Doxiciclina em pacientes com linfangioleiomiomatose: segurança e eficácia no bloqueio de metaloproteinases / Doxycycline use in patients with lymphangioleiomyomatosis: safety and efficacy in metalloproteinase blockade

OBJETIVO: A linfangioleiomiomatose (LAM) é caracterizada pela presença de cistos pulmonares, cuja formação está associada à hiperreatividade de metaloproteinases de matriz (MMP), principalmente MMP-2 e MMP-9. Objetivamos comparar os níveis dessas MMPs entre pacientes com LAM e controles saudáveis, assim como avaliar, nas pacientes com LAM, a segurança e a eficácia do tratamento com doxiciclina, um potente inibidor de MMPs. MÉTODOS: Estudo clínico prospectivo no qual as pacientes com LAM receberam doxiciclina (100 mg/dia) por seis meses, coletando-se amostras de urina e sangue para a dosagem de MMP-2 e MMP-9 antes e ao final do período. Foram ainda obtidas amostras de 10 mulheres saudáveis. RESULTADOS: De 41 pacientes com LAM que iniciaram o tratamento, 34 concluíram o protocolo. Os níveis de MMP-9 sérica e urinária foram significativamente inferiores no grupo controle (p < 0,0001). Comparando-se os valores antes e após o tratamento, a mediana do nível sérico da MMP-9 reduziu de 919 ng/mL para 871 ng/mL (p = 0,05), enquanto a mediana da dosagem urinária de MMP-9 diminui de 11.558 pg/mL para 7.315 pg/mL (p = 0,10). A mediana da MMP-2 sérica apresentou um decréscimo significativo após o tratamento (p = 0,04). Não foram detectados níveis de MMP-2 urinária. Epigastralgia, náuseas e diarreia foram os efeitos adversos mais prevalentes, e geralmente autolimitados. Apenas 1 paciente interrompeu o tratamento devido a efeitos colaterais. CONCLUSÕES: Pela primeira vez, conseguiu-se evidenciar em pacientes com LAM a redução dos níveis séricos e urinários de MMPs após o uso de doxiciclina, que se mostrou uma medicação segura, com efeitos colaterais leves e toleráveis.

OBJECTIVE: Lymphangioleiomyomatosis (LAM) is characterized by lung cysts, whose development is associated with matrix metalloproteinase (MMP) hyperactivity, principally that of MMP-2 and MMP-9. Our objective was to compare LAM patients and controls in terms of the levels of these MMPs, as well as to determine the safety and efficacy of treatment with doxycycline, a potent MMP inhibitor. METHODS: Prospective clinical study involving female LAM patients who received doxycycline (100 mg/day) for six months. Urine and blood samples were collected for the quantification of MMP-2 and MMP-9 before and after the treatment period. Samples from 10 healthy women were also collected. RESULTS:Of the 41 LAM patients who started the treatment, 34 completed the protocol. Serum and urinary MMP-9 levels were significantly lower in the controls than in the LAM patients (p < 0.0001). Comparing pre- and post-treatment values, we found that the median level of MMP-9 in serum decreased from 919 ng/mL to 871 ng/mL (p = 0.05), whereas that of MMP-9 in urine decreased from 11,558 pg/mL to 7,315 pg/mL (p = 0.10). After treatment, the median level of MMP-2 in serum was significantly lower (p = 0.04) and urinary MMP-2 levels were undetectable. Nausea, diarrhea, and epigastric pain were the most prevalent adverse affects and were often self-limiting. There was only one case in which the patient discontinued the treatment because of side effects. CONCLUSIONS: We have demonstrated, for the first time, a decrease in serum and urine levels of MMPs in LAM patients treated with doxycycline, which proved to be a safe medication, with mild and well-tolerated side effects.

Inibição das metaloproteinases da matriz como nova estratégia para prevenção da erosão dentinária / Matrix metalloproteinases inhibition as a new strategy to prevent dentin erosion

A degradação da dentina pelas metaloproteinases da matriz (MMPs) pode facilitar a progressão de lesões erosivas. Objetivos: Avaliar, por meio de uma série de 5 subprojetos: 1) A atividade de MMPs em dentina bovina e humana; 2) O efeito do chá verde contra erosão/abrasão de dentina; 3 e 4) Géis contendo inibidores de MMPs (epigallocatechin-3-galatte-EGCG, clorexidina-CHX e sulfato ferroso-FeSO4) sobre a prevenção de erosão de dentina sozinha ou associada à abrasão, respectivamente, e 5) Sobre a degradação de colágeno e desgaste. Material e Métodos: 1) Extração proteica de dentina de coroa e raiz bovina e humana foi realizada (ácido cítrico a 0,87 M, pH 2,3) e testada por zimografia e atividades gelatinolíticas; 2) Voluntários (n=10) bochecharam chá verde ou água (1 min, 10 mL) entre os desafios erosivos (Coca-Cola, pH 2,6, 4x/dia/5 min, extraoralmente) e abrasivos. A abrasão (escova elétrica + dentifrício não fluoretado) foi realizada imediatamente ou 30 min depois da erosão por 30 s. O desgaste da dentina foi analisado por perfilometria (µm); 3 e 4) Voluntários (n=10-13) utilizaram dispositivos palatinos contendo 12 blocos de dentina e distribuídos aleatoriamente para 6 grupos, de acordo com o tipo de gel aplicado ou não (não tratado-NT). Os géis tinham composição idêntica, exceto pela presença de EGCG (400 µM), CHX (0,012%), FeSO4 (1 mM), flúor (NaF-1,23%) ou sem ativo (Placebo-P). Os géis foram ou não aplicados sobre os espécimes em fina camada e removidos depois de 1 min. A erosão (Coca-Cola, pH 2,6, 4x5min/dia, extraoralmente) foi realizada por 5 ou 10 dias, respectivamente. A cada dia, depois do primeiro e último desafios erosivos, os blocos eram (ERO) ou não escovados (ERO+ABR) por 15 s (escova elétrica + solução de dentifrício não fluoretado). O desgaste da dentina foi avaliado por perfilometria (µm) depois de 5 e 10 dias. 5) Dentina (n=45/grupo) foi desmineralizada com ácido cítrico (0,87 M, pH 2,3) por 36 h. Na sequência, foi ou não tratada (NT)...

The dentin degradation by matrix metalloproteinases (MMPs) can increase the progression of erosive lesions. Objectives: Five studies were conducted to evaluate: 1) The activity of MMPs in bovine and human dentin; 2) The effect of green tea rinse against dentin erosion/abrasion; 3 and 4) Gels containing MMP inhibitors (epigallocatechin-3-galatte-EGCG, chlorexidine-CHX and ferrous sulphate-FeSO4) to prevent dentin erosion alone or associated with abrasion, respectively, and 5) On collagen degradation and wear. Material and Methods: 1) Protein extraction from crown and root of bovine and human dentin was performed (0.87 M citric acid, pH 2.3) and tested by zymography and gelatinolytic activities; 2) Volunteers (n=10) rinsed with green tea or water (1 min, 10 mL) between the erosive (Coke, pH 2.6, 4x5min/day, extraorally) and abrasive challenges. The abrasion (electric toothbrush + fluoride-free toothpaste) was performed immediately or 30 min after erosion for 30 s. Dentin wear was analyzed by profilometry (µm); 3 and 4) Volunteers (n=10-13) wore palatal devices containing 12 bovine dentin blocks randomly allocated to 6 groups, according to the type of gels applied or not (not treated-NT). The gels had identical composition, except for the presence of EGCG (400 µM), CHX (0.012%), FeSO4 (1 mM), fluoride (NaF-1.23%) or not (Placebo-P). Gels were applied or not on specimens once in a thin layer and removed after 1 min. Erosion (Coke, pH 2.6, 4x5 min/day, extraorally) was performed for 5 or 10 days, respectively. Each day, after first and last erosive challenges, blocks were (ERO) or not brushed (ERO+ABR) for 15 s (electric toothbrush + fluoride-free toothpaste slurry). Dentin wear was evaluated by profilometry (µm) after days 5 and 10. 5) Demineralization of dentin (n=45/group) was performed with 0.87 M citric acid, pH 2.3, for 36 h. In sequence, dentin was or not (NT) treated with the same gels described above. Specimens were stored in artificial saliva (5 days,...

Dose-dependent in vitro inhibition of rabbit corneal matrix metalloproteinases by an extract of Pothomorphe umbellata after alkali injury

The in vitro ability of Pothomorphe umbellata ethanolic crude extract to inhibit matrix metalloproteinase (MMP) in normal cornea and in cornea after alkali injury was demonstrated. Corneas of albino rabbits were injured with 1 N NaOH for 20 s. After 48 h the corneas were excised, homogenized and analyzed for MMP-9 (92 kDa), pro-MMP-2 (72 kDa) and MMP-2 (67 kDa) activity by gelatin zymography. The activity was also measured in untreated corneas. After electrophoresis of 20 æg protein, gels were incubated with 50, 100, or 250 µg/mL lyophilized hydroethanolic (1:1) root crude extract of P. umbellata standardized for 4-nerolidylcatechol (7.09 percent). The activity of the enzymes was compared with that of untreated gel. At 48 h after injury, the activity of all MMPs was increased compared with untreated eyes. When the gels were incubated with P. umbellata extract the activity of MMP-2, pro-MMP-2 and MMP-9 decreased in a dose-dependent manner. MMP-9 activity decreased by approximately 50 percent after incubation with 50 µg/mL and was completely abolished at 100 and 250 µg/mL of the extract. After incubation with 50 µg/mL the activity of pro-MMP-2 and MMP-2 also decreased by 50 percent. The activity of pro-MMP-2 was almost completely abolished after incubation with 250 µg/mL of the extract. For MMP-2 the incubation with 100 or 250 µg/mL of the extract of P. umbellata promoted a 10-fold decrease in activity. In conclusion, P. umbellata root crude extract can be useful as an alternative therapy to control MMP activity after corneal injury.