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Cell-controlled and spatially arrayed gene delivery from fibrin hydrogels.

Pedro Lei1, Roshan M Padmashali, Stelios T Andreadis

  • 1Bioengineering Laboratory, Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, Amherst, NY 14260, USA.

Biomaterials
|April 28, 2009
PubMed
Summary

Fibrin hydrogels enable efficient gene transfer by controlling the local cell-plasmid DNA (pDNA) microenvironment. This biomaterial allows for cell-controlled, spatially localized gene delivery for in vitro or in vivo applications.

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Area of Science:

  • Biomaterials Science
  • Gene Therapy
  • Tissue Engineering

Background:

  • Gene delivery systems are crucial for therapeutic applications.
  • Fibrin hydrogels offer a promising biomaterial scaffold for cell-based therapies.
  • Controlling the microenvironment is key for efficient gene transfection.

Purpose of the Study:

  • To investigate fibrin-mediated gene transfer using plasmid DNA (pDNA) embedded within hydrogels.
  • To compare gene transfection efficiency in 2D (surface) versus 3D (embedded) cell cultures.
  • To explore the potential of fibrin hydrogels for spatially localized gene delivery.

Main Methods:

  • Embedding pDNA within fibrin hydrogels during polymerization.
  • Utilizing 2D and 3D cell culture models for gene transfection.
  • Assessing cell viability and gene transfer efficiency under varying conditions (cell density, fibrinogen/pDNA concentrations).
  • Investigating the role of fibrin degradation using fibrinolytic inhibitors.
  • Demonstrating spatially localized gene delivery using cell-transfection microarrays.

Main Results:

  • Cell transfection efficiency was highly dependent on the 2D vs. 3D context, cell type/density, and concentrations of fibrinogen and pDNA.
  • Embedding cells within the fibrin matrix significantly reduced lipofectamine-induced cell death, particularly at low cell densities.
  • Fibrin degradation appears necessary for efficient gene transfer, as indicated by dose-dependent inhibition with fibrinolytic inhibitors.
  • Fibrin hydrogels enabled strictly confined gene transfer in a microarray format, preventing cross-contamination between adjacent spots.

Conclusions:

  • Fibrin hydrogels serve as an effective biomaterial for gene delivery.
  • Gene transfer efficiency is modulated by the cell-pDNA microenvironment within the hydrogel.
  • Fibrin-mediated gene delivery offers precise spatial control, suitable for applications like cell-transfection microarrays.
  • This approach holds potential for efficient, cell-controlled, and localized gene delivery in both in vitro and in vivo settings.