Lorenzo Leonardi

Near infrared spectroscopic assessment of hemodynamic changes in the early post-burn period

Near infrared reflectance spectroscopy and imaging was used to assess non-invasively the hemodynamic changes that occur in the early post-burn period in cutaneous burn injuries of varying depth. An acute porcine model was used to demonstrate the potential of near infrared spectroscopy and imaging to accurately determine the change in tissue oxygenation, blood volume and tissue water content following a thermal injury. Near infrared spectroscopy was used to monitor tissue at discrete locations, while spectroscopic imaging was able to survey large areas of tissue. Both methods were rapid and non-invasive. Tissue hemoglobin oxygen saturation, total hemoglobin and tissue water content were all affected by thermal injury and changed significantly over a 3 h post-burn monitoring period. Burns that ranged in severity between superficial and full thickness displayed a significantly different hemodynamic response. When the early post-burn profiles (1-3 h) of tissue hemoglobin oxygen saturation, total hemoglobin and tissue water content were considered jointly, injuries leading to superficial, intermediate partial thickness, deep partial thickness and full thickness burns could all be differentiated at high statistical significance. These results suggest that non-invasive hemodynamic monitoring in the early post-burn period using near infrared spectroscopy may be of value in the early assessment of burn injury.

Ex vivo detection and characterization of early dental caries by optical coherence tomography and Raman spectroscopy.

Early dental caries detection will facilitate implementation of nonsurgical methods for arresting caries progression and promoting tooth remineralization. We present a method that combines optical coherence tomography (OCT) and Raman spectroscopy to provide morphological information and biochemical specificity for detecting and characterizing incipient carious lesions found in extracted human teeth. OCT imaging of tooth samples demonstrated increased light backscattering intensity at sites of carious lesions as compared to the sound enamel. The observed lesion depth on an OCT image was approximately 290 microm matching those previously documented for incipient caries. Using Raman microspectroscopy and fiber-optic-based Raman spectroscopy to characterize the caries further, spectral changes were observed in PO4 (3-) vibrations arising from hydroxyapatite of mineralized tooth tissue. Examination of various ratios of PO4 (3-) nu2, nu3, nu4 vibrations against the nu1 vibration showed consistent increases in carious lesions compared to sound enamel. The changes were attributed to demineralization-induced alterations of enamel crystallite morphology and/or orientation. OCT imaging is useful for screening carious sites and determining lesion depth, with Raman spectroscopy providing biochemical confirmation of caries. The combination has potential for development into a new fiber-optic diagnostic tool enabling dentists to identify early caries lesions with greater sensitivity and specificity.

Clinical utilization of near-infrared spectroscopy devices for burn depth assessment.

The diagnosis of burn depth is based on a visual assessment and can be subjective. Near‐infrared (NIR) spectroscopic devices were used preclinically with positive results. The purpose of this study was to test the devices in a clinical setting using easily identifiable burn wounds. Adult patients with acute superficial and full‐thickness burns were enrolled. NIR point spectroscopy and imaging devices were used to collect hemodynamic data from the burn site and an adjacent unburned control site. Oxy‐hemoglobin and deoxy‐hemoglobin concentrations were extracted from spectroscopic data and reported as oxygen saturation and total hemoglobin. Sixteen patients (n=16) were included in the study with equal numbers in both burn wound groups. Point spectroscopy data showed an increase in oxygen saturation (p<0.0095) and total hemoglobin (<0.0001) in comparison with the respective control areas for superficial burn wounds. The opposite was true for full‐thickness burns, which showed a decrease in oxygenation (p<0.0001) and total hemoglobin (p<0.0147) in comparison with control areas. NIR imaging technology provides an estimate of hemodynamic parameters and could easily distinguish superficial and full‐thickness burn wounds. These results confirm that NIR devices can successfully distinguish superficial and full‐thickness burn injuries.

Classification of burn injuries using near-infrared spectroscopy

Early surgical management of those burn injuries that will not heal spontaneously is critical. The decision to excise and graft is based on a visual assessment that is often inaccurate but yet continues to be the primary means of grading the injury. Superficial and intermediate partial-thickness injuries generally heal with appropriate wound care while deep partial- and full-thickness injuries generally require surgery. This study explores the possibility of using near-infrared spectroscopy to provide an objective and accurate means of distinguishing shallow injuries from deeper burns that require surgery. Twenty burn injuries are studied in five animals, with burns covering <1% of the total body surface area. Carefully controlled superficial, intermediate, and deep partial-thickness injuries as well as full-thickness injuries could be studied with this model. Near-infrared reflectance spectroscopy was used to evaluate these injuries 1 to 3 hours after the insult. A probabilistic model employing partial least-squares logistic regression was used to determine the degree of injury, shallow (superficial or intermediate partial) from deep (deep partial and full thickness), based on the reflectance spectrum of the wound. A leave-animal-out cross-validation strategy was used to test the predictive ability of a 2-latent variable, partial least-squares logistic regression model to distinguish deep burn injuries from shallow injuries. The model displayed reasonable ranking quality as summarized by the area under the receiver operator characteristics curve, AUC = 0.879. Fixing the threshold for the class boundaries at 0.5 probability, the model sensitivity (true positive fraction) to separate deep from shallow burns was 0.90, while model specificity (true negative fraction) was 0.83. Using an acute porcine model of thermal burn injuries, the potential of near-infrared spectroscopy to distinguish between shallow healing burns and deeper burn injuries was demonstrated. While these results should be considered as preliminary and require clinical validation, a probabilistic model capable of differentiating these classes of burns would be a significant aid to the burn specialist.

Noninvasive Measurement of Edema in Partial Thickness Burn Wounds

A lack of noninvasive tools to quantify edema has limited our understanding of burn wound edema pathophysiology in a clinical setting. Near-infrared spectroscopy (NIR) is a new noninvasive tool able to measure water concentration/edema in tissue. The purpose of this study was to determine whether NIR could detect water concentration changes or edema formation in acute partial-thickness burn injuries. Adult burn patients within 72 hours postinjury, thermal etiology, partial-thickness burn depth, and <20% TBSA were included. Burn wounds were stratified into partial-thickness superficial or deep wounds based on histology and wound healing time. NIR devices were used to quantify edema in a burn and respective control sites. The sample population consisted of superficial (n = 12) and deep (n = 5) partial-thickness burn injuries. The patients did not differ with respect to age (40 ± 15 years), TBSA (5 ± 4%), and mean time for edema assessment (2 days). Water content increased 15% in burned tissue compared with the respective control regions. There were no differences in water content at the control sites. At 48 hours, deep partial-thickness injuries showed a 23% increase in water content compared with 18% superficial partial-thickness burns. NIR could detect differences in water content or edema formation in partial-thickness burns and unburned healthy regions. NIR holds promise as a noninvasive, portable clinical tool to quantify water content or edema in burn wounds.

OCT of early dental caries: a comparative study with histology and Raman spectroscopy

Early dental caries result from destruction of the tooths outer mineral matrix by acid-forming bacteria found in dental plaques. Early caries begin as surface disruptions where minerals are leached from the teeth resulting in regions of decreased mineral matrix integrity. Visually, these early carious regions appear as white spots due to the higher backscattering of incident light. With age these areas may become stained by organic compounds. Optical coherence tomography (OCT) examination of human teeth demonstrates a difference in penetration depth of the OCT signal into the carious region in comparison with sound enamel. However, while OCT demonstrates a structural difference in the enamel in the region of the caries, this technique provides little insight into the source of this difference. Raman spectroscopy provides biochemical measures derived from hydroxyapatite within the enamel as well as information on the crystallinity of the enamel matrix. The differences in the biochemical and morphological features of early caries and intact sound enamel are compared. Histological thin sections confirm the observations by OCT morphological imaging while Raman spectroscopy allows for biochemical identification of carious regions by a non-destructive method. Visual examination and conventional radiographic imaging of the intact tooth are used in clinical assessment prior to optical measurements. The combination of OCT, Raman spectroscopy and thin section histology aid in determining the changes that give rise to the visual white spot lesions.

Detecting intestinal ischemia using near infrared spectroscopy

Blood supply to the intestine can suddenly be interrupted. Acute mesenteric intestinal ischemia often requires invasive surgery to restore blood supply to the intestine. Early correction of vascular insufficiency is the most important factor in improving patient survival when confronted with acute mesenteric intestinal ischemia. A prolonged loss of blood flow results in irreversible damage to the intestine that can lead to death. It is also imperative that dead segments of the intestines be removed. Several subjective criteria are relied upon to differentiate viable from non-viable tissue, unfortunately, these criteria can lead to an inaccurate assessment. A porcine model of intestinal ischemia was used to determine the efficacy of using near infrared (NIR) spectroscopy to find ischemic segments of the intestine and detect the onset of reperfusion following resolution of vascular occlusion. Nine segments of intestine were identified and six were assigned to three treatment groups; (1) segments undergoing no vascular manipulations, (2) segments undergoing arterial/venous occlusion and (3) segments undergoing arterial/venous occlusion followed by reperfusion. The remaining segments were used as spacers and interposed between each of the ischemia segments. A classification model, using partial least square discriminant analysis, was built on the spectra collected from the segments with no vascular manipulations and the segments that were solely subjected to arterial/venous occlusion. The spectra collected from the intestinal segments that experienced both occlusion and reperfusion were used to test the classification model. The model was able to detect and distinguish ischemic intestinal tissue with a specificity and sensitivity exceeding 80% with an overall classification accuracy of 89%. The method appears to be well suited as an intra-operative assessment method when intestinal ischemia is a concern.

Evaluating the health of compromised tissues using a near-infrared spectroscopic imaging system in clinical settings: lessons learned

The present and accepted standard for determining the status of tissue relies on visual inspection of the tissue. Based on the surface appearance of the tissue, medical personnel will make an assessment of the tissue and proceed to a course of action or treatment. Visual inspection of tissue is central to many areas of clinical medicine, and remains a cornerstone of dermatology, reconstructive plastic surgery, and in the management of chronic wounds, and burn injuries. Near infrared spectroscopic imaging holds the promise of being able to monitor the dynamics of tissue physiology in real-time and detect pathology in living tissue. The continuous measurement of metabolic, physiological, or structural changes in tissue is of primary concern in many clinical and biomedical domains. A near infrared hyperspectral imaging system was constructed for the assessment of burn injuries and skin flaps or skin grafts. This device merged basic science with engineering and integrated manufacturing to develop a device suitable to detect ischemic tissue. This device has the potential of providing measures of tissue physiology, oxygen delivery and tissue hydration during patient screening, in the operating room or during therapy and post-operative/treatment monitoring. Results from a pre-clinical burn injury study will be presented.

Near-infrared Spectroscopy, In Vivo Tissue Analysis by

In vivo near-infrared (NIR) spectroscopy has the potential of becoming an important tool in a number of areas in clinical medicine. Technological developments in photonics that have been spurred on by the communication revolution have set the stage for rapid advancement of optical and NIR spectroscopy based on noninvasive or minimally invasive medical diagnostic techniques. The goal of this article is to review the current capabilities and limitations of in vivo NIR spectroscopy and highlight the impact of these capabilities and limitations in selected areas where NIR spectroscopy is being used to address clinical problems. The optical properties of tissues are briefly reviewed, as are the instrumental methods available to the experimentalist. These properties and methods largely dictate the feasibility of an in vivo spectroscopic diagnostic approach and constrain the scope of problems that can be tackled using optical–NIR spectroscopy. Some of the more successful applications are described, including studies of tissue oxygenation, ischemia, and viability. A number of factors that can confound interpretation of in vivo NIR results are discussed. The number and magnitude of confounding influences that arise in in vivo spectroscopy can be daunting to the experimentalist and may represent the largest barrier in transforming in vivo spectroscopic measurements into clinically meaningful and reliable information. In vivo NIR spectroscopy abounds with opportunity and challenge.

Applications of visible near-infrared spectroscopy and imaging in burn injury assessment

The major objective of the project is to develop a noninvasive method to assess thermal burns. Currently, the diagnosis relies primarily upon visual assessment of the injury by a burn specialist and/or plastic surgeon. The diagnosis is based on the surface appearance of the wound to determine the type or depth of the burn. Near IR spectroscopic measurements of injured tissue provide an objective means of distinguishing between surface and subsurface changes related to the tissue injury. An acute porcine model is employed to investigate the potential of near IR spectroscopy to accurately distinguish between burns of varying severity in the early postburn period. Parallel factor analysis is used to investigate the spectral changes related to burns of varying severity. Burn injuries drastically alter the physical and optical properties of the tissue. Thermal destruction of cutaneous vasculature disrupts perfusion and oxygen delivery to the affected tissue. Tissue blood oxygenation decreases with increased severity of the burn. The result demonstrate that near IR spectroscopy may provide a new tool for objective clinical assessment of burn injuries.