Skin Regeneration Using Dermal Substrates that Contain Autologous Cells and Silver Nanoparticles to Promote Antibacterial Activity: In Vitro Studies

Abstract

We hypothesized that the addition of silver nanoparticles (AgNP) to a dermal substrate would impart antibacterial properties without inhibiting the proliferation of contained cells. Our in vitro model was based on the commercial substrate, Integra. The substrate was prepared by simple immersion into 0 to 1% suspension of AgNP (75 or 200 nm diameter) followed by rinsing for 20 minutes and sterilization under an ultraviolet C lamp. A total of 107 human adipose stem cells per cubic centimeter were injected and after 1 hour, 6 × 105 keratinocytes/cm2 were seeded and cultured for up to 14 days. Constructs were evaluated using a metabolic assay (WST-1), and hematoxylin and eosin and immunoperoxidase staining. Bactericidal activity was measured using a log reduction assay against bacteria that are prevalent in burns. The presence of AgNP did not significantly change the metabolic activity of constructs after 14 days of culture, and the distribution of cells within the substrate was unchanged from the controls that did not have AgNP. Antibacterial activity of Integra containing AgNP (75 nm diameter) was concentration dependent. In conclusion, the addition of AgNP to the dermal substrate suppressed bacterial growth but did not significantly affect cell proliferation, and may represent an important property to incorporate into a future clinical skin regeneration system.