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Area of Science:

  • Biotechnology
  • Immunology
  • Biomaterials Science

Background:

  • The immune cell microenvironment critically regulates immune cell function and fate.
  • Three-dimensional (3D) culture systems offer insights into immune responses within tissue-mimicking environments.
  • Current 3D models often lack high-throughput capabilities, hindering broad application in host-pathogen interaction studies.

Purpose of the Study:

  • To establish a high-throughput macrophage-bacteria co-culture model mimicking lung tissue stiffness.
  • To investigate how varying microenvironment stiffness affects macrophage behavior and response to bacterial infection.
  • To provide a platform for studying host-pathogen interactions and developing therapeutic strategies.

Main Methods:

  • Utilized bioprinting to encapsulate and differentiate human THP-1 monocytes into macrophages within synthetic extracellular matrices (ECMs).
  • Fabricated ECMs with defined polymer and peptide bioinks in a 96-well plate format to mimic healthy (compliant) and diseased (stiff) lung tissue stiffness.
  • Assessed macrophage viability, immunocompetence (phenotype, phagocytosis, response to stimuli), and gene expression.
  • Studied the immune response of macrophages to *Pseudomonas aeruginosa* infection in different stiffness conditions.

Main Results:

  • Macrophages maintained viability and immunocompetence in the 3D culture system.
  • Macrophages in stiffer (fibrosis-inspired) ECMs showed higher basal expression of inflammation and fibrosis genes compared to compliant (healthy lung-inspired) ECMs.
  • In response to *P. aeruginosa*, macrophages in stiff microenvironments exhibited altered cytokine secretion (decreased IL-6/IL-1β, increased IL-10/TNF-α) compared to compliant environments.

Conclusions:

  • The developed high-throughput 3D model effectively mimics lung tissue stiffness and allows for controlled study of host-pathogen interactions.
  • Microenvironment stiffness significantly influences macrophage immune responses, including gene expression and cytokine profiles during bacterial infection.
  • This platform facilitates understanding of bacterial infections and aids in identifying potential therapeutic strategies.