Tissue Oxygenation within Diabetic Wounds can be Monitored Using Difluoroboron β– Diketonate Polylactide Nanoparticles

Abstract

OBJECTIVERoughly 6.5 million diabetic patients in the United States suffer from chronic wounds that exhibit impaired healing response1. These wounds often do not respond to standard treatment due to insufficient oxygenation in the wound bed2. Given the heterogeneity of oxygen levels in wounds and the importance of oxygen in tissue repair, it is compelling to develop a reliable way to non‐invasively monitor oxygen levels within chronic wounds over time. Previously, we demonstrated the ability of topically‐delivered BF2nbm(I)PLA boron dye nanoparticles (BNPs) that phosphoresce differentially at varying oxygen levels, to provide a spatial map of oxygen levels throughout the wound bed of a murine full‐thickness cutaneous wound3. The objective of this work is to validate the ability of BNP imaging to detect impaired tissue oxygenation during wound healing in diabetic individuals compared to healthy individuals.METHODSWe have developed a novel model of wound healing in the murine ear which allows measurements of oxygen levels in both tissue and blood vessels using BNP imaging and photoacoustic microscopy (PAM), respectively. PAM non‐invasively measures structural and hemodynamic changes, including vessel diameter, hemoglobin concentration, blood flow speed, and hemoglobin oxygen saturation levels in individual blood vessels down to the capillary level during microvascular remodeling within wound beds4. In this ear wound model, the epidermis and a portion of the dermis, including the center feeder arteriole and its main branches, are excised from the center of the ear lobe. The wound is covered with Tegaderm bandage to prevent drying, and the mouse is fitted with an Elizabethan collar to prevent it from removing the bandage. At regular intervals until the wound heals, BNPs are topically applied to the wound and excited by UV light for two minutes during which the fluorescence (F) and phosphorescence (P) emission signals are recorded using a camera. A custom Matlab program calculates the ratio of F:P emission intensities at each pixel in the wound bed image, which provides a relative mapping of oxygen levels throughout the wound. Following BNP imaging, PAM imaging is performed in order to calculate the percent sO2 in each blood vessel in/around the wound bed. BNP and PAM measurements are compared to one another for wounds in both healthy and diabetic mice.RESULTSDiabetic db/db mice exhibit muted wound oxygenation increases over time compared to non‐diabetic control mice. BNP fluorescence:phosphorescence emission ratios from wound beds emulate sO2 trends generated via PAM, confirming the ability of BNPs to assess tissue oxygenation over time in both diabetic and healthy wounds.CONCLUSIONSBNPs can be used to monitor tissue oxygenation over time in diabetic and healthy wounds. Given their low cost and potential for clinical monitoring of these wounds in humans, BNPs warrant further study for non‐invasively monitoring wound healing over time.Support or Funding InformationUVA School of Medicine Pinn Scholars ProgramThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.