ATR-FTIR spectroscopy imaging of bone repair in mandibular laser-osteotomy.

The aim of this study was to verify the effectiveness of attenuated total reflectance-fourier transform infrared (ATR-FTIR) spectroscopy in the characterization of bone repair in mandibular osteotomy using erbium, chromium-doped yttrium, scandium, gallium and garnet (Er,Cr:YSGG) laser and multilaminate drill on each side. Two mandible bone fragments were removed from 30 rabbits, and the process of bone repair was studied immediately, 3, 7, 15, 21, and 28 days after the surgery. The histological analysis allowed detecting differences in the early stages of tissue repair after bone cutting performed with the Er,Cr:YSGG laser or multilaminate drill. The ATR-FTIR spectroscopy technique was sensitive to changes in the organic content of bone tissue repair process.

Bioactive glass/poloxamer 407 hydrogel composite as a drug delivery system: The interplay between glass dissolution and drug release kinetics.

Since patients suffer pain in the post-surgery of bone repair interventions, bioactive glass/hydrogel drug delivery systems containing local anesthetics, such as ropivacaine, could improve patient life quality by combining bone regeneration with anesthetics. However, poloxamer-based hydrogel properties are sensitive to ions, temperature, and water contents and could be structurally influenced by the ionic dissolution products from bioactive glasses of different compositions. Therefore, this study evaluated the interplay between bioactive glass dissolution kinetics and poloxamer 407 properties, establishing a correlation between changes in the hydrogel and drug release kinetics. Three glass compositions were produced, yielding Ca-rich, Na-rich, and an intermediate glass composition. The influence of Ca/Na ratios on the glass structure and dissolution was investigated. Further, the glasses and ropivacaine were incorporated in the poloxamer hydrogel, and the self-assembly ability of poloxamer, the degradation rate, and the drug release kinetics of the composites were evaluated. The results suggested that glass connectivity affected the early-stage of glass dissolution, while sodium mobility influenced the long-term. The dissolution products from the glasses interact with the supramolecular structure of the poloxamer, causing structural changes responsible for hydrogel degradation. Consequently, by changing the Ca/Na ratio in the glasses, it is possible to modulate glass dissolution that, in turn, influences the ropivacaine release. Thus, we propose that the Ca/Na ratio in quaternary bioactive glasses can be used to modulate drug-delivery properties from systems based on bioactive glasses and poloxamer 407.

Using Fourier transform infrared spectroscopy to evaluate biological effects induced by photodynamic therapy.

BACKGROUND AND OBJECTIVE: Vibrational spectroscopic methods associated with multivariate statistical techniques have been succeeded in discriminating skin lesions from normal tissues. However, there is no study exploring the potential of these techniques to assess the alterations promoted by photodynamic effect in tissue. The present study aims to demonstrate the ability of Fourier Transform Infrared (FTIR) spectroscopy on Attenuated total reflection (ATR) sampling mode associated with principal component-linear discriminant analysis (PC-LDA) to evaluate the biochemical changes caused by photodynamic therapy (PDT) in skin neoplastic tissue. MATERIALS AND METHODS: Cutaneous neoplastic lesions, precursors of squamous cell carcinoma (SCC), were chemically induced in Swiss mice and submitted to a single session of 5-aminolevulinic acid (ALA)-mediated PDT. Tissue sections with 5 µm thickness were obtained from formalin-fixed paraffin-embedded (FFPE) and processed prior to the histopathological analysis and spectroscopic measurements. Spectra were collected in mid-infrared region using a FTIR spectrometer on ATR sampling mode. Principal Component-Linear Discriminant Analysis (PC-LDA) was applied on preprocessed second derivatives spectra. Biochemical changes were assessed using PCA-loadings and accuracy of classification was obtained from PC-LDA . RESULTS: Sub-bands of Amide I (1,624 and 1,650 cm(-1) ) and Amide II (1,517 cm(-1) ) indicated a protein overexpression in non-treated and post-PDT neoplastic tissue compared with healthy skin, as well as a decrease in collagen fibers (1,204, 1,236, 1,282, and 1,338 cm(-1) ) and glycogen (1,028, 1,082, and 1,151 cm(-1) ) content. Photosensitized neoplastic tissue revealed shifted peak position and decreased ß-sheet secondary structure of proteins (1,624 cm(-1) ) amount in comparison to non-treated neoplastic lesions. PC-LDA score plots discriminated non-treated neoplastic skin spectra from post-PDT cutaneous lesions with accuracy of 92.8%, whereas non-treated neoplastic skin was discriminated from healthy tissue with 93.5% accuracy and post-PDT cutaneous lesions was discriminated from healthy tissue with 89.7% accuracy. CONCLUSION: PC-LDA was able to discriminate ATR-FTIR spectra of non-treated and post-PDT neoplastic lesions, as well as from healthy skin. Thus, the method can be used for early diagnosis of premalignant skin lesions, as well as to evaluate the response to photodynamic treatment. Lasers Surg. Med. 48:538-545, 2016. © 2016 Wiley Periodicals, Inc.

ATR-FTIR spectroscopy for the assessment of biochemical changes in skin due to cutaneous squamous cell carcinoma.

Nonmelanoma skin cancers represent 95% of cutaneous neoplasms. Among them, squamous cell carcinoma (SCC) is the more aggressive form and shows a pattern of possible metastatic profile. In this work, we used Fourier transform infrared spectroscopy (FTIR) spectroscopy to assess the biochemical changes in normal skin caused by squamous cell carcinoma induced by multi-stage chemical carcinogenesis in mice. Changes in the absorption intensities and shifts were observed in the vibrational modes associated to proteins, indicating changes in secondary conformation in the neoplastic tissue. Hierarchical cluster analysis was performed to evaluate the potential of the technique to differentiate the spectra of neoplastic and normal skin tissue, so that the accuracy obtained for this classification was 86.4%. In this sense, attenuated total reflection (ATR)-FTIR spectroscopy provides a useful tool to complement histopathological analysis in the clinical routine for the diagnosis of cutaneous squamous cell carcinoma.

Noninvasive monitoring of photodynamic therapy on skin neoplastic lesions using the optical attenuation coefficient measured by optical coherence tomography.

Photodynamic therapy (PDT) has become a promising alternative for treatment of skin lesions such as squamous cell carcinoma. We propose a method to monitor the effects of PDT in a noninvasive way by using the optical attenuation coefficient (OAC) calculated from optical coherence tomography (OCT) images. We conducted a study on mice with chemically induced neoplastic lesions and performed PDT on these lesions using homemade photosensitizers. The response of neoplastic lesions to therapy was monitored using, at the same time, macroscopic clinical visualization, histopathological analysis, OCT imaging, and OCT-based attenuation coefficient measurement. Results with all four modalities demonstrated a positive response to treatment. The attenuation coefficient was found to be 1.4 higher in skin lesions than in healthy tissue and it decreased after therapy. This study shows that the OAC is a potential tool to noninvasively assess the evolution of skin neoplastic lesions with time after treatment.

Evaluation of two quantitative analysis methods of optical coherence tomography for detection of enamel demineralization and comparison with microhardness.

BACKGROUND AND OBJECTIVE: We aimed to evaluate in the same study two quantitative methods for quantification of incipient caries in human dental enamel by using optical coherence tomography (OCT): the optical attenuation coefficient and the area under the A-scan signal, and to compare their results with those obtained from microhardness analysis. STUDY DESIGN/MATERIALS AND METHODS: One hundred and sixty samples were obtained from 40 sound human third molars, which had their crowns sectioned. Simulated caries were created by a pH cycling method. OCT measurements were performed on the samples, before and after the induced demineralization. We determined the total optical attenuation coefficient from the OCT signal in each site and evaluated the sensitivity and specificity of this approach to the detection of the demineralization. Also, the areas under the OCT curves (AUC-OCT) and those from sectional microhardness tests (AUC-MH) were compared. RESULTS: Both the analysis of the optical attenuation coefficient and the AUC-OCT were adequate to efficiently distinguish sound and demineralized samples with sensitivity of 0.93 and specificity of 0.96. The AUC-MH and the AUC-OCT data presented linear relationship and correlation of 0.99. CONCLUSION: Both methods for signal analysis from OCT allowed detection of demineralization with good performance. The AUC-OCT approach enables obtaining a linear relation with the microhardness results, for a quantitative assessment of mineral loss in human teeth.

Effect of dental tissue conditioners and matrix metalloproteinase inhibitors on type I collagen microstructure analyzed by Fourier transform infrared spectroscopy.

This study aimed to evaluate the chemical interaction of collagen with some substances usually applied in dental treatments to increase the durability of adhesive restorations to dentin. Initially, the similarity between human dentin collagen and type I collagen obtained from commercial bovine membranes of Achilles deep tendon was compared by the Attenuated Total Reflectance technique of Fourier Transform Infrared (ATR-FTIR) spectroscopy. Finally, the effects of application of 35% phosphoric acid, 0.1M ethylenediaminetetraacetic acid (EDTA), 2% chlorhexidine, and 6.5% proanthocyanidin solution on microstructure of collagen and in the integrity of its triple helix were also evaluated by ATR-FTIR. It was observed that the commercial type I collagen can be used as an efficient substitute for demineralized human dentin in studies that use spectroscopy analysis. The 35% phosphoric acid significantly altered the organic content of amides, proline and hydroxyproline of type I collagen. The surface treatment with 0.1M EDTA, 2% chlorhexidine, or 6.5% proanthocyanidin did not promote deleterious structural changes to the collagen triple helix. The application of 6.5% proanthocyanidin on collagen promoted hydrogen bond formation.

Bond strength of self-etching primer to bur cut, Er,Cr:YSGG, and Er:YAG lased dental surfaces.

OBJECTIVE: The purpose of this study was to evaluate the tensile bond strength of a self-etching primer system to enamel and dentin surfaces treated with Er:YAG and Er,Cr:YSGG lasers. BACKGROUND DATA: The recently introduced self-etching primer systems have been shown to adhere better to dental surfaces with thin or no smear layers. Moreover, there have been no previous reports on the bond strength of these adhesives to Er,Cr:YSGG laser-irradiated enamel and dentin, which have been shown to be free of a smear layer. METHODS: Thirty samples of enamel and thirty of dentin were divided into three groups. The first group of each substrate served as a control with a standardized bur cut, and the other two groups were conditioned with Er:YAG (350 mJ, 10 Hz, 20 J/cm(2) for enamel; 300 mJ, 6 Hz, 17 J/cm(2) for dentin) and Er,Cr:YSGG laser (125 mJ, 20 Hz, 16 J/cm(2) for both substrates). After the bonding procedure, samples were restored with composite resin, and the tensile bond strength test was performed. RESULTS: The ANOVA two-way analysis and the Tukey test at 5% significance level showed that for enamel and dentin, the bond strength values were statistically higher in Er:YAG-laser treated than in Er,Cr:YSGG-laser treated surfaces (p = 0.0001). However, bond strength means for both laser-irradiated groups were statistically lower than for the bur cut group (Er:YAG: p = 0.0281 and Er,Cr:YSGG: p < 0.0001). SEM observation of laser-irradiated surfaces revealed a roughened aspect and absence of smear layer. CONCLUSIONS: The self-etching system adhesion was influenced by the type of erbium laser used, and the bond strength was higher in the Er:YAG-laser irradiated than in the Er,Cr:YSGG-laser irradiated surfaces.

Cultured epithelial cells response to phototherapy with low intensity laser.

BACKGROUND AND OBJECTIVES: Little is known about the intracellular response of epithelial cells to phototherapy. The aim of this in vitro study was to analyze the effect of phototherapy with low-energy lasers with different wavelengths and powers on cultured epithelial cell growth under different nutritional conditions. STUDY DESIGN/MATERIALS AND METHODS: Epithelial cell cultures (Vero cell line) grown in nutritional deficit in culture medium supplemented with 2% fetal bovine serum (FBS) were irradiated with low-energy laser from one to three times with a GaAlAs laser (660 nm) and InGaAlP (780 nm), 40 and 70 mW, respectively, with 3 or 5 J/cm2. Cell growth was indirectly assessed by measuring the cell mitochondrial activity. RESULTS: Nonirradiated cell cultures grown in nutritional regular medium supplemented with 10% FBS produced higher cell growth than all cultures grown in nutritional deficit irradiated or not. The overall cell growth of cultures grown under nutritionally deficit conditions was significantly improved especially when irradiated with 780 nm for three times. CONCLUSIONS: Phototherapy with the laser parameters tested increases epithelial cell growth rate for cells stressed by growth under nutritionally deficient states. This cell growth improvement is directly proportional to the number of irradiations; however, was not enough to reach the full cell growth potential rate of Vero epithelial cell line observed when growing under nutritional regular condition.

Ultrastructural and autoradiographical analysis show a faster skin repair in He-Ne laser-treated wounds.

There are evidences that low-intensity red laser radiation is capable to accelerate wound healing. Nowadays, this therapy has been gradually introduced in clinical practice although mechanisms underlying laser effects are poorly understood. To better understand the photobiological effects of laser radiation, this study investigated by electron microscopy, immunohistochemistry and autoradiography the morphological and functional features of irradiated and none irradiated injured mice skin. Full-thickness skin lesions were created on the back of mice and irradiated on days 1, 5, 8, 12, and 15 post-wounding with a He-Ne laser (lambda=632.8nm), dose 1J/cm(2), exposure time 3min. Non-irradiated lesions were used as a control. The mice were inoculated with (3)H-proline and sacrificed one hour after on the 8th, 15th and 22nd days to histological and radioautographical analysis. The irradiated-lesions showed a faster reepithelization compared with control lesions. The irradiated dermis contained a higher number of activated fibroblasts compared to control group and, most of them showed several cytoplasmic collagen-containing phagosomes. In irradiated-lesions, smooth muscle alpha-actin positive cells predominated, which correspond to a higher number of myofibroblasts observed in the electron microscope. Moreover, laser radiation reduced the local inflammation and appears to influence the organization of collagen fibrils in the repairing areas. Quantitative autoradiography showed that the incorporation of (3)H-proline was significantly higher in irradiated-dermis on the 15th day post-wounding (p<0.05). These results suggest that laser radiation may accelerate cutaneous wound healing in a murine model.

Chemical, morphological and thermal effects of 10.6-microm CO2 laser on the inhibition of enamel demineralization.

Studies have shown that enamel can be modified by pulsed CO2 laser to form a more acid-resistant substrate. This study evaluated the effects of a 10.6-microm CO2 laser on enamel surface morphology and chemical composition as well as monitored intrapulpal temperature changes during irradiation. Human teeth were irradiated with fluences of 1.5-11.5 J/cm2, and pulpal thermal as well as chemical and morphological modifications on enamel were assessed. The teeth were submitted to a pH-cycling model, and the mineral loss was determined by means of cross-sectional microhardness. For all irradiated groups, intrapulpal temperature changes were below 3 degrees C. FT-Raman spectroscopy and scanning electron microscopy indicated that fluences as low as 6.0 J/cm2 were sufficient to induce chemical and morphological changes in enamel. Then, for fluences reaching or exceeding 10.0 J/cm2, laser-induced inhibitory effects on demineralization were observed. It was thus concluded that laser energy density in the range of 10.0 and 11.5 J/cm2 could be applied to dental enamel in order to produce chemical and morphological changes and reduce the acid reactivity of enamel without compromising the pulp vitality.

Effect of ER:YAG and diode laser irradiation on the root surface: morphological and thermal analysis.

BACKGROUND: The aim of the present study was to compare the effects of Er:YAG and diode laser treatments of the root surface on intrapulpal temperature after scaling and root planing with hand instruments. METHODS: Fifteen extracted single-rooted teeth were scaled and root planed with hand instruments. The teeth were divided into 3 groups of 5 each and irradiated on their buccal and lingual surfaces: group A: Er:YAG laser, 2.94 microm/100 mJ/10 Hz/30 seconds; group B: diode laser, 810 nm/1.0 W/0.05 ms/30 seconds; group C: diode laser, 810 nm/1.4 W/0.05 ms/30 seconds. The temperature was monitored by means of a type T thermocouple (copper-constantan) positioned in the pulp chamber to assess pulpal temperature during and before irradiation. Afterwards, the specimens were longitudinally sectioned, and the buccal and lingual surfaces of each root were analyzed by scanning electron microscopy. RESULTS: In the Er:YAG laser group, the thermal analysis revealed an average temperature of -2.2 +/- 1.5 degrees C, while in the diode laser groups, temperatures were 1.6 +/- 0.8 degrees C at 1.0 W and 3.3 +/- 1.0 degrees C at 1.4 W. Electronic micrographs revealed that there were no significant morphological changes, such as charring, melting, or fusion, in any group, although the specimens were found to be more irregular in the Er:YAG laser group. CONCLUSIONS: The application of Er:YAG and diode lasers at the utilized parameters did not induce high pulpal temperatures. Root surface irregularities were more pronounced after irradiation with an Er:YAG laser than with a diode laser.