Protocol to assess extravasation of fluorescent molecules in mice after ultrasound-mediated blood-brain barrier opening.

Blood-brain barrier disruption (BBBD) using focused ultrasound (FUS) and microbubbles (MBs) is an effective tool for therapeutic delivery to the brain. Here, we present an optimized protocol for quantifying fluorescent molecules extravasation in mice. We describe steps for ultrasound treatment, injection of MBs and fluorescent dyes, brain harvesting, microscopy imaging, and image postprocessing algorithm. Our protocol has proven to successfully conduct a diameter-dependent analysis that measures vascular leakage following FUS-mediated BBBD at a single blood vessel resolution. For complete details on the use and execution of this protocol, please refer to Katz et al.1.

Diameter-dependent assessment of microvascular leakage following ultrasound-mediated blood-brain barrier opening.

Blood brain barrier disruption (BBBD) using focused ultrasound (FUS) and microbubbles (MB) is an effective tool for therapeutic delivery to the brain. BBBD depends to a great extent on MB oscillations. Because the brain vasculature is heterogenic in diameter, reduced MB oscillations in smaller blood vessels, together with a lower number of MBs in capillaries, can lead to variations in BBBD. Therefore, evaluating the impact of microvasculature diameter on BBBD is of great importance. We present a method to characterize molecules extravasation following FUS-mediated BBBD, at a single blood vessel resolution. Evans blue (EB) leakage was used as marker for BBBD, whereas blood vessels localization was done using FITC labeled Dextran. Automated image processing pipeline was developed to quantify the extent of extravasation as function of microvasculature diameter, including a wide range of vascular morphological parameters. Variations in MB vibrational response were observed in blood vessel mimicking fibers with varied diameters. Higher peak negative pressures (PNP) were required to initiate stable cavitation in fibers with smaller diameters. In vivo in the treated brains, EB extravasation increased as a function of blood vessel diameter. The percentage of strong BBBD blood vessels increased from 9.75% for 2-3 µm blood vessels to 91.67% for 9-10 µm. Using this method, it is possible to conduct a diameter-dependent analysis that measures vascular leakage resulting from FUS-mediated BBBD at a single blood vessel resolution.

Physiological measurements of facial skin response under personal protective equipment.

Medical device-related pressure ulcers (MDRPUs) were traditionally associated with skin-contacting medical devices applied to patients, eventually causing tissue damage. The coronavirus-2019 pandemic has brought a new variant of MDRPUs: facial skin irritation or damage associated with extended use of protective personal equipment (PPE), e.g. facemasks and respirators. In this context, we report here a comprehensive experimental evaluation including facial contact forces, skin temperatures and sub-epidermal moisture (SEM) measurements pre/post-PPE usage, to determine how these physiological parameters change under the effects of surgical facemasks and KN95 respirators and whether such potential changes can explain the commonly reported skin irritation or damage. We found that a surgical mask is potentially less irritating to facial skin than the KN95 respirator, as it applies lower forces and facilitates faster return of facial temperatures to their basal levels. Further, we demonstrated that use of dressing cuts for padding under a KN95 respirator considerably reduced localized forces and did not worsen the thermal and SEM readings at the skin-device contact sites. This study provides a basis for improvement of PPE designs, as it describes physiological measurement methodologies for quantitative comparisons of the effects of different PPE types on facial skin status.

Ultrasound elastography reveals the relation between body posture and soft-tissue stiffness which is relevant to the etiology of sitting-acquired pressure ulcers.

OBJECTIVE: Sitting-acquired pressure ulcers (PUs) are common in wheelchair users. These PUs are often serious and may involve deep tissue injury (DTI). Investigating the mechanical properties of the tissues susceptible to DTI may help in guiding the prevention and early detection of PUs. In this study, shear wave elastography (SWE) was used to measure the normative mechanical properties of the soft tissues of the buttocks, i.e. skeletal muscle and subcutaneous fat, under the ischial tuberosities, in a convenient sample of healthy adults without weight bearing and with weight bearing of different times. APPROACH: We compared the stiffness properties of these soft tissues between the lying prone and sitting postures, to determine whether there are detectable property changes that may be associated with the type of posture. We hypothesized that muscle contractions and 3D tissue configurations associated with the posture may influence the measured tissue stiffnesses. MAIN RESULTS: Our results have shown that indeed, SWE values differed significantly across postures, but not over time in a specific posture or for the right versus left sides of the body. SIGNIFICANCE: We have therefore demonstrated that soft-tissue stiffness increases when sitting with weight bearing and may contribute to increasing the potential PU risk in sitting compared to lying prone, given the stiffer behavior of tissues observed in sitting postures.

Protecting prone positioned patients from facial pressure ulcers using prophylactic dressings: A timely biomechanical analysis in the context of the COVID-19 pandemic.

Prone positioning is used for surgical access and recently in exponentially growing numbers of coronavirus disease 2019 patients who are ventilated prone. To reduce their facial pressure ulcer risk, prophylactic dressings can be used; however, the biomechanical efficacy of this intervention has not been studied yet. We, therefore, evaluated facial soft tissue exposures to sustained mechanical loads in a prone position, with versus without multi-layered silicone foam dressings applied as tissue protectors at the forehead and chin. We used an anatomically realistic validated finite element model of an adult male head to determine the contribution of the dressings to the alleviation of the sustained tissue loads. The application of the dressings considerably relieved the tissue exposures to loading. Specifically, with respect to the forehead, the application of a dressing resulted in 52% and 71% reductions in soft tissue exposures to effective stresses and strain energy densities, respectively. Likewise, a chin dressing lowered the soft tissue exposures to stresses and strain energy densities by 78% and 92%, respectively. While the surgical context is clear and there is a solid, relevant need for biomechanical information regarding prophylaxis for the prone positions, the projected consequences of the coronavirus pandemic make the present work more relevant than ever before.

The microclimate under dressings applied to intact weight-bearing skin: Infrared thermography studies.

BACKGROUND: When a patient is lying in a hospital bed (e.g. supine or prone), bodyweight forces distort soft tissues by compression, tension and shear, and may lead to the onset of pressure ulcers in those who are stationary and insensate, especially at their pelvic region. Altered localized microclimate conditions, particularly elevated skin temperatures leading to perspiration and resulting in skin moisture or wetness, are known to further increase the risk for pressure ulcers, which is already high in immobile patients. METHODS: We have used infrared thermography to measure local skin temperatures at the buttocks of supine healthy subjects, to quantitatively determine, for the first time in the literature, how skin microclimate conditions associated with a weight-bearing Fowler's position are affected by application of dressings. Our present methodology has been applied to compare a polymeric membrane dressing versus placebo foam, with a no-dressing case used as reference. FINDINGS: One hour of lying in a Fowler's position was already enough to cause considerable heat trapping (~3 °C rise) between the weight-bearing body and the support surface. Analyses of normalized local skin temperatures and entropy of the temperature distributions indicated that the polymeric membrane dressing material allowed better and more homogenous clearance of locally accumulated body-heat with respect to simple foam. INTERPRETATION: Infrared thermography is suitable for characterizing skin microclimate conditions under different dressings, and, accordingly, is effective in developing and evaluating pressure ulcer prevention and treatment strategies - both of which require adequate skin microclimate.