Relevance of bi-functionalized polyelectrolyte multilayers for cell transfection

Area of Science:

• Biomaterials Science
• Surface Chemistry
• Cell Biology

Background:

• Polyelectrolyte multilayers (PEMs) are powerful tools for creating biologically active surfaces.
• Controlled multi-functionalization of surfaces remains a significant challenge in biomaterial development.

Purpose of the Study:

• To demonstrate the simultaneous, controlled functionalization of PEMs with two distinct molecules: plasmid DNA (pDNA) and a signaling peptide (NDPMSH).
• To investigate the impact of this dual functionalization on gene delivery and cell transfection, particularly in melanoma cells.

Main Methods:

• Fabrication of poly-(allylamine hydrochloride)/poly-(sodium 4-styrene sulfonate) multilayers.
• Embedding of peptide-grafted poly(L-glutamic acid) (PGA-NDPMSH) within the multilayers.
• Deposition of pDNA complexed with poly(ethyleneimine) (PEI) on top of the functionalized films.
• Transfection assays using melanoma cells (B16-F1) and Huh-7 cells.
• Flow cytometry analysis to assess transfection efficiency and cellular response.

Main Results:

• Melanoma cells (B16-F1) showed efficient transfection after 24 hours of contact with the functionalized films.
• The functionalized films significantly enhanced transfection rates in Huh-7 cells, indicating receptor-independent enhancement.
• Transfected cells produced melanin, confirming both peptide signaling pathway activation and successful cell transfection.

Conclusions:

• Simultaneous functionalization of PEMs with pDNA-PEI complexes and PGA-NDPMSH is achievable and controllable.
• Incorporating specific peptides into multilayer films can enhance gene delivery and transfection efficiency in a receptor-independent manner.
• These functionalized biomaterial coatings show promise for advanced gene therapy and cell engineering applications.