Assessment of the hemodynamic profile in periodontal tissues of diabetic subjects with periodontitis by optical spectroscopy.

BACKGROUND AND OBJECTIVE: The influence of diabetes mellitus (DM) on the hemodynamics of periodontal tissues has not been assessed previously. The primary objective of this study was to validate optical spectroscopy as a periodontal diagnostic tool for subjects with type 2 DM and chronic periodontitis. MATERIAL AND METHODS: Using a portable optical near-infrared spectrometer, optical spectra were obtained from healthy (n = 127), gingivitis (n = 115), and periodontitis (n = 109) sites of 65 subjects with type 2 DM and chronic periodontitis. Healthy (n = 65) sites of 15 nondiabetic subjects without periodontitis were used as controls. A modified Beer-Lambert unmixing model that incorporates a nonparametric scattering-loss function was used to determine the relative contribution of deoxygenated hemoglobin and oxygenated hemoglobin (HbO2 ) to the overall spectrum. The balance between tissue oxygen delivery and oxygen utilization in periodontal tissues was assessed. RESULTS: In diabetic subjects, tissue oxygen saturation and HbO2 concentration were significantly decreased in the periodontitis sites (p < 0.01) compared with the healthy and gingivitis sites. Furthermore, tissue oxygenation in healthy sites of control subjects was significantly higher than that in sites of diabetic subjects (p < 0.01). CONCLUSION: In summary, the results of this study suggest that optical spectroscopy can monitor the hemodynamic profile in diabetic subjects with chronic periodontitis. Furthermore, healthy sites of diabetic subjects presented lower tissue oxygenation than did those of nondiabetic subjects.

On site noninvasive assessment of peri-implant inflammation by optical spectroscopy.

BACKGROUND AND OBJECTIVE: Optical spectroscopy has been proposed to measure regional tissue hemodynamics in periodontal tissue. The objective of this study was to further evaluate the diagnostic potential of optical spectroscopy in peri-implant inflammation in vivo by assessing multiple inflammatory parameters (tissue oxygenation, total tissue hemoglobin, deoxyhemoglobin, oxygenated hemoglobin and tissue edema) simultaneously. MATERIAL AND METHODS: A cross-sectional study was performed in a total of 64 individuals who presented with dental implants in different stages of inflammation. In brief, visible-near-infrared spectra were obtained, processed and evaluated from healthy (n = 151), mucositis (n = 70) and peri-implantitis sites (n = 75) using a portable spectrometer. A modified Beer-Lambert unmixing model that incorporates a nonparametric scattering loss function was employed to determine the relative contribution of each inflammatory component to the overall spectrum. RESULTS: Tissue oxygenation at peri-implantitis sites was significantly decreased (p < 0.05) when compared with that at healthy sites, which was largely due to an increase in deoxyhemoglobin and a decrease in oxyhemoglobin at the peri-implantitis sites compared with the mucositis and healthy sites. In addition, the tissue hydration index derived from the optical spectra in mucositis was significantly higher than that in other groups (p < 0.05). CONCLUSION: In summary, the results of this study revealed that hemodynamic alterations can be detected around diseased peri-implant sites by optical spectroscopy, and this method may be considered an alternative and feasible approach for the monitoring and diagnosis of peri-implant diseases.

Added soft tissue contrast using signal attenuation and the fractal dimension for optical coherence tomography images of porcine arterial tissue.

Optical coherence tomography (OCT) images of left-descending coronary tissues harvested from three porcine specimens were acquired with a home-build swept-source OCT setup. Despite the fact that OCT is capable of acquiring high resolution circumferential images of vessels, many distinct histological features of a vessel have comparable optical properties leading to poor contrast in OCT images. Two classification methods were tested in this report for the purpose of enhancing contrast between soft-tissue components of porcine coronary vessels. One method involved analyzing the attenuation of the OCT signal as a function of light penetration into the tissue. We demonstrated that by analyzing the signal attenuation in this manner we were able to differentiate two media sub-layers with different orientations of the smooth muscle cells. The other classification method used in our study was fractal analysis. Fractal analysis was implemented in a box-counting (fractal dimension) image-processing code and was used as a tool to differentiate and quantify variations in tissue texture at various locations in the OCT images. The calculated average fractal dimensions had different values in distinct regions of interest (ROI) within the imaged coronary samples. When compared to the results obtained by using the attenuation of the OCT signal, the method of fractal analysis demonstrated better classification potential for distinguishing amongst the tissue ROI.

In vivo determination of multiple indices of periodontal inflammation by optical spectroscopy.

BACKGROUND AND OBJECTIVE: Visible, near-infrared (optical) spectroscopy can be used to measure regional tissue hemodynamics and edema and therefore may represent an ideal tool with which to study periodontal inflammation in a noninvasive manner. The study objective was to evaluate the ability of optical spectroscopy to determine simultaneously multiple inflammatory indices (tissue oxygenation, total tissue hemoglobin, deoxyhemoglobin, oxygenated hemoglobin and tissue edema) in periodontal tissues in vivo. MATERIAL AND METHODS: Spectra were obtained, processed and evaluated from healthy, gingivitis and periodontitis sites (n = 133) using a portable optical, near-infrared spectrometer. A modified Beer-Lambert unmixing model that incorporates a nonparametric scattering loss function was used to determine the relative contribution of each inflammatory component to the overall spectrum. RESULTS: Optical spectroscopy was harnessed to generate complex inflammatory profiles of periodontal tissues. Tissue oxygenation at periodontitis sites was significantly decreased (p < 0.05) compared to sites with gingivitis and healthy controls. This was largely the result of an increase in deoxyhemoglobin in the periodontitis sites compared with healthy (p < 0.01) and gingivitis (p = 0.05) sites. Tissue water content per se showed no significant difference between the sites, but a water index associated with tissue electrolyte levels and temperature differed significantly between periodontitis sites and both healthy and gingivitis sites (p < 0.03). CONCLUSION: This study established that optical spectroscopy can simultaneously determine multiple inflammatory indices directly in the periodontal tissues in vivo. Visible, near-infrared spectroscopy has the potential to be developed into a simple, reagent-free, user-friendly, chairside, site-specific, diagnostic and prognostic test for periodontitis.

The effect of elevated dietary cholesterol on pulmonary surfactant function in adolescent mice.

It has been established that phospholipids and cholesterol interact in films of pulmonary surfactant (PS). Generally it is thought that phospholipids increase film stability whereas cholesterol increases film fluidity. To study this further, we modified dietary cholesterol in mice which received either standard rodent lacking cholesterol (sd), or high cholesterol (2%) diet (hc) for 1 month. Phospholipid stability was investigated by a capillary surfactometer (CS), which measures airflow resistance and patency. PS was collected by bronchiolar lavage and centrifuged to obtain the surface-active film (SAF). Results showed that the hc-SAF had significantly more cholesterol than sd-SAF. CS analyses at 37 degrees C showed no significance differences in airflow resistance between hc-SAF and sd-SAF. However, at 37 degrees C, sd-SAF showed greater ability to maintain patency compared to hc-SAF, whereas at 42 degrees C hc-SAF showed patency ability similar to sd-SAF. The results suggested that increased cholesterol in hc-SAF induced less stability in the SAF possibly due to cholesterol's fluidizing effect on phospholipids at physiological temperatures.

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.

DNA fragmentation in developing lung fibroblasts exposed to Stachybotrys chartarum (atra) toxins.

Stachybotrys chartarum (atra) is a toxic mold that grows on water-damaged cellulose-based materials. Research has revealed also that inhalation of S. chartarum spores caused marked changes in respiratory epithelium, especially to developing lungs. We analyzed the epigenetic potential of S. chartarum spore toxins on developing rat lung fibroblasts using single cell gel electrophoresis (comet assay). Isolated fetal lung fibroblasts were exposed to S. chartarum spore toxins for 15 min, 3, 14, or 24 hr and control cells were exposed to saline under the same conditions. Cells were embedded in agarose, electrophoresed under alkaline conditions and silver stained. DNA damage was assessed in terms of fragmentation as measured by comet tail length (DNA migration) and intensity (% DNA contained within head and tail). Upon visual inspection, control fibroblasts showed no DNA fragmentation whereas S. chartarum-treated cells had definable comets of various degrees depending upon the time-course. Analyses of the comets revealed that exposure to S. chartarum spore toxins for at least 15 min to 14 hr, induced increased DNA fragmentation in a time-dependent manner. The fact that exposure to toxins for 24 hr showed less damage suggested that developing lung fibroblasts may have the capability of repairing DNA fragmentation.

Infrared spectroscopy: shedding light on synovitis in patients with rheumatoid arthritis.

OBJECTIVES: It is difficult to determine the extent of synovial involvement early in the course of rheumatoid arthritis. A spectroscopic technique was used to characterize the synovium of the small finger joints in both early and late rheumatoid arthritis. This synovium was also compared against normal joints. METHODS: Near-infrared spectroscopy assesses the absorption of near-infrared light by specific joints, giving a characteristic "fingerprint" of the properties of the underlying tissues. Triple measurements by infrared spectroscopy were taken at the bilateral second and third metacarpophalangeal joints. Multivariate analysis was applied. RESULTS: Analysis was able to demonstrate relationships between the specific sources of spectral variation and joint tenderness or swelling as well as radiographic damage. Further use of multivariate analysis allowed recognition of the spectral patterns seen in early disease vs late rheumatoid arthritis and correct classification of over 74% of the joints. CONCLUSIONS: The spectral regions where differences occurred were in the absorption bands related to tissue oxygenation status, allowing the provocative implication that this technique could be detecting ischaemic changes within the joint. Near-infrared spectroscopy may thus be able to provide us with some information about the biochemical changes associated with synovitis.

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.

Eliminating the issue of skin color in assessment of the blanch response.

OBJECTIVE: The high melanin concentration in dark skin prevents the observation of a blanch response to light finger pressure. The objective of this study was to determine the ability of visible and near-infrared spectroscopy (the technique used in pulse oximetry) to monitor a blanch response from in vivo spectra in individuals with light and dark skin, based on changes in blood volume. DESIGN: A quasi-experimental repeated measures design was employed. A stepper motor with an attached spectrophotometer probe was used to deliver controlled pressure to the participants' forearms, mimicking the finger-blanching test. Visible and near-infrared spectra were acquired throughout the blanching cycle. SETTING: The In Vivo Tissue Optics Lab at the Institute for Biodiagnostics, Winnipeg, Manitoba, Canada. PARTICIPANTS: A convenience sample of 10 healthy light-skinned individuals and 10 healthy dark-skinned individuals. RESULTS: Determined by analysis of the spectra, the 2 groups differed in pigmentation in both the visible (P<.01) and near-infrared (P<.01) regions of the absorbance spectrum. There was a significant difference in total hemoglobin at high and low pressure in both the visible (P<.01) and near-infrared (P<.05) regions. CONCLUSIONS: The observation of a significant difference in total hemoglobin at high and low pressure in both light- and dark-skinned groups in this study demonstrates the ability of visible and near-infrared spectroscopy to monitor blood volume changes associated with a blanch response. These findings support the potential use of this technology as the basis of a clinically useful blanch response tool that is insensitive to skin color.

Near-infrared spectroscopic assessment of tissue hydration following surgery.

Near-infrared reflectance spectroscopy and imaging were used to assess tissue hydration following elevation of reverse McFarlane dorsal rat skin flaps (N = 6). A quantitative measure of tissue water content was derived using the integrated intensities of the strong absorption bands of water centered at 970 and 1450 nm. Near-infrared spectroscopy monitored tissue hydration at discrete locations, while imaging provided hydration maps of cutaneous tissues. Both methods were rapid and noninvasive. The near-infrared results were compared to laser Doppler flux measurements. Significant changes in tissue hydration were observed upon surgical elevation with substantial regional variation along the skin flap. These results indicate the potential of near-infrared spectroscopy and imaging to monitor tissue hydration changes in the skin following surgery.

Visible-near infrared multispectral imaging of the rat dorsal skin flap.

Visible-near infrared multispectral reflectance image sets were acquired from the dorsal surface of rats both before and after elevation of reversed McFarlane skin flaps. Raw images were dominated by uneven surface illumination and shadowing along with the variation associated with instrument response. These interfering features obscured variation associated with a change in tissue reflectance, which is related to the degree of flap perfusion. Logarithmic residual preprocessing followed by principal component analysis of multispectral images could clearly detect a difference in the optical properties between the base and distal section of the flap. The difference in the reflectance properties correlates with the varying degree of tissue perfusion. Principal component analysis detected this optical difference between the well-perfused base of the skin flap and the compromised distal section of the flap immediately following surgery. The first visual signs of compromised tissue perfusion appeared only 6 or more hours after surgery. The results from this study indicate that the application of principal component analysis to discrete wavelength near infrared multispectral reflectance images of skin flaps can effectively distinguish reflectance changes related to the degree of tissue perfusion immediately following surgical elevation of the reversed McFarlane skin flap. © 1999 Society of Photo-Optical Instrumentation Engineers.

Assessment of tissue viability using near-infrared spectroscopy.

Prolonged and severe tissue hypoxia results in tissue necrosis in pedicled flaps. We demonstrate the potential of near-infrared spectroscopy for monitoring of skin flaps. This approach clearly identifies tissue regions with low oxygen supply, and also the severity of this challenge, in a rapid and non-invasive manner with a high degree of reproducibility. Tissue haemoglobin oxygen saturation and water content of pre-selected dorsal sites were monitored for 72 h prior to, and 72 h following elevation of a reversed McFarlane rat dorsal skin flap (n = 9). Oxygen delivery to flap tissue dropped immediately upon flap elevation. This was most pronounced in the distal half of the flap and least pronounced in the region nearest its base. Haemoglobin oxygen saturation of tissue proximal to the vascular base of the flap recovered, exceeding pre-elevation saturation values, within 6 h of raising the flap. Typically, this higher haemoglobin oxygen saturation persisted for the full 72 h post-elevation observation period. At a distance greater than 2 cm from the vascular pedicle, the tissue remained hypoxic over the post-elevation monitoring period. Tissues remaining below a certain haemoglobin oxygen saturation threshold (oxygen saturation index < 1) for prolonged periods (> 6 h) became increasingly dehydrated, eventually becoming visibly necrotic. Tissues above this threshold (oxygen saturation index > 1), despite being significantly hypoxic, relative to the pre-elevation saturation values, remained viable over the 72 h post-elevation monitoring period.

Tissue viability by multispectral near infrared imaging: a fuzzy C-means clustering analysis.

Clinically, skin color, temperature, and capillary perfusion are used to assess tissue viability following microvascular tissue transfer. However, clinical signs that arise as a consequence of poor perfusion become evident only after several hours of compromised perfusion. This study demonstrates the potential usefulness of optical/infrared multispectral imaging in the prognosis of tissue viability immediately post-surgery. Multispectral images of a skin flap model acquired within 1 h of surgical elevation are analyzed in comparison to the final 72-h clinical outcome with a high degree of correlation. Regional changes in tissue perfusion and oxygenation present immediately following surgery are differentiated using fuzzy clustering and image processing algorithms. These methodologies reduce the intersubject variability inherent in infrared imaging methods such that the changes in perfusion are reproducible and clearly distinguishable across all subjects. Clinically, an early prognostic indicator of viability such as this would allow for a more timely intervention following surgery in the event of compromised microvasculature.

Fuzzy C-means clustering and principal component analysis of time series from near-infrared imaging of forearm ischemia.

Fuzzy C-means clustering and principal components analysis were used to analyze a temporal series of near-IR images taken of a human forearm during periods of venous outflow restriction and complete forearm ischemia. The principal component eigen-time course analysis provided no useful information and the principal component eigen-image analysis gave results that correlated poorly with anatomical features. The fuzzy C-means clustering analysis, on the other hand, showed distinct regional differences in the hemodynamic response and scattering properties of the tissue, which correlated well with the anatomical features of the forearm.

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.

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.

FT-IR photoacoustic depth profiling spectroscopy of enamel.

Photoacoustic Fourier transform infrared (PA-FT-IR) depth profiling spectra of the enamel of an intact human tooth are obtained in a completely nondestructive fashion. The compositional and structural changes in the tissue are probed from the enamel surface to a depth of about 200 microns. These changes reflect the state of tissue development. The subsurface carbonate gradient in the enamel could be observed over the range of about 10-100 microns. The carbonate-to-phosphate ratio increases in the depth profile. The depth profile also reveals changes in the substitutional distribution of carbonate ions. Type A carbonates (hydroxyl substituted) increase relative to type B carbonates (phosphate substituted) with increasing thermal diffusion length. In addition to the changes in the carbonate ion distribution and content, the PA-FT-IR depth profile clearly indicates a dramatic increase in the protein content relative to the phosphate content with increased depth. The changes in the carbonate content and distribution, along with the changes in the protein content, may be responsible for the changes observed in the apatitic structure in the depth profile of the enamel.