Biological modulation of corneal epithelial wound healing / Modulação biológica do processo de cicatrização epitelial da córnea

ABSTRACT The transparency and maintenance of corneal epithelial integrity are essential for its optical properties and, to preserve these characteristics, the epithelium undergoes continuous renewal. This renewal depends on the control of cell proliferation and differentiation mediated by mitogenic factors responsible for increasing mitoses and stimulating cellular migration. Cell-cell communication plays a pivotal role in epithelial healing process, and several cytokines and growth factors are involved in this process. Understanding the cross-talk and paracrine effects of these cytokines and growth factors released can help in the search for new therapeutic strategies to treat ocular surface diseases.

RESUMO A transparência e a manutenção da integridade epitelial da córnea são essenciais para suas propriedades ópticas e, para preservar tais características, o epitélio sofre renovação contínua. Essa renovação depende do controle da proliferação e diferenciação celular mediadas por fatores mitogênicos responsáveis pelo aumento das mitoses e estímulo à migração celular. A comunicação célula-célula desempenha um papel fundamental no processo de cicatrização epitelial, e várias citocinas e fatores de crescimento estão envolvidos neste processo. Compreender os efeitos cruzados e paracrinos dessas citocinas e fatores de crescimento liberados pode ajudar na busca de novas estratégias terapêuticas para o tratamento de doenças da superfície ocular.

Quemerina e fatores relacionados ao risco cardiovascular em crianças e adolescentes: uma revisão sistemática / Chemerin and factors related to cardiovascular risk in children and adolescents: a systematic review

RESUMO Objetivo: Sintetizar os achados sobre a quemerina e os fatores relacionados ao risco cardiovascular em crianças e adolescentes. Fontes de dados: Realizou-se uma revisão sistemática de acordo com os itens propostos pela diretriz PRISMA nas bases de dados PubMed, Science Direct e Lilacs. Utilizaram-se os descritores chemerin de forma associada a children e adolescent, sem limite de tempo. A pesquisa limitou-se a artigos originais realizados com seres humanos, em língua inglesa, excluindo-se a população adulta e idosa, assim como os artigos de revisão, comunicação breve, cartas e editoriais. Síntese dos dados: Após análise dos estudos por dois revisores, de forma independente, segundo os critérios de elegibilidade, permaneceram na revisão sete artigos, publicados entre 2012 e 2016. Foram incluídos estudos de delineamento transversal, prospectivo, coorte e caso-controle. A importância da adipocina quemerina nos fatores de risco para doenças cardiovasculares é demonstrada por meio de sua associação com obesidade e diabetes melito, assim como com parâmetros clínicos, antropométricos e bioquímicos. Entretanto, a força da evidência dos estudos é relativamente baixa, em função da heterogeneidade das publicações, destacando-se como limitações o número reduzido das amostras e sua ausência de representatividade, a falta de padronização dos métodos de dosagem, o delineamento transversal de grande parte dos estudos e a impossibilidade de extrapolação dos resultados. Conclusões: A desregulação da quemerina provocada pelo aumento de tecido adiposo pode contribuir para o aparecimento de doenças cardiovasculares, sugerindo que tal adipocina tem papel relevante na identificação precoce de indivíduos em risco.

ABSTRACT Objective: To review findings on chemerin and factors related to cardiovascular risk in children and adolescents. Data source: A systematic review was performed, according to the standards proposed by the PRISMA guideline, on PubMed, Science Direct, and Lilacs databases. The descriptor "chemerin" was used in combination with "children" and "adolescent", no time limit applied. The research encompassed only original articles written in English, conducted with human subjects - the adult and elderly populations excluded -, as well as literature reviews, brief communications, letters, and editorials. Data synthesis: After independent analyses of the studies by two reviewers, seven articles meeting the eligibility criteria, published between 2012 and 2016, remained for the review. Cross-sectional, prospective, cohort, and case-control studies were included. The importance of chemerin adipokines on the risk factors for cardiovascular disease is demonstrated by its association with obesity and diabetes mellitus, as well as clinical, anthropometric, and biochemical parameters. However, the strength of evidence from these studies is relatively low, due to their heterogeneity, with several limitations such as small samples and consequent lack of representativeness, lack of standardization in dosage methods, cross-sectional design of most studies, and impossibility of extrapolating results. Conclusions: The deregulation of chemerin caused by increased adipose tissue may contribute to the development of cardiovascular diseases, suggesting that this adipokine may play a significant role in early identification of individuals at risk.

Role of vitamin K-dependent protein Gas6 in the expression of endothelial cell adhesion molecule-1 and chemokines induced by Porphyromonas gingivalis lipopolysaccharide / 北京大学学报(医学版)

OBJECTIVE@#Growth-arrest-specific protein 6 (Gas6) is a vitamin K-dependent protein and involved in cell proliferation, survival, adhesion and migration . Also it has been shown to play an important role in the inflammatory response .The aim of present study was to investigate the role of Gas6 in the process of the expression of adhesion molecules and chemokines of human umbilical vein endothelial cells (HUVECs) induced by Porphyromonas gingivalis lipopolysaccharide(P.g-LPS).@*METHODS@#After up-regulation and down-regulation of the expression of Gas6, the vascular endothelial cells were stimulated with 1 mg/L P.g-LPS for 3 h and 24 h. Real-time quantitative polymerase chain reaction(real-time PCR) was taken to detect the expression of the cell adhesion molecules:intercellular adhesion molecule-1 (ICAM-1) and E-selectin, as well as chemokines:interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1). Wound healing assay was taken to observe the migration ability of endothelium cells in different groups.@*RESULTS@#After 3 h of P.g-LPS stimulation, the expression of adhesion molecules and chemokine in the down-regulation group was not significantly different from that in the control group,while in the up-regulation group the decrease of E-selectin, ICAM-1, IL-8 and MCP-1 was 81%±0%, 47%±3%, 76% ± 3%, 26% ± 6% respectively. After 24 h of P.g-LPS stimulation, the expression of adhesion molecules and chemokine in down-regulation group was significantly higher than that in control group (2.06±0.07, 1.99±0.11, 3.14±0.15, 1.84±0.03 flod), while these molecules in the down-regulation group was significantly lower than in the control group (29%±1%, 62%±3%, 69%±1%, 41%±2%). Differences were statistically significant (P<0.01). Wounding healing assay showed that down-regulation of Gas6 enhanced migration ability of endothelial cells while up-regulation of Gas6 weakened this ability,which was consistent with the trend of real-time PCR result.@*CONCLUSION@#Down-regulation of the Gas6 gene enhanced the expression of ICAM-1, E-selectin, IL-8 and MCP-1 in HUVECs after P.g- LPS stimulating, while up-regulaiton of the Gas6 gene weakened the expression of ICAM-1, E-selectin, IL-8 and MCP-1 in HUVECs after P.g-LPS stimulating,suggesting that Gas6 may play a role in the process of endothelial cell adhesion.

Effects of concentrated growth factors on the angiogenic properties of dental pulp cells and endothelial cells: an in vitro study

Abstract The aim was to investigate the angiogenic effects of concentrated growth factors on human dental pulp cells and human umbilical vein endothelial cells. Cells were treated with concentrated growth factor extracts. The CCK-8 assay and cell cycle assay were conducted to evaluate cell growth. Cell migration was evaluated by the Transwell migration assay. Angiogenesis-associated mRNA and protein expression levels were determined using quantitative real-time PCR and Western blotting, respectively. A tube formation assay was conducted to evaluate the angiogenic capacity in vitro. The data showed that compared with the control, concentrated growth factor extracts significantly promoted dental pulp cell proliferation and differentiation and endothelial cell proliferation and migration in a dose-dependent manner (p < 0.05). Concentrated growth factor extracts also promoted the tube-like structure formation of endothelial cells in vitro. The RT-PCR and Western blot results showed that concentrated growth factor extracts upregulated the expression of angiogenesis-related genes - chemokine receptor-4, platelet-derived growth factor, and vascular endothelial growth factor - in dental pulp cells. In conclusion, concentrated growth factors showed proangiogenic effects on dental pulp cells and endothelial cells and have good application potential for dental pulp revascularization.

Growth factors and cox2 in wound healing: an experimental study with ehrlich tumors / Avaliação dos fatores de crescimento e cox-2 na cicatrização de camundongos com neoplasia de ehrlich

ABSTRACT Background: Healing is an innate biological phenomenon, and carcinogenesis acquired, but with common humoral and cellular elements. Carcinogenesis interferes negatively in healing. Aim: To evaluate the histological changes in laparotomy scars of healthy Balb/c mice and with an Ehrlich tumor in its various forms of presentation. Methods: Fifty-four mice were divided into three groups of 18 animals. First group was the control; the second had Ehrlich tumor with ascites; and the third had the subcutaneous form of this tumor. Seven days after tumor inoculation, all 54 mice were submitted to laparotomy. All of the animals in the experiment were operated on again on 7th day after surgery, with resection of the scar and subsequent euthanasia of the animal. The scars were sent for histological assessment using immunohistochemical techniques to evaluate Cox-2 (cyclooxygenase 2), VEGF (vascular endothelial growth factor) and FGF (fibroblast growth factor). Semi-quantitatively analysis was done in the laparotomy scars and in the abdominal walls far away from the site of the operation. Results: Assessing the weight of the animals, the correct inoculation of the tumor and weight gain in the group with tumoral ascites was observed. The histological studies showed that groups with the tumor showed a statistically significant higher presence of Cox-2 compared to the control. In the Cox-2 analysis of the abdominal wall, the ascites group showed the most significant difference. VEGF did not present any significant differences between the three groups, regardless of the site. The FGF showed a significant increase in animals with the tumor. Conclusion: Histological findings in both laparotomy scar and the abdominal wall showed that with Ehrlich's neoplasia there was an exacerbated inflammatory response, translated by more intense expression of Cox-2 and greater fibroblast proliferation, translated by more intense expression of FGF, that is, it stimulated both the immediate inflammatory reactions, observed with Cox-2 reactions, and late scarring by fibroblasts and FGF.

RESUMO Racional: A cicatrização é fenômeno biológico inato, e a carcinogênese adquirido, mas com elementos humorais e celulares comuns. A carcinogênese interfere de forma negativa na cicatrização. Objetivo: Avaliar as modificações histológicas nas cicatrizes laparotômicas de camundongos Balb/c sadios como controles, e com a neoplasia de Ehrlich, em suas diferentes formas de apresentação. Métodos: Foram utilizados 54 camundongos, divididos em três grupos de 18 animais cada um. O primeiro era controle; o segundo com a neoplasia de Ehrlich em sua forma ascítica; e o terceiro na forma subcutânea. Sete dias após a inoculação do tumor, todos os 54 camundongos foram submetidos à laparotomia e reoperados no sétimo dia de pós-operatório, com ressecção da cicatriz e posterior eutanásia. As cicatrizes foram encaminhadas para estudo histológico com técnicas imunoistoquímicas para avaliar Cox-2 (ciclo-oxigenase 2), VEGF (fator de crescimento do endotélio vascular) e FGF (fator de crescimento dos fibroblastos) e analisadas de forma semiquantitativana tanto na cicatriz laparotômica como na parede abdominal mais distante do local operado. Resultados: Avaliando o peso, observou-se a correta inoculação do tumor e o aumento de peso no grupo com a neoplasia na modalidade ascítica. Os estudos histológicos mostraram que os grupos com a neoplasia apresentaram maior presença da Cox-2 em relação ao controle, estatisticamente significante. No estudo da Cox-2 da parede abdominal foi o local em que o grupo ascítico apresentou a diferença mais expressiva. O VEGF não apresentou diferenças significantes entre os três grupos, independentemente do local estudado. O FGF teve aumento significante nos animais com neoplasia. Conclusão: Os achados histológicos encontrados tanto na cicatriz das laparotomias quanto na parede abdominal mostraram que com a neoplasia de Ehrlich houve resposta inflamatória exacerbada, traduzida por expressão mais intensa da Cox-2 e maior proliferação fibroblástica, traduzida por expressão mais intensa do FGF, ou seja, estimulou tanto as reações inflamatórias imediatas, observadas nas reações da Cox-2, como nas cicatriciais tardias com os fibroblastos e o FGF.

Orthodontic movement in deciduous teeth

Deciduous teeth exfoliate as a result of apoptosis induced by cementoblasts, a process that reveals the mineralized portion of the root while attracting clasts. Root resorption in deciduous teeth is slow due to lack of mediators necessary to speed it up; however, it accelerates and spreads in one single direction whenever a permanent tooth pericoronal follicle, rich in epithelial growth factor (EGF), or other bone resorption mediators come near. The latter are responsible for bone resorption during eruption, and deciduous teeth root resorption and exfoliation. Should deciduous teeth be subjected to orthodontic movement or anchorage, mediators local levels will increase. Thus, one should be fully aware that root resorption in deciduous teeth will speed up and exfoliation will early occur. Treatment planning involving deciduous teeth orthodontic movement and/or anchorage should consider: Are clinical benefits relevant enough as to be worth the risk of undergoing early inconvenient root resorption?.

O dente decíduo é esfoliado graças à apoptose em seus cementoblastos, que desnuda a parte mineralizada da raiz e atrai os clastos. A rizólise é lenta, pois faltam mediadores em quantidade para acelerar o processo, mas ela se acelera e unidireciona quando se aproxima um folículo pericoronário de dente permanente rico em EGF e outros mediadores da reabsorção óssea - os responsáveis pelas reabsorções óssea na erupção e dentária decídua na rizólise e esfoliação. Se houver movimentação ortodôntica ou ancoragem em dentes decíduos, aumenta-se, também, o nível local desses mesmos mediadores, devendo-se estar bem consciente de que haverá uma aceleração da rizólise e, em decorrência, uma antecipação de sua esfoliação. No planejamento de casos em que dentes decíduos estejam envolvidos na movimentação ortodôntica e/ou ancoragem, deve-se ponderar: o benefício clínico para o paciente será relevante, a ponto de valer o risco de uma rizólise abreviada e inconveniente?.

Platelet-rich plasma in bone repair of irradiated tibiae of Wistar rats / Plasma rico em plaquetas na reparação óssea em tíbias irradiadas de ratos Wistar

PURPOSE: To evaluate the influence of PRP addition on bone repair of circular defects created in irradiated tibiae of rats by histometric analysis. METHODS: Sixty male Wistar rats had the right tibiae irradiated with 30 Gy. After 30 days monocortical defects were created and platelet-rich plasma was applied in 30 rats. In the control group defects were created but not filled. The animals were desanguinated after 4, 7, 14, 21, 56 and 84 days and the tibiae removed for histological processing. RESULTS: There was a tendency in the PRP group to increased bone neoformation from 14-days to 84-days; in the control group increased bone neoformation was not seen after 21 days or later. CONCLUSION: The addition of platelet-rich plasma had a beneficial effect in the initial cellular regeneration period and enhanced bone formation in later periods when compared to control.

OBJETIVO: Avaliar histometricamente a influência do PRP na reparação óssea de defeitos circulares criados em tíbia irradiada de ratos Wistar. MÉTODOS: Sessenta ratos machos tiveram a tíbia direita irradiada com 30Gy. Após 30 dias, defeitos ósseos monocorticais foram criados e PRP foi adicionado em 30 ratos. No grupo controle os defeitos foram criados, mas não preenchidos. Os animais foram sacrificados em 4, 7, 14, 21, 56 e 84 dias e a tíbia removida para processamento histológico. RESULTADOS: Houve uma tendência do grupo PRP mostrar uma neoformação óssea significativamente maior nos períodos de 14 a 84 dias; no grupo controle o aumento da reparação óssea não se manteve após 21 dias. CONCLUSÃO: A adição de PRP mostrou-se benéfica no período inicial de reparação celular e em períodos seqüentes foi estímulo à quantidade neoformação óssea, quando comparado ao controle.

A DNA enzyme targeting Egr-1 inhibits rat vascular smooth muscle cell proliferation by down-regulation of cyclin D1 and TGF-â1

We have demonstrated that a synthetic DNA enzyme targeting early growth response factor-1 (Egr-1) can inhibit neointimal hyperplasia following vascular injury. However, the detailed mechanism of this inhibition is not known. Thus, the objective of the present study was to further investigate potential inhibitory mechanisms. Catalytic DNA (ED5) and scrambled control DNA enzyme (ED5SCR) were synthesized and transfected into primary cultures of rat vascular smooth muscle cells (VSMCs). VSMC proliferation and DNA synthesis were analyzed by the MTT method and BrdU staining, respectively. Egr-1, TGF-â1, p53, p21, Bax, and cyclin D1 expression was detected by RT-PCR and Western blot. Apoptosis and cell cycle assays were performed by FACS. Green fluorescence could be seen localized in the cytoplasm of 70.6 ± 1.52 and 72 ± 2.73 percent VSMCs 24 h after transfection of FITC-labeled ED5 and ED5SCR, respectively. We found that transfection with ED5 significantly inhibited cultured VSMC proliferation in vitro after 24, 48, and 72 h of serum stimulation, and also effectively decreased the uptake of BrdU by VSMC. ED5 specifically reduced serum-induced Egr-1 expression in VSMCs, further down-regulated the expression of cyclin D1 and TGF-â1, and arrested the cells at G0/G1, inhibiting entry into the S phase. FACS analysis indicated that there was no significant difference in the rate of apoptosis between ED5- and ED5SCR-transfected cells. Thus, ED5 can specifically inhibit Egr-1 expression, and probably inhibits VSMC proliferation by down-regulating the expressions of cyclin D1 and TGF-â1. However, ED5 has no effect on VSMC apoptosis.

O sistema apelinérgico: papel na fisiologia e patologia humanas e potenciais aplicações terapêuticas / The apelinergic system: the role played in human physiology and pathology and potential therapeutic applications

A apelina é um peptídeo recentemente descoberto e identificado como o ligando endógeno do receptor APJ. A apelina e o receptor APJ são expressos numa grande variedade de tecidos, tais como coração, cérebro, rins e pulmões, onde a sua interação pode ter importantes efeitos fisiopatológicos. Com efeito, a última década foi fértil no esclarecimento de possíveis papéis desempenhados pela apelina na fisiologia humana, nomeadamente como peptídeo regulador dos sistemas cardiovascular, hipotálamo-hipófisário, gastrointestinal e imunitário. Um possível envolvimento da apelina na patogênese de doenças com elevada prevalência e co-morbilidades, como a hipertensão arterial, a insuficiência cardíaca e o diabete melito tipo 2, perspectivam-na como um possível alvo terapêutico a explorar no futuro. Este trabalho fornece uma visão geral dos efeitos fisiológicos da apelina e apresenta o possível papel desse peptídeo na patogênese de várias doenças, associado a implicações terapêuticas que poderão vir a ser, assim, exploradas.

Apelin is a recently discovered peptide, identified as an endogenous ligand of receptor APJ. Apelin and receptor APJ are expressed in a wide variety of tissues including heart, brain, kidneys and lungs. Their interaction may have relevant pathophysiologic effects in those tissues. In fact, the last decade has been rich in illustrating the possible roles played by apelin in human physiology, namely as a regulating peptide of cardiovascular, hypothalamus-hypophysis, gastrointestinal, and immune systems. The possible involvement of apelin in the pathogenesis of high prevalence conditions and comorbidities - such as hypertension, heart failure, and Diabetes Mellitus Type 2 (T2DM) - rank it as a likely therapeutic target to be investigated in the future. The present paper is an overview of apelin physiologic effects and presents the possible role played by this peptide in the pathogenesis of a number of conditions as well as the therapeutic implications that might, therefore, be investigated.

The Effect of Interleukin-4 and Amphiregulin on the Proliferation of Human Airway Smooth Muscle Cells and Cytokine Release

Airway smooth muscle (ASM) hyperplasia and angiogenesis are important features associated with airway remodeling. We investigated the effect of IL-4 and amphiregulin, an epidermal growth factor family member, on the proliferation of human ASM cells and on the release of vascular endothelial growth factor (VEGF) and monocyte chemotactic protein (MCP)-1 from human ASM cells. Human ASM cells were growth-arrested for 48 hr and incubated with platelet-derived growth factor (PDGF)- BB, interleukin (IL)-4, amphiregulin, and VEGF to evaluate cell proliferation. The cells were treated with PDGF, IL-4 and amphiregulin to evaluate the release of VEGF, MCP-1. IL-4 suppressed unstimulated and PDGF-stimulated ASM cell proliferation. Amphiregulin stimulated ASM cell proliferation in a dose-dependent manner. VEGF did not have any influence on ASM cell proliferation. IL-4 stimulated VEGF secretion by the ASM cells in a dose-dependent manner and showed added stimulatory effects when co-incubated with PDGF. Amphiregulin did not promote VEGF secretion. IL-4 and amphiregulin showed no stimulatory effects on MCP-1 secretion. The results of this study showed that IL-4 had bifunctional effects on airway remodeling, one was the suppression of the proliferation of the ASM cells and the other was the promotion of VEGF release by the ASM cells, and amphiregulin can promote human ASM cell proliferation.

Effect of the human follicle-stimulating hormone-binding inhibitor and its N-terminal fragment on follicle-stimulating hormone-induced progesterone secretion by granulosa cells in vitro.

Intrafollicular factors play an important role in folliculogenesis. The follicle-stimulating hormone (FSH)-binding inhibitor (FSHBI), purified by our laboratory from human ovarian follicular fluid, has been shown to suppress ovulation and induce follicular atresia/apoptosis in mice as well as impair fertility in marmosets, the New World monkeys. The octapeptide, a peptide corresponding to the N-terminal region of human FSHBI (hFSHBI), has been synthesized and also shows FSHBI activity in vitro. In the present study, we have attempted to identify the mechanism of action of the peptide in granulosa cell cultures. Rat granulosa cell cultures were treated with varying concentrations of the octapeptide or partially purified hFSHBI (gel chromatography fraction hGF 2) in the presence or absence of human FSH (hFSH) and the amount of progesterone (P;4) secreted in the culture supernatants after 3 h/48 h was estimated. Both hGF2 and the octapeptide failed to alter basal levels as well as 8-bromo cAMP-induced P;4 production, while FSH-induced P 4 secretion was inhibited in a dose-dependent manner. These studies reveal that the octapeptide, a fragment of FSHBI, and the native protein have similar activity in vitro and both compounds alter FSH action at the receptor level upstream of cAMP formation.

Blastocyst-endometrium interaction: intertwining a cytokine network

The successful implantation of the blastocyst depends on adequate interactions between the embryo and the uterus. The development of the embryo begins with the fertilized ovum, a single totipotent cell which undergoes mitosis and gives rise to a multicellular structure named blastocyst. At the same time, increasing concentrations of ovarian steroid hormones initiate a complex signaling cascade that stimulates the differentiation of endometrial stromal cells to decidual cells, preparing the uterus to lodge the embryo. Studies in humans and in other mammals have shown that cytokines and growth factors are produced by the pre-implantation embryo and cells of the reproductive tract; however, the interactions between these factors that converge for successful implantation are not well understood. This review focuses on the actions of interleukin-1, leukemia inhibitory factor, epidermal growth factor, heparin-binding epidermal growth factor, and vascular endothelial growth factor, and on the network of their interactions leading to early embryo development, peri-implantatory endometrial changes, embryo implantation and trophoblast differentiation. We also propose therapeutical approaches based on current knowledge on cytokine interactions.

Fisiopatologia do transtorno afetivo bipolar: o que mudou nos últimos 10 anos? / Physiopathology of bipolar disorders: what have changed in the last 10 years?

Apesar dos crescentes esforços para o entendimento da neurobiologia do transtorno afetivo bipolar (TAB), sua exata fisiopatologia permanece indeterminada. Inicialmente, a pesquisa estava voltada para o estudo das aminas biogênicas, devido aos efeitos dos diversos agentes psicofarmacológicos. Mais recentemente, evidências apontam que disfunções nos sistemas de sinalização intracelular e de expressão gênica podem estar associadas ao TAB. Estas alterações podem estar associadas a interrupções nos circuitos reguladores do humor, como sistema límbico, estriado e córtex pré-frontal, sendo que os efeitos neuroprotetores do uso crônico dos estabilizadores de humor podem reverter este processo patológico. Este artigo tem como objetivo trazer uma atualização dos achados recentes sobre a neuroquímica do TAB.

The application of vascular endothelial growth factor in forensic science / 法医学杂志

Vascular Endothelial Growth Factor (VEGF) is a potent angiogenic factor with a key role in several pathological processes, including wound repair as well as a effective vascular permeability factor. This article review the present study of VEGF in molecular biology, the connection with repair and expression regulation and so on.