Method for promoting in-vitro and in-vivo degradation and cell extension adherence of alginate-based 3D printed bioink

A technology of alginate and bio-ink, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of alginate limitations and achieve the effect of promoting adhesion, changing porosity and compressive modulus

Inactive Publication Date: 2019-08-27
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows create tiny structures called algae within an organism's own structure during its growth period without causing harm themselves. By adding certain substances like alkaline earth metal salt into these alga particles, they gradually break down over time while still being able to keep their shape when placed under specific environmental factors such as temperature, pH level, etc., resulting in improved stability and performance compared to other methods previously described. Additionally, the algal material contains various types of polysaccharides which help promote blood clotting and wound healing processes through fibers. Overall, this new technique helps improve the properties of artificially made objects produced from alginate alone.

Problems solved by technology

Technologies described earlier include techniques like casting extracellular matrices onto substrates called stents, crushing necrosis cores from patients' bodies, culturing human epitheliutrophils, growing monoclinal tracts containing cells, injecting these cells into recipients, and delivering them back inside the patient's own body overcomes issues associated with current approaches including rejection responses due to inflammatory processes during surgery, poor wound healing mechanisms, reduced cell motility postoperatively, and potential negative side effectiveness related to prosthetic devices. Additionally, there may exist concerns about how effective therapy requires sufficient engraftment and growth of new ones undergoing natural metabolism. To address these challenges, research efforts aim towards developing improved solutions involving enzyme activation, cytokines release stimulation, matrix resin synthesis, and gene transfer technologies.

Method used

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  • Method for promoting in-vitro and in-vivo degradation and cell extension adherence of alginate-based 3D printed bioink
  • Method for promoting in-vitro and in-vivo degradation and cell extension adherence of alginate-based 3D printed bioink
  • Method for promoting in-vitro and in-vivo degradation and cell extension adherence of alginate-based 3D printed bioink

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Example 1: Using alginate lyase to promote the in vivo and in vitro degradation of alginate-based bio-3D printing blocks.

[0045] Alginate / gelatin-mixed bioink using alginate lyase, as shown in Figure 2 and Figure 3, Figure 2APrints cultured in vitro can be observed, and the degradation time is correlated with the alginate lyase concentration. Figure 2B Printed blocks with different concentrations of sodium alginate lyase, H&E staining results, the shaded part is alginate, and the fragmentation degree of alginate skeleton is significantly different with the change of enzyme concentration (scale bar: 200 μm). Such as Figure 3B As shown, the mouse dermal fibroblasts with DiI fluorescent signal were mixed into the bio-ink, and the printed block was implanted subcutaneously in nude mice, and traced detection was carried out using a living small animal instrument. Figure 3A Shown is the result of sampling tissue sections after the print blocks were implanted subcutane...

Embodiment 2

[0058] Example 2: Using alginate lyase to promote cell expansion and migration in alginate-based bio-3D printing blocks.

[0059] Under physiological conditions, cells are generally polygonal and have the characteristics of stretching and adhesion. Conventional alginate lacks cell attachment sites, and although it has good biocompatibility, cells stretch less in alginate print blocks. Using alginate lyase, as shown in Figure 4, can significantly increase cell stretching in the printed block. Such as Figure 4A As shown, in the print block without adding alginate lyase, most of the cells were oval, and in the print block added with alginate lyase, the formation of cell pseudopodia and cell extension could be observed (scale bar: 50 μm). Figure 4B This is the state of GFP fluorescent mouse dermal fibroblasts in alginate-based bioink. In the print block added with alginate lyase, the proportion of cell extension to the total cells was significantly increased, and the ratio of ...

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Abstract

The invention discloses a method for promoting in-vitro and in-vivo degradation and cell extension adherence of alginate-based 3D printed bioink. The method comprises the following steps: step 1, adopting alginate as an extracellular matrix main body skeleton and constructing celliferous 3D bio-printed bioink; step 2, mixing alginate lyase with alginate-based bioink; step 3, adding cell suspensioninto the obtained bioink, choosing print parameters and printing out a 3D printing block through a 3D bio-printer; step 4, regulating and controlling in-vitro and after-transplantation-in-vivo degradation time and cell extension adherence degree of the 3D printing block through adjusting the concentration of the alginate lyase according to actual demands. The method disclosed by the invention canpromote the degradation and cell extension adherence of the bioink with the alginate as the skeleton under in-vivo and in-vitro environments, and enables the alginate 3D printed bioink to be adaptedto more in-vivo and in-vitro experiments and be better applied to soft tissue repairing and transplantation.

Description

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Claims

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Application Information

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Owner GENERAL HOSPITAL OF PLA
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