Assessing Wound Healing in Vivo Using a Dual-Function Phosphorescent Probe Sensitive to Tissue Oxygenation and Regenerating Collagen

Levels of tissue oxygenation and collagen regeneration are critical indicators in the early evaluation of wound healing. Traditionally, these factors have been assessed using separate instruments and different methodologies. Here, we adopt the spatially averaged phosphorescence lifetime approach using Re^I-diimine complexes (Re^I-probe) to enable simultaneous quantification of these two critical factors in healing wounds. The topically applied, biocompatible Re^I-probe penetrates wound tissue effectively and selectively binds to collagen fibers. During collagen regeneration, the phosphorescence lifetimes of the collagen-bound probe significantly extend from an initial range of 4.5-6.5 μs on day 0 to 5.5-8.5 μs by day 7. Concurrently, unbound probes in the tissue interstitial spaces exhibit a phosphorescence lifetime of 4.5-5.2 μs, revealing the oxygenation states. Using phosphorescence lifetime imaging microscopy (PLIM) and a frequency domain phosphorescence lifetime measurement (FD-PLM) system, we validated the dual-functionality of this Re^I-probe in differentiating healing stages in chronic wounds. With its noninvasive, quantitative measurement capabilities for cutaneous wounds, this Re^I-probe-based approach offers promising potential for early wound healing diagnosis.