Defocused high-power diode laser accelerates skin repair in a murine model through REDOX state modulation and reepithelization and collagen deposition stimulation.

Skin wounds represent a burden in healthcare. Our aim was to investigate for the first time the effects of defocused high-power diode laser (DHPL) on skin healing in an animal experimental model and compare it with gold standard low-level laser therapy. Male Wistar rats were divided into 5 groups: Negative control; Sham; 0.1 W laser (L0.1 W); DHPL Dual 1 W (DHPLD1 W); and DHPL Dual 2 W (DHPLD2 W). Rats were euthanized on days 3, 5, 10, 14 and 21. Clinical, morphological, PicroSirus, oxidative stress (MDA, SOD and GSH) and cytokines (IL-1ß, IL-10 and TNF-α) analyses were performed. A faster clinical repair was observed in all laser groups at D10 and D14. DHPLD1 W exhibited lower inflammation and better reepithelization compared to other groups at D10. DHPL protocols modulated oxidative stress by decreasing MDA and increasing SOD and GSH. Collagen maturation was triggered by all protocols tested and L0.1 W modulated cytokines release (IL-1ß and TNF-α) at D3. In conclusion, DHPL, especially DHPL1 W protocol, accelerated skin healing by triggering reepithelization and collagen maturation and modulating inflammation and oxidative stress.

Lycium barbarum polysaccharide fraction associated with photobiomodulation protects from epithelium thickness and collagen fragmentation in a model of cutaneous photodamage.

Ultraviolet radiation (UVR) is the major etiologic agent of cutaneous photoaging, and different strategies are used to prevent and treat this condition. The polysaccharide fraction (LBPF) isolated from Lycium Barbarum fruits (goji berry) contains several active ingredients with antioxidant, immune system modulation, and antitumor effects. In addition, the photobiomodulation (PBM) is widely applied in photoaging treatment. This study investigated the effects of LBPF and PBM against the UVR-induced photodamage in the skin of hairless mice. The mice were photoaged for 6 weeks in a chronic and cumulative exposure regimen using a 300-W incandescent lamp that simulates the UVR effects. From the third to the sixth week of photoaging induction, the animals received topical applications of LBPF and PBM, singly or combined, in different orders (first LBPF and then PBM and inversely), three times per week after each session of photoaging. After completion of experiments, the dorsal region skin was collected for the analysis of thickness, collagen content, and metalloproteinases (MMP) levels. A photoprotective potential against the increase of the epithelium thickness and the fragmentation of the collagen fibers was achieved in the skin of mice treated with LBPF or PBM singly, as well as their combination. All treatments maintained the skin collagen composition, except when PBM was applied after the LBPF. However, no treatment protected against the UVR-induced MMP increase. Taken together, we have shown that the LBPF and PBM promote a photoprotective effect in hairless mice skin against epidermal thickening and low collagen density. Both strategies, singly and combined, can be used to reduce the UVR-induced cutaneous photoaging.

Effect of topical 5-ALA mediated photodynamic therapy on proliferation index of keratinocytes in 4-NQO-induced potentially malignant oral lesions.

Fractionation can improve photodynamic therapy (PDT) efficacy for potentially malignant oral lesion treatment. The aim of this study was to demonstrate the apoptosis/proliferation index of oral keratinocytes after two sessions of topical 5-ALA-mediated PDT in 4-Nitroquinoline-1-oxide-induced potentially malignant oral lesion, and to suggest the ideal interval between PDT sessions. Immuno-histochemical tests for proliferating cell nuclear antigen and caspase-3, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay were performed at 6h, 24h, 48h, and 72h time intervals after PDT. The number of positive cells showing caspase-3 expression was significantly higher, mainly at 6h after PDT. In the first cycle of PDT, the highest frequency of positive cells for TUNEL was found at 24h. At 72h after PDT, proliferating cell nuclear antigen positive cells increased significantly, indicating that there was an epithelial response in direction towards DNA repair and cell proliferation at this time. Because cell proliferation increases and cell death index decreases at 72h after PDT, it is recommended that the interval between the PDT sessions must not be longer than 2days up to total lesion remission.

Histomorphological and angiogenic analyzes of skin epithelium after low laser irradiation in hairless mice.

It is not well-understood how low-laser therapy affects the skin of the applied area. This study analyzes skin of the masseteric region of mice from the HRS/J strain after three different application regimens (three, six or ten applications per regimen) of low intensity laser at 20 J/cm(2) and 40 mW for 20 sec on alternate days. Three experimental groups according to the number of laser applications (three, six or ten) and three control groups (N = 5 animals for each group) were used. On the third day after the last irradiation, all animals were sacrificed and the skin was removed and processed to analyze the relative occupation of the test area by each epithelial layer and the aspects of neovascularization. Data were submitted to statistical analyzes. The irradiated groups compared to their respective controls at each period of time, showed no significant difference in relative occupation of the test area by the layers and epithelium areas for three and six applications, but for ten applications, a significant decrease (P < 0.05) in the basal and granulosum layers, and epithelium areas were found. From the comparisons of the three irradiated groups together, the group with six laser applications showed statistical difference (P < 0.05) in total epithelium and on the layers. Vascular endothelial growth factor (VEGF) and VEGFR-2 immunoreactivities were similar for the control and irradiated groups. Results suggested a biostimulatory effect with low risks associated with superficial tissues, when the treatment aims the deeper layers after six applications.

Histological analysis of healthy epithelium of Wistar rats in vivo irradiated with different intensities of therapeutic ultrasound.

BACKGROUND: Ultrasound (US) is a widely used and studied resource for physical therapy treatments. Given the scarcity of studies on the effects of US on healthy tissue, many physical therapy professionals make unfounded decisions regarding its methods and parameters of application. OBJECTIVES: The possible histological and morphometric changes in the healthy tissue of Wistar rats in vivo irradiated with different intensities of US were evaluated. METHODS: Thirty Wistar rats, randomly distributed among five groups of six animals each, were treated on the right side of the dorsal region, over an area of 4 cm(2). The left side served as a control. The treatment was applied over a four-day period, with two mins. of daily irradiation. The output intensity was checked using a precision dosimeter before the applications. Histological and morphometric analyses were performed using the Image Tool software. RESULTS: There were slight inflammatory infiltration and thinning of the dermis fibers, particularly in the groups irradiated with 1.5 and 2 W/cm(2). There was also thickening of the epidermis in the samples from the irradiated animals. To evaluate the quantitative results, the statistical analyses consisted of one-way ANOVAs with the post-hoc Tukey tests. There were significant differences in epidermis thicknesses between the control group and the groups irradiated with 1.0, 1.5 and 2.0 W/cm(2). CONCLUSIONS: Higher doses of US produced changes in the epidermis and dermis, i.e. increased thickness and collagen fiber thinning and proliferation, respectively. These results serve as a warning of the possible implications of therapeutic ultrasound use in esthetics.

Análise histológica em tecido epitelial sadio de ratos Wistar (in vivo) irradiados com diferentes intensidades do ultrassom / Histological analysis of healthy epithelium of Wistar rats in vivo irradiated with different intensities of therapeutic ultrasound

CONTEXTUALIZAÇÃO: O Ultrassom (US) é um dos recursos físicos amplamente utilizado e pesquisado nos tratamentos de fisioterapia. Sabe-se que diante de uma escassa literatura sobre efeitos do US em tecidos sadios, muitos profissionais fisioterapeutas acabam realizando aplicações infundadas de métodos e parâmetros. OBJETIVO: Avaliar possíveis alterações histológicas e morfométricas do tecido sadio in vivo de ratos Wistar irradiados com diferentes intensidades de US. MÉTODOS: Trinta ratos da linhagem Wistar, distribuídos aleatoriamente em cinco grupos de seis animais cada foram tratados na região dorsal do lado direito numa área de 4cm². O lado esquerdo serviu como controle. O tratamento foi feito durante quatro dias com 2 minutos de irradiação. Verificou-se a intensidade de saída com dosímetro de precisão antes das aplicações. Analisou-se a histologia e a morfometria por meio do software Image Tool. RESULTADOS: Observou-se um discreto infiltrado inflamatório e adelgaçamento das fibras da derme, principalmente dos grupos irradiados com 1.5 e 2W/cm². Notou-se também um aumento na espessura da epiderme nas amostras dos animais irradiados. Para avaliar os resultados quantitativos, utilizou-se como análise estatística ANOVA one way e o teste post hoc de Tukey. Na espessura da epiderme, obtiveram-se diferenças significativas entre grupo controle e os grupos irradiados com 1.0, 1.5 e 2.0W/cm². CONCLUSÃO: Sob ação do US nas doses maiores houve alterações na epiderme e derme, respectivamente, o aumento da espessura e proliferação com adelgaçamento das fibras colágenas, o que alerta para possíveis implicações do uso do US em estética.

BACKGROUND: Ultrasound (US) is a widely used and studied resource for physical therapy treatments. Given the scarcity of studies on the effects of US on healthy tissue, many physical therapy professionals make unfounded decisions regarding its methods and parameters of application. OBJECTIVES: The possible histological and morphometric changes in the healthy tissue of Wistar rats in vivo irradiated with different intensities of US were evaluated. METHODS: Thirty Wistar rats, randomly distributed among five groups of six animals each, were treated on the right side of the dorsal region, over an area of 4 cm². The left side served as a control. The treatment was applied over a four-day period, with two mins. of daily irradiation. The output intensity was checked using a precision dosimeter before the applications. Histological and morphometric analyses were performed using the Image Tool software. RESULTS: There were slight inflammatory infiltration and thinning of the dermis fibers, particularly in the groups irradiated with 1.5 and 2 W/cm². There was also thickening of the epidermis in the samples from the irradiated animals. To evaluate the quantitative results, the statistical analyses consisted of one-way ANOVAs with the post-hoc Tukey tests. There were significant differences in epidermis thicknesses between the control group and the groups irradiated with 1.0, 1.5 and 2.0 W/cm². CONCLUSIONS: Higher doses of US produced changes in the epidermis and dermis, i.e. increased thickness and collagen fiber thinning and proliferation, respectively. These results serve as a warning of the possible implications of therapeutic ultrasound use in esthetics.

The severity of epithelial dysplasia is associated with loss of maspin expression in actinic cheilitis.

BACKGROUND: Prolonged exposure of the lip to sunlight may cause actinic cheilitis (AC) and squamous cell carcinoma (SCC). Maspin is a serpin with tumor suppressor functions. This work analyzed the presence and distribution of maspin in AC and lip SCC. METHODS: Sections from 36 cases diagnosed as AC (18 cases with mild epithelial dysplasia, 11 with moderate and 7 with severe), 18 cases diagnosed as lip SCC and 7 specimens containing normal lip vermillion epithelium were submitted for immunohistochemical analysis to detect maspin. RESULTS: All AC cases with mild and two cases with moderate dysplasia were scored 3. The remaining nine cases with moderate dysplasia were identified as score 2, whereas all cases with severe dysplasia were scored 1. Positive staining for maspin decreased from the basal layer to the surface. Among the 18 lip SCCs studied, 15 cases showed abundant staining for maspin. Epithelium adjacent to the SCCs also showed intense positive staining in all cells. CONCLUSIONS: Our results suggest that the loss of maspin expression occurs from the basal layer to the surface. Lip SCCs related to solar radiation show an intense presence of maspin protein in almost all tumor cells as well as the neighboring epithelium.

A study of actinic cheilitis treatment by two low-morbidity CO2 laser vaporization one-pass protocols.

Therapeutic approaches to chronic actinic cheilitis focus on the removal or destruction of diseased epithelium. The CO(2) laser has become an important therapeutic alternative, achieving clinical resolution in around 90% of patients. Although many laser physical parameters have been reported, some are known for their low potential for scar induction without compromising the success of the results. The aim of this clinicohistological study was to compare the therapeutic responses to two low-morbidity protocols involving a single laser pass. A total of 40 patients with chronic multicentric and microscopically proven disease were randomly submitted to two conservative CO(2) laser protocols using a bilateral comparative model. The degree of histological atypia of the epithelium was determined in 26 patients both pre- and postoperatively for both protocols. Other histological phenomena were assessed in addition to this central analysis parameter. Clinical recurrence occurred in 12.5% of patients for each protocol, together with a significant reduction in the degree of epithelial atypia (p < 0.001), which was occasionally complete. However, no difference was found between the protocols (p > 0.05). Using these morphological parameters it was not possible to determine whether postoperative epithelial atypias in part of the sample were reactive or residual in nature. A few patients may show minor postoperative lesions. Due to their potential to achieve clinical and importantly microscopic resolution, the studied protocols may be used for mild through moderate dysplastic epithelium and clinically diffuse disease.

Histomorphometry and immunohistochemical features of grade I (WHO) oral radiomucositis.

AIMS: The aims of this study were to describe the immunohistopathological and morphometric features of oral mucositis grade I (WHO). MATERIAL AND METHODS: Ten samples of oral mucositis were biopsied and submitted to histopathological, morphometric and immunohistochemical analyses (CD68, Ki-67 and p53). The samples were compared with the buccal mucosa of head and neck cancer patients before radiotherapy (NMCP), normal buccal mucosa (NM) and oral dysplasia (OD). RESULTS: Epithelial thickness, area and perimeter were decreased in oral mucositis and inflammatory components, increased when compared with NMCP. CD68 immunoreactivity, near to the epithelium, was more evident in oral mucositis than in NMCP (P = 0.01). The Ki-67 counts were higher in oral mucositis than in NM and NMCP (P = 0.001 and P = 0.043, respectively), but without any difference with OD (P = 0.284). The p53 staining was present in all cases of mucositis and oral dysplasia, but negative in NMCP and NM. CONCLUSIONS: Oral mucositis grade I (WHO) presented epithelial atypia and atrophy, increased inflammatory response, with relevant Ki-67 count and positiveness for p53.

Morphology and morphometry of mice's terminal ileum wall exposed to x-ray

Foram estudados os aspcectos morfológicos da parede do íleo de camundongos da linhagem C58BL expostos aos raios X, corpo inteiro, na dose única de 154,8 mC/Kg (600 R) e também a morfometria das figuras de mitose e das células caliciformes à microscopia de luz comum. Observamos alteraçöes morfológicas intensas no epitélio da mucosa ileal no período de 24 horas após irradiaçäo, tendência a recuperaçäo às 72 horas e normalizaçäo às 144 horas. A análise morfométrica demonstrou o índice mitótico significantemente baixo (quase nulo) no período significantemente baixo (quase nulo) no período de 24 hotas depois da irradiaçäo, rebote às 72 horas e normalizaçäo do mesmo às 144 horas. O índice de células caliciformes evidenciou diminuiçäo significante no período de 72 horas após irradiaçäo e recuperaçäo às 144 horas

Alteraçöes morfológicas e morfométricas do epitélio da mucosa retal de camundongos expostos a dose única de raios x / Morphological and morphometric epithelium alterations of rectal mucosa in mice exposed to a single dose of x-rays

O objetivo deste trabalho, acompanhando a linha de pesquisa da Disciplina de Gastroenterologia Círurgica da Escola Paulista de Medicina, foi o de estudar as alteraçöes agudas morfológicas e morfométricas do epitélio da mucosa retal de 60 camundongos Mus musculus, da linhagem C57BL, dos quais 45 foram expostos aos raios X, dose única de 154,8mC/Kg (600R), corpo inteiro, sendo que os 15 restantes utilizados como controle. Os camundongos foram sacrificados após 24h, 72h e 144h, sendo o reto ressecado e submetido a estudo histológico morfológico e morfométrico de seu epitélio. A análise dos resultados permitiu as seguintes conclusöes: a) as alteraçöes morfológicas das células do epitélio das criptas foram hipertrofia nuclear, picnose, criólise, cariólise, cariorréxis e hipertrofia das células califormes; b) o número das criptas diminuiu às 24h, diminuiçäo que se acentuou às 72h, havendo, no entanto, um aumento significante às 144h; c) tanto o número de mitoses quanto o índice mitótico sofreram acentuada diminuiçäo às 24h, recuperando-se a partir das 72h, e mantendo-se normal às 144h

Nucleolar organizer regions as markers of incipient cellular alterations in squamous epithelium.

Transcriptionally active nucleolar organizer regions identified by silver staining (AgNOR) vary in number with cellular activity and/or malignant transformation and have been used as a diagnostic tool. A morphometric study of AgNORs was performed in an experimental model of irradiated squamous epithelium (Wistar rat sole skin) 4, 8, and 14 hours and 1, 2, 5, and 7 days post-irradiation with 50 Gy of x-rays. A statistically significant and progressive rise in AgNOR average volume of up to 238% and reduction in AgNOR number/nucleus of up to 40% were detected as a function of post-irradiation time. A statistically significant 46% increase in AgNOR volume was detected as early as 8 h post-irradiation, when no histological changes were observable in routine preparations. These results suggest that AgNORs may be useful as a quantitative marker of incipient changes in cellular activity and caution against the indiscriminate use of AgNORs in the follow-up of lesions which may have been exposed to radiotherapy. Furthermore, this study suggests the possibility of using AgNORs as a sensitive biological dosimeter in cases of uncontrolled exposure to radiation.

Herpes simplex virus in postradiation cervical smears. A morphologic and immunocytochemical study.

From January 1987 to August 1988, cytomorphologic criteria of both herpes simplex virus (HSV) and radiation effects were observed in Papanicolaou smears from 3 of 1,340 patients who had received radiotherapy for squamous cell carcinoma of the cervix. Avidin-biotin immunoperoxidase staining, using a rabbit IgG polyclonal HSV antibody, confirmed the presence of HSV antigen in those three postradiation smears. Both multinucleated molded cells and epithelial cells that lacked cytopathic effects were positive for HSV. Three other postradiation smears from these cases were similarly positive for HSV antigen; the one preradiation smear was negative. In situ hybridization and immunoperoxidase studies on sections from the preradiation biopsies were negative: severely altered neoplastic cells showed no reactivity. The absence of HSV markers in the preradiation specimens suggests that the HSV infections were secondary to the radiotherapy; further studies are needed to prove this association and to assess the possible mechanisms. These cases clearly indicate that the overlapping features of radiation and viral effects (such as multinucleation) may be present simultaneously.