Temporal changes in microvessel leakiness during wound healing discriminated by in vivo fluorescence recovery after photobleaching

摘要

Non-technical summaryWound closure depends on vascular ingrowth into the injured area and this process has traditionally been studied in tissue sections of animals ex vivo. Following the creation of a wound within a surgically implanted dorsal window chamber, we captured microscopic images of the ingrowing vasculature on successive days after injury. Using a combination of surgical, confocal microscopic and mathematical techniques we quantified the flux of plasma into and around vessels including: newly formed vessel sprouts, nascent flowing vascular segments and pre-existing vessels within the same wound. From these analyses we are able to discriminate: (1) vessels with differing maturity, (2) that vascular sprouts get progressively less leaky and (3) TNP-470 (an anti-angiogenic agent), reduces leakiness in sprouts with co-incident secondary effects on pre-existing vessels. These techniques can be used to assess both functional maturity and the effects of therapeutics on the vasculature of healing wounds.