RESUMEN
PURPOSE: Stratifin is a potent anti-fibrogenic factor that stimulates the expression of matrix metalloproteinase-1 (MMP-1) in dermal fibroblasts. The propose of this work was to develop a controlled release delivery system for stratifin that can be applied at the time of wound closure to release stratifin and stimulate the expression of MMP-1 in a sustained manner over the late stages of wound healing (after 3 days). METHODS: Stratifin was complexed to chitosan particles, which were then encapsulated in PLGA microspheres and blended into crosslinked hyaluronic acid films. In vitro release was assessed using fluorescent-tagged stratifin, cytotoxicity by MTT assay and bioactivity by measuring the levels of MMP-1 expression in cultured fibroblasts. RESULTS: The release of stratifin was delayed for 3 days and then controlled for 30 days so that 60% of the total stratifin loaded was released. The released protein significantly stimulated the expression of MMP-1 in cultured fibroblasts without compromising cell viability. By complexing to chitosan, the initial burst release was reduced, so that only 5% of stratifin was released in 3 days. CONCLUSION: This stratifin delivery system has the potential to be used as an anti-fibrogenic factor-associated wound insert for improving post-surgical scarring in closed wound.
Asunto(s)
Biomarcadores de Tumor/administración & dosificación , Exonucleasas/administración & dosificación , Hidrogeles , Metaloproteinasa 1 de la Matriz/metabolismo , Proteínas de Neoplasias/administración & dosificación , Piel/efectos de los fármacos , Proteínas 14-3-3 , Biomarcadores de Tumor/farmacología , Quitosano , Exonucleasas/farmacología , Exorribonucleasas , Fibroblastos/efectos de los fármacos , Fibroblastos/enzimología , Humanos , Ácido Láctico , Proteínas de Neoplasias/farmacología , Tamaño de la Partícula , Ácido Poliglicólico , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/farmacología , Piel/citología , Piel/enzimologíaRESUMEN
BACKGROUND & OBJECTIVE: p53 gene is the main regulator of 14-3-3sigma. Activated p53 could induce the expression of 14-3-3sigma, while 14-3-3sigma stabilizes the expression of p53 and enhances its transcriptional activity. p63 and p73, the members of p53 family, also have some functions similar to p53. This study was to investigate the effect of 14-3-3sigma on the transcriptional activity of p73. METHODS: Luciferase reporter assay, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot were used to evaluate the effect of 14-3-3sigma on the transcriptional activity of p73 in p53-deficient human lung carcinoma cell line H1299. Colony formation test was used to evaluate the effect of 14-3-3sigma on the transcriptional activity of p73 in p53-mutant human breast cancer cell line MDA-MB-436. RESULTS: The luciferase activities induced by bax and p21WAF1 promotors were significantly higher in p73-transfected H1299 cells than in control H1299 cells (P<0.01), and were further increased by the transfection of p73 (25 ng) and 14-3-3sigma (100, 200, and 400 ng) in a dose-dependent manner (P<0.01). The expression of bax and p21WAF1 were higher in p73-transfected H1299 cells than in control H1299 cells, and were significantly higher in p73-and 14-3-3sigma-transfected H1299 cells than in p73-transfected H1299 cells (P<0.01). The number of colonies was fewer in p73-transfected MDA-MB-436 cells than in control MDA-MB-436 cells, and the colonies were significantly smaller in p73-and 14-3-3sigma-transfected H1299 cells than in p73-transfected H1299 cells (P<0.01). CONCLUSION: 14-3-3sigma can enhance the transcriptional activity of p73 in a dose-dependent manner.
Asunto(s)
Biomarcadores de Tumor/farmacología , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/biosíntesis , Proteínas de Unión al ADN/metabolismo , Exonucleasas/farmacología , Neoplasias Pulmonares/metabolismo , Proteínas de Neoplasias/farmacología , Proteínas Nucleares/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Proteína X Asociada a bcl-2/biosíntesis , Proteínas 14-3-3 , Biomarcadores de Tumor/administración & dosificación , Western Blotting , Neoplasias de la Mama/patología , Línea Celular Tumoral , Proliferación Celular , Ensayo de Unidades Formadoras de Colonias , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Proteínas de Unión al ADN/genética , Relación Dosis-Respuesta a Droga , Exonucleasas/administración & dosificación , Exorribonucleasas , Femenino , Humanos , Luciferasas/metabolismo , Neoplasias Pulmonares/patología , Proteínas de Neoplasias/administración & dosificación , Proteínas Nucleares/genética , Plásmidos , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteína Tumoral p73 , Proteínas Supresoras de Tumor/genética , Proteína X Asociada a bcl-2/genéticaRESUMEN
AIDS: Presentation highlights are provided from the Nineteenth International Congress on Chemotherapy in Montreal. Some of the new data on anti-HIV therapeutics include phase I/II data on Agouron's protease inhibitor AG1343; information on the new protease inhibitor palinavir PRO2000 (a gp120/CD4 binding inhibitor); and liposomal (fat-enclosed) formulations of both foscarnet and exonuclease (a cellular enzyme that destroys viral DNA). Two additional studies from Brazil on the efficacy and safety of roxithromycin for cryptosporidiosis (the Food and Drug Administration (FDA) has not approved roxithromycin for sale in the U.S.) are discussed, as is the use of imiquimod treatment for genital warts.^ieng
