezoelectric properties. MRS Advances 10, 1385–1391 (2025).） successfully prepared flexible piezoelectric devices with shrinkage as low as 10% at room temperature using solvent evaporation of polar solvents to aid direct ink writing to fabricate flexible piezoelectric devices. Guo et al （Guo SZ, Gosselin F, Guerin N, et al. Solvent-cast three-dimensional printing of multifunctional microsystems. Small, 9: 4118-4122.） prepared a complex free-form spiral using methylene chloride as a solvent and solvent casting direct writing of microstructures using thermoplastic polymer solution inks. However, most of the organic solvents adopted by the method are toxic, the cell compatibility is poor, the size collapse is obvious after the fiber is solidified, the printing precision is greatly reduced, in addition, the method still needs post-treatment and is difficult to co-print with the cell-carrying hydrogel, and a micropore structure is not formed in the printed fiber, so that the proliferation and migration of cells are not facilitated. Therefore, there is a need for a room temperature printing process for a polymer with both printing precision and biocompatibility, so as to realize high-precision collaborative co-printing of polymer frame ink and cell-loaded gel ink under mild conditions, thereby constructing a heterogeneous tis