Michael G. Sowa

Infrared spectroscopy: a new frontier in medicine

As we enter the second half of the nineties, one of the major challenges for biological infrared spectroscopists is to transfer the knowledge we have gained from studies on isolated molecules to the complex world of medicine. That it is possible to meet this challenge is suggested by comparison with the development of other biophysical techniques, such as magnetic resonance spectroscopy and imaging, which have already found their place in medical research and practice. The Spectroscopy Group in Winnipeg is developing and evaluating a variety of new IR techniques for the analysis of body fluids and tissues, both in vitro and in vivo. Herein, we review these methodologies, which comprise both instrumental (imaging and spatially localized IR spectroscopy) and interpretational procedures aimed at optimizing the measurements and their conversion to biodiagnostic information.

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.

Modification of the extracellular matrix following myocardial infarction monitored by FTIR spectroscopy

Comparison of mid- and near-infrared spectra of control and infarcted rat ventricular tissue reveals the presence of absorptions in infarcted tissue which are highly characteristics of collagen, indicating large scale deposition of type I collagen in the myocardium following infarction. These results demonstrate that IR spectroscopy may be used to rapidly monitor the modifications of the extracellular matrix associated with myocardial infarction.

Visualization of cutaneous hemoglobin oxygenation and skin hydration using near-infrared spectroscopic imaging.

Background/aims: The visualization of skin hemodynamics and tissue water content has important implications in a number of areas of dermatology, plastic surgery, and clinical skin evaluation. The aim of this study was to develop instrumentation and techniques for infrared spectroscopic imaging, and to evaluate whether they can be used to make objective assessments of skin health, perhaps even before clinical signs are evident.

Detection of early dental caries using polarized Raman spectroscopy

A new technique based on polarized Raman spectroscopy is demonstrated for detecting early dental caries on extracted human teeth. Sound tooth enamel exhibited strong Raman polarization anisotropy whereas early caries consistently showed a lower degree of Raman polarization anisotropy. In particular, for sound enamel, the Raman peak arising from the symmetric nu1 vibration of PO(4) (3-) at 959 cm(-1) is strongly polarized. This is in contrast to the spectra of carious lesions that displayed weaker polarization dependence at 959 cm(-1). Such difference in the degree of Raman polarization anisotropy allows for discrimination between early dental caries and sound enamel.

An Update on Novel Non-Invasive Approaches for Periodontal Diagnosis

For decades there has been an ongoing search for clinically acceptable methods for the accurate, non-invasive diagnosis and prognosis of periodontitis. There are several well-known inherent drawbacks with current clinical procedures. The purpose of this review is to summarize some of the newly emerging diagnostic approaches, namely, infrared spectroscopy, optical coherence tomography (OCT), and ultrasound. The history and attractive features of these new approaches are briefly illustrated, and the interesting and significant inventions related to dental applications are discussed. The particularly attractive aspects for the dental community are that some of these methods are totally non-invasive, do not impose any discomforts to the patients during the procedure, and require no tissue to be extracted. For instance, multiple inflammatory indices withdrawn from near infrared spectra have the potential to identify early signs of inflammation leading to tissue breakdown. Morphologically, some other non-invasive imaging modalities, such as OCT and ultrasound, could be employed to accurately measure probing depths and assess the status of periodontal attachment, the front-line of disease progression. Given that these methods reflect a completely different assessment of periodontal inflammation, if clinically validated, these methods could either replace traditional clinical examinations for the diagnosis of periodontitis or at least serve as attractive complementary diagnostic tools. However, the potential of these techniques should be interpreted more cautiously given the multifactorial character of periodontal disease. In addition to these novel tools in the field of periodontal inflammatory diseases, other alternative modalities like microbiologic and genetic approaches are only briefly mentioned in this review because they have been thoroughly discussed in other comprehensive reviews.

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.

Assessment of early demineralization in teeth using the signal attenuation in optical coherence tomography images

Optical coherence tomography imaging is used to improve the detection of incipient carious lesions in dental enamel. Measurements of signal attenuation in images acquired with an 850-nm light source were performed on 21 extracted molars from eight human volunteers. Stronger attenuation was observed for the optical coherence tomography (OCT) signal in healthy enamel than in carious lesions. The measured attenuation coefficients from the two groups form distinct statistical populations. The coefficients obtained from sound enamel fall within the range of 0.70 to 2.14 mm(-1) with a mean value of 1.35 mm(-1), while those in carious regions range from 0.47 to 1.88 mm(-1), with a mean value of 0.77 mm(-1). Three values are selected as the lower threshold for signal attenuation in sound enamel: 0.99, 0.94, and 0.88 mm(-1). These thresholds were selected to provide detection of sound enamel with fixed specificities of 90%, 95%, and 97.5%, respectively. The corresponding sensitivities for the detection of carious lesions are 92.8%, 90.4%, and 87%, respectively, for the sample population used in this study. These findings suggest that attenuation of OCT signal at 850 nm could be an indicator of tooth demineralization and could be used as a marker for early caries detection.

Collagen morphology and texture analysis: from statistics to classification

In this study we present an image analysis methodology capable of quantifying morphological changes in tissue collagen fibril organization caused by pathological conditions. Texture analysis based on first-order statistics (FOS) and second-order statistics such as gray level co-occurrence matrix (GLCM) was explored to extract second-harmonic generation (SHG) image features that are associated with the structural and biochemical changes of tissue collagen networks. Based on these extracted quantitative parameters, multi-group classification of SHG images was performed. With combined FOS and GLCM texture values, we achieved reliable classification of SHG collagen images acquired from atherosclerosis arteries with >90% accuracy, sensitivity and specificity. The proposed methodology can be applied to a wide range of conditions involving collagen re-modeling, such as in skin disorders, different types of fibrosis and muscular-skeletal diseases affecting ligaments and cartilage.