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Braz. j. biol ; 84: e253616, 2024. tab, graf
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1355880


Abstract This study evaluated the effect of the volatile oil of Alpinia zerumbet (VOAz) on caveolin-1 gene expression and muscular fibrosis. The rats were immobilized to induce fibrosis of the gastrocnemius muscle, and they were treated with VOAz. Collagen quality was assessed by histology and the expression of the caveolin-1 (CAV-1) gene was evaluated using qPCR. Histomorphological analysis indicated a significant reduction in the perimeter, width, and intensity of collagen in the treated groups, thus showing that the oil was effective in regulating the quality of collagen at the three concentrations. The results of expression levels suggested a decrease in the lesioned group and in two treatment groups (0.0115 µg/g and 0.009 µg/g). However, with the lowest concentration (0.0065 µg/g), no significant difference was observed, with levels similar to those found in healthy tissue. Therefore, the results showed that VOAz has the potential to be a non-invasive and low-cost alternative to aid in the treatment of muscular fibrosis.

Resumo Este estudo avaliou o efeito do óleo volátil de Alpinia zerumbet (OVAz) na expressão do gene da caveolina-1 e na fibrose muscular. Os ratos foram imobilizados para induzir a fibrose do músculo gastrocnêmio, e foram tratados com OVAz. A qualidade do colágeno foi avaliada com histologia e à expressão do gene caveolina-1 (CAV-1) foi avaliada usando qPCR. A análise histomorfológica indicou uma redução significativa no perímetro, largura e intensidade do colágeno nos grupos tratados. Os resultados dos níveis de expressão sugeriram diminuição nos grupos de lesão e em dois grupos de tratamento (0,0115 µg/g e 0,009 µg/g). No entanto, com a menor concentração (0,0065 µg/g), não foi observada diferença significativa, apresentando níveis semelhantes aos encontrados em tecido saudável. O uso do OVAz foi eficaz para reverter as alterações do colágeno causadas pela fibrose, e sua menor concentração apresentou uma possível tendência de aumento na expressão do CAV-1. Portanto, os resultados mostraram que o OVAz tem potencial para ser uma alternativa não invasiva e de baixo custo para auxiliar no tratamento da fibrose muscular.

Animals , Rats , Oils, Volatile/pharmacology , Collagen/metabolism , Alpinia/chemistry , Caveolin 1/metabolism , Muscles/drug effects , Fibrosis , Plant Oils/pharmacology , Brazil , Rats, Wistar , Disease Models, Animal , Muscles/pathology
Rev. Assoc. Med. Bras. (1992) ; 68(2): 159-164, Feb. 2022. tab, graf
Article in English | LILACS | ID: biblio-1365364


SUMMARY OBJECTIVE: The objective of this study was to explore the molecular mechanism underlying the occurrence of benign bile duct stricture and the target of low-dose paclitaxel in the prevention of benign bile duct stricture. METHODS: Under the stimulation of transforming growth factor beta 1, the expression of collagen type I and connective tissue growth factor were detected on isolated primary fibroblasts. The phosphorylation levels of JNK and Smad2L were detected using Western blot. The effect of low-dose paclitaxel on the transforming growth factor beta 1-induced inhibition of type I collagen and connective tissue growth factor expression and JNK and Smad2L phosphorylation was also observed. RESULTS: Transforming growth factor beta 1 induced the secretion of type I collagen and connective tissue growth factor as well as JNK phosphorylation in biliary fibroblasts. The JNK inhibitor or siRNA-Smad2 inhibited the transforming growth factor beta 1-induced secretion of type I collagen and connective tissue growth factor. Low-dose paclitaxel inhibited the expression of type I collagen induced by transforming growth factor beta 1 and may inhibit the secretion of collagen in biliary fibroblasts. CONCLUSION: The activation of JNK/Smad2L induced by transforming growth factor beta 1 is involved in the occurrence of benign bile duct stricture that is mediated by the overexpression of type I collagen and connective tissue growth factor, and low-dose paclitaxel may inhibit the phosphorylation of JNK/Smad2L.

Humans , Paclitaxel/pharmacology , Collagen , MAP Kinase Signaling System , Collagen Type I/metabolism , Collagen Type I/pharmacology , Smad2 Protein , Fibroblasts/metabolism
J. venom. anim. toxins incl. trop. dis ; 28: e20210056, 2022. tab, graf, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1360567


The association of scaffolds to repair extensive bone defects can contribute to their evolution and morphophysiological recomposition. The incorporation of particulate biomaterials into three-dimensional fibrin bioproducts together with photobiomodulation therapy (PBM) has potential and can improve regenerative medicine procedures. The objective of this experiment was to evaluate the effects of PBM therapy on critical size defects filled with xenogenic bone substitute associated with fibrin biopolymer. Methods: A critical defect of 8 mm was performed in 36 Wistar male adult rats that were divided into four groups. Groups BC and BC-PBM were defined as controls with defects filled by a clot (without or with PBM, respectively) and groups XS and XS-PBM that comprised those filled with biocomplex Bio-OssTM in association with fibrin biopolymer. PBM was applied immediately after the surgery and three times a week every other day, with the parameters: wavelength of 830 nm, energy density 6.2 J/cm2, output power 30 mW, beam area of 0.116 cm2, irradiance 0.258,62 W/cm2, energy/point 0.72 J, total energy 2.88 J. Fourteen and 42 days after the surgery, animals were euthanatized and subjected to microtomography, qualitative and quantitative histological analysis. Results: The BC-PBM and XS-PBM groups had a similar evolution in the tissue repair process, with a higher density of the volume of new formed bone in relation to the groups without PBM (p = 0.04086; p = 0.07093, respectively). Intense vascular proliferation and bone deposition around the biomaterial particles were observed in the animals of the groups in which biocomplex was applied (XS and XS-PBM). Conclusion: PBM therapy allowed an improvement in the formation of new bone, with a more organized deposition of collagen fibers in the defect area. Biocomplex favored the insertion and permanence of the particulate material in bone defects, creating a favorable microenvironment for accelerate repair process.(AU)

Biocompatible Materials , Biopolymers , Collagen , Low-Level Light Therapy
Rev. ADM ; 78(6): 314-331, nov.-dic. 2021. ilus, tab, graf
Article in Spanish | LILACS | ID: biblio-1354416


La dentina se compone de un mineral de fosfato de calcio identificado como dahllita, que se dispone en pequeños cristales de hidroxiapatita carbonatada con dimensiones de 36 × 25 × 4 nm, y por una fase orgánica cuyo principal componente es el colágeno tipo 1 en 90%, que se orienta en forma de malla. Esta conformación corresponde a los dientes permanentes. Dentro de las estructuras, encontramos túbulos dentinarios que miden, aproximadamente, entre 0.5-1 µm de diámetro en la periferia y hasta 3-5 µm cerca de la pulpa. En el presente estudio, realizado en dentina de dientes temporales, el lumen de dichos túbulos es más grande cuando se encuentra cerca de la pulpa dental. Asimismo, se encontraron cambios elementales importantes de acuerdo con las diferentes profundidades en las que se observó, encontrando un aumento en el peso porcentual de carbono cuando se encuentra a mayor profundidad, lo que indica una composición orgánica mayor en la dentina pulpar. En estudios de dientes permanentes esta composición es disminuida y con mayor concentración en la dentina cercana a la unión amelodentinaria. En dentina de dientes temporales se encontraron diferencias en el recuento de túbulos dentinarios por mm2, comparado a la dentina de dientes permanentes, donde el número de túbulos no varía mucho (AU)

Dentin is composed of a calcium phosphate mineral identified as dahllite, which is arranged in small crystals of carbonated hydroxyapatite with dimensions of 36 × 25 × 4 nm, and by an organic phase whose main component is type l collagen in 90%, which is oriented in the form of a mesh. This conformation corresponds to permanent teeth. Within the structures, we find dentin tubules that measure approximately 0.5-1 µm in diameter at the periphery and up to 3-5 µm near the pulp. In the present study, carried out in dentin of primary teeth, the lumen of these tubules is larger when it is close to the dental pulp. Likewise, important elemental changes were found according to the different depths in which it was observed, finding an increase in the percentage weight of carbon when it is at a greater depth, indicating a greater organic composition in the pulp dentin. In studies of permanent teeth, this composition is decreased and with a higher concentration in the dentin near the amelodentinal junction. In dentin of primary teeth, differences were found in the count of dentin tubules per mm2, compared to dentin of permanent teeth, where the number of tubules did not vary much (AU)

Humans , Tooth, Deciduous , Dentin/ultrastructure , Dentinogenesis , Phosphates , Phosphoric Acids , Acid Etching, Dental , Microscopy, Electron, Scanning , Calcium , Statistical Analysis , Collagen , Durapatite , Dentition, Permanent , Collagen Type I , Minerals
An. bras. dermatol ; 96(6): 730-734, Nov.-Dec. 2021. graf
Article in English | LILACS | ID: biblio-1355621


Abstract Lipoid proteinosis is a rare autosomal recessive disease, characterized by hyaline deposits of PAS-positive material in tissues due to mutations in the ECM1 gene. This study evaluated the ultrastructure of the skin of a 6-year-old child affected by this condition. The light microscopy identified PAS-positive hyaline deposits, which were more intense in the papillary dermis. Scanning electron microscopy of the dermis showed a compact papillary dermis and fibrillar deposits in the middle dermis. Transmission electron microscopy clearly showed the deposition of fibrillar material in the dermis, forming clusters adherent to elastic fibers, between the collagen bundles and the collagen fibers, and also filling up the cytoplasm of dermal fibroblasts.

Humans , Child , Lipoid Proteinosis of Urbach and Wiethe , Skin , Extracellular Matrix Proteins , Collagen , Hyalin , Microscopy
Rev. Círc. Argent. Odontol ; 79(230): 17-20, dic. 2021. ilus, graf
Article in Spanish | LILACS | ID: biblio-1358178


Luego de una extracción dental ocurren inevitablemente procesos de reabsorción y remodelación ósea, donde la dimensión y morfología de la cresta alveolar se ve modificada, representando un problema para la rehabilitación de la zona. Estudios clínicos han documentado un promedio de 4,0 a 4,5 mm de reabsorción ósea horizontal luego de una exodoncia, como así también cambios dimensionales significativos en los alrededores del hueso alveolar. El propósito fue evaluar y comparar clínica y tomográficamente los procesos de reparación y conservación del reborde alveolar post-extracción de paredes intactas, con y sin la utilización de esponjas de colágeno intraalveolar como relleno y placa termoformada como barrera física, durante el proceso de cicatrización. Se seleccionaron pacientes con indicación de exodoncia de elementos dentarios normalmente implantados y clínicamente aceptables, aplicando los criterios de exclusión, se realiza toma de impresión del terreno para la confección de una placa de protección rígida 0,8 termoformada para ser colocada posterior a la extracción durante la masticación por un período de 30 días. Se indica tomografía cone-beam post operatoria inmediata y a los tres meses para analizar, medir y comparar alto y ancho de crestas alveolares residuales. En la evaluación clínica y tomográfica de los casos estudiados, el grupo control donde se usó únicamente placa de protección alveolar arrojó mejores resultados que el grupo donde se colocaron esponja de colágeno en el interior del alvéolo. Palabras clave: Regeneración ósea, esponja de colágeno, cicatrización ósea, alvéolo postextracción, placa de protección alveolar (AU)

After a tooth extraction, bone resorption and remodeling processes inevitably occurs, where size and morphology of the alveolar crest is modified, representing a problem for the rehabilitation of the area. Clinical studies have documented an average of 4.0 to 4.5 mm of horizontal bone resorption after an extraction, us well us substantial dimensional changes around the alveolar bone. The purpose was to evaluate and compare clinical and tomographically both repair and preservation of post extraction alveolar ridge of intact walls processes, with and without the use of intraalveolar collagen sponges as filler and a thermoformed protective plaque, us physica? barrier, during healing process. Patients with normally implanted and clinically acceptable tooth with extraction indication were selected, applying the exclusion criteria, impression of the field is taken to build a 0.8 rigid thermoformed protective plaque in order to be placed after extraction and used during chewing for a period of 30 days. Immediate and three months post-operative cone beam tomography are indicated to analyze, measure and compare height and width of residual alveolar crests. In the clinical and tomographic evaluation of the cases treated, control group where only alveolar protective plaque was used, showed better results than the group with intraalveolar collagen sponge (AU)

Humans , Male , Female , Bone Regeneration , Alveolar Bone Loss , Collagen , Argentina , Schools, Dental , Tooth Extraction , Wound Healing , Tomography, X-Ray Computed , Statistical Analysis , Cone-Beam Computed Tomography
Fisioter. Bras ; v.22(4): 597-608, Nov 2, 2021.
Article in Portuguese | LILACS | ID: biblio-1353441


A flacidez tissular abdominal é uma disfunção dermatológica que incomoda principalmente as mulheres. A radiofrequência e o microagulhamento são recursos utilizados para minimizar essa flacidez. Objetivo: Investigar os efeitos do microagulhamento associado a radiofrequência na flacidez tissular abdominal. Métodos: Trata-se de um estudo experimental, controlado e randomizado, com amostra de 20 mulheres, faixa etária entre 18 e 35 anos, dispostas em dois grupos: Grupo 1 (G1) foi aplicada 1 sessão de microagulhamento, após 15 dias reavaliação utilizando a plicometria e perimetria e Grupo 2 (G2) 1 sessão de microagulhamento, após 15 dias realizaram-se 4 sessões de radiofrequência com intervalo de 1 dia entre as sessões. Resultados: O G2 apresentou diminuição de flacidez do músculo reto abdominal direito apresentando p = 0,009, flanco direito p = 0,001 e flanco esquerdo p = 0,004, assim como a redução da circunferência abdominal. A avaliação de satisfação corporal do G2 teve escore final p = 0,029. Conclusão: O microagulhamento associado a radiofrequência promoveram uma melhora clínica da flacidez tissular abdominal e flancos. (AU)

Female , Adult , Cutis Laxa , Dry Needling , Radio Waves , Collagen , Elastin , Cell Proliferation , Fibroblasts
Rev. Assoc. Med. Bras. (1992) ; 67(9): 1342-1348, Sept. 2021. graf
Article in English | LILACS | ID: biblio-1351459


SUMMARY OBJECTIVE: This study aimed to assess the effect of the collagen/silk fibroin scaffolds seeded with human umbilical cord-mesenchymal stem cells on functional recovery after acute complete spinal cord injury. METHODS: The fibroin and collagen were mixed (mass ratio, 3:7), and the composite scaffolds were produced. Forty rats were randomly divided into the Sham group (without spinal cord injury), spinal cord injury group (spinal cord transection without any implantation), collagen/silk fibroin scaffolds group (spinal cord transection with implantation of the collagen/silk fibroin scaffolds), and collagen/silk fibroin scaffolds + human umbilical cord-mesenchymal stem cells group (spinal cord transection with the implantation of the collagen/silk fibroin scaffolds co-cultured with human umbilical cord-mesenchymal stem cells). Motor evoked potential, Basso-Beattie-Bresnahan scale, modified Bielschowsky's silver staining, and immunofluorescence staining were performed. RESULTS: The BBB scores in the collagen/silk fibroin scaffolds + human umbilical cord-mesenchymal stem cells group were significantly higher than those in the spinal cord injury and collagen/silk fibroin scaffolds groups (p<0.05 or p<0.01). The amplitude and latency were markedly improved in the collagen/silk fibroin scaffolds + human umbilical cord-mesenchymal stem cells group compared with the spinal cord injury and collagen/silk fibroin scaffolds groups (p<0.05 or p<0.01). Meanwhile, compared to the spinal cord injury and collagen/silk fibroin scaffolds groups, more neurofilament positive nerve fiber ensheathed by myelin basic protein positive structure at the injury site were observed in the collagen/silk fibroin scaffolds + human umbilical cord-mesenchymal stem cells group (p<0.01, p<0.05). The results of Bielschowsky's silver staining indicated more nerve fibers was observed at the lesion site in the collagen/silk fibroin scaffolds + human umbilical cord-mesenchymal stem cells group compared with the spinal cord injury and collagen/silk fibroin scaffolds groups (p<0.01, p< 0.05). CONCLUSION: The results demonstrated that the transplantation of human umbilical cord-mesenchymal stem cells on a collagen/silk fibroin scaffolds could promote nerve regeneration, and recovery of neurological function after acute spinal cord injury.

Humans , Animals , Rats , Spinal Cord Injuries , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Fibroins , Spinal Cord , Umbilical Cord , Collagen , Recovery of Function , Tissue Scaffolds
Rev. Ciênc. Méd. Biol. (Impr.) ; 20(2): 301-306, set 29, 2021. tab, fig
Article in Portuguese | LILACS | ID: biblio-1354493


Introdução: o tabagismo é uma das principais causas evitáveis de mortes no mundo representando um problema de saúde pública. Objetivo: investigar a relação da exposição passiva à fumaça principal do cigarro e as possíveis alterações histomorfométricas das células gliais, arteríolas e da matriz extracelular do nervo olfatório de ratas. Metodologia: trata-se de um estudo experimental, analítico e quantitativo. Vinte ratas randomizadas divididas em dois grupos, controle e tabaco, foram expostas à inalação da fumaça principal do cigarro por 60 dias utilizando dispositivo validado na literatura. Resultados: a exposição à inalação da fumaça principal do cigarro resultou em alterações significativas no grupo tabaco, tais como, elevação nos níveis de cotinina no plasma sanguíneo, aumento na espessura da parede dos vasos sanguíneos, aumento na porcentagem do colágeno total do tecido, diminuição no número total de astrócitos e aumento no número total de micróglias. Conclusão: a exposição à fumaça principal do cigarro resulta em alterações histomorfométricas que poderiam causar alterações funcionais no nervo olfatório como perda sensorial olfativa. Os achados constatados são fortes o suficiente para servir como alerta a toda a população e às autoridades de saúde, no que se refere às leis antifumo, principalmente em ambientes fechados.

Introduction: smoking is one of the main preventable causes of death in the world and represents a worldwide public health problem. Objective: to investigate the relationship of second hand tobacco smoke and possible histomorphometric changes of glial cells, arterioles and extracellular matrix of the olfactory nerve in rats. Methodology: experimental, analytical and quantitative study, twenty wistar animals randomized into two control and tobacco groups, were exposed to inhalation of main cigarette smoke for 60 days using a device validated in the literature. Results: exposure to inhalation of main cigarette smoke resulted in changes in the tobacco group, such as increased levels of cotinine in the blood plasma, increased thickness of the blood vessel wall, increased percentage of total tissue collagen, decreased in the total number of astrocytes and increase in the total number of microglia. Conclusion: exposure to main cigarette smoke results in histomorphometric changes that can cause changes in the olfactory nerve such as sensory olfactory loss. Our findings are strong enough to serve as a warning to the entire population and to health authorities in relation to smokefree laws especially in closed environments.

Animals , Male , Female , Olfactory Nerve , Rats , Tobacco Use Disorder , Neuroglia , Collagen , Tobacco Products , Anatomy , Analytical Methods , Epidemiology, Experimental
Rev. bras. ortop ; 56(3): 333-339, May-June 2021. tab, graf
Article in English | LILACS | ID: biblio-1288682


Abstract Objective To evaluate clinically and radiologically the results of the treatment of chondral lesions using collagen membrane - autologous matrix-induced chondrogenesis (AMIC). Methods This is a series of observational cases, in which 15 patients undergoing AMIC were analyzed. The clinical evaluation was made by comparing the Lysholm and International Knee Document Commitee (IKDC) scores in the pre- and postoperative period of 12 months, and radiological evaluation using the Magnetic Resonance Observation of Cartilage Repair Tissue (MOCART) score in the same postoperative period. Results The mean age of the patients was 39.2 years old, and the mean size of the chondral lesions was 1.55cm2. There was a significant improvement in clinical scores, with a mean increase of 24.6 points on Lysholm and of 24.3 on IKDC after 12 months. In the radiological evaluation, MOCART had a mean of 65 points. It was observed that the larger the size of the lesion, the greater the improvement in scores. Conclusion Evaluating subjective clinical scores, the treatment of chondral lesions with the collagen membrane showed good results, as well as the evaluation of MOCART, with greater benefit in larger lesions.

Resumo Objetivo Avaliar clínica e radiologicamente os resultados do tratamento das lesões condrais com a membrana de colágeno - condrogênese autóloga induzida por matriz. Métodos Trata-se de uma série de casos observacional, na qual foram analisados 15 pacientes submetidos a condrogênese autóloga induzida por matriz. A avaliação clínica foi feita comparando os escores de Lysholm e International Knee Document Commitee (IKDC, na sigla em inglês) no pré- e pós-operatório de 12 meses, e avaliação radiológica através do escore de Magnetic Resonance Observation of Cartilage Repair Tissue (MOCART, na sigla em inglês) no mesmo período de pós-operatório. Resultados A média de idade dos pacientes foi 39,2 anos, e a média do tamanho das lesões condrais foi de 1,55cm2. Houve uma melhora significativa nos escores clínicos, com média de aumento de 24,6 pontos no Lysholm e de 24,3 no IKDC, após 12 meses. Na avaliação radiológica, o MOCART teve média de 65 pontos. Observou-se que quanto maior o tamanho da lesão, maior foi a melhora nos escores. Conclusão Avaliando escores clínicos subjetivos, o tratamento das lesões condrais com a membrana de colágeno mostrou bons resultados, assim como a avaliação de MOCART, com maior benefício em lesões maiores.

Humans , Male , Female , Adolescent , Adult , Middle Aged , Postoperative Period , Magnetic Resonance Spectroscopy , Cartilage, Articular , Collagen , Chondrogenesis , Knee Injuries
Int. braz. j. urol ; 47(2): 287-294, Mar.-Apr. 2021. tab, graf
Article in English | LILACS | ID: biblio-1154463


ABSTRACT Purpose: Despite high success rates in the treatment of urinary incontinence, complications related to the use of polypropylene (PP) meshes are still a concern, especially in vaginal prolapses surgeries. The objective of this study was to assess the effect of autologous platelet-rich plasma (PRP) coating on the integration of PP meshes implanted in the vaginal submucosa of rabbits. Materials and Methods: Thirty adult New Zealand rabbits were randomly divided into two groups (n=15): PP, implanted with conventional PP meshes; and PRP, implanted with autologous PRP coated PP meshes. Animals in both groups (n=5) were euthanized at 7, 30 and 90 days postoperatively, the vaginas extracted and sent to immunohistochemical analysis for the assessment of the pro-inflammatory agent TNF-α, anti-inflammatory agents TGF-β and IL-13, collagen metabolism marker MMP-2, and angiogenesis marker CD-31. AxioVision™ image analysis was used for the calculation of the immunoreactive area and density. Statistical analysis was performed with ANOVA followed by Tukey test (p <0.05). Results: Animals in the PRP group showed significantly increased expression of the angiogenesis agent CD-31 at all experimental times when compared to the PP group (p <0.0001). However, no differences concerning the expression of the other markers were observed between the groups. Conclusion: The addition of autologous PRP gel to PP meshes can be simply and safely achieved and seems to have a positive effect on implantation site angiogenesis. Further investigations are required to ascertain PPR coated meshes clinical efficacy in prolapses and stress urinary incontinence surgeries.

Animals , Female , Polypropylenes , Platelet-Rich Plasma , Rabbits , Surgical Mesh , Vagina/surgery , Collagen
Journal of Biomedical Engineering ; (6): 1103-1110, 2021.
Article in Chinese | WPRIM | ID: wpr-921851


China is the country with high incidence of high myopia in the world. High myopia can cause severe vision impairment. So far, there is no effective treatment for high myopia in clinic. Scleral collagen cross-linking surgery has been proven to be effective in preventing animal eye axial elongation

Animals , Collagen , Cross-Linking Reagents , Finite Element Analysis , Photosensitizing Agents , Riboflavin , Sclera
Article in Chinese | WPRIM | ID: wpr-921824


The effect of parasitic ions on the results of ultraviolet A (UVA) cross-linking in iontophoresis was still not clear. In this work, the porcine sclera was cross-linked by riboflavin lactate Ringer's solution (group A) and riboflavin normal saline (group B)

Animals , Collagen , Cross-Linking Reagents , Ions , Iontophoresis , Permeability , Photosensitizing Agents/pharmacology , Riboflavin , Sclera , Swine , Ultraviolet Rays
Chinese Journal of Biotechnology ; (12): 3231-3241, 2021.
Article in Chinese | WPRIM | ID: wpr-921420


The source of recombinant collagen is clean, and it has the advantages of flexible sequence design, high yield and high purity, so it has a wide application prospect as biomaterials in tissue engineering and other fields. However, how to promote the cross-linking of recombinant collagen molecules and make them form a more stable spatial structure is the difficulty to be overcome in the design of recombinant collagen nanomaterials. Unnatural amino acid O-(2-bromoethyl)-tyrosine was incorporated into collagen by two-plasmid expression system. The results showed that high-purity collagen incorporated with unnatural amino acid could be obtained by induction with final concentration of 0.5 mmol/L IPTG and 0.06% arabinose at 25 °C for 24 hours. The intermolecular cross-linking through thioether bond was formed between collagen molecule incorporated with unnatural amino acid and collagen molecule with cysteine mutation in pH 9.0 NH4HCO3 buffer, which formed aggregates with the largest molecular size up to 1 micrometre. The results pave the way for the design of recombinant collagen biomaterials.

Amino Acids , Biocompatible Materials , Collagen/genetics , Sulfides
Article in English | WPRIM | ID: wpr-880634


OBJECTIVES@#Silence of SET domain containing lysine methyltransferase 7 (SET7) alleviates myocardial tissue injury caused by ischemia-reperfusion. But the effects of SET7 on angiotensin II (Ang II)-induced myocardial fibroblast proliferation and the collagen synthesis are not clear. The purpose of this study was to explore the effect of SET7 on the proliferation and collagen synthesis of myocardial fibroblasts and its mechanisms.@*METHODS@#Myocardial fibroblasts were isolated and identified by immunofluorescence. Myocardial fibroblasts were randomly divided into 4 groups: a control group (cells were normally cultured), an Ang II group (cells were treated with 100 nmol/L Ang II for 24 h), a siCtrl group (cells were transfected with siRNA control and were then treated with 100 nmol/L Ang II for 24 h), and a siSET7 group (cells were transfected with siRNA SET7 and were then treated with 100 nmol/L Ang II for 24 h). Cell counting kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) assay were used to evaluate cell proliferation. Real-time PCR was used to detect the mRNA levels of SET7, collagen I, collagen III, and α-smooth muscle actin (α-SMA). Western blotting was used to detect the protein expression of SET7, collagen I, collagen III, α-SMA, sonic hedgehog (Shh), ptched1 (Ptch1), and glioma-associated oncogene homolog 1 (Gli1).@*RESULTS@#Fluorescence microscopy showed positive vimentin staining, and myocardial fibroblasts were in good condition. As compared to the control group, the mRNA and protein levels of SET7 in the Ang II group were significantly upregulated; cell proliferation rate and EdU fluorescence intensity in the Ang II group were significantly increased; the mRNA and protein levels of collagen I, collagen III, and α-SMA were significantly upregulated (all @*CONCLUSIONS@#Silence of SET7 gene inhibits Ang II-induced proliferation and collagen synthesis of myocardial fibroblasts. Shh signaling pathway may be involved in this process.

Angiotensin II/pharmacology , Cell Proliferation , Cells, Cultured , Collagen/genetics , Fibroblasts , Hedgehog Proteins
Chinese Journal of Biotechnology ; (12): 646-654, 2021.
Article in Chinese | WPRIM | ID: wpr-878589


The high performance liquid chromatography (HPLC) and enzyme-linked immunoassay (ELISA) were used to investigate the changes of collagen and matrix metalloproteinase-1 (MMP-1) in liver, lung and kidney during growth process of mice. The mice from 0 to 18 weeks were used as the research objects. The contents and proportions of hydroxyproline (Hyp), which were used to calculate the collagen contents, in liver, lung and kidney of different weeks were analyzed with HPLC. The contents and activity of MMP-1 in liver, lung and kidney of different weeks were analyzed with ELISA. The results showed that the collagen contents in liver, lung, and kidney were different (Lung(COL)>Kidney(COL)>Liver(COL)), and they all increased first and then decreased with weeks. The collagen contents in liver, lung, and kidney reached the highest level in the ninth (5.52 ng/mg), sixth (54.10 ng/mg) and ninth (19.20 ng/mg) week, respectively. Then it declined slowly from 9 to 18 weeks. The result of ELISA showed that the MMP-1 contents in liver, lung and kidney decreased first and then increased with weeks, and the trend of MMP-1 activity was opposite. It indicated that the increase of collagen contents in the tissues will inhibit the secretion of MMP-1.

Animals , Collagen , Kidney , Liver , Lung , Matrix Metalloproteinase 1 , Matrix Metalloproteinase 2 , Mice
Rev. bras. oftalmol ; 80(3): e0005, 2021. tab, graf
Article in English | LILACS | ID: biblio-1280120


ABSTRACT Objective: The objective was to evaluate the efficacy of gabapentin in the management of neuropathic pain in patients with keratoconus, who were treated with fast (10 minutes) epi-off corneal crosslinking (CXL). Methods: This was a prospective, double-blind, randomized study. The sample comprised patients with bilateral progressive keratoconus, aged 12 years or older, who underwent a bilateral epi-off corneal CXL (fast - 10 minutes) procedure. One group was given placebo orally, and the other group received gabapentin 600 mg orally, both preoperatively. The visual analogue scale (VAS) was applied to record postoperative pain up to 48 hours after procedure. The study was conducted at the Belotto Stock Centro Oftalmológico, in the city of Joaçaba, Santa Catarina, Brazil, from June 2018 to September 2019. Results: At no point in the study significant differences were observed between groups, in terms of pain intensity measured by means of the VAS questionnaire, or of opioid use (Paco®), though opioid consumption was 21% lower in the group receiving gabapentin. Conclusion: We concluded gabapentin has no efficacy in postoperative pain control after epi-off corneal CXL (fast - 10 minutes). Although there was no statistically significant difference, the group that received gabapentin suffered less pain, resulting in lower opioid consumption. UTN number: U1111-1256-0330.

RESUMO Objetivo: Avaliar a eficácia do uso da gabapentina no manejo da dor neuropática em pacientes portadores de ceratocone submetidos ao tratamento de crosslinking corneano epi-off fast de 10 minutos. Métodos: Tratou-se de pesquisa prospectiva, duplo-cega, randomizada. A amostra foi composta de pacientes com ceratocone progressivo bilateral, a partir dos 12 anos de idade, submetidos ao procedimento de crosslinking corneano acelerado epi-off fast de 10 minutos bilateral. Um grupo recebeu placebo via oral e o outro, gabapentina 600mg, via oral, ambos no pré-operatório. A Escala Visual Analógica foi aplicada para registrar a dor pós-operatória até 48 horas após o procedimento. A pesquisa foi realizada no período de junho de 2018 a setembro de 2019 em um centro oftalmológico. Resultados: Não foram observadas diferenças estatísticas significativas para ambos os grupos, tanto na intensidade da dor medida pela Escala Visual Analógica, como na redução do uso do opioide (Paco®), em qualquer horário analisado durante um período de 48 horas. No entanto, houve redução de 21% no consumo de opioides pelo grupo que fez uso da gabapentina. Conclusão: A gabapentina não demonstrou eficácia no controle da dor no pós-operatório do crosslinking corneano epi-off fast de 10 minutos. No entanto, observou-se que, mesmo não havendo diferença estatisticamente significativa, houve diminuição da dor no grupo em que foi usada a gabapentina, resultando na redução do consumo de opioides. Número UTN: U1111-1256-0330.

Humans , Male , Female , Adolescent , Adult , Pain, Postoperative/drug therapy , Cross-Linking Reagents/therapeutic use , Keratoconus/therapy , Riboflavin/therapeutic use , Ultraviolet Rays , Pain Measurement , Double-Blind Method , Administration, Oral , Prospective Studies , Surveys and Questionnaires , Collagen/metabolism , Photosensitizing Agents/therapeutic use , Cornea/metabolism , Eye Pain/drug therapy , Gabapentin/administration & dosage , Gabapentin/therapeutic use , Analgesics/therapeutic use
Article in English | LILACS, BBO | ID: biblio-1155012


ABSTRACT Objective: To investigate the difference of chemical bonds between urethane dimethacrylate (UDMA) bonding agents with ethanol solvent and acetone solvent on dentin collagen. Material and Methods: This experimental comparison study used three groups: G1 (Control): UDMA and collagen; G2: UDMA, collagen and ethanol; and G3: UDMA, collagen and acetone. The groups were then pelleted and analysed with FTIR, then the peak value of carbonyl absorption band from each study group was calculated. The result of FTIR analysis and the peak of carbonyl absorption band (P) was calculated using the formula: P = (BC / AB) X 100; AB. BC is measured in centimeters. The study of chemical bond differences between ethanol-solvent UDMA agents compared with acetone-solvent on dentin collagen resulted in a graph of peak of carbonyl absorption bands of UDMA and dentin collagen groups. To determine the chemical bonds of UDMA from the top of the carbonyl ester absorption bands with wavenumber absorption in range 1700-1750 cm-1, the decreasing peak of the carbonyl absorption bands is assumed as more chemical bonds that formed. Data were analysed using Anova one way and Tukey HSD test. Results: There were significant differences between the three study groups (p<0.05). Conclusion: UDMA bonding agents' chemical bonds with acetone solvent are much higher than the chemical bonds between UDMA bonding agents with ethanol solvent on dentin collagen.

Dental Bonding/instrumentation , Dental Materials , Dentin , Ethanol/chemistry , Solvents/chemistry , Epidemiology, Experimental , Analysis of Variance , Collagen/chemistry , Statistics, Nonparametric , Indonesia
Braz. j. med. biol. res ; 54(4): e10692, 2021. tab, graf
Article in English | LILACS | ID: biblio-1153536


Fibrosis caused by the increase in extracellular matrix in cardiac fibroblasts plays an important role in the occurrence and development of atrial fibrillation (AF). The aim of this study was to investigate the role of hsa-miR-4443 in AF, human cardiac fibroblast (HCFB) proliferation, and extracellular matrix remodeling. TaqMan Stem-loop miRNA assay was used to measure hsa-miR-4443 expression in patients with persistent AF (n=123) and healthy controls (n=100). Patients with AF were confirmed to have atrial fibrosis by late gadolinium enhancement. At the cellular level, after hsa-miR-4443 mimic and inhibitor were transfected with HCFBs, proliferation, apoptosis, migration, and invasion were analyzed. Lastly, hsa-miR-4443-targeted gene and transforming growth factor (TGF)-β1/α-SMA/collagen pathway were evaluated by dual-luciferase reporter assay and western blot, respectively. In patients with AF, hsa-miR-4443 decreased significantly and collagen metabolism level increased significantly. Logistic regression analysis showed that low hsa-miR-4443 level was a risk factor of AF (P<0.001). The receiver operating characteristic curve revealed that hsa-miR-4443 was useful for predicting AF (area under the curve: 0.828, sensitivity: 0.71, specificity: 0.78, P<0.001). In HCFBs, hsa-miR-4443 targeted thrombospondin-1 (THBS1) and downregulated TGF-β1/α-SMA/collagen pathway. The inhibition of hsa-miR-4443 expression promoted HCFB proliferation, migration, invasion, myofibroblast differentiation, and collagen production. The significant reduction of hsa-miR-4443 can be used as a biomarker for AF. hsa-miR-4443 protected AF by targeting THBS1 and regulated TGF-β1/α-SMA/collagen pathway to inhibit HCFB proliferation and collagen synthesis.

Humans , Atrial Fibrillation , MicroRNAs/genetics , Fibrosis , Collagen , Contrast Media , Thrombospondin 1/genetics , Cell Proliferation , Transforming Growth Factor beta1 , Fibroblasts , Gadolinium
Rev. bras. ortop ; 55(6): 778-782, Nov.-Dec. 2020. graf
Article in English | LILACS | ID: biblio-1156183


Abstract Objectives The present paper aims to evaluate and compare the histological features of fresh and frozen menisci stored in a tissue bank for 1 month and for 5 years. Methods The meniscal grafts were subjected to a histological study. A total of 10 menisci were evaluated; 2 were frozen for 5 years, 4 were frozen for 1 month, and 4 were fresh, recently harvested specimens. Histological properties were evaluated in sections stained with hematoxylin and eosin and Masson trichrome methods. Results The menisci frozen for 1 month showed partially preserved collagen fiber structure and no significant hydropic tissue degeneration. The menisci frozen for 5 years presented an evident dissociation of collagen fibers and multiple foci of hydropic degeneration. Discussion Degeneration was much more significant in menisci stored for 5 years, indicating that a long freezing period results in substantial progression of tissue deterioration. This may suggest that the 5-year period, considered the maximum time for graft storage before transplant, is too long. Conclusion Grafts stored for 1 month showed a slight degenerative change in collagen fibers, whereas menisci frozen for 5 years presented significant tissue degeneration.

Resumo Objetivos Avaliar e comparar as características histológicas de meniscos frescos e meniscos congelados armazenados em banco de tecidos por 1 mês e por 5 anos. Métodos Foi feito um estudo histológico com enxertos meniscais. Avaliamos 10 meniscos, sendo 2 que ficaram armazenados sob congelamento por 5 anos, 4 armazenados congelados por 1 mês, e 4 frescos, recém captados. Foram feitos cortes histológicos corados com hematoxilina e eosina e Tricrômico de Masson, para avaliação das propriedades histológicas. Resultados Os meniscos congelados por 1 mês apresentaram preservação parcial da estrutura das fibras colágenas, sem degeneração hidrópica significativa do tecido. Nos meniscos congelados por 5 anos, observamos dissociação evidente das fibras colágenas, com presença de múltiplos focos de degeneração hidrópica. Discussão Encontramos degeneração bem mais significativa nos meniscos armazenados por 5 anos, o que indica que o longo período de congelamento leva à progressão significativa da degeneração do tecido. Isto pode sugerir que o período de 5 anos, considerado período máximo que o enxerto pode permanecer armazenado antes de ser transplantado, é um período muito longo. Conclusão Nos enxertos armazenados por 1 mês, existiu apenas discreta alteração degenerativa das fibras colágenas, enquanto que nos meniscos com 5 anos de congelamento foi observada degeneração significativa do tecido. Tibiais

Tissue Banks , Wounds, Penetrating , Collagen , Eosine Yellowish-(YS) , Transplants , Meniscus , Freezing , Goals , Hematoxylin