Design And Test Of Targeted Lipid-nanoparticles In Burn Wound Care
Subhadip Ghatak, Jilong Li, Mohamad S. El Masry, Amitava Das, Yang Liu, Sashwati Roy, Robert J. Lee, Chandan K. Sen.
The Ohio State University, Columbus, OH, USA.
Background - Active-targeted lipid nanocarriers minimize off-target effects. Transferrin-conjugation strategy serves that purpose but may not be well suited for non-cancer applications such as wound. In wounds, although skin barrier function is breached, abundance of inflammatory cells at the site of injury poses a major challenge for delivery. Phagocytic clearance of nanoparticles is a threat. Methods - Novel lyophilized keratinocyte-targeted nanocarriers (TLNκ) were designed and loaded with anti-miR to test efficacy in treating cutaneous burn injury. TLNκ employed DOTAP/DODAP combination pH-responsive lipid components to improve endosomal escape. Keratinocyte-targeting was achieved using the peptide sequence ASKAIQVFLLAG. To minimize interference of clearance by non-targeted cells, especially immune cells in the acute wound microenvironment, surface charge was neutralized. Lyophilization extended shelf life of these nanoparticles. Results - Encapsulation efficiency of anti-miR in lyophilized TLNκ was 96.54%. Cargo stability of lyophilized TLNκ was tested. After 9 days of loading with anti-miR-210, TLNκ was effective in lowering abundance of the hypoxamiR miR-210 in keratinocytes challenged with hypoxia. Keratinocyte uptake of DiD-labelled TLNκ/anti-miR-107 was selective and exceeded 90% within 4h. Topical application of hydrogel-dispersed lyophilized TLNκ/anti-miR-107 encapsulating LNA anti-miR-107 twice a week effectively sequestered keratinocyte miR-107. On day 24, the wound area in TLNκ/anti-miR-107 treated group was reduced to 4% of the initial wound area. Barrier function of the skin, a functional measure of wound closure, as measured by trans-epidermal water loss was restored. Application of TLNκ/anti-miR-107 depleted miR-107 and upregulated dicer expression causing differentiation of keratinocytes. Expression of junctional proteins such as claudin, loricrin, filaggrin, ZO-1 and ZO-2 were significantly upregulated following TLNκ/anti-miR-107 treatment. Conclusion - The nanoparticles reported herein are promising as topical therapeutic agents in the management of burn injury. A translational advantage of TLNκ is that all material used for its formulation has prior history of FDA approval for human use.
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