Profile for Cristina Kurachi

STATEMENT OF THE PROBLEM: Fluorescence is an optical signal that is present in natural teeth and some dental restorative materials as a consequence of its molecules energetic decrease. Restorative materials need to match the fluorescence properties of surrounding tooth structure to achieve the best esthetics and appear undetectable. PURPOSE: The fluorescence of 10 commercial composites (shade A2 or equivalent) was tested against that of tooth structure using contrast differences. MATERIALS AND METHODS: Thirty-three standard preparations (3.0-mm wide and 2.00-mm depth) were done on mean maxillaries incisors and divided into 10 groups containing three test samples each. High-definition images of the restored areas and adjacent tooth structure were obtained both under white light of the visible spectrum (control) and ultraviolet light (UV-A=300/ 400 nm). The contrast between composites and tooth structure, expressed in absolute values, was analyzed through digital processing Matlab and Origin softwares and by one-way analysis of variance and Tukey's test (p< (TPH Spectrum [A2] < (Charisma [A2]=Filtek Supreme [A2B]) < (Filtek Supreme [A2E] =Z250 [A2]=Z100 [A2]). CONCLUSION: There is a considerable variation of fluorescence between the composites and the natural tooth structure. CLINICAL SIGNIFICANCE Ideal restorative materials should have fluorescence similar to that of natural teeth. Therefore, it is important to select a composite that possesses adequate fluorescence.

BACKGROUND AND OBJECTIVE: This work evaluates the application of a 810 nm diode laser operating in the range of 400-1,200 mW for bacterial reduction at periodontal treatment. The aim of this study is to examine the immediate effect of the diode medium power laser in reducing the bacterial concentration at periodontal pockets induced in Wistar rats. STUDY DESIGN/MATERIALS AND METHODS: Two bacterial collections were performed on each animal. Microbiological samples were collected before and immediately after laser irradiation. In each group of laser power, eight animals were used, totaling 40 animals. RESULTS: The initial and the final bacterial count revealed that laser irradiation induces considerable bacterial elimination, especially for Prevotella sp, Streptococcus beta-hemolitico, Fusobacterium sp, Pseudomonas sp. CONCLUSIONS: Our results indicate that this laser can constitute an alternative device to traditional infrared systems for bacterial reduction, with some advantage when economical and practical standpoints are considered. (c) 2004 Wiley-Liss, Inc.

Recent advances in optical techniques have created a great range of possibilities for diagnosis and therapeutics in liver related diseases. With the uses of efficient light sources like lasers and LEDs (Light Emitting Diodes) it is possible to employ the light-tissue interaction to promote hepatic tissue regeneration after partial hepatectomy, to detect hepatocarcinoma and steatosis by utilizing optical fluorescence, to evaluate the metabolism of the liver during hepatic transplantation as well as to treat liver tumors. We present here an overview of the technique presently in development at the Ribeirâo Preto Faculty of Medicine-USP in cooperation with the Physics Institute of São Carlos-USP. The results obtained so far have been the subject of a list of publications and are here presented as an overview. A new perspective for modern application of optical techniques in different medical practices related to the liver is presented.

OBJECTIVE: In vivo studies of antimicrobial PDT in animal models of oral candidosis are scarce and the association of porphyrin and LED light has not been evaluated for in vivo photoinactivation of Candida. In this study the effectiveness of photodynamic therapy (PDT) on the inactivation of Candida albicans in vivo was evaluated. Study design. Seventy-one 6-week-old female Swiss mice were immunosuppressed, provided tetracycline to their drinking water, then orally swabbed with a suspension of C. albicans (10(7) CFU/mL). Four days after oral inoculation, PDT was performed on the dorsum of the tongue after topical administration of Photogem at 400, 500, or 1000 mg/L and followed 30 minutes later by illumination with LED light (305 J/cm(2)) at 455 or 630 nm (n = 5 each). After swabbing to recover yeast from the tongue, the number of surviving yeast cells was determined (CFU/mL) and analyzed by ANOVA and Holm-Sidak tests (P < .05). Animals were humanely killed, and the tongues surgically removed and processed for histological evaluation of presence of yeast and inflammatory reaction. Results. PDT resulted in a significant reduction in C. albicans recovered from the tongue (P < .001) when compared with mice from the positive control group. There was no difference between the concentrations of Photogem and LED light wavelengths used. Histological evaluation of the tongue revealed that PDT causes no significant adverse effects to the local mucosa. Conclusion. PDT promoted significant reduction in the viability of C. albicans biofilm without harming the tongue tissue. Copyright © 2009 Mosby, Inc. All rights reserved.

In this study we have used fluorescence spectroscopy to determine the post-mortem interval. Conventional methods in forensic medicine involve tissue or body fluids sampling and laboratory tests, which are often time demanding and may depend on expensive analysis. The presented method consists in using time-dependent variations on the fluorescence spectrum and its correlation with the time elapsed after regular metabolic activity cessation. This new approach addresses unmet needs for post-mortem interval determination in forensic medicine, by providing rapid and in situ measurements that shows improved time resolution relative to existing methods.

The purpose of this research was to evaluate the severity of renal ischemia/reperfusion injury as determined by histology and by laser-induced fluorescence (LIF) with excitation wavelengths of 442 nm and 532 nm. Wistar rats (four groups of six animals) were subjected to left renal warm ischemia for 20, 40, 60 and 80 min followed by 10 min of reperfusion. Autofluorescence was determined before ischemia (control) and then every 5-10 min thereafter. Tissue samples for histology were harvested from the right kidney (control) and from the left kidney after reperfusion. LIF and ischemia time showed a significant correlation (p0.05, and IFF at 532 nm p>0.05. After reperfusion LIF tended to return to preischemic basal levels which occurred in the presence of histological damage. This suggests that factors other than morphological alterations may have a more relevant effect on changes observed in LIF. In conclusion, renal ischemia/reperfusion changed tissue fluorescence induced by laser. The excitation light of 442 nm showed a better correlation with the ischemia time and with the severity of tissue injury.

Although the oral cavity is easily accessible to inspection, patients with oral cancer most often present at a late stage, leading to high morbidity and mortality. Autofluorescence imaging has emerged as a promising technology to aid clinicians in screening for oral neoplasia and as an aid to resection, but current approaches rely on subjective interpretation. We present a new method to objectively delineate neoplastic oral mucosa using autofluorescence imaging. Autofluorescence images were obtained from 56 patients with oral lesions and 11 normal volunteers. From these images, 276 measurements from 159 unique regions of interest (ROI) sites corresponding to normal and confirmed neoplastic areas were identified. Data from ROIs in the first 46 subjects were used to develop a simple classification algorithm based on the ratio of red-to-green fluorescence; performance of this algorithm was then validated using data from the ROIs in the last 21 subjects. This algorithm was applied to patient images to create visual disease probability maps across the field of view. Histologic sections of resected tissue were used to validate the disease probability maps. The best discrimination between neoplastic and nonneoplastic areas was obtained at 405 nm excitation; normal tissue could be discriminated from dysplasia and invasive cancer with a 95.9% sensitivity and 96.2% specificity in the training set, and with a 100% sensitivity and 91.4% specificity in the validation set. Disease probability maps qualitatively agreed with both clinical impression and histology. Autofluorescence imaging coupled with objective image analysis provided a sensitive and noninvasive tool for the detection of oral neoplasia.

BACKGROUND: Optical spectroscopy is a noninvasive technique with potential applications for diagnosis of oral dysplasia and early cancer. In this study, we evaluated the diagnostic performance of a depth-sensitive optical spectroscopy (DSOS) system for distinguishing dysplasia and carcinoma from non-neoplastic oral mucosa. METHODS: Patients with oral lesions and volunteers without any oral abnormalities were recruited to participate. Autofluorescence and diffuse reflectance spectra of selected oral sites were measured using the DSOS system. A total of 424 oral sites in 124 subjects were measured and analyzed, including 154 sites in 60 patients with oral lesions and 270 sites in 64 normal volunteers. Measured optical spectra were used to develop computer-based algorithms to identify the presence of dysplasia or cancer. Sensitivity and specificity were calculated using a gold standard of histopathology for patient sites and clinical impression for normal volunteer sites. RESULTS: Differences in oral spectra were observed in: (1) neoplastic versus nonneoplastic sites, (2) keratinized versus nonkeratinized tissue, and (3) shallow versus deep depths within oral tissue. Algorithms based on spectra from 310 nonkeratinized anatomic sites (buccal, tongue, floor of mouth, and lip) yielded an area under the receiver operating characteristic curve of 0.96 in the training set and 0.93 in the validation set. CONCLUSIONS: The ability to selectively target epithelial and shallow stromal depth regions appeared to be diagnostically useful. For nonkeratinized oral sites, the sensitivity and specificity of this objective diagnostic technique were comparable to that of clinical diagnosis by expert observers. Thus, DSOS has potential to augment oral cancer screening efforts in community settings.

BACKGROUND: Steatosis is diagnosed on the basis of the macroscopic aspect of the liver evaluated by the surgeon at the time of organ extraction or by means of a frozen biopsy. Aim: In the present study, the applicability of laser-induced fluorescence (LIF) spectroscopy was investigated as a method for the diagnosis of different degrees of steatosis experimentally induced in rats. MATERIAL AND METHODS: Rats received a high-lipid diet for different periods of time. The animals were divided into groups according to the degree of induced steatosis diagnosis by histology. The concentration of fat in the liver was correlated with LIF by means of the steatosis fluorescence factor (SFF). RESULTS: The histology classification, according to liver fat concentration was, Severe Steatosis, Moderate Steatosis, Mild Steatosis and Control (no liver steatosis). Fluorescence intensity could be directly correlated with fat content. It was possible to estimate an average of fluorescence intensity variable by means of different confidence intervals (P=95%) for each steatosis group. SFF was significantly higher in the Severe Steatosis group (P<0.001) compared with the Moderate Steatosis, Mild Steatosis and Control groups. CONCLUSION: The various degrees of steatosis could be directly correlated with SFF. LIF spectroscopy proved to be a method capable of identifying the degree of hepatic steatosis in this animal model, and has the potential of clinical application for non-invasive evaluation of the degree of steatosis.

OBJECTIVE: The aim of this study was the evaluation of two different photosensitizers activated by red light emitted by light-emitting diodes (LEDs) in the decontamination of carious bovine dentin. MATERIALS AND METHODS: Fifteen bovine incisors were used to obtain dentin samples which were immersed in brain-heart infusion culture medium supplemented with 1% glucose, 2% sucrose, and 1% young primary culture of Lactobacillus acidophilus 10(8) CFU/mL and Streptococcus mutans 10(8) CFU/mL for caries induction. Three different concentrations of the Photogem solution, a hematoporphyrin derivative (1, 2, and 3 mg/mL) and two different concentrations of toluidine blue O (TBO), a basic dye (0.025 and 0.1 mg/mL) were used. To activate the photosensitizers two different light exposure times were used: 60 sec and 120 sec, corresponding respectively to the doses of 24 J/cm(2) and 48 J/cm(2). RESULTS: After counting the numbers of CFU per milligram of carious dentin, we observed that the use of LED energy in association with Photogem or TBO was effective for bacterial reduction in carious dentin, and that the greatest effect on S. mutans and L. acidophilus was obtained with TBO at 0.1 mg/mL and a dose of 48 J/cm(2). It was also observed that the overall toxicity of TBO was higher than that of Photogem, and that the phototoxicity of TBO was higher than that of Photogem. CONCLUSION: Based on our data we propose a mathematical model for the photodynamic effect when different photosensitizer concentrations and light doses are used.

The efficacy of fluorescence spectroscopy to detect squamous cell carcinoma is evaluated in an animal model following laser excitation at 442 and 532 nm. Lesions are chemically induced with a topical DMBA application at the left lateral tongue of Golden Syrian hamsters. The animals are investigated every 2 weeks after the 4th week of induction until a total of 26 weeks. The right lateral tongue of each animal is considered as a control site (normal contralateral tissue) and the induced lesions are analyzed as a set of points covering the entire clinically detectable area. Based on fluorescence spectral differences, four indices are determined to discriminate normal and carcinoma tissues, based on intraspectral analysis. The spectral data are also analyzed using a multivariate data analysis and the results are compared with histology as the diagnostic gold standard. The best result achieved is for blue excitation using the KNN (K-nearest neighbor, a interspectral analysis) algorithm with a sensitivity of 95.7% and a specificity of 91.6%. These high indices indicate that fluorescence spectroscopy may constitute a fast noninvasive auxiliary tool for diagnostic of cancer within the oral cavity.

A multispectral digital microscope (MDM) is designed and constructed as a tool to improve detection of oral neoplasia. The MDM acquires in vivo images of oral tissue in fluorescence, narrow-band (NB) reflectance, and orthogonal polarized reflectance (OPR) modes, to enable evaluation of lesions that may not exhibit high contrast under standard white light illumination. The device rapidly captures image sequences so that the diagnostic value of each modality can be qualitatively and quantitatively evaluated alone and in combination. As part of a pilot clinical trial, images are acquired from normal volunteers and patients with precancerous and cancerous lesions. In normal subjects, the visibility of vasculature can be enhanced by tuning the reflectance illumination wavelength and polarization. In patients with histologically confirmed neoplasia, we observe decreased blue/green autofluorescence and increased red autofluorescence in lesions, and increased visibility of vasculature using NB and OPR imaging. The perceived lesion borders change with imaging modality, suggesting that multimodal imaging has the potential to provide additional diagnostic information not available using standard white light illumination or by using a single imaging mode alone.

OBJECTIVE: The aim of this study was to compare the angiogenic effects of laser and light-emitting diode (LED) illumination on wounds induced in rats, with varied fluence. BACKGROUND DATA: The LED is an alternative light source that accelerates wound healing, and its efficiency concerning the angiogenic effect was compared to low-level laser therapy (LLLT). METHODS: The experimental model consisted of a circular wound inflicted on the quadriceps of 120 rats, using a 15-mm-diameter "punch." Animals were divided randomly into five groups: two groups of laser, with dosages of 5 and 20 J/cm(2), respectively, two groups of LED, also with dosages of 5 and 20 J/cm(2), and a control group. Six hours after wound infliction, the treated groups received the diverse applications accordingly and were irradiated every 24 h. Angiogenesis was studied through histomorphometry on days 3, 7, 14, and 21 after the wounds were inflicted. RESULTS: On days 3, 7, and 14, the proliferation of blood vessels in all irradiated groups was superior in comparison to those of the control group (p < 0.05). Treatment with fluence of 5 J/cm(2) was better than the laser group with 20 J/cm(2) on day 21. CONCLUSION: Red LLLT and LED demonstrated expressive results in angiogenesis. Light coherence was shown not to be essential to angiogenesis. However, further studies are needed in order to investigate the photobiomodulatory effects of LED in relation to LLLT in various biological tissues.

BACKGROUND: The thickness and depth of invasion of skin tumors may be limiting factors for topical photosensitizer-based photodynamic therapy (PDT). The use of PDT with systemic photosensitizer needs to be further explored as a modality of treatment for nonmelanoma skin cancer (NMSC). OBJECTIVE: The objective was to present six patients with multiple, nodular, and/or pigmented NMSC treated successfully with purified hematoporphyrin derivative (PHD) and PDT using prior debulking. METHODS: After 24 hours of systemic PHD (1.5 mg/kg), 12 lesions of NMSC were selected for PHD-PDT alone and 6 nodular/elevated lesions for PHD-PDT following a debulking procedure. The tumor area was illuminated in one single-dose session of 300 J/cm(2), at an intensity range of 130 to 150 mW/cm(2), with a 630-nm-wavelength diode laser. RESULTS: The prior curettage provided significant reduction in volume and/or pigmentation of lesions. After the session of PHD-PDT with prior curettage and additional topical 20% ALA-PDT in two lesions or PHD-PDT alone, 83% (5/6) of lesions and 58% (7/12) of lesions, respectively, maintained a complete clinical response, 22.2+/-8.9 months of follow-up. CONCLUSIONS: The combination of prior debulking with systemic agents-PDT appears to be a good option for multiple, pigmented, and/or nodular lesions of NMSC and can allow the improvement of clinical results.

OBJECTIVE: The aim of this study was to evaluate temperature variation induced by a diode laser in periodontal repair. BACKGROUND DATA: A diode laser operating with medium power can constitute an alternative device for bacterial reduction at periodontal pockets. This procedure will probably avoid any kind of undesirable thermal damage to the irradiated tissues. METHODS: The temperature variation induced by a 810-nm diode laser was investigated in an in vitro study, varying the soft tissue thickness, and in an in vivo study for soft periodontal and bone tissues. The laser powers used were 600 mW, 800 mW, 1.0 W, and 1.2 W, and the light was delivered by a 300-microm fiber. RESULTS: The laser parameters and irradiation time used did not induce a temperature variation high enough to cause thermal irreversible damage to the periodontal tissues investigated. CONCLUSIONS: This study contributes to the establishment of thermally safe working parameters of a diode medium power laser.