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The objective of this study was to investigate the therapeutic wound healing potential and molecular mechanisms of shikonin as small molecules in vitro. A mouse burn model was used to explore the potential therapeutic effect of shikonin; we traced proliferating cells in vivo to locate the active area of skin cell proliferation. Through the results of conventional pathological staining, we found that shikonin has a good effect on the treatment of burned skin and promoted the normal distribution of skin keratin at the damaged site. At the same time, shikonin also promoted the proliferation of skin cells at the damaged site; importantly, we found a significant increase in the number of fibroblasts at the damaged site treated with shikonin. Most importantly, shikonin promotes fibroblasts to repair skin wounds by regulating the PI3K/AKT signaling pathway. This study shows that shikonin can effectively promote the proliferation of skin cell, and local injection of fibroblasts in burned skin can play a certain therapeutic role. [ABSTRACT FROM AUTHOR]

Introducción: la enfermedad de Pompe (EP) o glucogenosis tipo II es una enfermedad autosómica recesiva causada por mutaciones en el gen GAA que codifica para la proteína alfa-1,4-glucosidasa. Su deficiencia lleva a un almacenamiento anormal de glucógeno en los lisosomas de varias células, a través de los diferentes tejidos, lo que causa un compromiso musculoesquelético predominante. Contenidos: los fenotipos de la enfermedad dependen de las variantes genéticas y de los niveles de la actividad enzimática residual. La enfermedad se presenta como EP de inicio infantil, EP de inicio tardío y EP intermedio, por lo que es de suma importancia su diagnóstico temprano, por medio de estudios moleculares como la secuenciación de Sanger y la secuenciación de nueva generación. Conclusiones: se ha demostrado, mediante diferentes estudios, que las variaciones genéticas pueden diferir entre etnias, y es importante su caracterización molecular para determinar el tratamiento más adecuado, de acuerdo con el estado del material inmunológico de reacción cruzada (CRIM).

Introducción. El tumor miofibroblástico inflamatorio es una enfermedad proliferativa rara, de etiología incierta, caracterizada por la proliferación de miofibroblastos epitelioides o fusionados mezclados con células inflamatorias, predominantemente mononucleares. En general se considera una lesión benigna, aunque en algunos casos esta neoplasia ha mostrado un comportamiento agresivo en cuanto a recidiva local y metástasis. El tratamiento definitivo es la resección quirúrgica completa. Caso clínico. Paciente de 67 años con dos meses de evolución de fiebre y masa abdominal, en quien se realizó una tomografía computarizada de abdomen que identificó una lesión de aspecto infiltrativo tumoral, comprometiendo la grasa retroperitoneal en la transcavidad de los epiplones. Por vía percutánea se tomó una biopsia que informó un pseudotumor inflamatorio retroperitoneal. Fue llevado a cirugía radical abdominal, con patología quirúrgica final que describió un tumor miofibroblástico inflamatorio de compromiso multifocal, adherido a la serosa del estómago e intestino delgado, sin compromiso muscular. Discusión. El tumor inflamatorio miofibroblástico es una entidad rara, de etiología por esclarecer y difícil diagnóstico. Presentamos el caso clínico de un paciente con tumor inflamatorio miofibroblástico gastrointestinal. Conclusión. Se describe el caso clínico de un paciente con un tumor inflamatorio miofibroblástico gastrointestinal, de presentación rara en nuestro medio. Es importante la comparación con casos similares para poder hacer conclusiones útiles en la práctica clínica.

Background and objective. Advances in Tissue Engineering promoted the development of skin substitutes. One of the main drawbacks of skin bioengineering is the requirement of a considerable quantity of viable cells in short periods of time, which leads to a challenging biofabrication process. This article aims to analyse the effects of the application of three different types of human mesenchymal stem cell-derived secretome in fibroblast cultures. The final goal is to achieve new strategies to promote the scalability of artificial skin manufacturing processes. Methods. The effects of the three different types of human mesenchymal stem cell-derived secretome were analyzed at 24, 48 and 72 hours. To study cell viability, cell proliferation and cell migration we exposed human fibroblasts to different secretome concentrations (C1 and C2). An in vivo study was proposed to corroborate in vitro results. Results. The use of mesenchymal stem cell-derived secretome improved the currently available fibroblast culture protocols. The use of secretome was associated with increased cell proliferation and cell migration while maintaining high viability values. Data were especially positive when secretome from dental pulp mesenchymal stem cells and adipose tissue mesenchymal stem cells were applied. Conclusions. The application of mesenchymal stem cell-derived secretome maintained high cell viability data and increased fibroblast proliferation. In vivo and in vitro studies showed no short-term adverse effects. [ABSTRACT FROM AUTHOR]

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Carnosine is known as a natural dipeptide, which inhibits the proliferation of tumor cells throughout its action on mitochondrial respiration and cell glycolysis. However, not much is known about its effects on the metabolism of healthy cells. We explored the effects of Karnozin EXTRA® capsule with different concentrations of L-carnosine, on the cell viability and the expressions of intermediate filament vimentin (VIM) and superoxide dismutase (SOD2) in normal fibroblasts BHK-21/C13. Furthermore, we investigated its action on the energy production of these cells. Cell viability was quantified by the MTT assay. The Clark oxygen electrode (Oxygraph, Hansatech Instruments, England) was used to measure the "intact cell respiration rate", state 3 of ADP-stimulated oxidation, maximum oxidation capacity and the activities of complexes I, II and IV. Results showed that Karnozin EXTRA® capsule in concentrations of 2 and 5 mM of L-carnosine did not induce toxic effects and morphological changes in treated cells. Our data revealed a dose-dependent immunofluorescent signal amplification of VIM and SOD2 in the BHK-21/C13 cell line. This supplement substantially increased the recorded mitochondrial respiration rates in the examined cell line. Due to the stimulation of mitochondrial energy production in normal fibroblasts, our results suggested that Karnozin EXTRA® is a potentially protective dietary supplement in the prevention of diseases with altered mitochondrial function. [ABSTRACT FROM AUTHOR]

Background: Cutaneous fibroma is a benign neoplasm affecting the fibroblasts and collagen matrix that develops in the dermis or subcutaneous tissue. This neoplasm is uncommon in cattle, and few reports have described the treatment and resolution of this neoplasm. Despite its benign character, a veterinarian should consider cutaneous fibroma in the differential diagnosis of skin tumors. This report aims to describe a rare case of large fibroma in the scapular region in a cow, with emphasis on the clinical-surgical and anatomopathological aspects of the condition. Case: A 3-year-old Girolando 3/4 cow was attended to at a rural property in Lagamar-MG, Brazil. According to the owner, the animal presented with a small mass in the right scapular region that grew progressively over 1 year and 6 months. Clinical examination revealed an exuberant and painless increase in volume on palpation in the proximal region of the right thoracic limb, which, in its vertical axis, extended from the proximal end of the scapula to near the olecranon tuberosity, and, in its horizontal axis, extended from the 6th intercostal space to the scapulohumeral joint, reaching the dimensions 66 cm and 62 cm, respectively. It presented with multiple nodules that were firm in consistency with extensive areas of ulceration. Neoplasia was suspected, and surgical excision was decided upon. The cow was sedated and restrained in the left lateral decubitus position. Trichotomy and antisepsis of the operative field were performed followed by an infiltrative anesthetic block around the tumor. The tumor was excised maintaining a safety margin of 1 cm. Dermorrhaphy was not possible, and healing by secondary intention was awaited. In the postoperative period, antibiotic therapy with benzathine penicillin, analgesia with meloxicamand dipyrone and daily dressing of the wound were performed. There were no postoperative complications and complete healing occurred approximately 100 days after surgery. One year after the surgical procedure, the owner reported that the cow did not present with recurrence of the neoplasm. The resected tumor weighed 11.2 kg, and, when cut, presented with solid conformation and whitish coloration. Tumor fragments were harvested, fixed in 10% formalin, and sent for histopathological examination, which revealed neoproliferation of remarkable cellular density composed of dense, well vascularized fibrocollagenous connective tissue arranged in multidirectional bundles and undulating pattern. Mild cellular pleomorphism was identified, and no mitosis figures were observed. Alcian blue staining was negative for mucopolysaccharides, differing from Masson’s trichrome staining, which widely stained the fibrocollagenous tissue blue. In view of these findings, the diagnosis of cutaneous fibroma was confirmed. Discussion: Cutaneous fibromas are benign neoplasms of fibrous tissue, and they are uncommon in cattle and may be associated with bovine papillomavirus and/or trauma. Although the origin of cutaneous fibroma is not clear, the present report stands out due to the large size of the tumor mass. The complete healing of the surgical wound, the absence of recurrence one year after surgery and the return of the animal to dairy production demonstrate that the surgical treatment was adequate. The macro- and microscopic characteristics of the cutaneous fibroma in this case corroborate with other cases reported in the literature. Large cutaneous fibroma is uncommon in bovines, and may hinder surgical excision and prolong healing time, as well as the complete recovery of the animal. Moreover, the differential diagnosis with other neoplasms of fibroblastic origin is relevant, especially for those with malignant biological behavior, such as fibrosarcoma and myxosarcoma. [ABSTRACT FROM AUTHOR]

RESUMO Cicatrização de ferida é um processo dinâmico, que tem por objetivo restaurar a continuidade do tecido lesionado. No entanto, em alguns casos, é necessário favorecer condições adequadas para viabilizar o processo fisiológico. Neste estudo foram utilizados ratos Wistar, divididos aleatoriamente entre cinco grupos, com 12 animais cada, sendo eles: grupo P (Bidens pilosa L.), grupo mel, grupo Co1 (pomada comercial alopática), grupo Co2 (pomada comercial homeopática) e grupo CT (controle). As lesões foram geradas por incisão com punch de 8mm, sendo tratadas diariamente de forma tópica. Foram eutanasiados quatro animais por grupo, no terceiro, sétimo e 14º dias do experimento, e o material coletado foi armazenado em formalina 10% e encaminhado para processamento histológico. Posteriormente, realizou-se a contagem de leucócitos mononucleares, fibroblastos e neovasos e avaliou-se a arquitetura de fibras colágenas. Os resultados da contagem foram analisados pela ANOVA, seguida pelo teste de Tukey (P

El agrandamiento gingival (AG) es el incremento excesivo del volumen del tejido gingival. Comúnmente el AG se asocia con enfermedades sistémicas, ingesta prolongada de medicamentos y factores locales como el tratamiento por ortodoncia. El propósito de este trabajo fue identificar y describir los aspectos morfológicos in vivo e in vitro de los tejidos y fibroblastos gingivales en pacientes con AG por ortodoncia e individuos sanos. La identificación de fibras de colágeno y elásticas de la encía de cuatro pacientes con AG por ortodoncia y cinco individuos sanos fueron realizados mediante las tinciones Masson-Goldner y Orceina, respectivamente. A partir de cultivos celulares, la morfología de fibroblastos gingivales se estudió por inmunofluorescencia. Para este fin, fueron empleados los anticuerpos anti-actina y anti-vimentina. En los pacientes con AG por ortodoncia fue observado tejido epitelial con acantosis e hiperqueratinizado, acompañado de red compacta de fibras de colágeno. Las fibras elásticas de pacientes con AG tuvieron una disposición difusa y aleatoria en el tejido conjuntivo. Los fibroblastos gingivales in vitro de pacientes con AG por ortodoncia e individuos sanos fueron morfológicamente similares.

Copyright of Fisioterapia Brasil is the property of Atlantica Editora and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Odontológica Colombiana is the property of Universidad Nacional de Colombia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: To observe clinically, in rabbits, the side effects of topical injection of subconjunctival cyclophosphamide, studying its role as an antifibrotic drug. Methods: Prospective study in 20 albino rabbits of New Zealand race. All rabbits were treated with cyclophosphamide, 10mg/ml in a volume of 0.3 ml, in the left eye through subconjunctival injection. They were evaluated for 1, 7, 30, and 60 days after the procedure. All the animals were examined for the detection of ocular reactions such as necrosis, hyperemia, chemosis, secretion, opacity, and iritis. Other side effects as changes in the behavior, in the feed, and the water consumption were also evaluated. Results: It was observed that from the 20 rabbits studied, three rabbits (15%) showed side effects only at the 24 hours analysis. One rabbit (5%) presented hyperemia, one rabbit (5%) had hyperemia associated with iritis, and one rabbit (5%) presented hyperemia associated with secretion. These reactions were not observed at 1, 7, 30, and 60 days. Conclusion: Cyclophosphamide subconjunctival injection induces minor side effects on the conjunctiva of rabbits such as hyperemia, associated with iritis and secretion. [ABSTRACT FROM AUTHOR]

The establishment of primary keloid fibroblast culture has always been a fundamental measure for studying mechanisms of keloid disease. The quality of the primary cell culture can directly affect the results of further experiments. This study was performed to investigate the optimal growth conditions, including the optimal storage time and collagenase treatment time, for in vitro cell culture models and the suitable methods for epidermis-dermis separation in different tissues. Keloid tissues, keloid-surrounding tissues, and normal skin tissues were collected from patients, for primary fibroblast culture. Two methods, tissue explant and collagenase digestion, were deployed and compared. Expression levels of the keloid-related genes a-SMA, Col1, and Col3 were assessed in cells cultured using both methods, to verify the qualities of the primary cells. A comparative analysis was conducted between the two methods and among the three different tissues used. Bacterial and lipid contamination was immediately minimized after the samples were processed. Different methods of epidermis removal and different durations of collagenase digestion were required in different tissues to generate optimal results. Real-time PCR results showed that the mRNA expression levels of keloid-related genes in cultured fibroblasts correlated to their in vivo expression profile, as previously reported in other studies. The results of this study have revealed several key points in the culture of primary keloid fibroblasts and demonstrated the correlation in gene expression between in vivo keloid fibroblasts and in vitro primary keloid fibroblasts. [ABSTRACT FROM AUTHOR]

Introdução: Dermatomiosite juvenil (DMJ) é doença sistêmica que afeta a musculatura proximal e a pele de crianças. A doença ulcerada é um desafio terapêutico. Objetivo: Avaliar a melhora da doença ulcerada na DMJ, pelo uso de terapia celular. Métodos: Realização de cocultura de fibroblastos e queratinócitos autólogos e aplicação dessas células nas úlceras juntamente com cola de fibrina e colocação de membrana de quitosana-alginato ou quitosana-xantana sobre as lesões. Resultados: Menos de 12 horas após a terapia, o paciente referiu completa eliminação da dor e, dentro de dois dias, estava presente tecido de cicatrização. Algumas das úlceras estavam quase completamente cicatrizadas no final da primeira semana, e algumas das calcinoses desapareceram. Essa técnica não cura a doença, mas melhora a qualidade de vida, sendo possível criopreservar as células saudáveis do paciente para tratar novas lesões. Sendo as células de origem autóloga, elimina-se o risco de rejeição. Além disso, esse procedimento não necessita de debridamento das lesões nem hospitalização. Conclusões: A aplicação de culturas autólogas de fibroblastos e queratinócitos em úlceras já é considerada tratamento efetivo em pacientes com queimaduras e outras feridas cutâneas e, agora mostrou-se também eficaz no tratamento de feridas na DMJ.

Introduction: The Vismia genus belongs to the Hypericaceae family and comprises around 57 species of which 17 have been located in Venezuela. Previous investigations have been carried out in extracts as well as pure isolated compounds, revealing antimicrobial, antioxidant and anti-HIV, among other, biological activities. Objective: This investigation aims to determine the cytotoxic activity of essential oils from leaves of Vismia baccifera Triana & Planch (VBJ and VBV) and Vismia macrophylla Kunth (VM) collected in three different locations of the Venezuelan Andean region. Methods: Essential oils obtained by hydrodistillation were analyzed using gas chromatography-mass spectrometry (GC-MS) and their cytotoxic activity was analyzed following the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Human tumor cell lines from SKBr3, MCF-7 and PANC-1, two breast carcinomas and one pancreatic adenocarcinoma of ductal type, were tested with the oil samples and human dermis fibroblasts were used as non-tumor cells. Results: β-caryophyllene and trans-caryophyllene were present as major components in VBJ and VBV, respectively, while γ-bisabolene was the main component in the VM sample. Anticancer activity was observed on V. baccifera essential oil against SKBr3, MCF-7 and PANC-1. The selectivity index showed that VBV is highly selective against the SKBr3 cell line and has no activity against non-tumor cells. Conclusions: These results are considered a contribution to natural products research and may provide supportive data for future studies on cancer. [ABSTRACT FROM AUTHOR]

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RESUMO Objetivo: avaliar os efeitos da administração tópica do metronidazol na diferenciação de fibroblastos e na contração da ferida durante cicatrização experimental por segunda intenção em ratos. Métodos: cento e oito animais foram submetidos a uma ferida circular no dorso, com 2cm de diâmetro e divididos em seis grupos: grupo controle, com aplicação de solução salina sobre a ferida e cinco grupos experimentais divididos de acordo com a concentração da solução do metronidazol utilizada (4%, 6%, 8%,10% e 12%). Curativos foram realizados diariamente durante todo o período do experimento, que foi subdividido em três momentos de análise: três, sete e 14 dias. A contração da ferida foi avaliada por planimetria digital e os miofibroblastos e protomiofibroblastos foram identificados usando técnicas de imuno-histoquímica CD34 e a-SMA. Resultados: a contração da ferida não apresentou diferença entre os grupos e o controle. Os protomiofibroblastos foram significativamente mais numerosos aos sete dias (p=0,022) nos grupos metronidazol de 4%, 6% e 8%. Após 14 dias, nos mesmos grupos, os miofibroblastos predominaram significativamente (p=0,01). Conclusão: a administração tópica de solução de metronidazol em feridas de pele com cicatrização por segunda intenção foi capaz de melhorar a diferenciação de fibroblastos. A fase de contração da cicatrização de feridas permaneceu inalterada, sem redução significativa da contração avaliada pela planimetria digital. Estes resultados podem ser utilizados em favor do processo de cicatrização de feridas.

Copyright of RGO: Revista Gaúcha de Odontologia is the property of RGO: Revista Gaucha de Odontologia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Introducción: Las resinas dentales basadas en siloranos, son el resultado de la unión química de siloxanos y oxiranos. Dentro del proceso de evolución de las resinas, se han modificado sus formulaciones para optimizar sus propiedades físicas, mecánicas y estéticas. Sin embargo, aún presentan problemas como la contracción de polimerización, problemas alérgicos y toxicológicos. Objetivo: Determinar el potencial efecto citotóxico de las resinas a base de siloranos sobre la viabilidad celular en un cultivo de fibroblastos de la línea L929. Metodología: Se estudió la exposición de fibroblastos de ratón L929 a resinas dentales a base de siloranos Filtek p90 y metacrilatos Filtek p60 en tres tiempos de fotocurado durante 6 días. Se evaluó el efecto sobre la viabilidad celular mediante pruebas de exclusión vital, micrografías electrónicas y composición elemental de superficie (Mg, Na, K y Ca) por ESEM/EDAX. Resultados: La resina Filtek p90 mostró un mayor efecto citotóxico a tiempos de fotocurado prolongados (20 segundos o más) respecto a la resina Filtek p60. El efecto resultó más marcado para 20 segundos de fotocurado, ya que las concentraciones de Ca y K en la superficie celular fueron las más afectadas. Conclusiones: La resina dental Filtek p90 ejerce un efecto citotóxico relativamente bajo sobre las células de fibroblastos L929 (p

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Nowadays, thermal water benefits for the health of skin cells has been widely broadcasting and its use is recommended for therapeutic and esthetic purposes. In this work, the study of the effect of thermal water was conducted in cultures of fibroblastic cells (Vero E6), a type of cells found in the dermis layer and essential components of the skin. The cells were cultivated using thermal, deionized (ultrapure) and tap water. A growth cell curve for each condition was made to compare which type of water promotes a higher growth rate. Cells cultivated with thermal water exhibited a higher and similar growth rate than cells cultivated with deionized water with 48 and 76 hours of culture, respectively. The mineral contents of the three types of water, culture mediums and cells were also studied. The mineral content analysis showed that thermal water contains significant amounts of Ca, Na and Mg, but the higher contents found do not enhanced the uptake of these elements by the cells. [ABSTRACT FROM AUTHOR]

Copyright of Acta Veterinaria Brasilica is the property of Acta Veterinaria Brasilica and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Introduction: Continuous exposition of the peritoneal membrane to conventional dialysis solutions is an important risk factor for inducing structural and functional alterations. Objective: To compare in vitro mouse fibroblast NIH-3T3 cell viability after exposition to a neutral pH dialysis solution in comparison to cells exposed to a standard solution. Methods: Experimental study to compare the effects of a conventional standard or a neutral-pH, low-glucose degradation products peritoneal dialysis solution on the viability of exposed fibroblasts in cell culture. Both solutions were tested in all the commercially available glucose concentrations. Cell viability was evaluated with tetrazolium salt colorimetric assay. Results: Fibroblast viability was significantly superior in the neutral pH solution in comparison to control, in all three glucose concentrations (Optical density in nm-means ± SD: 1.5% 0.295 ± 0.047 vs. 0.372 ± 0.042, p < 0.001; 2.3% 0.270 ± 0.036 vs. 0.337 ± 0.051, p < 0.001; 4.25% 0.284 ± 0.037 vs. 0.332 ± 0.032, p < 0.001; control vs. neutral pH respectively, Student t Test). There was no significant difference in cell viability between the three concentrations of glucose when standard solution was used (ANOVA p = 0.218), although cell viability was higher after exposition to neutral pH peritoneal dialysis fluid at 1.5% in comparison to 2.3 and 4.25% glucose concentrations (ANOVA p = 0.008: Bonferroni 1.5% vs. 2.3% p = 0.033, 1.5% vs. 4.25% p = 0.014, 2.3% vs. 4.25% p = 1.00). Conclusion: Cell viability was better in neutral pH dialysis solution, especially in the lower glucose concentration. A more physiological pH and lower glucose degradation products may be responsible for such results.

Copyright of Biomédica: Revista del Instituto Nacional de Salud is the property of Instituto Nacional de Salud of Colombia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Resumen Objetivo: Determinar la efectividad de un injerto basado en el cocultivo de fibroblastos gingivales y queratinocitos en membrana de colágeno comercial Mem-Lok® (BioHorizons, Alabama, Estados Unidos) en el tratamiento de recesiones gingivales. Materiales y métodos: Esta investigación fue descriptiva y de diseño experimental. La muestra se conformó de 10 ratas Sprague Dawley a las que se indujeron recesiones gingivales. A 8 de ellas se les aplicó el injerto, y las 2 restantes no recibieron tratamiento. Resultados: El análisis descriptivo de los resultados de- terminó la posibilidad de obtener un cocultivo celular. Luego de la aplicación del injerto, las características clínicas periodontales indicaron salud, consistencia firme, textura a manera de puntillado, contorno festoneado, biotipo grueso, sondaje periodontal de 1 mm, y la posición de la encía a nivel del límite amelocementario. Conclusiones: El injerto aplicado logró una cobertura radicular del 100% en todos los casos. No se observó sangrado ni contracción cicatrizal. Palabras clave: Fibroblastos, mallas de colágeno, queratinocitos, recesión gingival. Abstract Aim: To determine the effectiveness of a graft based on co-cultivation of gingival fibroblast and keratinocytes in commercial collagen membrane Mem-Lok® (BioHorizons, Alabama, USA) in the treatment of gingival recessions. Materials and methods: This research was descriptive and experimental in design. The sample was composed of 10 Sprague Dawley rats which were induced gingival recession; two of them were not treated and the graft was applied in eight of them. Results: A descriptive analysis of the results was performed, which showed that it was possible to obtain a cell co-culture. After the application of the graft, clinical periodontal characteristics were observed that indicated health: the consistency was firm, the texture resembled dots, scalloped and knife edge margin, a thick biotype and the depth of gingival sulcus was 1 mm. Conclusions: The applied graft achieved a 100% radicular coverage in all cases and no bleeding or scar contraction was observed. Key words: Collagen membrane, fibroblast, gingival recession, keratinocytes.

Objetivo: Comparar in vitro el efecto citotóxico de tres agentes quelantes sobre fibroblastos del ligamento periodontal humano. Materiales y métodos: Se utilizaron cultivos de fibroblastos de ligamento periodontal humano, los cuales fueron colocados en contacto con los agentes quelantes a evaluar (RCPrep, Glyde, EDTA al 17%) a intervalos de 15, 30 y 60 minutos. Se midió la absorbancia para cada uno de los grupos, para determinar el grado de actividad enzimática, que es indicador de muerte celular. Previo a la cuantificación de la absorbancia se corroboró microscópicamente la formación de los cristales de formazán, los cuales se forman alrededor de los fibroblastos, y su presencia es indicador de integridad de la membrana y de la actividad metabólica. Por microscopia se verificó la formación de cristales de formazan, después de agregar azul de tripán. Resultados: El Glyde mostró mayor grado de citotoxicidad, con una diferencia estadísticamente significativa, al compararlo con EDTA 17% y el RC-PREP. El EDTA presentó mayor citotoxicidad que el RC-PREP a los 15minutos, evento que cambió a los 30 y 60 minutos. Conclusiones: Los agentes quelantes RC-Prep, Glyde y EDTA tienen un efecto citotóxico a nivel de los fibroblastos del ligamento periodontal, siendo el EDTA el de menor efecto citotóxico a los 30 y 60 minutos comparado con RC-Prep y Glyde.

BACKGROUND: The possible participation of keratinocytes in wound remodeling has been widely studied. This study investigated the impact of keratinocytes in wound contraction. METHODS: Murine type I collagen gels populated by human fibroblasts and seeded with human keratinocytes on the surface to form a dermo-epidermal equivalent were used as the study group. Collagen gels populated by only fibroblasts were used as the control group. The criteria for the preparation and storage of gels were similar for both groups. RESULTS: An evident and statistically significant increase in gel contraction was observed in samples populated by keratinocytes compared to the control group. CONCLUSIONS: These results suggest that keratinocytes not only modulate fibroblast proliferation but also play an active role in wound contraction per se. Further research on the mechanisms involved in the communication pathways between cells and between cells and the matrix shall be assessed from the perspective of keratinocyte participation in wound healing and pathologic scarring.INTRODUÇÃO: A eventual participação de queratinócitos na remodelagem da ferida tem sido estudada há muito tempo. Este trabalho investigou o impacto dos queratinócitos na contração da ferida. MÉTODO: Foi utilizado gel de colágeno tipo I murino povoado por fibroblastos humanos com queratinócitos humanos semeado na superfície (grupo estudo), formando um equivalente dermoepidérmico. Géis de colágeno povoado apenas por fibroblastos foram utilizados como grupo controle. Os critérios de confecção e armazenagem dos géis foram iguais para ambos os grupos. RESULTADOS: Houve aumento evidente e estatisticamente significante na contração de gel das amostras povoadas por queratinócitos, em comparação ao grupo controle. CONCLUSÕES: Esses resultados sugerem que os queratinócitos não só podem modular a proliferação de fibroblastos, mas também, por si só, desempenhar papel ativo na contração da ferida. Novas investigações sobre mecanismos envolvidos nas vias de comunicação entre células e entre célula e matriz devem ser avaliadas sob o ponto de vista de participação dos queratinócitos na cicatrização de feridas e formação de cicatrizes patológicas.

INTRODUCTION: Fetal calf serum (FCS) is commonly used as a supplement in the culture medium for fibroblast cells. This supplementation is far from ideal as sample quality varies from batch to batch and the composition of FCS is not completely known. In addition, FCS may be contaminated with viruses and/or prions and may also cause adverse immunologic responses in humans. Due to these facts, a worldwide effort is being made to find alternatives for xenobiotic elements in cell cultures. Human serum could be a safer alternative, especially for clinical application. METHODS: We investigated human serum as a substitute for FCS in human fibroblast culture. Fresh human serum was obtained from 10 healthy volunteers. Fibroblasts were cultivated in multiwell plates containing either Dulbecco's modified Eagle's medium (DMEM) plus 10% FCS (D10) or DMEM plus 10% human serum (D10H). Cell counts were obtained between 24 and 264 hours of cultivation; results were expressed as the mean number of cells ± standard error of the mean to create cell proliferation curves. RESULTS: There was no statistical difference in fibroblast proliferation between the two groups. Human serum supported human fibroblast growth and proliferation, suggesting that it may be a potential substitute for FCS in human cell culture. Cells cultivated with human serum presented a different morphology, appearing smaller and more rounded as compared to cells cultivated in D10. CONCLUSIONS: These results demonstrate that human serum can be substituted for FCS in human fibroblasts culture and that fibroblasts cultivated in the presence of human serum have a morphology that is similar to in vivo fibroblasts.INTRODUÇÃO: Soro bovino fetal (SBF) é comumente usado como suplemento no meio de cultura para cultivar fibroblastos. Essa forma de suplementação, porém, não é ideal, pois a qualidade das amostras de SBF é variada e sua composição não é completamente conhecida. Além disso, o SBF pode apresentar contaminação por vírus e príons ou causar complicações imunológicas. Assim, a comunidade científica tem buscado alternativas ao uso de elementos xenobióticos em cultura celular. O soro humano pode ser uma dessas alternativas, principalmente para aplicação clínica. MÉTODO: Soro humano, obtido de sangue de 10 voluntários saudáveis submetidos a avaliação sorológica prévia, foi testado como substituto do SBF em cultura de fibroblastos humanos. As células foram cultivadas em placas multipoços, contendo Dulbecco's Modified Eagle's Medium (DMEM) mais 10% de SBF (D10) ou DMEM mais 10% de sroro humano (D10H). Entre 24 e 264 horas de exposição aos meios testados, as células foram contadas e os resultados foram expressos em média ± erro padrão da média, para obtenção de curvas de proliferação celular. RESULTADOS: Não houve diferença estatística entre os grupos de proliferação. Fibroblastos na presença de soro humano aparentavam ser menores e mais arredondados em comparação àqueles mantidos em D10. CONCLUSÕES: Os resultados permitem inferir que o soro humano pode substituir o SBF em cultura de fibroblastos e que fibroblastos cultivados em meio suplementado por soro humano apresentam morfologia mais semelhante àqueles in vivo.

Purpose: To quantify keloid fibroblasts after irradiation with 470nm blue LED, in vitro. Methods: Fibroblasts from keloid and adjacent skin have been obtained from 6 patients. Cells have been cultivated and maintained in DMEM culture medium. In Petri dishes, they were irradiated with energy doses of 6J, 12J and 18J. After 24 h, counting was done by the average of the triplicates for each sample. Results: There were no significant differences in the number of irradiated keloid fibroblasts at the studied doses (p=0.261). In adjacent skin fibroblasts, differences were observed (p=0.025) concerning the doses of 18 J and 6 J (p=0.03). Conclusions: There was a reduction in the number of adjacent skin fibroblasts irradiated with 470nm blue LED at the energy dose of 18 J compared to the ones irradiated at the energy dose of 6 J. There were no changes in keloid fibroblasts counting at any of the doses applied, 24 h after irradiation.Objetivo: Quantificar fibroblastos de quelóide após irradiação com LED azul de 470nm, in vitro. Métodos: Foram obtidos fibroblastos de quelóide e pele adjacente, de seis pacientes. As células foram cultivadas e mantidas em meio de cultura DMEM. Em placas de Petri, receberam irradiação com doses de energia de 6J, 12J e 18J. Após 24 horas a contagem foi feita pela média da triplicata para cada amostra. Resultados: Não houve diferença na quantidade de fibroblastos de quelóide irradiados nas doses estudadas (p=0,261). Observou-se diferença nos fibroblastos de pele adjacente (p=0,025), com relação às doses de 18 J e 6 J (p=0,03). Conclusões: Houve redução dos fibroblastos de pele adjacente irradiados com LED azul de 470 nm na dose de energia de 18 J em relação à dose de 6 J. Não houve alteração na quantidade de fibroblastos de quelóide nas doses aplicadas após 24 horas da irradiação.

O Dermatomiofibroma está incluído no grupo de lesões neoplásicas mesenquimais benignas de linhagem fibroblástica e miofibroblástica da pele. É uma doença rara, havendo aproximadamente 100 casos descritos na literatura mundial até o momento. Este artigo relata o caso de uma mulher jovem com apresentação clínica típica e diagnóstico histopatológico de dermatomiofibroma. Foram realizadas colorações especiais que mostraram preservação das fibras colágenas e a imunohistoquímica revelou positividade para vimentina e negatividade para actina e S100. Por se tratar de doença rara, os achados histopatológicos são de grande importância, mas a supeição clínica é possível em casos típicos como este.Dermato myofibroma is included in the group of benign cutaneous mesenchymal neoplastic lesions of fibroblastic and myofibroblastic lineage. It's a rare disease and there are approximately only one hundred cases described worldwide in the medical literature up to now. The present study reports the case of a young woman with typical clinical cutaneous lesion and histopathological diagnosis of dermato myofibroma. Special stains were carried out which showed preserved collagen fibers and immunohistochemistry was positive for vimentin and negative for actin and S100. As it is a rare disease, the histopathological findings are of great importance but clinical suspicion is possible in typical cases such as this one.

Objetivou-se com este trabalho avaliar o efeito do número da passagem e do sexo das células doadoras de núcleo no desenvolvimento embrionário e fetal após transferência nuclear. Para isso, oócitos bovinos foram maturados, enucleados e reconstruídos com células somáticas de animal adulto. Após a fusão e ativação química, os zigotos reconstituídos foram cultivados em Charles Rosenkranz 2 (CR2) com monocamada de células da granulosa a 38,8ºC em atmosfera umidificada a 5% de CO2 em ar, durante sete dias, e transferidos para receptoras sincronizadas. As taxas de clivagem e desenvolvimento a blastocisto de embriões reconstruídos com células cultivadas por tempo maior foram inferiores às obtidas com os demais tempos de cultivo. Além disso, os blastocistos produzidos não resultaram no desenvolvimento de uma gestação a termo. Embora a taxa de clivagem em embriões fêmeas tenha sido maior, o número de embriões que atingiram o estádio de blastocisto foi maior nos embriões machos. No período gestacional, fêmeas apresentaram maior taxa de aborto entre 90 e 120 dias de gestação. Esses resultados indicam que células doadoras de núcleos cultivados por longos períodos dificultam a produção de blastocistos e aumentam as chances de perdas durante a gestação. Embriões clonados machos têm maior competência para se desenvolver a blastocisto e resultam em menor taxa de perda gestacional.The objective of this study was to evaluate the effects of culture time and sex of nuclei donor cells on embryo and fetal development after nuclear transfer. Thus, bovine oocytes were matured, enucleated and reconstructed with somatic cells from an adult animal. After fusion and chemical activation, the reconstituted zygotes were cultured in Charles Rosenkranz 2 (CR2) on a granular monolayer cell at 38.8ºC in a humidified atmosphere 5% CO2 in air for seven days, and transferred to synchronized receptors. Cleavage rates and development to blastocyst of embryo reconstructed with cells cultured for a longer time were lower than rates obtained with other culture times. Moreover, these produced blastocysts did not result in the development of full term pregnancy. Although cleavage rates were higher in female embryos, the number of embryos that reached blastocyst stage was higher in male embryos. During gestation period, females showed higher abortion rates from 90 to 120 days of gestation. These results indicate that cells donnors of nuclei cultured for long periods make the production of blastocysts difficult and increase the chances of losses during pregnancy. Cloned male embryos are more succesful in becoming blastocysts and result in lower gestational loss rate.

PURPOSE: To study the collagen density and the population of fibroblasts in cutaneous injuries in rats under the influence of nicotine. METHODS: The scars of abdominal wounds in rats were analyzed. 2 mg/kg/d of nicotine was administered to the animals in the experiment group and the solution used as a vehicle for the animals in the control group. Treatment was begun seven days prior to surgery and maintained for seven or fourteen days following surgery. The removed scars were prepared for histopathological study. Histological cuts were stained by Sirius Supra red F3BA for collagen analysis and submitted to the examination using the immunohistochemical technique, which enabled us to recognize the population of fibroblasts. RESULTS: No significant difference was found in type I collagen density after seven days (p=0.912), nor after fourteen days (p=0.211). The control group had more type III collagen after seven days (p=0.004), but after fourteen days there was no significant difference (p=0,720). The total quantification of collagen, although higher in the control group, was not significantly so at any time during the study (p=0.103 after seven days and p=0.549 after fourteen days). The average of fibroblasts per field was lower after seven days (p=0.0001) and after fourteen days (p=0.0000). CONCLUSION: Under the conditions of this experiment, nicotine reduced the fibroblast population without modifying collagen density significantly.OBJETIVO: Estudar a densidade de colágeno e a população de fibroblastos em feridas cutâneas de ratos sob a influência da nicotina. MÉTODOS: Analisaram-se as cicatrizes de feridas abdominais de ratos. Foi administrada 2 mg/kg/d de nicotina, por via subcutânea aos animais do grupo experimento e a solução usada como veículo para os animais do grupo controle. O tratamento foi iniciado sete dias antes do ato operatório e mantido por sete ou quatorze dias no pós-operatório. As cicatrizes ressecadas foram preparadas para estudo histo-patológico. Cortes histológicos foram corados pelo Sirius Supra red F3BA para a análise do colágeno e submetidos à exame pela técnica imuno-histoquímica permitiram reconhecer a população de fibroblastos. RESULTADOS: Verificou-se que não existiu diferença significante na densidade de colágeno tipo I, na avaliação feita com sete dias (p=0,912) e com 14 dias (p=0,211). O grupo controle mostrava mais colágeno tipo III com sete dias (p=0,004), mas aos 14 dias não havia diferença significante (p=0,720). A quantificação do colágeno total, embora fosse maior no grupo controle, não o foi de forma significante em nenhum dos tempos estudados (p=0,103 aos sete dias e p=0,549 aos 14 dias). A média de fibroblastos por campo esteve diminuída na avaliação de sete dias (p=0,0001) e na de quatorze dias (p=0,0000). CONCLUSÃO: Nas condições do experimento, a nicotina promoveu diminuição da população de fibroblastos sem modificar a densidade do colágeno de forma significante.