Aqueous two-phase deposition and fibrinolysis of fibroblast-laden fibrin micro-scaffolds

Stephen Robinson, Jonathan Chang, Eric Parigoris, Louise Hecker, Shuichi Takayama

Research output: Contribution to journalArticlepeer-review

Abstract

This paper describes printing of microscale fibroblast-laden matrices using an aqueous two-phase approach that controls thrombin-mediated enzymatic crosslinking of fibrin. Optimization of aqueous two-phase formulations enabled polymerization of consistent sub-microliter volumes of cell-laden fibrin. When plasminogen was added to these micro-scaffolds, the primary normal human lung fibroblasts converted it to plasmin, triggering gradual degradation of the fibrin. Time-lapse live-cell imaging and automated image analysis provided readouts of time to degradation of 50% of the scaffold as well as maximum degradation rate. The time required for degradation decreased linearly with cell number while it increased in a dose-dependent manner upon addition of TGF-β1. Fibroblasts isolated from idiopathic pulmonary fibrosis patients showed similar trends with regards to response to TGF-β1 stimulation. Addition of reactive oxygen species (ROS) slowed fibrinolysis but only in the absence of TGF-β1, consistent with published studies demonstrating that pro-fibrotic cellular phenotypes induced by TGF-β1 are mediated, at least in part, through increased production of ROS. FDA-approved and experimental anti-fibrosis drugs were also tested for their effects on fibrinolysis rates. Given the central role of fibrinolysis in both normal and pathogenic wound healing of various tissues, the high-throughput cell-mediated fibrinolysis assay described has broad applicability in the study of many different cell types and diseases. Furthermore, aqueous two-phase printing of fibrin addresses several current limitations of fibrin bio-inks, potentially enabling future applications in tissue engineering and in vitro models.

Original languageEnglish (US)
Article number035013
JournalBiofabrication
Volume13
Issue number3
DOIs
StatePublished - Jul 2021
Externally publishedYes

Keywords

  • aqueous two-phase system
  • fibrin
  • fibrinolysis
  • fibrosis
  • phenotypic assay
  • wound healing

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Biomaterials
  • Biomedical Engineering

Fingerprint

Dive into the research topics of 'Aqueous two-phase deposition and fibrinolysis of fibroblast-laden fibrin micro-scaffolds'. Together they form a unique fingerprint.

Cite this