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A Multilayer Microfluidic Platform for the Conduction of Prolonged Cell-Free Gene Expression
Published on: October 6, 2019
Cross-linked bioreducible layer-by-layer films for increased cell adhesion and transgene expression
Jenifer Blacklock1, Torsten K Sievers, Hitesh Handa
1Department of Biomedical Engineering, Wayne State University, Detroit, Michigan 48202, USA. blacklock@mpikg.mpg.de
The Journal of Physical Chemistry. B
|April 8, 2010
Summary
Cross-linking bioreducible polymer and DNA films with DIP enhances rigidity and cell adhesion. This improves gene delivery duration, showing potential for localized gene therapy applications.
Area of Science:
- Biomaterials Science
- Polymer Chemistry
- Gene Delivery Systems
Background:
- Layer-by-layer (LbL) films offer tunable properties for biomedical applications.
- Bioreducible polymers and DNA are promising for gene delivery.
- Controlling film properties like rigidity and cell interaction is crucial for effective transfection.
Purpose of the Study:
- To investigate the impact of 1,5-diiodopentane (DIP) cross-linking on the properties and performance of rPDMAEMA/DNA LbL films.
- To evaluate the effects of cross-linking on film rigidity, biodegradability, cell adhesion, and in vitro transfection activity.
- To determine the cross-linking density and understand the changes in film hydrophobicity.
Main Methods:
- Fabrication of rPDMAEMA/DNA LbL films.
- Cross-linking using 1,5-diiodopentane (DIP).
- Characterization using Atomic Force Microscopy (AFM), ellipsometry, and Quartz Crystal Microbalance with Dissipation (QCM-D).
- Analysis using rubber elasticity theory and Flory-Rehner theory.
- In vitro cell adhesion and transfection assays with mouse fibroblast and smooth muscle cells using SEAP DNA.
Main Results:
- Cross-linking with DIP significantly increased film rigidity (Young's modulus > 4x).
- Cross-linking increased hydrophobicity and altered film swelling, with a calculated cross-linking density of ~0.82 mmol/cm³.
- Film disassembly by DTT was unaffected by cross-linking.
- Enhanced cell adhesion and prolonged in vitro transfection activity (up to 7 days) were observed.
Conclusions:
- DIP cross-linking effectively modifies the physical and chemical properties of rPDMAEMA/DNA LbL films.
- Cross-linked films demonstrate improved performance for cell adhesion and sustained gene transfection.
- These findings support the development of cross-linked bioreducible polymer coatings for advanced gene delivery applications.
