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.

Latin America Optics and Photonics 2022: introduction to the feature issue.

The 2022 Latin America Optics and Photonics Conference (LAOP 2022), the major international conference sponsored by Optica in Latin America, returned to Recife, Pernambuco, Brazil, after its first edition in 2010. Held every two years since (except for 2020), LAOP has the explicit objective to promote Latin American excellence in optics and photonics research and support the regional community. In the 6th edition in 2022, it featured a comprehensive technical program with recognized experts in fields critical to Latin America, highly multidisciplinary, with themes from biophotonics to 2D materials. The 191 attendees of LAOP 2022 listened to five plenary speakers, 28 keynotes, 24 invited talks, and 128 presentations, including oral and posters.

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.

Morphological, optical, and elemental analysis of dental enamel after debonding laminate veneer with Er,Cr:YSGG laser: A pilot study.

Laminate veneer removal is becoming a routine procedure at the dental clinic and the use of laser can facilitate its removal. This work aimed to evaluate the morphological, elemental, and optical changes in the remaining enamel after veneer removal using Er,Cr:YSGG laser. Forty-four enamel slabs were prepared and randomly distributed into nine experimental groups, for bonding using lithium disilicate laminates with three different luting agents (Variolink Veneer, RelyX U200, and RelyX Veneer). Then each agent was debonded using Er,Cr:YSGG laser (2.78 µm) using two different protocols:3.5 W, 48.14 J/cm2 , 20 Hz non-contact and 3.0 W, 48.14 J/cm2 , 20 Hz non-contact. The morphological, optical, and elemental analysis of enamel was performed before cementation and after laser debonding, using scanning electron microscopy (SEM), optical coherence tomography (OCT), and energy-dispersive X-ray spectroscopy (EDS). The level of statistical significance adopted was 5%. The EDS analysis of enamel after debonding revealed a significant increase in silane and carbon, as well as a decrease in calcium and phosphate contents. Analysis showed the presence of residual cement in most experimental groups but the morphological analysis showed alteration of the enamel's prisms only in the groups that used RelyX Veneer and Variolink Veneer cements. There was no evidence of deleterious morphological changes resulting from irradiation. However, an increase in the optical attenuation coefficient by the OCT was observed due to the presence of the remaining cement. It can be concluded that the Er,Cr:YSGG laser, in the mean powers used, is efficient for veneer removal without causing deleterious effects for the enamel.

Monitoring the Progress and Healing Status of Burn Wounds Using Infrared Spectroscopy.

Burns are one of the leading causes of morbidity worldwide and the most costly traumatic injuries. A better understanding of the molecular mechanisms in wound healing is required to accelerate tissue recovery and reduce the health economic impact. However, the standard techniques used to evaluate the biological events associated to wound repair are laborious, time-consuming, and/or require multiple assays/staining. Therefore, this study aims to evaluate the feasibility of Fourier transform infrared (FT-IR) spectroscopy to monitor the progress and healing status of burn wounds. Burn injuries were induced on Wistar rats by water vapor exposure and biopsied for further histopathological and spectroscopic evaluation at four time-points (3, 7, 14, and 21 days). Spectral data were preprocessed and compared by principal component analysis. Pairwise comparison of post-burn groups to each other revealed that metabolic activity induced by thermal injury decreases as the healing progresses. Higher amounts of carbohydrates, proteins, lipids, and nucleic acids were evidenced on days 3 and 7 compared to healthy skin and reduced amounts of these molecular structural units on days 14 and 21 post-burn. FT-IR spectroscopy was used to determine the healing status of a wound based on the biochemical information retained by spectral signatures in each phase of healing. Our findings demonstrate that FT-IR spectroscopy can monitor the biological events triggered by burn trauma as well as to detect the wound status including full recovery based on the spectral changes associated to the biochemical events in each phase.

Surface Evaluation of Enamel Etched by Er,Cr:YSGG Laser for Orthodontic Purpose.

AIM: To compare the effect of erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser at different irradiation parameters and acid etching on the shear bond strength (SBS) of orthodontic brackets to enamel. MATERIALS AND METHODS: Forty bovine incisors were randomly distributed into groups (n = 10): GI: 37% phosphoric acid etching; GII: Er,Cr:YSGG laser etching 19.1 J/cm2; GIII: Er,Cr:YSGG, 29.3 J/cm2; and GIV: Er,Cr:YSGG, 42.4 J/cm2. After treatments, metallic brackets were bonded using Transbond XT adhesive system. After light curing, the samples were subjected to 500 thermal cycles, debonded with a universal testing machine, and the SBS values were recorded. After debonding, surface morphology was evaluated using scanning electron microscopy (SEM) and optical coherence tomography (OCT). The values of SBS testing were analyzed by one-way analysis of variance (ANOVA) and Tukey post hoc test, at 5% significance level. RESULTS: The mean SBS values of GI, GII, GIII, and GIV groups were 6.2 ± 1.7 MPa, 4.6 ± 2.5 MPa, 7.0 ± 2.2 MPa, and 8.0 ± 3.6 MPa, respectively. Laser irradiation promoted rough surfaces in all parameters used, and the OCT analysis revealed higher optical changes on lased groups when compared with phosphoric acid. CONCLUSION: Er,Cr:YSGG laser irradiation operated at 42.4 J/cm2 and 29.3 J/cm2 is a better alternative for etching enamel prior to the orthodontic treatment than the phosphoric acid. CLINICAL SIGNIFICANCE: Er,Cr:YSGG laser irradiation is better than the phosphoric acid for etching enamel prior to the orthodontic treatment because laser irradiation promotes similar SBSs and preventing demineralization around orthodontic brackets.

General model for depth-resolved estimation of the optical attenuation coefficients in optical coherence tomography.

We present the proof of concept of a general model that uses the tissue sample transmittance as input to estimate the depth-resolved attenuation coefficient of tissue samples using optical coherence tomography (OCT). This method allows us to obtain an image of tissue optical properties instead of intensity contrast, guiding diagnosis and tissues differentiation, extending its application from thick to thin samples. The performance of our method was simulated and tested with the assistance of a home built single-layered and multilayered phantoms (~100 µm each layer) with known attenuation coefficient on the range of 0.9 to 2.32 mm-1 . It is shown that the estimated depth-resolved attenuation coefficient recovers the reference values, measured by using an integrating sphere followed by the inverse adding doubling processing technique. That was corroborated for all situations when the correct transmittance value is used with an average difference of 7%. Finally, we applied the proposed method to estimate the depth-resolved attenuation coefficient for a thin biological sample, demonstrating the ability of our method on real OCT images.

Variation on Molecular Structure, Crystallinity, and Optical Properties of Dentin Due to Nd:YAG Laser and Fluoride Aimed at Tooth Erosion Prevention.

This in vitro study evaluated the compositional, crystalline, and morphological effects promoted by Nd:YAG laser on root dentin, and verified the effects of laser and topical acidulated phosphate fluoride application (APF-gel) on dentin erosion. 180 bovine dentin slabs were randomized into 4 groups (n = 45): G1-untreated, G2-APF-gel (1.23% F-, 4 min), G3-Nd:YAG (1064 nm, 84.9 J/cm², 10 Hz), and G4-APF-gel application followed by Nd:YAG laser irradiation. The compositional, crystalline, and morphological effects promoted by treatments were investigated on five samples of each experimental group. The other samples were submitted to a 5-day, 10-day, or 15-day erosive and abrasive demineralization and remineralization cycling in order to create erosion lesions. The area and depth of lesions, as well as the optical attenuation coefficient, were assessed, and all data were statistically analysed (p < 0.05). Nd:YAG laser promoted the reduction of carbonate, the formation of tetracalcium phosphate, as well as the melting and recrystallization of the dentin surface. Laser significantly decreased the area and depth of erosion lesions and altered the optical attenuation coefficient when compared to untreated and APF-gel groups, but the association of APF-gel and laser did not promote an additional effect. Nd:YAG laser irradiation can be a promissory treatment to prevent dentin erosion and the abrasion process.

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.

Effects of a Low-Intensity Laser on Dental Implant Osseointegration: Removal Torque and Resonance Frequency Analysis in Rabbits.

The objective of this study was to investigate how a low-intensity laser affects the stability and reverse torque resistance of dental implants installed in the tibia of rabbits. Thirty rabbits received 60 dental implants with the same design and surface treatment, one in each proximal metaphysis of the tibia. Three groups were prepared (n = 10 animals each): conventional osseointegration without treatment (control group), surgical sites irradiated with a laser beam emitted in the visible range of 680 nm (Lg1 group), surgical sites irradiated with a laser beam with a wavelength in the infrared range of 830 nm (Lg2 group). Ten irradiation sessions were performed 48 hours apart; the first session was during the immediate postoperative period. Irradiation energy density was 4 J/cm(2) per point in 2 points on each side of the tibias. The resonance frequency and removal torque values were measured at 2 time points after the implantations (3 and 6 weeks). Both laser groups (Lg1 and Lg2) presented a significant difference between resonance frequency analysis values at the baseline and the values obtained after 3 and 6 weeks (P > .05). Although the removal torque values of all groups increased after 6 weeks (P < .05), both laser groups presented greater mean values than those of the control group (P < .01). Photobiomodulation using laser irradiation with wavelengths of 680 and 830 nm had a better degree of bone integration than the control group after 6 weeks of observation time.

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.

The characterization of normal thyroid tissue by micro-FTIR spectroscopy.

In this paper, we report the spectral patterns of normal human thyroid tissue and methodology to interpret hyperspectral imaging data and protein conformational changes observed therein. Raw image datasets were imported into software written in-house in the MATLAB environment and processed to yield pseudo-color images of the tissue sections. All spectra were vector normalized, noise-filtered, and corrected for water-vapour contributions and scattering effects before being subjected to Hierarchical Cluster Analysis (HCA) and correlated with histological structures obtained from images of H&E-stained parallel tissue sections. We successfully identified a protein structural heterogeneity that can be correlated with the spatially resolved amount of iodine in the thyroglobulin structure of colloids and follicular cells.

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.

Exploring bacterial diversity of endodontic microbiota by cloning and sequencing 16S rRNA.

INTRODUCTION: The characterization of microbial communities infecting the endodontic system in each clinical condition may help on the establishment of a correct prognosis and distinct strategies of treatment. The purpose of this study was to determine the bacterial diversity in primary endodontic infections by 16S ribosomal-RNA (rRNA) sequence analysis. METHODS: Samples from root canals of untreated asymptomatic teeth (n = 12) exhibiting periapical lesions were obtained, 16S rRNA bacterial genomic libraries were constructed and sequenced, and bacterial diversity was estimated. RESULTS: A total of 489 clones were analyzed (mean, 40.7 ± 8.0 clones per sample). Seventy phylotypes were identified of which six were novel phylotypes belonging to the family Ruminococcaceae. The mean number of taxa per canal was 10.0, ranging from 3 to 21 per sample; 65.7% of the cloned sequences represented phylotypes for which no cultivated isolates have been reported. The most prevalent taxa were Atopobium rimae (50.0%), Dialister invisus, Prevotella oris, Pseudoramibacter alactolyticus, and Tannerella forsythia (33.3%). CONCLUSIONS: Although several key species predominate in endodontic samples of asymptomatic cases with periapical lesions, the primary endodontic infection is characterized by a wide bacterial diversity, which is mostly represented by members of the phylum Firmicutes belonging to the class Clostridia followed by the phylum Bacteroidetes.

Influence of the additional Er:YAG laser conditioning step on the microleakage of class V restorations.

The purpose of this study was to evaluate the influence of an additional Er:YAG laser conditioning step after laser cavity preparations, on the microleakage of class V composite restorations. Forty-eight bovine incisors were randomly divided into four groups: G1(control) cavities prepared with bur, G2- cavities prepared with laser (400 mJ/2 Hz), G3- cavities prepared and subsequently conditioned with Er:YAG laser (60 mJ/2 Hz); G4-idem for G3, but the laser conditioning was carried out without water-spray. All the cavities were restored using Clearfill SE Bond and Z-250 composite resin. The samples were thermal cycled for 700 cycles and then immersed in 50% silver nitrate solution. The sectioned restorations were exposed to a photoflood lamp to reveal silver nitrate penetration. The Kruskal-Walis one-way analyses of variance test and post hoc Wilcoxon pair-wise comparison were used to compare microleakage degrees. At the gingival margin G2 showed a lower microleakage mean than the control bur-prepared cavities (p = 0.0003). At occlusal margins there were no statistically significant differences between the groups (p = 0.28). It may be concluded that Er:YAG laser class V cavity preparations do not need to be followed by an additional laser conditioning step to result in levels of microleakage similar to or lower than those obtained after bur preparations.

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.