Improving Wound Healing with DICER1-Modified Keratinocytes

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Regenerative Medicine

Background:

• Impaired wound healing, common in conditions like diabetes, stems from disruptions in the complex healing process.
• DICER1 (a key RNA-processing enzyme) levels are reduced in stalled diabetic wounds, suggesting a role in impaired closure.

Purpose of the Study:

• To investigate the role of DICER1 in wound closure.
• To develop and evaluate a novel, less toxic gene delivery system for DICER1 for therapeutic applications in wound healing.

Main Methods:

• HaCaT cells were transfected with DICER1 overexpression plasmids using Lipofectamine 2000 and a novel peptide-based nanocarrier (M9-DICER1-CS-A).
• Characterization of M9-DICER1-CS-A nanocomplexes (size, PDI, surface charge, nucleic acid condensation/release).
• Assessed cell viability, wound closure rates, and expression of wound-healing genes post-transfection. Engineered cells were also embedded in a hydrogel matrix.

Main Results:

• DICER1 overexpression in HaCaT cells significantly increased wound closure and upregulated wound-healing genes compared to controls.
• The M9-DICER1-CS-A nanocarrier demonstrated good characteristics (size, homogeneity, charge) and effective gene delivery.
• M9-DICER1-CS-A transfection resulted in comparable wound closure to Lipofectamine 2000 but with significantly improved cellular viability.
• Engineered HaCaT cells in hydrogels showed sustained viability and proliferation, indicating potential for sustained therapeutic effect.

Conclusions:

• DICER1 acts as a critical promoter of wound closure, particularly in the context of diabetes.
• The M9-DICER1-CS-A nanocarrier represents a safer and effective alternative for DICER1 gene delivery in wound healing.
• DICER1-engineered cells within hydrogels offer a promising cell-based therapeutic strategy for treating impaired wound closure.