Captopril inhibits thioacetamide-induced chronic liver injury associated with the suppression of inflammation / hypoxia- inducible factor 1-alpha / profibrogenic axis-mediated hepatotoxicity in rats / Captopril inhibe la lesión hepática crónica inducida por tioacetamida asociada con la supresión de la inflamación/factor 1-alfa inducible por hipoxia/hepatotoxicidad mediada por el eje profibrogénico en rata

SUMMARY: Liver transplantation is the only available method to treat liver failure induced by chronic liver injury. We sought to determine whether the angiotensin-converting enzyme inhibitor, captopril, can inhibit the development of chronic liver injury induced by the hepatotoxic agent thioacetamide (TAA) in association with the suppression of inflammation (hsCRP, TNF-α, and IL-6) / hypoxia- inducible factor 1-alpha (HIF-1α) / profibrosis (TIMP-1, MMP-9, and α-SMA) axis that mediates liver injury. Therefore, the model group of rats was injected for eight weeks with 200 mg/kg TAA starting at week two. The protective group was pretreated with 150 mg/ kg captopril daily for two weeks prior to TAA injections and continued receiving both capropril and TAA agents until being humanely scrificed at week 10. We observed a substantial damage to liver tissue in the model group as demonstrated by a significant (p<0.0001) increase in blood and hepatic tissue levels of high sensitivity C-reactive protein (hsCRP), tumor necrosis factor-a (TNF-α), interleukin- 6 (L-6), HIF-1α, tissue inhibitor of metalloproteinases-1 (TIMP-1), matrix metalloproteinase-9 (MMP-9), alpha-smooth muscle actin (α-SMA), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). All these parameters were significantly (p<0.0244) protected by captopril. Also, a significant (p<0.0001) positive correlation was observed between a-SMA (profibrosis) and the serum and tissue levels of hsCRP, TNF-α, HIF-1α, TIMP-1, MMP-9, and ALT. Thus, these findings suggest that the induction of chronic liver injury by the hepatotoxic compound, TAA is associated with the upregulation of inflammation/HIF-1α/profibrosis, with captopril exhibiting beneficial hepatic pleotropic effects.

El trasplante de hígado es el único método disponible para tratar la insuficiencia hepática inducida por una lesión hepática crónica. Buscamos determinar si el inhibidor de la enzima convertidora de angiotensina, captopril, puede inhibir el desarrollo de lesión hepática crónica inducida por el agente hepatotóxico tioacetamida (TAA) en asociación con la supresión de la inflamación (hsCRP, TNF-α e IL-6) / factor inducible por hipoxia 1-alfa (HIF-1α) / profibrosis (TIMP-1, MMP-9 y α- SMA) eje que media la lesión hepática. Por lo tanto, al grupo modelo de ratas se le inyectó durante ocho semanas 200 mg/kg de TAA a partir de la semana dos. El grupo protector fue pretratado con 150 mg/kg de captopril al día durante dos semanas antes de las inyecciones de TAA y continuó recibiendo capropril y agentes TAA hasta que fue sacrificado en la semana 10. Observamos un daño sustancial en el tejido hepático en el grupo modelo, como lo demuestra un aumento significativo (p<0,0001) de los niveles en sangre y tejido hepático de proteína C reactiva de alta sensibilidad (hsCRP), factor de necrosis tumoral-α (TNF-a), interleucina-6 (L-6), HIF-1α, inhibidor tisular de metaloproteinasas-1 (TIMP-1), metaloproteinasa de matriz-9 (MMP-9), actina de músculo liso alfa (α-SMA), alanina aminotransferasa (ALT) y aspartato aminotransferasa (AST). Todos estos parámetros estaban significativamente (p<0,0244) protegidos por captopril. Además, se observó una correlación positiva significativa (p<0,0001) entre α-SMA (profibrosis) y los niveles séricos y tisulares de hsCRP, TNF-α, HIF-1α, TIMP- 1, MMP-9 y ALT. Por lo tanto, estos hallazgos sugieren que la inducción de daño hepático crónico por el compuesto hepatotóxico, TAA, está asociada con la regulación al alza de la inflamación/HIF-1α/profibrosis, con captopril exhibiendo efectos pleotrópicos hepáticos beneficiosos.

Terapêutica adjuvante na modulação da resposta imune no tratamento da doença periodontal: uma revisão de literatura / Adjuvant therapeutic in the modulation of the immune response in the treatment of periodontal disease: a literature review

Introdução: A Doença Periodontal tem caráter multifatorial, já que depende de condições microbiológicas, imunogenéticas e sistêmicas do hospedeiro. Representa inflamação crônica das estruturas de suporte e proteção dental. Desencadeia uma complexa estimulação imunológica, bem como a produção de citocinas inflamatórias, que mediam a destruição óssea e de tecido conjuntivo, provocando perda dental e complicações à distância. A compreensão da etiopatogênese, permitiu os conceitos de modulação, que referem-se às modificações dos aspectos danosos da resposta inflamatória. Objetivo: O presente artigo tem como objetivo realizar uma revisão dos estudos sobre as principais terapêuticas adjuvantes na modulação da resposta imune frente à doença periodontal. Revisão de Literatura: Foi realizada uma revisão da literatura, onde foram selecionados artigos científicos em inglês, publicados entre os anos 2005 a 2020, por meio das bases de dados PubMed e ScienceDirect. No decorrer das buscas, foram utilizadas as palavraschaves "Inflamation", "Periodontal Disease", "Subantimicrobial Dose of Doxycycline", "Periodontal Disease", "Host Response Modulation". Resultados e Conclusão: A literatura é bem promissora em relação à terapia de controle complementar da doença periodontal. Dessa forma, novas pesquisas nessa área podem trazer inúmeros beneficos aos pacientes, sendo, assim, um novo caminho para o contorno da resistência bacteriana(AU)

Introduction: Periodontal disease has a multifactorial character, depending on the host's microbiological, immunogenetic and systemic conditions. It represents chronic inflammation of dental support and protection structures. It triggers a complex immune stimulation, as well as the production of inflammatory cytokines, which mediate bone and connective tissue destruction, causing tooth loss and complications at a distance. The understanding of etiopathogenesis allowed the concepts of modulation, which refers to the modifications of the harmful aspects of the inflammatory response. This article has the escape of conducting a review of studies on the main mechanisms of modulation against periodontal disease. Objective: This article aims to rev iew the studies on the main modulation mechanisms in the face of periodontal disease. Literature Review: A literature review was carried out in which scientific articles were selected in English, published between 2005 and 2020, through the PubMed and ScienceDirect databases. During the searches, the keywords "Inflammation", "Periodontal Disease", "Subantimicrobial Dose of Doxycycline", "Periodontal Disease", "Host Response Modulation". Results and Conclusion: The literature is very promising with complementary control therapy for periodontal disease. Thus, new research in this area can bring countless benefits to patients, thus being a new way to bypass bacterial resistance(AU)

Efeito de soluções contendo hexametafosfato de sódio e quercetina, associados ou não ao fluoreto, sobre o desgaste erosivo dentinário in vitro / Effect of sodium hexametaphosphate and quercetin, associated or not with fluoride, on dentin erosion in vitro

O objetivo do presente estudo foi investigar o efeito de soluções contendo fluoreto (F), hexametafosfato de sódio (HMP) e quercetina (QC), sozinhos ou em associação, sobre a erosão dentinária e sobre a inibição de metaloproteinases da matriz (MMPs) -2 e -9, em protocolos in vitro. Blocos de dentina radicular bovina (4 × 4 × 2 mm; n = 96), selecionados por dureza superficial, foram aleatoriamente divididos em 8 grupos experimentais (n = 12/grupo) e tratados 2×/dia (um minuto) com as seguintes soluções: (1) água deionizada (controle negativo); (2) 1100 ppm F ("F"); (3) 1,0% HMP ("HMP"); (4) 0,03% QC ("QC"); (5) F+HMP; (6) F+QC; (7) HMP+QC; e (8) F+HMP+QC. Os blocos foram submetidos a desafios erosivos 4×/dia, por 5 dias (exposição dinâmica a ácido cítrico 50 mmol.l-1 , pH 3,2, 90 s). Em seguida, foram analisados quanto à perda dentinária (perfilometria) e à perda de dureza integrada em profundidade (área sob a curva, ∆KHN). O potencial antiproteolítico das soluções contendo F, HMP e/ou QC foi analisado por zimografia. Os dados de perda dentinária (log10) foram submetidos a ANOVA um critério, seguido do teste de Tukey. Os resultados de ∆KHN (dados brutos) foram submetidos a ANOVA dois critérios, medidas repetidas, seguido do teste HolmSidak (p< 0,05). O menor desgaste erosivo foi observado no grupo F+HMP+QC. Nas menores profundidades (5-30 µm), os blocos tratados com a solução contendo F+HMP+QC apresentaram os maiores valores de ∆KHN. A análise zimográfica mostrou que todos os tratamentos promoveram atividade antiproteolítica total da MMP-2, com exceção da QC administrada sozinha (inibição de 77%). Para MMP-9, todas as soluções contendo HMP e a associação de F+QC apresentaram atividade antiproteolítica total. Conclui-se que a adição de HMP e QC a soluções contendo F levou a uma maior proteção contra a erosão dentinária, tanto em superfície (perda dentinária) quanto em relação ao conteúdo mineral do tecido remanescente (∆KHN), além de promover uma completa inibição da atividade de MMPs -2 e -9 in vitro(AU)

The aim of the present study was to investigate the effect of solutions containing fluoride (F), sodium hexametaphosphate (HMP) and quercetin (QC), alone or in association, on dentin erosion and on the inhibition of matrix metalloproteinases (MMPs) - 2 and -9, using in vitro protocols. Bovine root dentin blocks (4 × 4 × 2 mm; n = 96), selected by surface hardness, were randomly divided into 8 experimental groups (n = 12/group) and treated 2×/day (one minute) with the following solutions: (1) deionized water (negative control); (2) 1100 ppm F ("F"); (3) 1.0% HMP ("HMP"); (4) 0.03% QC ("QC"); (5) F+HMP; (6) F+QC; (7) HMP+QC; and (8) F+HMP+QC. Blocks were submitted to erosive challenges 4×/day for 5 days (dynamic exposure to 50 mmol.l-1 citric acid, pH 3.2, 90 s). They were then analyzed for dentin loss (profilometry) and integrated hardness loss in depth (area under the curve, ∆KHN). The antiproteolytic potential of solutions containing F, HMP and/or QC was analyzed by zymography. Dentin loss results (log10 transformed) were submitted to one-way ANOVA, followed by Tukey's test. ∆KHN data (raw) were submitted to two-way, repeated-measures ANOVA, followed by the Holm-Sidak test (p< 0.05). The lowest dentin erosive wear was promoted by F+HMP+QC. At the lowest depths (5-30 µm), blocks treated with F+HMP+QC showed the highest values of ∆KHN. Zymography analysis showed that all treatments completely inhibited MMP-2 activity, except for QC administered alone (77% inhibition). For MMP-9, all the solutions containing HMP or the association of F+QC promoted total antiproteolytic activity. It was concluded that the addition of HMP and QC to F solutions led to greater protection against dentin erosion, both at the surface (dentin loss) and in relation to the mineral content of the remaining tissue (∆KHN), in addition to promoting a complete inhibition of MMPs -2 and -9 activity in vitro(AU)

Planejamento, síntese, modelagem molecular e avaliação biológica de novos potenciais inibidores duais HDAC/ MMP não-hidroxamatos / Design, synthesis, molecular modeling and biological evaluation of novel potential non-hydroxamate dual HDAC/ MMP inhibitors

A inibição de alvos específicos como metaloproteinase de matriz (MMP) e histona desacetilase (HDAC) é amplamente estudada para impedir o progresso do câncer. Foi estabelecido que a inibição concomitante de MMP e HDAC é eficaz no combate de tumores sólidos e hematológicos. Ambos os alvos possuem um íon Zn2+ em seu sítio ativo, fundamental para a atividade destas enzimas. A alta afinidade dos inibidores conhecidos de MMP e de HDAC é conferida, principalmente, por um potente grupo ligante de zinco (ZBG). O ácido hidroxâmico é o ZBG mais potente conhecido atualmente, entretanto, este apresenta instabilidade farmacocinética, levando a ineficácia e genotoxicidade em testes clínicos. Frente a este contexto, o presente trabalho teve como objetivo o planejamento, síntese, modelagem molecular e avaliação biológica de novos inibidores duais MMP/HDAC não-hidroxamatos. Os compostos foram planejados utilizando estratégias de hibridação molecular, a partir de arcabouços provenientes inibidores de HDAC e MMP, gerando compostos arilsulfonamídicos com variações no tipo de ZBG inserido e na sua respectiva posição relativa na estrutura geral. Foram sintetizados sete análogos, em duas a três etapas reacionais, utilizando métodos de sulfonilação e acoplamento com agentes condensantes, partindo dos ésteres para e meta aminobenzoicos. Os rendimentos globais variaram de 25% a 55% e os produtos obtidos foram caracterizados por RMN 1H e 13C, LC/MS, CLAE e ponto de fusão. Os compostos tiveram sua atividade citotóxica avaliada em células HOG (oligodendroma) e T98G (glioblastoma), dentre os quais o 6a, que possui o ZBG 2-amino anilida, foi o mais promissor, apresentando atividade nas duas linhagens na casa de nM. Ensaios de coordenação com Fe2+ comprovaram a capacidade quelante dos análogos contendo ácido hidroxâmico e dos demais compostos citotóxicos, 4a e 4b (ZBG-2, salicilal-hidrazona), o que não foi observado para o composto 6a. Os estudos de ancoramento molecular permitiram sugerir um modo de interação para todos os ZBG propostos frente aos respectivos alvos (HDAC e MMP), sendo observado que o ZBG 4 (2-amino anilida) faria a interação de modo monodentado com a HDAC, enquanto não seria possível o encaixe no sítio catalítico da MMP. Conclui-se, portanto, que o planejamento proposto permitiu a obtenção de compostos promissores como antitumorais, e que a substituição do ácido hidroxâmico por outros ZBG fornece moléculas ativas frente a células tumorais. Entretanto, a avaliação biológica frente à MMP e HDAC é necessária para confirmar o mecanismo de ação proposto

Inhibition of specific targets such as matrix metalloproteinase (MMP) and histone deacetylase (HDAC) is extensively studied regarding arrest cancer growth. Particularly, concomitant inhibition of MMP and HDAC is effective against solid and hematologic tumors. Both targets have an ion Zn2+ at their catalytic site, which is essential for respective enzymatic activity. High affinity of known MMP and HDAC inhibitors is mainly provided by a potent zinc binding group (ZBG). Hydroxamic acid is the most potent ZBG currently known; however, it presents low pharmacokinetics stability, which results in its ineffectiveness and genotoxicity along clinical trial. So, the aim of this work comprised the design, synthesis, molecular modeling and biological evaluation of novel potential non-hydroxamate dual HDAC/ MMP inhibitors. Compounds were designed by molecular hybridation, employing scaffolds from HDAC and MMP inhibitors, which provided arylsulfonamides with variation about the ZBG type and its respective relative position in the general structure. Seven compounds were synthesized, in two to three reaction steps, through methods that comprise sulfonilation and coupling with condensing agents, using para and meta-aminobenzoic esters as starting material. Compounds showed global yields around 25-55 % and were characterized by 1H and 13C NMR, LC/MS, HPLC and melting point. Compounds were evaluated about their cytotoxicity against HOG (oligodendroma) and T98G (glioblastoma) cells, which 6a, with ZBG 2-aminobenzamide, was the most promising molecules, presenting activity against both cell lines at nM range. Coordination assays with Fe2+ proved the chelating capacity of hydroxamate analogues as well as the cytotoxic compounds, 4a and 4b (ZBG-2, salicylal-hydrazone), which was not observed about 6a. Molecular docking allowed to suggest an interaction model for all proposed ZBG with the respective targets (MMP and HDAC), showing that (ZBG-4) 2-aminobenzamide interacts with HDAC by monodentate way, but does not docks at MMP catalytic site. We conclude that the proposed design allowed obtaining promising compounds as antitumors agents, and the replacement of hydroxamic acid by other ZBG provide active molecules against tumor cells. However, biological evaluation against MMP and HDAC is necessary to confirm the proposed action mechanism

Resin-dentin bond stability of etch-and-rinse adhesive systems with different concentrations of MMP inhibitor GM1489

Abstract Enzymatic degradation of the hybrid layer can be accelerated by the activation of dentin metalloproteinases (MMP) during the bonding procedure. MMP inhibitors may be used to contain this process. Objective To evaluate the degree of conversion (DC%), dentin bond strength (µTBS) (immediate and after 1 year of storage in water), and nanoleakage of an experimental (EXP) and a commercial (SB) adhesive system, containing different concentrations of the MMP inhibitor GM1489: 0, 1 µM, 5 µM and 10 µM. Methodology DC% was evaluated by FT-IR spectroscopy. Dentin bond strength was evaluated by µTBS test. Half of beams were submitted to the µTBS test after 24 h and the other half, after storage for 1 year. From each tooth and storage time, 2 beams were reserved for nanoleakage testing. Data were analyzed using ANOVA and Tukey's test to compare means (α=0.05). Results All adhesive systems maintained the µTBS after 1 year of storage. Groups with higher concentrations of inhibitor (5 µM and 10 µM) showed higher µTBS values than groups without inhibitor or with 1 µM. The nanoleakage values of all groups showed no increase after 1 year of storage and values were similar for SB and EXP groups, in both storage periods. The inhibitor did not affect the DC% of the EXP groups, but the SB5 and SB10 groups showed higher DC% values than those of SB0 and SB1. Conclusions The incorporation of GM1489 in the adhesive systems had no detrimental effect on DC%. The concentrations of 5 µM GM1489 for SB and 5 µM or 10 µM for EXP provided higher μTBS than groups without GM1489, in the evaluation after 1 year of storage; whereas the concentration of inhibitor did not affect adhesive systems nanoleakage.

Influência de inibidores de metaloproteinases na degradação da camada híbrida / Influence of metalloproteinase inhibitors on hybrid layer degradation

Objetivo: realizar uma revisão de literatura acerca da eficácia de utilização da clorexidina (CHX) e de outros tipos de inibidores de metaloproteinases (MMPs) na resistência de união da camada híbrida. Métodos: a busca bibliográfica foi realizada na base de dados PubMed, nos meses de novembro e dezembro de 2018. A pesquisa ocorreu em três fases, com os descritores previamente selecionados. Foram incluídas publicações dos últimos 10 anos no formato de pesquisas científicas realizadas in vitro ou in vivo. Após análise, obedecendo aos critérios de inclusão e exclusão, foram incluídos sete estudos na presente revisão. Resultados/Revisão de literatura: na interface adesiva, os estudos mostram que as MMPs são ativadas durante a etapa de ataque ácido realizada nos protocolos de aplicação de sistemas adesivos, podendo ser ativada tanto por procedimentos adesivos com condicionamento ácido prévio como por sistemas adesivos autocondicionantes. Além da CHX, outras substâncias foram pesquisadas e se mostraram eficazes na inibição de MMPs. Considerações finais: por meio da inibição da atividade das MMPs, é possível obter uma maior durabilidade da interface adesiva e uma menor degradação hidrolítica do colágeno presente na camada híbrida. (AU)

Objective: to perform a literature review on the efficacy of chlorhexidine (CHX) and other types of metalloproteinase inhibitors (MMPs) on hybrid layer bond strength. Methods: the bibliographic search was performed in PubMed, in the months of november and december of 2018. The research was carried out in three phases with the previously selected descriptors. Publications have been included in the last 10 years in the form of scientific research conducted in vitro or in vivo. After analysis, following the inclusion and exclusion criteria, 7 studies were included in the present review. Results / Literature review: in the adhesive interface, the studies show that the MMPs are activated during the acid attack stage carried out in the application protocols of adhesive systems, and can be activated either by adhesive procedures with prior acid conditioning or self-etching adhesive systems. In addition to CHX, other substances were investigated and shown to be effective in inhibiting MMPs. Final considerations: through the inhibition of the MMPs activity it is possible to obtain a greater durability of the adhesive interface and lower hydrolytic degradation of the collagen present in the hybrid layer. (AU)

Papel da MMP-9 e do TGF-ß na expressão de componentes da Matriz Extracelular por fibroblastos gengivais de camundongos normais e diabéticos com doença periodontal in vitro / Role of MMP-9 and TGF-ß on extracellular matrix components expression by in vitro fibroblasts from normal and diabetic mice with periodontal disease

Na presença de Diabetes Mellitus, a doença periodontal (DP) pode acentuar o aparecimento de complicações e aumentar sua prevalência. O objetivo desta pesquisa foi avaliar se o Fator de Crescimento Transformador -ß do tipo 1 (TGFß1, sigla em inglês) e Metaloproteinase de matriz -9 (MMP-9, sigla em ingês) estão modulando a formação de componentes de matriz extracelular em fibroblastos gengivais (FG) de camundongos diabéticos e com DP. Os 24 animais foram separados em 4 grupos, e então foram tratados com estreptozotocina para indução de diabetes e foi realizado indução da DP através de ligadura com fio de seda. Após a eutanásia e coleta da gengiva, FG dos camundongos foram cultivados. A expressão gênica de RNAm e produção de proteína para MMP-9, TGF-ß1 e colágeno tipo 1 (Col1a1) foram avaliadas. Eles também foram tratados com inibidores de MMP-9 e TGF-ß1, e dexametasona (Dexa) por 24h. FG de animais normais e diabéticos com DP induzida tiveram expressão aumentada de MMP-9 e TGF-ß1 em 6 e 24h, diminuindo em 48h. Expressão gênica de Col1a1 foi diminuída em FG com DP em 6, 24 e 48h. Inibidor de MMP-9 (MMP-9i) bloqueou a expressão gênica de MMP-9 e de TGF-ß1 em FG normais e diabéticos em 24h. A expressão de col1a1 foi inibida pelo MMP-9i somente em FG normais, mas não em diabéticos. A expressão de receptor de TGF-ß do tipo 1 (TGF-ß R1) foi aumentada em FG normais e diabéticos, porém MMP-9i diminuiu esta expressão somente em FG diabéticos. Inibidor de TGF-ß (SB431542) e Dexa interromperam a expressão gênica de MMP-9, TGF-ß1 e col1a1 em FG normais e diabéticos com DP em 24h. TGF-ß R1 também foi inibido em FG normais e diabéticos com DP em 24h de tratamento com SB431542. Juntos, esses achados sugerem que indivíduos diabéticos têm síntese aumentada de TGF-ß e MMP-9 em FG durante a DP devido a ativação do TGF-ß R1 via ALK5 e presença de MMP-9. Por outro lado, o aumento na expressão de colágeno é devido a ativação de TGF-ß R1 em FG normais e diabéticos, e presença de MMP-9 em FG normais, somente(AU)

In presence of Diabetes Mellitus, periodontal disease (PD) can accentuate complication appearance and increase prevalence. The aim of this study was to evaluate whether Transforming Growth factor-ß 1 (TGF-ß1) and Matrix Metaloproteinase-9 (MMP-9) are modulating the extracellular matrix components in gingival fibroblasts (GF) from diabetic mice (D) with PD. 24 animals were divided in 4 groups then they were treated with streptozotocin to diabetes induction and DP was inducted through a ligature with silk thread. After euthanasia and gingiva harvest, mice GF were cultured. MMP-9, TGF- ß1, and type 1 Collagen (col1a1) mRNA expression and protein production were evaluated. They were also treated with MMP-9 and TGF-ß inhibitors, and dexamethasone (Dexa) for 24h. Normal and diabetic GF from PD-induced mice had increased MMP-9 and TGF-ß1 expression at 6 and 24 h decreasing at 48 h. Col1a1 gene expression was decreased in normal GF with PD at 6, 24 and 48 h. In diabetic GF col1a1 expression was inhibited at 6, 24 and 48 h. MMP-9 inhibitor (MMP-9i) blocked the MMP-9 expression, and TGF-ß1 gene expression in normal and diabetic GF at 24 h. The col1a1 gene expression was inhibited by MMP-9i only in normal GF but not in diabetic. TGF-ß R1 was increased in normal and diabetic GF with PD, but MMP9i decreased this expression only in diabetic GF. TGF-ß inhibitor (SB431542) and Dexa abrogated MMP-9, TGF-ß1 and col1a1 expression in normal and diabetic GF with PD at 24 h. TGF-ß R1 was also inhibited in normal and diabetic GF with PD after 24 h of treatment with SB431542. Taken together, these findings suggests that diabetic individuals has increased synthesis of TGF-ß and MMP-9 in GF during PD due to TGF-ß R1 activation via Alk5 and presence of MMP-9. On the other hand, increased in the col1a1 expression is due to TGF-ß R1 activation in normal and diabetic GF and presence of MMP-9 in normal GF(AU)

Updating the role of matrix metalloproteinases in mineralized tissue and related diseases

Abstract Bone development and healing processes involve a complex cascade of biological events requiring well-orchestrated synergism with bone cells, growth factors, and other trophic signaling molecules and cellular structures. Beyond health processes, MMPs play several key roles in the installation of heart and blood vessel related diseases and cancer, ranging from accelerating metastatic cells to ectopic vascular mineralization by smooth muscle cells in complementary manner. The tissue inhibitors of MMPs (TIMPs) have an important role in controlling proteolysis. Paired with the post-transcriptional efficiency of specific miRNAs, they modulate MMP performance. If druggable, these molecules are suggested to be a platform for development of "smart" medications and further clinical trials. Thus, considering the pleiotropic effect of MMPs on mammals, the purpose of this review is to update the role of those multifaceted proteases in mineralized tissues in health, such as bone, and pathophysiological disorders, such as ectopic vascular calcification and cancer.

La alteración de la reparación pulmonar durante la neutropenia puede revertirse mediante metaloproteinasa de matriz-9 / The alteration of pulmonary repair during neutropenia can be reverted by metaloproteinase matrix-9

ANTECEDENTES: Los neutrófilos pueden causar la interrupción del tejido durante la migración y la liberación de moléculas citotóxicas. Sin embargo, los beneficios del agotamiento de los neutrófilos observados en modelos experimentales de lesión pulmonar no se corresponden con el mal resultado de los pacientes neutropénicos. MÉTODOS: Para aclarar el papel de los neutrófilos durante la reparación, los ratones con lesión pulmonar inducida porventilador(VILI) se volvieron neutropénicos después del daño y se les siguió durante 48 horas de respiración espontánea. Se recogieron los pulmones y se midieron los mediadores inflamatorios y las metaloproteinasas de la matriz. Se recolectó líquido de lavado broncoalveolar (BALF) de pacientes ventilados con síndrome de dificultad respiratoria aguda, con o sin neutropenia, se midieron los mismos mediadores y se estudiaron sus efectos en un modelo ex vivo de reparación alveolar. Finalmente, los ratones neutropénicos fueron tratados después de VILI con metaloproteinasa-9 de matriz exógena (MMP-9). RESULTADOS: Los pulmones de animales neutropénicos mostraron una reparación retardada y mostraron niveles más altos de factor de necrosis tumoral H, interferón H y proteína inflamatoria de macrófagos 2, y ausencia de MMP-9. El BALF de pacientes neutropénicos ventilados con síndrome de dificultad respiratoria aguda mostró resultados similares. Los BALF de pacientes neutropénicos produjeron una tasa de cierre tardío de las heridas epiteliales ex vivo, que se mejoró mediante la eliminación de colágeno o la adición de MMP-9 exógeno. Por último, el tratamiento de ratones neutropénicos con MMP-9 exógena después de VILI redujo el daño tisular sin modificar las concentraciones de citocina. CONCLUSIÓN: La liberación de MMP-9 de los neutrófilos es necesaria para el procesamiento de la matriz adecuada y la reparación del pulmón.

MMP Inhibitor Ilomastat Improves Survival of Mice Exposed to γ-Irradiation / 生物医学与环境科学（英文）

There is still a need for better protection against or mitigation of the effects of ionizing radiation following conventional radiotherapy or accidental exposure. The objective of our current study was to investigate the possible roles of matrix metalloproteinase inhibitor, ilomastat, in the protection of mice from total body radiation (TBI), and the underlying protective mechanisms. Ilomastat treatment increased the survival of mice after TBI. Ilomastat pretreatment promoted recovery of hematological and immunological cells in mice after 6 Gy γ-ray TBI. Our findings suggest the potential of ilomastat to protect against or mitigate the effects of radiation.

Relationship between matrix metalloproteinases and the occurrence and development of ovarian cancer

Ovarian cancer is one of the most malignant genital cancers, with a high mortality rate. Many researchers have suggested that matrix metalloproteinases (MMPs) have remarkably high expression in ovarian cancer tissues. MMPs are considered to be related to the occurrence, development, invasion and metastasis of ovarian cancer. Moreover, some studies have discovered that the unbalance between MMPs and tissue inhibitor of metalloproteinases (TIMPs) are associated with the malignant phenotype of tumors. This review summarizes the latest research progress of MMPs in ovarian cancer. The investigation of MMP mechanism in ovarian cancer will facilitate the development of effective anti-tumor drugs, and thereby improve the survival rate of patients with ovarian cancer.

Research progress on the animal models and treatment strategies of diabetic foot ulcer / 浙江大学学报·医学版

The suitable experimental animal model is important in research of pathogenesis and therapeutic strategies of diabetic foot ulcer, and the murine model is the most commonly used one at present. It can be divided into two types: the animal model simulating pathological conditions and the model simulating clinical symptoms. This article reviews the current research progress on the mechanisms of diabetic ulcer pathogenesis, and relevant treatment strategies, including the inhibition of matrix metalloproteinases (MMPs) expression, promotion of angiogenesis and anti-inflammatory therapy.

Progress on matrix metalloproteinase inhibitors / 华西口腔医学杂志

Continuing advances in dentin bonding technology and adhesives revolutionized bonding of resin-based composite restorations. However, hybrid layers created by contemporary dentin adhesives present imperfect durability, and degradation of collagen matrix by endogenous enzymes is a significant factor causing destruction of hybrid layers. Bond durability can be improved by using enzyme inhibitors to prevent collagen degradation and to preserve integrity of collagen matrix. This review summarizes progress on matrix metalloproteinase inhibitors (including chlorhexidine, ethylenediaminetetraacetic acid, quaternary ammonium salt, tetracycline and its derivatives, hydroxamic acid inhibitors, bisphosphonate derivative, and cross-linking agents) and suggests prospects for these compounds.

The use of sodium trimetaphosphate for matrix metalloproteinase inhibition, remineralization and bonding to dentin / O uso de trimetafosfato de sódio sobre a inibição de metaloproteinases e na remineralização e adesão à dentina

The adhesive process to dentin substrate depends on the condition determined by the combined action of the mineral loss and the endogenous enzymes activity. Thus, considering a more complete therapeutic approach, sodium trimetaphosphate (STMP) may be a novel strategy that conciliates the remineralization potential to the promotion of dentin strengthening and its stability, possibly directing mineral nucleation and controlling the rate of biodegradation. In this study, the effect of STMP was evaluated in 2 studies. In study 1, different concentrations of STMP (0.5, 1.5, 3.5 and 5%) were investigated to assess their anti-proteolytic capacity on human purified MMPs-2 and -9 by zymography. Afterwards, only the concentrations (1.5, 3.5 and 5%) that showed total inhibition of both MMPs were used to evaluate their remineralizing capacity in dentin substrate submitted to artificial cariogenic challenge, through surface hardness (SH) and cross-sectional hardness (CSH). In study 2, based on the previous results, the capacity of the 1.5% STMP associated or not with NaF or Ca(OH)2 solutions in improving the dentin bond strength of a universal adhesive system was evaluated by the microtensile test . Thus, these studies suggest that 1.5% STMP is an effective inhibitor of collagen degradation mediated by purified human MMPs-2 and -9. In addition, demineralized and treated dentin with 1.5% STMP supplemented with Ca(OH)2 may induce remineralization. Thus, the use of STMP can be introduced as a new strategy that combines enzymatic inhibition and remineralization potential, reestablishing favorable conditions to affected dentin. These evidences support perspectives of therapies to restructure dentin and propose feasible and promising clinical strategies.(AU)

O processo adesivo ao substrato dentinário depende da condição determinada pela ação combinada da perda mineral e atividade de enzimas endógenas. Deste modo, considerando uma abordagem terapêutica mais completa, o trimetafosfato de sódio (STMP) pode ser uma estratégia inovadora que concilia o potencial remineralizador à promoção do fortalecimento da dentina e sua estabilidade, possivelmente direcionando a nucleação mineral e controlando a taxa de biodegradação. Neste trabalho, o efeito do STMP foi avaliado em 2 estudos. No estudo 1, diferentes concentrações de STMP (0,5; 1,5; 3,5 e 5%) foram investigadas para avaliar sua capacidade anti-proteolítica sobre as MMPs-2 e -9 purificadas humanas, por zimografia. Posteriormente, somente as concentrações (1,5; 3,5 e 5%) que apresentaram capacidade de inibição total de ambas MMPs foram utilizadas para avaliar sua capacidade remineralizadora em substrato dentinário submetido ao desafio cariogênico artificial, através da dureza de superfície (DS) e longitudinal (DL). No estudo 2, baseado nos resultados anteriores, foi avaliada a capacidade do STMP à 1,5% associado ou não a soluções de NaF ou Ca(OH)2 em melhorar a resistência de união à dentina de um sistema adesivo universal pelo teste de microtração. Desta forma, estes estudos sugerem que o STMP à 1,5% apresenta-se como um inibidor eficaz da degradação do colágeno mediada por MMPs-2 e -9 humanas purificadas. Além disso, a dentina humana desmineralizada e tratada com STMP à 1,5% suplementada com Ca(OH)2 pode induzir à remineralização. Assim, o uso de STMP pode ser introduzido como uma nova estratégia que combina inibição enzimática e potencial de remineralização, reestabelecendo condições favoráveis a partir de uma dentina afetada. Estas evidências sustentam perspectivas de terapias para reestruturar a dentina e propor estratégias clínicas factíveis e promissoras.(AU)

Expression of TIMP-2 in HPV-16 infected oral squamous cell carcinoma in patients in Iraq

Aim: To determine the expression of tissue inhibitors of metalloproteinases (TIMP-2) in oralsquamous cell carcinoma (OSCC) and the difference in its expression level between positiveand negative HPV-16 (human papilloma virus- 16) OSCC patients. Methods: This study wasconducted on 33 biopsies obtained from patients with OSCC and 10 normal oral mucosa ascontrols. In situ hybridization (ISH) was used to investigate the presence of HPV-16, whileimmunohistochemistry (IHC) was used to estimate the expression level of TIMP-2. Results: TheTIMP-2 was expressed in 27 (81.8%) of OSCC sections with no significant difference betweenits expression level in HPV-16 positive and HPV-16 negative OSCC cases (p=0.058). TIMP-2was found to be highly expressed in OSCC sections, and the presence of HPV was not relatedto its overexpression. Conclusions: The percentage of samples that appeared to accommodatedetectable HPV-16 was high, but no significant difference was observed in relation to TIMP-2expression level. Future studies with a larger number of patients are highly recommended toaddress the possible association between TIMp-2 and OSCC positive HPV-16.

Preventive effect of toothpastes with MMP inhibitors on human dentine erosion and abrasion in vitro

ABSTRACT The use of gels and mouthrinses with MMP inhibitors (chlorhexidine, and green tea extract) was shown to prevent erosive wear. The aim of this study was to analyze the protective effect of toothpastes containing MMP inhibitors on dentine loss induced by erosion in vitro. Material and Methods Five groups each containing 12 specimens of human root dentine were prepared. The specimens were subjected to 1 min erosion by immersion in a cola drink, 4 times a day, for 5 d. Each day, after the first and last erosive challenges, the specimens were brushed for 15 s with a slurry of dentifrice and water (1:3) containing placebo, 1,100 ppm fluoride, 0.61% green tea extract, 0.12% chlorhexidine or 0.004% chlorhexidine (commercial toothpaste). Between the acid challenges, the specimens were stored in artificial saliva with remineralizing potential until the next treatment. Dentine loss was determined using profilometry. Data were analyzed using one-way ANOVA after log transform (p<0.05). Results The mean wear values (μm) were as follows: placebo 1.83±0.53; 0.61% green tea extract 1.00±0.21; fluoride 1.27±0.43; 0.12% chlorhexidine 1.19±0.30; and 0.004% chlorhexidine 1.22±0.46. There was a significant difference in wear between placebo and all the treatment toothpastes, which did not differ from each other. Conclusion The results suggest that toothpastes containing MMP inhibitors are as effective as those based on NaF in preventing dentine erosion and abrasion.

Effect of calcium channel blockers on primary cultured human urethra scar fibroblasts / 中南大学学报(医学版)

To investigate effects of verapamil on primary cultured human urethral scar fibroblasts (USFs) and to provide basis for protecting the formation of urethra scar.
 Methods: The cell proliferation was evaluated with the cell counting kit (CCK)-8 method after USFs were incubated various verapamil concentrations (50, 100, 150, 200, or 250 μmol/L) or solvent for 12, 24, or 48 h. The protein level of matrix metalloproteinase (MMP) was evaluated with ELISA after cells were incubated with verapamil (100 μmol/L) or solvent (control cells) for 24 h.
 Results: The proliferation of USFs was obviously suppressed after verapamil treatment, which was in a dose-dependent and time-dependent manner. Meanwhile, the protein levels of MMP-2 and MMP-9 in the verapamil treatment group increased obviously compared with those of the control groups (P<0.05).
 Conclusion: Calcium channel blockers may prevent the excessive formation of urethra scar by inhibiting the proliferation of urethral scar fibroblasts and enhancing the activity of MMP.

Influência de agentes antiproteolíticos na resistência de união da interface dentina, adesivo e cimento resinoso / Influence of antiproteolytic agents on dentin, adhesive and luting cement interface

O objetivo deste trabalho, foi avaliar a influência de agentes antiproteolíticos, imediatamente e após 12 meses, na cimentação adesiva. Foram utilizadas as porções coronárias de 64 molares humanos hígidos. As amostras foram divididas aleatoriamente em 4 grupos, de acordo com o tratamento da dentina: CT (grupo controle); CXL (grupo clorexidina); EGCG (grupo epigalocatequina-3-galato); AT (grupo antocianina). Para a restauração, foram confeccionados cilindros em resina composta Filtek Z350 XT. Após o condicionamento ácido da dentina, as soluções antiproteolíticas foram aplicadas e mantidas sobre a superfície por 60 s, em seguida as restaurações foram cimentadas às amostras com o adesivo Adper Single Bond 2 + cimento resinoso Variolink II. Todos os grupos foram subdivididos de acordo com o tempo de armazenamento em água destilada para o ensaio mecânico: I (Imediato = após 24 h); A (Armazenado = após 12 meses). As amostras foram cortadas em palitos e submetidas ao teste de microtração. Os dados obtidos foram submetidos ao teste de ANOVA a 2 fatores, seguido do teste de Tukey (α=5%). Foram encontradas diferenças para os fatores Tratamento (p=0,03) e Armazenamento (p=0,00). Os valores em MPa de média e desvio padrão para interação entre os fatores Tratamento X Armazenamento, foram: CT.I: 42.56 (±7,60) A; EGCG.I: 39.6 (±8,99) A; CLX.I: 39.37 (±9,44) A; AT.I: 38.69 (±8,65) A; CT.A: 31.85 (±6,96) A; AT.A: 31.29 (±4,89) AB; EGCG.A: 30.94 (±9,91) AB; CLX.A: 18.68 (±4,53) B. Foram confeccionadas duas amostras adicionais de cada grupo para avaliação descritiva da microdureza knoop de subsuperfície e avaliação qualitativa da interface adesiva em MEV. Conclui-se que as soluções de EGCG e AT foram capazes de manter a estabilidade da resistência de união após 12 meses de armazenamento e que a solução de CLX afetou negativamente os valores de resistência de união comparado aos demais subgrupos(AU)

The aim of this study was to evaluate immediately and longitunally the antiproteolytic agent influence to adhesive cementation. Were used 64 coronary portion of sound human molars. The samples were divided into 4 groups according to the dentin treatment: CG (Control group); CLX (chlorhexidine group); EGCG (epigallocatechin-3- gallate group); AT (anthocyanin group). Cylindrical composite blocks were prepared using a resin composite Filtek Supreme for the restorations. The solutions were applied on the acid-etched dentin and were kept for 60 s. The resin composite blocks were cemented with the adhesive system Adper Single Bond 2 + resin cement Variolink II. All groups were subdivided according to the storage time in distilled water for mechanical testing: I (Immediate = after 24 h); S (Stored = after 12 months). Samples were cut into beams and tested in a universal testing machine at a crosshead speed of 0.5 mm/min until failure. The data were submitted to two-way ANOVA, followed by the Tukey test (α = 5%). Significant differences were found for the Treatment factor (p = 0.03) and Storage factor (p = 0.00). The mean values and standard deviation for interaction between the factors Treatment X Storage, were CG.I: 42.56 (± 7.60) A; EGCG.I: 39.6 (± 8.99) A; CLX.I: 39.37 (± 9.44) A; AT.I: 38.69 (± 8.65) A; CG.S: 31.85 (± 6.96) A; AT.S: 31.29 (± 4.89) AB; EGCG.S: 30.94 (± 9.91) AB; CLX.S: 18.68 (± 4.53) B. Two additional samples from each group were confectioned to descriptive analyses of microhardness knoop of dentin subjacent to the bonding interface and a qualitative interface evaluation with SEM (scanning electron microscope). It can be conclude that EGCG and AT solutions were able to maintain the bond strength stability after 12 months of storage and CLX solution adversely affect the bond strength values compared to other subgroups(AU)

Influencia de los inhibidores de las metaloproteinasas, agentes reticuladores y remineralización biomimética en la longevidad de la unión adhesiva: parte 3: agentes reticuladores / Metalloproteinase inhibitors, cross-linking agents and biomimetic remineralization influence in the adhesive bond longevity: part 3: cross-linking agents

Las propiedades mecánicas del colágeno se deben a agentes intrínsecos de entrecruzamiento [Al-Ammar et al, 2009]. El aumento en el número de enlaces de la molécula de colágeno mejora su estabilidad e integridad, colaborando con el mantenimiento de propiedades adecuadas de la unión adhesiva a lo largo del tiempo [Bedran-Russo et al, 2009; Breschi et al, 2008]. Existen varios enfoques que permiten modificar el sustrato dentinario mediante la promoción de la formación de enlaces exógenos, con el objetivo de aumentar la resistencia de la red de colágeno [Bedran-Russo et al, 2007; Bedran-Russo et al, 2008]. Estos enfoques se dividen en métodos mecánicos [Bedran-Russo et al, 2007; Bedran-Russo et al, 2008; Castellan et al, 2010; Han et al, 2003] y método fotoxidativo [Cova et al, 2011]. Dentro de los primeros, los agentes reticuladores de origen natural son capaces de estabilizar el colágeno de la dentina [Bedran-Russo et al, 2007] sin afectar la resistencia de la unión adhesiva y sin generar toxicidad. Sin embargo, el uso de estos agentes por sí solo no permite asegurar la estabilidad de la unión adhesiva a lo largo del tiempo; porque como es sabido, la longevidad de dicha unión no depende únicamente de las características del sustrato, sino también de propiedades inherentes al sistema adhesivo por un lado, y de la presencia de humedad por otro.

The mechanical properties of collagen are due to intrinsic crosslinking agents [Al-Ammar et al, 2009]. The increase in the number of links of the collagen molecule improves its stability and integrity, collaborating with the maintenance of adequate adhesive bonding properties over time [Bedran-Russo et al, 2009; Breschi et al, 2008]. There are several approaches for modifying the dentin substrate by promoting the formation of exogenous links, in order to increase the strength of the collagen network [Bedran-Russo et al, 2007; Bedran-Russo et al, 2008]. These approaches are divided into mechanical methods [Bedran-Russo et al, 2007; Bedran-Russo et al, 2008; Castellan et al, 2010; Han et al, 2003] and foto oxidating method [Cova et al, 2011]. Within the first, the naturally occurring crosslinking agents are capable of stabilizing the dentin collagen [Bedran-Russo et al, 2007] without affecting the strength of the adhesive bond and without generating any toxicity. However, the use of these agents alone does not ensure the stability of the adhesive bond over time; because as it is known, the longevity of such binding does not only depend on the substrate’s characteristics, but also on the adhesive system’s properties on one side, and the presence of moisture on the other