Analysis of the distribution of microhardness, mineral, and organic content of human, bovine, and ovine teeth / Análisis de la distribución de microdureza, contenido mineral y orgánico de dientes humanos, bovinos y ovinos

Objetive. Human teeth have been commonly used for in vitro and in situ studies. Cu­rrently, other animals' teeth have been purposed for dental research to overcome human teeth' problematic availability. This study aimed to investigate the enamel and dentin from human, bovine, and ovine teeth concerning the microhardness, organic, and in­ organic contents via micro­Raman spectroscopy. Methods. Human, bovine, and ovine teeth were divided according to their type and age into seven groups: Ovine; Bovine­12 months; Bovine­24 months; Bovine­36 months; Bovine­48 months; Bovine­+60 months; Human (control). The enamel's microhardness (superficial and deep) and den­tin (superficial, middle, and deep) were analyzed. The calcium/phosphate ratio and am­ide contents were determined by micro­Raman spectroscopy. Results. Overall, the mi­crohardness of human enamel was superior to the other species. Dentin's microhardness was similar among groups. Ovine group showed lower values of calcium/phosphate ratio than human. Amide content was similar between bovine and human. The microhardness and calcium/phosphate ratio of enamel and dentin, respectively, decreased as the age of bovine teeth increased. Conclusions. Researchers must be aware and take into consider­ation the differences of ovine and bovine enamel compared to human enamel. Other al­ternatives that are more similar to the microhardness of human enamel should be sought. Bovine teeth of 12 and 24 months are suitable substitutes for dentin of human teeth. Researchers must also be aware of the age of the animals and specify it in the studies.

Objetivo. Los dientes humanos se han utilizado comúnmente para estudios in vitro e in situ. Actualmente, los dientes de otros animales se han destinado a la investiga­ción dental para superar la disponibilidad problemática de los dientes humanos. Este estudio tuvo como objetivo investigar el esmalte y la dentina de los dientes humanos, bovinos y ovinos en relación con la microdureza y los contenidos orgánicos e inor­gánicos a través de la espectroscopia micro­Raman. Métodos. Los dientes humanos, bovinos y ovinos se dividieron según su tipo y edad en siete grupos: Ovinos; Bovino­12 meses; Bovino­24 meses; Bovino­36 meses; Bovino­48 meses; Bovino­+60 meses; Hu­mano (control). Se analizó la microdureza del esmalte (superficial y profunda) y de la dentina (superficial, media y profunda). La relación calcio/fosfato y los contenidos de amida se determinaron mediante espectroscopía micro­Raman. Resultados. En general, la microdureza del esmalte humano fue superior a la de otras especies. La microdureza de la dentina fue similar entre los grupos. El grupo ovino mostró valores más bajos de la relación calcio/fosfato que el humano. El contenido de amida fue similar entre bovinos y humanos. La microdureza y la relación calcio/fosfato del esmalte y la dentina, respectiva­mente, disminuyeron a medida que aumentaba la edad de los dientes bovinos. Conclusiones. El esmalte de los dientes ovinos y bovinos no es un sustituto adecuado del de los dientes humanos. Se deben buscar otras alternativas que sean similares a la microdureza del esmalte humano. Sin embargo, los dientes bovinos de 12 y 24 meses son sustitutos adecuados de la dentina de los dientes humanos. Los investigadores deben conocer la edad de los animales y especificarla en los estudios.

Tissue repair capacity of bioceramic endodontic sealers in rat subcutaneous tissue

Abstract This study aimed to evaluate the tissue repair capacity of four bioceramic endodontic sealers by quantifying type I and III collagen fibers. The following sealers were tested: EndoSequence BC Sealer (Brasseler, Brasseler, Savannah, USA), Bio C Sealer (Angelus, Londrina, Brazil), Bioroot RCS (Septodont, Santa Catarina, Brazil), and Sealer Plus BC (MKLife, Porto Alegre, Brazil). Polyethylene tubes 1.5 mm in diameter and 1 cm in length containing the endodontic sealers were implanted in the subcutaneous tissue of five rats (Rattus norvegicus albinus, Wistar lineage). After 14 days, the animals were euthanized, and collagen fibers were quantified from the histological tissue sections. Given a non-normal distribution of the data, a gamma regression with log link function was employed and implemented through the generalized linear models module, was used to test whether there was a significant difference between the sealers. The pairwise comparison was performed using Least significant difference. There were significant differences between the sealers for type I (p=0.001), type III (p=0.023), and total collagen (p=0.002). Overall, Bioroot sealer was statistically superior to the other sealers, except in the analysis of type III collagen, in which there was no difference between the Bioroot sealer and Bio C Sealer sealer and the control group (p>0.05). Bioroot RCS bioceramic endodontic sealer stimulates a greater production of collagen.

Resumo Este estudo visou avaliar a capacidade de reparação de tecidos de quatro cimentos endodônticos biocerâmicos através da quantificação de fibras colágenas de tipo I e III. Foram testados os seguintes cimentos: EndoSequence BC Sealer (Brasseler, Brasseler, Savannah, EUA), Bio C Sealer (Angelus, Londrina, Brasil), Bioroot RCS (Septodont, Santa Catarina, Brasil), e Sealer Plus BC (MKLife, Porto Alegre, Brasil). Foram implantados tubos de polietileno de 1,5 mm de diâmetro e 1 cm de comprimento contendo os cimentos endodônticos no tecido subcutâneo de cinco ratos (Rattus norvegicus albinus, linhagem Wistar). Após 14 dias, os animais foram eutanasiados e as fibras colágenas foram quantificadas a partir de cortes histológicos do tecido. Diante de uma distribuição não-normal dos dados, uma regressão gama com função de ligação log, implementada por meio do módulo de modelos lineares generalizados, foi empregada para testar se havia diferença significativa entre os cimentos. A comparação dois a dois foi realizada utilizando Least significant difference. Houve diferença significativa entre os cimentos para os colágenos tipo I (p=0,001), tipo III (p=0,023) e colágeno total (p=0,002). No geral, o cimento Bioroot foi estatisticamente superior aos demais cimentos, com exceção na análise do colágeno tipo III na qual não houve diferença entre o cimento Bioroot e o cimento Bio C Sealer e o grupo controle (p>0,05). O cimento endodôntico biocerâmico Bioroot RCS foi capaz de estimular uma maior produção de colágeno.

Experimental investigation of early assessment of corpora cavernosa fibrosis with two-dimensional shear wave elastography / 亚洲男科学杂志(英文版)

This study explored the usefulness of two-dimensional shear wave elastography (2D-SWE) in the early assessment of corpora cavernosa fibrosis (CCF). New Zealand male rabbits were randomly assigned to an experimental group or a control group. Recombinant human transforming growth factor beta 1 (TGF-β1) was injected into the dorsal penis tissue of rabbits in the experimental group. Conventional ultrasound and 2D-SWE examinations were performed before and 20 days after injection. Penile histological analysis was performed by hematoxylin-eosin staining, sirius red staining, and immunohistochemistry. Measurement of 2D-SWE examination results was performed using shear wave elastography quantitative measurement (SWQ). Histological analysis outcomes were the proportion of smooth muscle cells (SMCs), collagen fibers (CFs), collagen type I (Col I), and collagen type III (Col III), as well as the SMCs/CFs ratio, measured by sirius red staining. Other histological analysis outcomes were the positive area proportion (PAP) of TGF-β1 (PAPT), fibronectin (PAPF), and Col III (PAPC), measured by immunohistochemistry. After recombinant human TGF-β1 injection, SWQ was higher in the experimental group than that in the control group (P < 0.001); however, there were no differences in conventional ultrasound results. There were significant differences in histological outcomes between the two groups (all P < 0.05). These results indicated that 2D-SWE was superior for identifying early histological changes in CCF.

Alteración del colágeno en la Hipertrofia Gingival en pacientes con ortodoncia: caracterización histológica e inmunohistoquímica / Collagen alteration in gingival hypertrophy in orthodontic patients: histological and immunohistochemical characterization

Resumen Introducción: La hipertrofia gingival (HG) es el aumento del volumen de la encía asociado a ciertas enfermedades sistémicas, hereditarias (idiopático), ingesta de algunos medicamentos o a factores locales como el tratamiento ortodóntico, capaz de provocar cambios histológicos en el tejido conectivo gingival. Objetivo: Describir las características histológicas e identificar el colágeno tipo I y tipo III en tejidos gingivales de sujetos con hipertrofia gingival portadores de ortodoncia. Materiales y método: Se diseñó un estudio de casos y controles que incluyó el análisis de biopsias de tejido gingival de 12 pacientes sometidos a cirugías periodontales. La muestra se dividió en dos grupos: individuos sanos (control; n= 6) y pacientes con HG portadores de ortodoncia (pacientes; n= 6). Las muestras fueron procesadas e incluidas en parafina. Las tinciones Masson-Goldner y rojo sirius/verde rápido fueron empleadas. El colágeno tipo I y tipo III fueron identificados mediante inmunohistoquímica con anticuerpos monoclonales. Resultado: En los pacientes con HG portadores de ortodoncia se observó un epitelio hiperplásico y tejido conectivo denso con abundantes fibras de colágeno distribuidos aleatoriamente. La inmunodetención de colágeno tipo I indicó la presencia de abundantes fibras desorganizadas y el colágeno tipo III fue inmunolocalizado subyacente a la membrana basal, vasos sanguíneos y toda la extensión del tejido conectivo de los pacientes con HG con tratamiento ortodóntico. Conclusión: La acumulación de fibras de colágeno, particularmente del colágeno tipo I y tipo III, son hallazgos histológicos que caracterizan la HG en pacientes portadores de ortodoncia. Futuros estudios son necesarios para dilucidar el fenotipo de los fibroblastos gingivales y la probable pérdida homeostática entre la producción y degradación de colágeno en esta patología.

Abstract Introduction: Gingival hypertrophy (GH) is the increase in the volume of the gingiva associated with certain systemic, hereditary (idiopathic) diseases, the intake of some medications or local factors such as orthodontic treatment, capable of causing histological changes in the gingival connective tissue. Objective: To describe the histological characteristics and identify type I and type III collagen in gingival tissues of subjects with gingival hypertrophy wearing orthodontics. Method: A case-control study was designed that included the analysis of gingival tissue biopsies from 12 patients submitted to periodontal surgeries. The sample was divided into two groups: healthy individuals (Control; n= 6) and patients with GH wearing orthodontics (Patients; n= 6). The samples were processed and embedded in paraffin. Masson-goldner and sirius red/fast green stains were used. Type I and type III collagen were identified by immunohistochemistry with monoclonal antibodies. Result: A hyperplastic epithelium and dense connective tissue with abundant randomly distributed collagen fibers were observed in patients with orthodontic GH. Immunodetention of type I collagen indicated the presence of abundant disorganized fibers and type III collagen was inmunolocalized underlying the basement membrane, blood vessels and the entire extension of the connective tissue of patients with GH orthodontic. Conclusion: The accumulation of collagen fibers, particularly type I and type III collagen, are histological findings that characterize GH in orthodontic wearers. Future studies are necessary to elucidate the phenotype of gingival fibroblasts and the probable homeostatic loss between collagen production and degradation in this pathology.

Vacuum sealing drainage enhances wound healing by up-regulating collagen type I/III ratio in rats / 中国组织工程研究

BACKGROUND: Vacuum sealing drainage can enhance acute and chronic wound healing. The ratio of collagen type I/III play a critical role in the structural stability of skin tissue and skin repair, but its change during vacuum sealing drainage accelerating wound healing remains unclear. OBJECTIVE: To observe the effect of vacuum sealing drainage on the ratio of collagen type I/III during wound healing and to explore the potential mechanism underlying acute wound repair in rats. METHODS: A full-thickness wound, with a diameter of 20 mm, was created on the back of healthy male rats. All model rats were then randomized into two groups: blank control and vacuum sealing drainage groups. The wound surface was photographed at three observational time points (1, 3, 7 days after operation), and wound closure rate was calculated and compared. The mRNA and protein expression levels of type I collagen and type III collagen and ratio of collagen type I/III were detected by RT-qPCR and immunohistochemistry. The structure of granulation tissue and length of re-epithelialization were histologically detected. RESULTS AND CONCLUSION: Compared with the blank control group, treatment with vacuum sealing drainage significantly increased the expression of type I collagen and type III collagen at mRNA and protein levels (P < 0.05), enhanced wound healing rate (P < 0.05) as well as increasing the ratio of collagen type I/III starting from the 3rd day after operation (P < 0.05). To conclude, the vacuum sealing drainage can accelerate wound healing by up-regulating the protein expression of type I collagen and type III collagen, the ratio of collagen type I/III and increasing wound tensile strength.

Collagenofibrotic glomerulopathy- report of a rare renal disease with serial biopsies

@#Introduction: Collagenofibrotic glomerulopathy or collagen type-III glomerulopathy is a rare glomerular disease characterised by the deposition of type III collagen fibres in the subendothelial space and mesangium of the glomerulus. Case Report: Here, we present a case of collagenofibrotic glomerulopathy in a 49-year-old Indian female, the first to be reported from Singapore. Renal biopsy showed PAS (periodic acid-Schiff), silver and Congo red negative, amorphous extracellular material that expanded mesangial and subendothelial regions. Such materials were strongly positive for anti-collagen III immunofluorescent staining. Under electron microscopy, the mesangial and some subendothelial regions were greatly expanded by abundant collagen fibres which were different from normal collagen III fibres in both appearance and periodicity. Discussion: The availability of past renal biopsies for reference offered insight into disease progression. From the initial diagnosis of focal segmental glomerulosclerosis to eventually collagenofibrotic glomerulopathy over a time span of more than 10 years, this case highlights the gradual accumulation of collagen fibres in the glomeruli before classical features are apparent. It also emphasises the importance of electron microscopy in the diagnosis of this disease.

miR-27a-3p inhibited synthesis of Col I and Col III in pulmonary fibroblasts through Wnt3a/β-catenin signaling pathway / 中国药理学通报

Aim: To explore the effects of microRNA- 27a-3p(miR-27a-3p) on collagen type I (Col I) and collagen type III (Col III) synthesis in pulmonary fibroblasts and the underlying mechanisms. Methods: Human pulmonary fibroblasts MRC-5 were cultured and then transfected with miR-27a-3p mimic or its inhibitor. qPCR and Western blot were used to detect miR- 27a-3p levels and nuclear β-catenin content, respectively. The expression of Col I, Col III and Wnt3a was measured using qPCR and Western blot. Bioinformatics predicted the potential of miR-27a-3p bound to Wnt3a 3'-untranslated region (3'-UTR). Dual luciferase reporter gene analyzed the effects of miR-27a-3p mimic transfection on luciferase activity of wild type and mutant Wnt3a 3'-UTR. MRC-5 cells were treated with the Wnt3a/β-catenin signaling pathway inhibitor Dkkl, followed by transfection with miR-27a-3p mimic or its inhibitor. Col I and Col III expression was detected by qPCR and Western blot. Results: miR-27a- 3p mimic markedly increased miR-27a-3p levels and decreased Col I, Col III, Wnt3a and β-catenin expression (P 0. 05), revealing Wnt3a as a target of miR-27a-3p. In addition, Dkkl pretreatment almost fully reversed the inductive effect of miR-27a-3p inhibitor on Col I and Col III expression in MRC-5 cells (P < 0. 05). Conclusions: miR-27a-3p inhibits the biosynthesis of Col I and Col III in pulmonary fibroblasts, which is attributed to inactivated Wnt3a/β-catenin signaling pathway.

Collagen I and III in women with diastasis recti

OBJECTIVES: Interest in elucidating the etiology of hernias has encouraged countless studies of musculoaponeurotic structures in individuals with and without hernias. Studies of hernia patients have firmly demonstrated a correlation between hernias and collagen alterations in their fascia. Diastasis recti is an increased width of the abdominal midline that is exclusively composed of interlacing aponeurotic expansions of the anterolateral abdominal muscles. The condition is common among women undergoing abdominoplasty, and many factors, not only mechanical, play a role. The goal of this study is to evaluate and compare collagen type I and III levels in the midline fascia of women with and without diastasis recti to report their possible influence on this condition. METHODS: This is a case-control study nested within a surgical cohort of 18 women with diastasis recti and 18 women without the condition (cases and controls, respectively). Fascia from the midline of the abdominal wall was collected and analyzed through immunohistochemistry using polyclonal antibodies to collagen type I and III. RESULTS: Both type I and type III collagen were less abundant in women with diastasis recti than in those without the condition, and the difference was statistically significant (p<0.001). CONCLUSION: Low collagen type I and type III levels in the midline of the abdominal wall may play a key role in the development of diastasis recti.

Biomechanic and histologic analysis of fibroblastic effects of tendon to bone healing by transforming growth factor ß1 (TGF-ß1) in rotator cuff tears

Abstract Purpose: To evaluate the effect of transforming growth factor β1 (TGF-β1) on tendon-to-bone reconstruction of rotator cuff tears. Methods: Seventy-two rat supraspinatus tendons were transected and reconstructed in situ. At 8 and 16 weeks, specimens of three groups; that is control, L-dose (low dose), and H-dose (high dose) were harvested and underwent a biomechanical test to evaluate the maximum load and stiffness values. Histology sections of the tendon-to-bone interface were identified by hematoxylin-eosin or Masson trichrome stain. Collagen type III was observed by picric acid sirius red staining under polarized light. The level of insulin-like growth factor 1 (IGF-1) and vascular endothelial growth factor (VEGF) was measured by the enzyme-linked immunosorbent assay (ELISA) method. Results: Collagen type III of the H-dose group had a significant difference in histology structure compared with the L-dose group (P<0.05). The maximum load and stiffness decreased significantly in the control group compared with the values of the L-dose and H-dose groups. The stiffness among the three groups differed significantly at the same postoperative time (P<0.05). Interestingly, progressive reestablishment of collagen type III affected tendon-to-bone healing significantly in the later stages. Conclusion: The H-dose was associated with an increased collagen type III morphology stimulated by TGF-β1.

Comparative analysis of the effects of Copaifera multijuga oil-resin and nitrofurazona in the cutaneous wound healing process / Análise comparativa dos efeitos do óleo-resina de Copaifera multijuga e da nitrofurazona na cicatrização de ferida cutânea

ABSTRACT Objectives: to evaluate, histologically and macroscopically, the influence of Copaifera multijuga (Copaíba) oil-resin on the healing process of cutaneous wounds, comparing it with nitrofurazone. Methods: we divided 36 rats into three groups of 12 animals, according to the treatment to be administered. Group SL (control) received saline on the lesion; the Group OIL received topical treatment with Copaíba oil; and the Group NITRO was treated with Nitrofurazone. We inflicted a circular wound of 8mm in diameter on the back of each animal. We subdivided each of the three groups of 12 animals into three subgroups, according to treatment time and euthanasia (7, 14 and 21 days). All animals received the proposed treatment daily. We photographed the lesions for area measurement, as well as for evaluation of macroscopic aspects. We resected and stained the scars to quantify and qualify elastic fibers, collagen, degree of epithelization, neovascularization and inflammation. Results: although the saline solution provided a faster wound closure in its initial phase, after 14 days the wound size of the three groups tested was the same. Levels of inflammation and neovascularization were similar in all three groups. The amount of collagen and elastic fibers was higher in the Nitrofurazone and Copaíba oil groups. Conclusion: in male Wistar rats, Copaifera multijuga oil-resin positively influences the healing process, but it is less effective than nitrofurazone in healing by secondary intention.

RESUMO Objetivo: avaliar histologicamente e macroscopicamente a influência do óleo-resina de Copaifera multijuga no processo de cicatrização de feridas cutâneas, comparando com o grupo submetido ao uso da nitrofurazona. Métodos: foram utilizados 36 ratos, divididos em três grupos de 12 animais, conforme o tratamento a ser administrado. Grupo SF (controle, recebeu soro fisiológico sobre a lesão), Grupo ÓLEO (tratamento tópico com óleo de Copaíba), Grupo NITRO (tratamento tópico com Nitrofurazona). Foi confeccionada uma ferida circular de 8mm de diâmetro no dorso de cada animal. Cada um dos três grupos de 12 animais foi subdividido em três subgrupos, de acordo com o tempo de tratamento e de eutanásia (7, 14 e 21 dias). Todos os animais receberam o tratamento proposto diariamente. As lesões foram fotografadas para mensuração de sua área, bem como, avaliados aspectos macroscópicos. As cicatrizes foram ressecadas e coradas, para quantificar e qualificar as fibras elásticas, colágenas, grau de epitelização, neovascularização e inflamação. Resultados: embora o soro fisiológico tenha proporcionado um fechamento mais rápido da ferida em sua fase inicial, a partir de 14 dias o tamanho das feridas dos três grupos testados se equivaleu. Níveis de inflamação e neovascularização foram semelhantes nos três grupos. A quantidade de fibras colágenas e elásticas foi maior nos grupos Nitrofurazona e Óleo de Copaíba. Conclusão: em ratos machos da linhagem Wistar, o óleo-resina de Copaifera multijuga influencia positivamente no processo de cicatrização, porém é menos eficaz que a nitrofurazona na cicatrização por segunda intenção.

Type III collagen disorders: A case report and review of literature.

Collagen type III is a normal component of interstitium and blood vessels. Collagenofibrotic glomerulopathy (CG) and nail patella syndrome (NPS) are the diseases of abnormal type III collagen deposition. In spite of these curved frayed structures with a periodicity of 45–60 nm are deposited in subendothelium and mesangium in CG, they are found only in the basement membrane in NPS. The clinical features of CG are confined to the kidney, NPS has associated extra‑renal manifestations. Electron microscopy is essential to make the renal diagnosis in both these rare diseases. Both the entities considered to be systemic diseases evidence to suggest similar deposition in other organs, understanding etiopathogenesis and disease progression await research.

Avaliação da alteração do perfil de colágeno de dentes extraídos de pacientes submetidos a radioterapia de cabeça e pescoço

O comprometimento da saúde dental pela incidência de radiação é tido comoresultado da redução do fluxo salivar bem como de possíveis danos radiogênicosdiretos à estrutura dental. A natureza exata destes últimos ainda está por serelucidada. A partir deste contexto, o presente estudo objetiva analisar apredominância anatômica de colágeno tipo I e colágeno tipo III da dentina e doligamento periodontal assim como a quantidade de colágenos tipos I e III doligamento periodontal de dentes contidos nos campos de radiação extraídos depacientes submetidos a radioterapia (RT) para tratamento de neoplasias malignasde cabeça e pescoço. A amostra foi constituída de 20 dentes, sendo oito dentescariados submetidos à dose total de radiação menor que 60 Gy (grupo experimentalI - GE I), seis dentes cariados submetidos à dose total de radiação igual ou maiorque 60 Gy (grupo experimental II - GE II) e seis dentes hígidos erupcionados nãosubmetidos a radiação (grupo controle - GC). Foi realizado estudo histomorfométricoatravés da técnica de coloração Picrosirius Red com luz polarizada. Análisequalitativa: em relação à dentina, percebeu-se predominância de colágeno tipo IIInas regiões circunjacentes às lesões cariosas no GE I, predominância geral decolágeno tipo III em toda a dentina no GE II e predominância geral de colágeno tipo Ina dentina do GC...

Compromised dental health by incidence of radiation is taken as a result of thereduction in salivary flow as well as possible direct radiogenic damage to toothstructure. The exact nature of the latter remains to be elucidated. The aim of thispaper is to investigate the anatomical predominance of collagen type I and type III inthe dentin and periodontal ligament and the amount of these collagens in theperiodontal ligament of extracted teeth within the radiation fields of patients who haveunderwent radiotherapy (RT) for treatment of malignant head and neck tumors. Thesample consisted of 20 teeth, 8 carious teeth of patients who underwent total dose ofless radiation than 60 Gy (experimental group I - EG I), 6 carious teeth of patientswho underwent total radiation dose equal to or greater than 60 Gy (experimentalgroup II - EG II) and 6 healthy erupted teeth not subjected to radiation (control group- CG). A histomorphometrical study was conducted using the staining techniquePicrosirius Red with polarized light. Qualitative analysis of dentin noticed thepredominance of type III collagen in the surrounding regions to carious lesions in EGI, general predominance of type III collagen throughout the dentin of EG II andgeneral predominance of type I collagen in the dentin of the CG...

Influencia do tempo de exposicao a obesidade induzida por dieta hiperlipidica sobre os colagenos tipo I e III miocardico / Influence of Term of Exposure to High-Fat Diet-Induced Obesity on Myocardial Collagen Type I and III

FUNDAMENTO: A obesidade é um fator de risco para muitas complicações médicas; a pesquisa médica demonstrou que as alterações hemodinâmicas, morfológicas e funcionais estão correlacionadas com a duração e gravidade da obesidade. OBJETIVO: O presente estudo determinou a influência do tempo de exposição à obesidade induzida por dieta com alto teor de gordura no colágenos tipo I e III miocárdico. MÉTODOS: Ratos machos com trinta dias de idade, da raça Wistar, foram distribuídos aleatoriamente em dois grupos: um grupo de controle (C) alimentado com ração padrão e um grupo de ratos obesos (Ob) alternadamente alimentados com uma de quatro dietas palatáveis ricas em gordura. Cada dieta foi mudada diariamente, e os ratos foram mantidos em suas respectivas dietas por 15 (C15 e Ob15) e 30 (C30 e Ob30) semanas consecutivas. A obesidade foi determinada pelo índice de adiposidade. RESULTADOS: O grupo Ob15 foi similar ao grupo C15 em relação à expressão de colágeno miocárdico tipo I; contudo, a expressão no grupo Ob30 foi menor do que no grupo C30. O tempo de exposição à obesidade foi associado com uma redução de colágeno do tipo I no grupo Ob30, quando comparado com o Ob15. A obesidade não afetou a expressão do colágeno tipo III. CONCLUSÃO: Este estudo mostrou que o tempo de exposição à obesidade por 30 semanas induzida por uma dieta rica em gordura insaturada causou uma redução na expressão do colágeno miocárdico tipo I em ratos obesos. No entanto, nenhum efeito foi observado em relação à expressão do colágeno miocárdico tipo III .

BACKGROUND: Obesity is a risk factor for many medical complications; medical research has shown that hemodynamic, morphological and functional abnormalities are correlated with the duration and severity of obesity. OBJECTIVE: Present study determined the influence of term of exposure to high-fat diet-induced obesity on myocardial collagen type I and III. METHODS: Thirty-day-old male Wistar rats were randomly distributed into two groups: a control (C) group fed a standard rat chow and an obese (Ob) group alternately fed one of four palatable high-fat diets. Each diet was changed daily, and the rats were maintained on their respective diets for 15 (C15 and Ob15) and 30 (C30 and Ob30) consecutive weeks. Obesity was determined by adiposity index. RESULTS: The Ob15 group was similar to the C15 group regarding the expression of myocardial collagen type I; however, expression in the Ob30 group was less than C30 group. The time of exposure to obesity was associated with a reduction in collagen type I in Ob30 when compared with Ob15. Obesity did not affect collagen type III expression. CONCLUSION: This study showed that the time of exposure to obesity for 30 weeks induced by unsaturated high-fat diet caused a reduction in myocardial collagen type I expression in the obese rats. However, no effect was seen on myocardial collagen type III expression. .

Non-cross-linked collagen type I/III materials enhance cell proliferation: in vitro and in vivo evidence

Objective: To analyze Mucograft®(MG), a recently introduced collagen matrix, in vitro and in vivo, and compare it with BioGide®(BG), a well-established collagen membrane, as control. Material and Methods: A detailed analysis of the materials surface and ultra-structure was performed. Cellular growth patterns and proliferation rates of human fibroblasts on MG and BG were analyzed in vitro. In addition, the early tissue reaction of CD-1 mouse to these materials was analyzed by means of histological and histomorphometrical analysis. Results: MG showed a three-fold higher thickness both in dry and wet conditions, when compared to BG. The spongy surface of BG significantly differed from that of MG. Cells showed a characteristic proliferation pattern on the different materials in vitro. Fibroblasts tended to proliferate on the compact layers of both collagens, with the highest values on the compact side of BG. In vivo, at day three both materials demonstrated good tissue integration, with a mononuclear cell sheet of fibroblasts on all surfaces, however, without penetrating into the materials. Conclusions: The findings of this study showed that MG and BG facilitate cell proliferation on both of their surfaces in vitro. In vivo, these two materials induce a comparable early tissue reaction, while serving as cell occlusive barriers. .

The effect of cycle dynamic tensile forces on tissue engineered tendon in vitro / 中华骨科杂志

Objective To investigate the tissue remodeling and cell alignment of TDBM scaffolds seeded with rabbit tenocytes under the cycle dynamic tensile force or static tension-free culture in vitro. Methods TDBM were made by ourselves, and uniaxial cyclic tendon stretching device was designed and manufactured on our own. Primary tenocytes were isolated from the Achilles tendon of three-day-old New Zealand white rabbits and seeded into scaffolds, and were cultured collectively in DMEM in vitro. Samples were divided into two groups:dynamic tension-loaded group, and static tension-free group. Fresh natural tendons were used to be positive control. The experiment's time was six weeks. The scaffold-cell complexes were harvested at 3 and 7 days of culture for Inverted microscope and scanning electron micrograph (SEM) analysis. The morphological characters of the samples, including the general view, HE and Masson's dyeing, were observed at 2, 4 and 6 weeks. In addition, the gene expression of the I-type collagen and III-type collagen of the samples was detected by using Real time PCR at every week. Set fresh natural tendon as control. Results The inverted microscope and SEM showed that it was nice compatible condition between the tendon cells and TD-BM scaffold. In addition, the tendon of tension-loaded group revealed a structure of longitudinally aligned collagen fi bers and dense structure of collagen fibers arranged in orderly form a unique corrugated structure. Tenocytes layer located between the col-lagen fibers and aligned longitudinally along the force axis, with increased matrix deposition after the 3th week showed by RT-PCR. The cell/matrix ratio decreases. When cultured to 6 weeks, the tissue structure was very similar to that of fresh natural ten-don pattern. By contrast, HE and Masson's staining revealed the collagen fibro-tissue structure in tension-free groups with disorga-nized matrix structure and randomly distributed cells. Collagen fibers were gradually degraded with time. In tension-loaded group, Real-time PCR showed that gene expression of I-type collagen and III-type collagen gradually increase, but in tension-free group, the relative gene expression of I-type collagen and III-type collagen was highest at 3rd week, and from that time the relative expres-sion gradually decrease. Conclusion Under the dynamic stress, the TDBM scaffolds seeded with rabbit tenocytes can promote extra-cellular matrix biosynthesis and tendon structure remodeling, which is a promising method for tendon tissue engineering.

Análise da expressão das metaloproteinases e seus inibidores teciduais no músculo detrusor de pacientes com obstrução infravesical por hiperplasia prostática benigna / Expression of metalloproteinases and their tissue inhibitors in the detrusor muscle of patients with bladder outlet obstruction due to benign prostatic hyperplasia

Introdução: A obstrução infravesical (OIV) de longo prazo secundária a hiperplasia prostática benigna (HPB) pode causar alterações funcionais e morfológicas na bexiga. Um dos principais eventos consiste no aumento da deposição de colágeno e perda de complacência vesical, levando a alteração de armazenamento e esvaziamento urinário. O aumento da deposição de colágeno na matriz extracelular (MEC) da musculatura detrusora é a principal razão para a diminuição da complacência vesical. Na bexiga, assim como em outros órgãos, este fenômeno depende da atividade equilibrada de enzimas proteolíticas, incluindo as metaloproteinases (MMP) e os seus inibidores endógenos (inibidores teciduais de metaloproteinases-TIMPs). Como estes fenômenos são desconhecidos na bexiga obstruída, o objetivo deste estudo foi avaliar a expressão gênica de colágeno, MMPs e seus inibidores na bexiga de pacientes com obstrução infravesical. Material e Métodos: Foi realizada uma análise prospectiva e controlada de 43 pacientes com OIV devido a HPB, que foram submetidos à ressecção transuretral da próstata (RTUP) entre 2011 e 2012. Como grupo controle foram selecionados espécimes de músculo detrusor de 10 pacientes que foram submetidos a prostatectomia radical retropúbica devido adenocarcinoma de próstata. Todos estes pacientes tinham idade menor que 60 anos, tamanho de próstata menor que 30 gramas ao ultra-som e escore internacional de sintomas prostáticos (IPSS) menor que 7. Todos os pacientes foram submetidos a estudo urodinâmico pré e pós operatório (após 6 meses). A biópsia de fragmento de músculo da bexiga foi realizada ao final da RTUP e colocada em solução estabilizadora de RNA para quantificação da expressão de colágenos I e III, metaloproteinases de matriz 1, 2 e 9, e inibidores de MMPs (TIMP1, TIMP2 e RECK) na bexiga de pacientes com HPB. Os genes descritos foram avaliados através da técnica de reação em cadeia da polimerase quantitativa em tempo real (qRT-PCR)....

Introduction: Long-term Bladder outlet obstruction (BOO) secondary to Benign prostatic Hyperplasia (BPH) can cause functional and morphological abnormalities in the bladder, such as increased collagen deposition and loss of compliance, leading to urinary storage and voiding symptoms. A decrease in bladder compliance is known to be correlated with deterioration of renal function. Increased deposition of collagen in the extracellular matrix (ECM) is the primary reason for a decreased compliance. In the bladder, as in other organs, this phenomenon is dependent on the balanced activity of proteolytic enzymes, including matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue inhibitors of metalloproteinases (TIMPs). The imbalance between MMPs and TIMPs is a key regulator in ECM turnover. Since these mechanisms are unknown in the obstructed bladder, the objective of this study was to evaluate gene expression of collagen, MMPs and their inhibitors in patients with bladder outlet obstruction due to BPH. Material and Methods: We performed a prospective and controlled analysis of 43 patients with BOO due to BPH who underwent transurethral resection of the prostate (TURP) from 2011 to 2012. The control group was comprised of 10 bladder specimens from patients with < 60 years who underwent radical prostatectomy with an International Prostatic Symptom Score (IPSS) < 8 and prostate volume < 30 grams. All patients underwent urodynamic analysis pre and post operatively after 6 months. A biopsy of the bladder muscle was performed at the end of TURP for analysis of collagen, metalloproteinases and TIMPs gene expressions. For this purpose we used the quantitative real time polymerase chain reaction method (qRT-PCR)...

Effect of curcumin on cardiac remodeling and heart function change induced by adriamycin in rats / 心血管康复医学杂志

Objective: To study the effect of curcumin on cardiac remodeling and heart function change induced by adriamycin in rats. Methods: A total of 50 SD rats were randomly divided into blank control group (n=10), adriamycin group (n=20) and curcumin intervention group (n=20). General conditions of rats, including weight, hair color, diet and activity capacity, were observed. Left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS) and left ventricular end-diastolic dimension (LVEDd) were observed by echocardiography. Rat heart was weighed and levels of cardiac collagen type Ⅰ and Ⅲ were observed using immunohistochemistry method. Results: Compared with blank control group after experiment, there were significant increase in heart weight [(168.53±26)mg vs. (208.34±31)mg], heart weight/body weight [(0.36±0.06) vs. (0.52±0.08)], and significant decrease in body weight [(469.27±34)g vs. (428.47±45)g]; significant increase in LVEDd [(4.87±0.19)mm vs. (7.49±0.83)mm], significant decrease in LVEF [(69.53±6.25)% vs. (41.46±4.38)%] and LVFS [(45.83±3.79)% vs. (17.25±2.92)%] in adriamycin group, P<0.05 all, significant increase in expression rates of collagen type Ⅰ and Ⅲ, P<0.01.Compared with adriamycin group, there were significant increase in body weight (452.46±38)g, significant decrease in heart weight (178.73±38)mg, heart weight/body weight (0.39±0.07); significant decrease in LVEDd (5.96±0.65)mm and significant increase in LVEF (53.12±5.43)% and LVFS (36.57±3.66)%, P <0.05 all; significant decrease in expression rates of collagen type Ⅰ and Ⅲ in curcumin group, P <0.01. Conclusion: Curcumin can improve cardiac remodeling and heart function induced by adriamycin in rats.

The Effects of Botulinum Toxin A on Collagen Synthesis, Expression of MMP (matrix metalloproteinases)-1,2,9 and TIMP (tissue inhibitors of metalloproteinase)-1 in the Keloid Fibroblasts

Keloids are characterized by excessive extracelluar matrix (ECM) deposition such as collagen, fibronectin, elastin, and proteoglycans in the dermis. Recently, the use of botulinum toxin A (BTXA) in the treatment of keloids have had good results. To investigate the therapeutic effect of BTXA on the keloids, we evaluated the mRNA expression of collagen type I, type III, MMP (matrix metalloproteinases)-1, and TIMP (tissue inhibitor of metalloproteinases)-1 on keloid fibroblasts (KFs, n=5) after administration of BTXA. We also evaluated the enzymatic activity of MMP-2 and 9 by using zymography with BTXA. The same process was repeated after administration of TGF-beta in addition to BTXA. Type III collagen mRNA expression was decreased significantly when BTXA was administrated on KFs regardless of the presence or absence of TGF-beta. MMP-1 mRNA expression in KFs was increased according to the BTXA concentration increment, however, not increased with TGF-beta. Moreover, MMP-2 enzymatic activity in KFs was increased when BTXA administrated regardless of the presence or absence of TGF-beta. These results suggest that the down regulation of collagen III expression, the up regulation of MMP-1, and increased MMP-2 enzymatic activity on KFs after BTXA administration are able to decrease the excess collagen deposition in keloids.

Influência da dieta hipercolesterolêmica na remodelação do colágeno da matriz extracelular da parede vesical em ratos / Influence of the hypercholesterolemic diet on the collagen remodeling of the bladder wall extracellular matrix in rats

Introdução: A bexiga é responsável em armazenar urina em volume adequado e de esvaziar seu conteúdo de forma plena. Suas propriedades miogênicas intrínsecas e viscoelásticas são as responsáveis por esta função. Disfunções vesicais podem ser decorrentes, dentre outras causas, de anormalidades intrínsecas da musculatura detrusora ou da composição de sua matriz extracelular (MEC). O colágeno corresponde a 50% do estroma vesical, possuindo importante papel na adaptação vesical a condições fisiopatológicas específicas. Os colágenos tipo I e III são os mais comuns, sendo o colágeno tipo III o primeiro a ser sintetizado em processos de reparação e fibrose. Diversas afecções como a obstrução infravesical (OIV) parcial crônica podem induzir estes processos através da remodelação da MEC e conseqüentemente alterar a função vesical. Acredita-se que a hipercolesterolemia também o faça, porém ainda não foi reproduzida tal associação a nível morfológico. O objetivo deste estudo é avaliar se dieta hipercolesterolêmica promove alterações estruturais vesicais em ratos, especialmente no que diz respeito à remodelação colágena. Método: Foram utilizadas 45 ratas da raça Wistar, de quatro semanas de idade, divididas em três grupos: 1) controle com dieta comum padrão para roedores (DN); 2) modelo de OIV com DN e 3) controle com dieta de alto teor lipídico (DATL 1,25% colesterol). Análise sérica do colesterol e fração LDL e medição do peso corporal foram realizadas em todos os animais inicialmente e no final do estudo. Com quatro semanas de estudo, as ratas dos grupos 1 e 3 foram submetidas à cirurgia simulada, enquanto os animais do grupo 2 foram efetivamente submetidos à cirurgia de OIV parcial. Após dissecção da uretra, fez-se uma ligadura parcial com Nylon 5-0, com um lúmen residual de aproximadamente 1 mm. Após seis semanas, todos os animais foram submetidos à remoção de suas bexigas e então sacrificados. Análise morfológica foi realizada através da coloração...

Purpose: Preserved bladder function is defined as the adequate storage and emptying of its urinary content. Compliance is an important factor for these functions and is directly related to the extracellular matrix composition. Its abnormalities can lead to bladder dysfunctions. The collagen represents 50% of bladder stroma, playing an important role in the bladder adaptation to specific pathologic conditions. Types I and III collagens are the most prevalent in bladder wall whereas type III collagen is the first synthesized in reparation and fibrosis processes. Bladder outlet obstruction (BOO) promotes this process and hypercholesterolemia is also believed to create conditions for it, although no morphologic association has already been demonstrated. In this study we aimed to verify if hypercholesterolemic diet promotes structural bladder wall modifications, regarding the collagen remodeling. Methods: Forty-five female heterogenic Wistar 4 weeks-old rats were divided into three groups: 1) control fed on a normal diet (ND); 2) BOO model fed on a ND and 3) control fed on a hypercholesterolemic diet (HCD 1.25% cholesterol). Initially, serum cholesterol, LDL-cholesterol and body weight were measured. Four weeks later groups 1 and 3 underwent a sham operation while group 2 underwent a partial BOO operation. After the urethra was dissected a 5-zero nylon suture was passed and tied loosely around the urethra with a 22G needle besides it. Six weeks later the bladders of all animals were removed, serum cholesterol and LDL-cholesterol analysis was performed, body weight was measured and then they were sacrificed. Morphological analysis was performed by Picrosirius red staining and immunohistochemistry for types I and III collagen. Statistical analysis was done comparing groups by the Oneway-Anova method and Tukey multiple comparisons when needed. Significance was considered when p < 0.05. Results: Wistar rats fed on a HC diet had a significant increase...

Relationship between serum concetrations of type III procollagen, hyluronic acid and histopathological findings in the liver of HCV-positive blood donors

BACKGROUND: Serologic markers have been proposed for monitoring hepatic fibrosis in chronic liver disease. Among fibrosis markers, type III procollagen (PIIIP) and hyaluronic acid have been studied in these patients. AIM: To evaluate the association between these serum markers with histological findings. METHODS: A prospective cross-sectional study was carried out with HCV-positive blood donors. The studied population included men and women whose age ranged from 18 to 60 years, with elevated liver function tests [ALT levels > 1.5 times the normal value and alterations of two or more of the following: any changes in the levels of ALT, aspartate aminotransferase, conjugated bilirrubin, gammaglobulin, gammaglutamyltranspeptidase, albumin, platelet count; alkaline phosphatase levels >1.5 times the normal value, or prothrombin time below 70 percent and above 60 percent]. Fourty-nine patients were submitted to liver biopsy, blood analysis of PIIIP, hyaluronic acid, besides liver function tests. RESULTS: Liver function tests were not associated with tissular fibrosis, as assessed by ALT (>1.5 times above normal, fibrosis risk=18.8 percent; <1.5 times, 11.8 percent). Elevated PIIIP was correlated with 66.7 percent chance of fibrosis, whereas normal levels, 9.3 percent. Hyaluronic acid, when elevated, gave a chance of 33.3 percent of fibrosis; when normal, 12.5 percent. CONCLUSION: There was no association between liver function tests, hyaluronic acid and fibrosis. However, PIIIP was related with liver fibrosis. Maybe, this marker should be useful to assess fibrosis in patients with chronic hepatitis C.

RACIONAL: Marcadores sorológicos têm sido propostos para monitorar fibrose hepática em doença crônica do fígado. Dentre os marcadores de fibrose, ácido hialurônico e procolágeno tipo III têm sido estudados nestes pacientes. OBJETIVO: Avaliar a associação de marcadores séricos de fibrose com achados histológicos. MÉTODOS: Foi realizado estudo transversal prospectivo em doadores de sangue anti-HCV positivos. A população estudada incluiu homens e mulheres com idade entre 18-60 anos com provas de função hepática alteradas (níveis de alanina aminotransferase >1.5 vezes do normal e alterações de dois ou mais dos seguintes: qualquer alteração nos níveis de alanina aminotransferase, aspartato aminotransferase, bilirrubina conjugada, gamaglobulina, gamaglutamiltranspeptidase, albumina, plaquetas, níveis de fosfatase alcalina >1,5 vezes o valor normal, tempo de protrombina abaixo de 70 por cento e acima de 60 por cento). Quarenta e nove pacientes foram submetidos a biopsia hepática e coleta de sangue para análise de procolágeno tipo III, ácido hialurônico e provas funcionais hepáticas. RESULTADOS: Não houve relação entre elevação de provas de função hepática e a presença de fibrose - ALT (>1,5 vezes acima do normal, risco de fibrose = 18,8 por cento; <1,5 vezes, 11,8 por cento). Procolágeno tipo III elevado foi correlacionado com 66,7 por cento chances de fibrose, enquanto nível normal, 9,3 por cento. Ácido hialurônico, quando elevado, demonstrou chance de 33,3 por cento de fibrose; quando normal, 12.5 por cento. CONCLUSÕES: Não houve associação entre provas de função hepática, ácido hialurônico e fibrose, mas houve entre esta última e procolágeno tipo III. Talvez este marcador possa ser útil para avaliar fibrose em pacientes com hepatite crônica pelo vírus C.