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Delivery and expression of pDNA embedded in collagen matrices
Hagit Cohen-Sacks1, Victoria Elazar, Jianchuan Gao
1Department of Pharmaceutics, Faculty of Medicine, School of Pharmacy, The Hebrew University of Jerusalem, Post Office Box 12065, 91120 Jerusalem, Israel.
Summary
Collagen matrices effectively deliver plasmid DNA (pDNA) for gene therapy. These biocompatible carriers enhance gene expression in vitro and in vivo, showing promise for tissue engineering and localized treatments.
Area of Science:
- Biomaterials Science
- Gene Therapy
- Tissue Engineering
Background:
- Non-viral gene carriers are crucial for safe and effective gene therapy.
- Collagen matrices offer biocompatibility and localized delivery potential.
- Enhancing plasmid DNA (pDNA) binding within matrices improves transfection efficiency.
Purpose of the Study:
- To develop and characterize collagen matrices for embedding and delivering plasmid DNA (pDNA).
- To evaluate the gene delivery and expression capabilities of these matrices in vitro and in vivo.
- To assess the potential of collagen-pDNA matrices for tissue engineering and localized gene therapy.
Main Methods:
- Formulation of collagen matrices with condensed pDNA using condensing agents.
- Characterization of pDNA release kinetics from the matrices.
- In vitro transfection assays using various cell lines (NIH 3T3, 293, MDA-MB-231, SMCs) with different pDNA formulations (naked pDNA, pDNA-liposome, pDNA-polyethylenimine).
- In vivo studies involving subdermal implantation in rats and perivascular treatment in balloon-injured rat carotid arteries.
Main Results:
- pDNA and condensed pDNA were released intact from collagen matrices within 1-2 days.
- Collagen matrices containing pDNA formulations (especially pDNA-liposome) showed significantly higher gene expression compared to naked pDNA in vitro.
- pDNA-liposome matrices demonstrated a dose-dependent transfection response across multiple cell types.
- Subdermal implantation and perivascular treatment with collagen-pDNA matrices resulted in significantly enhanced gene expression compared to non-condensed pDNA controls.
Conclusions:
- A straightforward method for incorporating cationic pDNA formulations into collagen matrices was successfully developed.
- These bioactive collagen matrices are effective non-viral gene carriers for localized gene therapy.
- The findings support the suitability of these matrices for tissue engineering applications and localized therapeutic strategies.