Abstract
Wound healing is a well‐coordinated process, necessitating efficient formation of new blood vessels. Vascularization defects are therefore a major risk factor for chronic, non‐healing wounds. The dynamics of mammalian tissue revascularization, vessel maturation, and remodeling remain poorly understood due to lack of suitable in vivo imaging tools. A label‐free large‐scale optoacoustic microscopy (LSOM) approach is developed for rapid, non‐invasive, volumetric imaging of tissue regeneration over large areas spanning up to 50 mm with a depth penetration of 1.5 mm. Vascular networks in dorsal mouse skin and full‐thickness excisional wounds are imaged with capillary resolution during the course of healing, revealing previously undocumented views of the angiogenesis process in an unperturbed wound environment. Development of an automatic analysis framework enables the identification of key features of wound angiogenesis, including vessel length, diameter, tortuosity, and angular alignment. The approach offers a versatile tool for preclinical research in tissue engineering and regenerative medicine, empowering label‐free, longitudinal, high‐throughput, and quantitative studies of the microcirculation in processes associated with normal and impaired vascular remodeling, and analysis of vascular responses to pharmacological interventions in vivo.
Highlights
Wound healing is a well-coordinated process, necessitating efficient formation of new blood vessels
The improved resolution was crucial to resolve fine capillary networks formed during wound healing, while the high DOF aided with imaging the rough topology of the dorsal www.advancedscience.com skin
Our large-scale optoacoustic microscopy (LSOM) method, based on volumetric imaging of optical absorption, was used to monitor vascular and structural characteristics coupled to the various phases of wound healing and tissue regeneration at capillary-level resolution
Summary
Wound healing is a well-coordinated process, necessitating efficient formation of new blood vessels. Vascular networks in dorsal mouse skin and full-thickness excisional wounds are imaged with capillary resolution during the course of healing, revealing previously trols blood flow to the site of injury This allows distribution of oxygen and nutrients and mediates the inflammatory response and the formation of new tissue.[1,2] Deficiencies in wound vascularization are a undocumented views of the angiogenesis process in an unperturbed wound main driving force of pathological healing environment. The approach offers a versatile tool for preclinical research in tissue engineering and regenerative medicine, empowering label-free, longitudinal, high-throughput, and quantitative chronic ulcers, which pose an enormous social and economic challenge.[1,3,4,5] Accurate evaluation of wound healing and tissue regeneration is paramount for devising and testing optimal treatment strategies, studies of the microcirculation in processes associated with normal and and non-invasive in vivo imaging provides impaired vascular remodeling, and analysis of vascular responses to pharmacological interventions in vivo. We developed a new imaging pipeline based on largescale optoacoustic microscopy (LSOM) tailored for intravital, non-invasive, longitudinal imaging of intact and wounded opaque murine skin with superb resolution and imaging speed, which allows for a quantitative analysis of vascular network alterations during wound healing
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