ABSTRACT
Background The incidence of posterior capsular opacification (PCO) is increasing with the growing of cataract surgery rate.Recent researches provend that disintegrin has inhibitory effect on PCO,and echistatin is one of the disintegrin prime families.Objective This study was to investigate the effects of disintegrin and echistatin on proliferation,adhestion and migration in human lens epithelial cells (LECs) line (SRA01/04).Methods Human LECs line at logarithmic growth phase was used in the study.Cells were cocultured with medium and different concentrations of echistatin (0,2.5,5.0,7.5,10.0,15.0,20.0 mg/L) for different time.The proliferative inhibitory rates of LECs were detected by MTT method 24,48 and 72 hours after cultured.Anti-adhesion effect of echistatin were analyzed by the same assay in 90 minutes.Cell scratching test was performed to evaluate the migration ability of LECs.The width of the scratch was recorded in the culture plate covered with cells under an inverted microscope.After being cultured for 24 hours and 48 hours with echistatin,cell migration distances was examined.Results Compared with the 0 mg/L echistatin group,cells proliferation was obviously inhibited.After cultured with 2.5,5.0,7.5,10.0,15.0,20.0 mg/L echistatin,the proliferation inhibitory rate was 2.6％,15.4％,21.2％,34.7％,46.1％,58.2％ at 24 hours;6.6％,21.9％,38.2％,50.0％,60.7％,76.9％ at 48 hours and 9.8％,29.0％,46.6％,63.4％,69.1％,92.4％ at 72 hours,respectively.The absorbance value (A) in the 5.0,7.5,10.0,15.0,20.0 mg/L groups were significantly lower than that in the 0 mg/L group (P＜ 0.05).With the prolongation of acting time of Ecs,the A value of the cells was gradually reduced,with statistically significant difference (P＜0.05).The adhesion inhibitory rate was 2.6％,15.0％,26.1％,35.3％,45.2％ and 54.5％ in the 2.5,5.0,7.5,10.0,15.0,20.0 mg/L group,respectively.Compared with the result in the 0 mg/L group,the A value in the 5.0,7.5,10.0,15.0,20.0 mg/L group was statistically significant (P＜0.05).After cultured for 24 hours and 48 hours,cell migration distance shortened in the 5.0,7.5,10.0,15.0,20.0 mg/L group,showing a statistically significant difference among them (P＜0.05).Cell migration distance was gradually shortened with the lapse of action time of Ecs with the significant difference (P ＜ 0.05).Conclusions echistatin has inhibitory effects on proliferation,adhestion and migration for human LECs in vitro in time-and dose-dependent manner.It is inferred that echistatin may play a role in the prevention and treatment of PCO.
ABSTRACT
O presente trabalho investigou microscopicamente o efeito da equistatina sobre o processo reabsortivo de dentes incisivos superiores de ratos, extraídos e reimplantados. Foram utilizados 42 animais, divididos em grupos com e sem equistatina. Os tempos extra-alveolares dos dentes foram de 30 e 60 minutos e os períodos experimentais pós-cirúrgicos foram 15, 60 e 90 dias. Os espécimes obtidos foram processados e corados em H.E. para observar os processos biológicos presentes. Os eventos microscópicos foram avaliados por dois examinadores, de modo quantitativo histomorfométrico e descritivo, de acordo com o tipo de infiltrado e intensidade da reação inflamatória; com o tipo, extensão e localização de reabsorção dentária; e presença de anquilose alveolodentária. O nível de concordância entre examinadores, determinado pelo índice Kappa, revelou-se quase perfeito para todas as variáveis avaliadas. A comparação entre os grupos testes e controle para as variáveis apresentadas foi feita por meio do teste Mann-Whitney e mostrou não haver diferença estatística entre a administração de equistatina e tempo extra-alveolar em relação à intensidade inflamatória nas diferentes porções radiculares. Quanto ao tipo de reabsorção estabelecida, notou-se que a presença de reabsorção inflamatória foi significativamente maior (p<0,05) no grupo controle nos tempos de 30 e 60 minutos no período pós-cirúrgico de 15 dias. A anquilose alveolodentária também esteve significativamente mais presente no grupo tratado com equistatina com 30 minutos extra-alveolar e 15 dias de período experimental. Houve, ainda, mais anquilose alveolodentária no grupo controle de 60 minutos extra-alveolar, com o período experimental de 60 dias. No período experimental de 90 dias todos os espécimes avaliados apresentaram processos reabsortivos inflamatórios e ausência de anquilose alveolodentária. O teste Kruskall-Wallis avaliou as diferenças significantes entre grupos, em relação à intensidade inflamatória por...
This study aimed to microscopically investigate the effect on the dental resorptive process of subgingival implants of ELVAX polymer with peptide echistatin in reimplantation of upper incisors in rats. For this purpose, 42 animals were used and divided into groups with and without echistatin. Extra alveolar socket period was 30 and 60 minutes and post-surgical experimental periods were 15, 60 and 90 days. Specimens were processed and stained with H.E. to observe the biological processes in the area. The microscopic events were evaluated by two examiners. Quantitative histomorphometric and descriptive evaluation of the events were performed according to the presence, type and location of the inflammatory response, incidence of resorptions or dental ankylosis. The level of agreement between examiners determined by the Kappa index, proved to be almost perfect for all variables. The comparison between the experimental and control groups for the variables presented was performed using the Mann-Whitney test and showed no statistical difference between the administration of echistatin and extra alveolar socket period relative to the inflammatory intensity in different portions of the root. Regarding the type of resorption established, it was noted that the presence of inflammatory resorption was significantly higher (p<0.05) in the control group on days 30 and 60 minutes in the postoperative period of 15 days. Dental ankylosis was also significantly more prevalent in the group treated with echistatin in extra alveolar socket period of 30 minutes and 15-day trial period. There was even more dental ankylosis in control group with 60 minutes of extra alveolar socket, with the trial period of 60 days. In the trial period of 90 days all specimens studied featured inflammatory resorptives processes and absence of dental ankylosis. The Kruskal-Wallis test assessed significant differences between groups in relation to the intensity of...
Subject(s)
Animals , Male , Rats , Tooth Avulsion/surgery , Platelet Aggregation Inhibitors/therapeutic use , Peptides/therapeutic use , Polyvinyls/therapeutic use , Root Resorption , Tooth Replantation/methods , Rats, Wistar , Time Factors , Treatment OutcomeABSTRACT
O presente trabalho investigou microscopicamente o efeito da equistatina sobre o processo reabsortivo de dentes incisivos superiores de ratos, extraídos e reimplantados. Foram utilizados 42 animais, divididos em grupos com e sem equistatina. Os tempos extra-alveolares dos dentes foram de 30 e 60 minutos e os períodos experimentais pós-cirúrgicos foram 15, 60 e 90 dias. Os espécimes obtidos foram processados e corados em H.E. para observar os processos biológicos presentes. Os eventos microscópicos foram avaliados por dois examinadores, de modo quantitativo histomorfométrico e descritivo, de acordo com o tipo de infiltrado e intensidade da reação inflamatória; com o tipo, extensão e localização de reabsorção dentária; e presença de anquilose alveolodentária. O nível de concordância entre examinadores, determinado pelo índice Kappa, revelou-se quase perfeito para todas as variáveis avaliadas. A comparação entre os grupos testes e controle para as variáveis apresentadas foi feita por meio do teste Mann-Whitney e mostrou não haver diferença estatística entre a administração de equistatina e tempo extra-alveolar em relação à intensidade inflamatória nas diferentes porções radiculares. Quanto ao tipo de reabsorção estabelecida, notou-se que a presença de reabsorção inflamatória foi significativamente maior (p<0,05) no grupo controle nos tempos de 30 e 60 minutos no período pós-cirúrgico de 15 dias. A anquilose alveolodentária também esteve significativamente mais presente no grupo tratado com equistatina com 30 minutos extra-alveolar e 15 dias de período experimental. Houve, ainda, mais anquilose alveolodentária no grupo controle de 60 minutos extra-alveolar, com o período experimental de 60 dias. No período experimental de 90 dias todos os espécimes avaliados apresentaram processos reabsortivos inflamatórios e ausência de anquilose alveolodentária. O teste Kruskall-Wallis avaliou as diferenças significantes entre grupos, em relação à intensidade inflamatória por...
This study aimed to microscopically investigate the effect on the dental resorptive process of subgingival implants of ELVAX polymer with peptide echistatin in reimplantation of upper incisors in rats. For this purpose, 42 animals were used and divided into groups with and without echistatin. Extra alveolar socket period was 30 and 60 minutes and post-surgical experimental periods were 15, 60 and 90 days. Specimens were processed and stained with H.E. to observe the biological processes in the area. The microscopic events were evaluated by two examiners. Quantitative histomorphometric and descriptive evaluation of the events were performed according to the presence, type and location of the inflammatory response, incidence of resorptions or dental ankylosis. The level of agreement between examiners determined by the Kappa index, proved to be almost perfect for all variables. The comparison between the experimental and control groups for the variables presented was performed using the Mann-Whitney test and showed no statistical difference between the administration of echistatin and extra alveolar socket period relative to the inflammatory intensity in different portions of the root. Regarding the type of resorption established, it was noted that the presence of inflammatory resorption was significantly higher (p<0.05) in the control group on days 30 and 60 minutes in the postoperative period of 15 days. Dental ankylosis was also significantly more prevalent in the group treated with echistatin in extra alveolar socket period of 30 minutes and 15-day trial period. There was even more dental ankylosis in control group with 60 minutes of extra alveolar socket, with the trial period of 60 days. In the trial period of 90 days all specimens studied featured inflammatory resorptives processes and absence of dental ankylosis. The Kruskal-Wallis test assessed significant differences between groups in relation to the intensity of...
Subject(s)
Animals , Male , Rats , Tooth Avulsion/surgery , Platelet Aggregation Inhibitors/therapeutic use , Peptides/therapeutic use , Polyvinyls/therapeutic use , Root Resorption , Tooth Replantation/methods , Rats, Wistar , Time Factors , Treatment OutcomeABSTRACT
Objective: To construct a polycistron tandem repeated Echistatin (Ecs) gene. Methods: Three Ecs genes with independent initiation and termination codon were ligated tandem through restriction enzyme sites after amplified with 3 pairs of primers using pMD18T-Ecs as template. The polycistron Ecs gene was inserted into pET30a and expressed in E.coli BL21(DE3) with IPTG induction. The expression results were identified by 18% SDS-PAGE and Western blot. Results: The expression of Ecs polycistron was accomplished with 18% expression level of total protein determined by SDS-PAGE and Western blot. Conclusion: The successful expression of Ecs polycistron provided a new method for the preparation of low molecular weight protein.