Abstract Background Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a rare inherited skin disease caused by variants in the COL7A1 gene, coding for type VII collagen (C7), an important component of anchoring fibrils in the basement membrane of the epidermis. RDEB patients suffer from skin fragility starting with blister formation and evolving into chronic wounds, inflammation and skin fibrosis, with a high risk of developing aggressive skin carcinomas. Restricted therapeutic options are limited by the lack of in vitro models of defective wound healing in RDEB patients. Results In order to explore a more efficient, non-invasive in vitro model for RDEB studies, we obtained patient fibroblasts derived from discarded dressings) and examined their phenotypic features compared with fibroblasts derived from non-injured skin of RDEB and healthy-donor skin biopsies. Our results demonstrate that fibroblasts derived from RDEB chronic wounds (RDEB-CW) displayed characteristics of senescent cells, increased myofibroblast differentiation, and augmented levels of TGF-β1 signaling components compared to fibroblasts derived from RDEB acute wounds and unaffected RDEB skin as well as skin from healthy-donors. Furthermore, RDEB-CW fibroblasts exhibited an increased pattern of inflammatory cytokine secretion (IL-1β and IL-6) when compared with RDEB and control fibroblasts. Interestingly, these aberrant patterns were found specifically in RDEB-CW fibroblasts independent of the culturing method, since fibroblasts obtained from dressing of acute wounds displayed a phenotype more similar to fibroblasts obtained from RDEB normal skin biopsies. Conclusions Our results show that in vitro cultured RDEB-CW fibroblasts maintain distinctive cellular and molecular characteristics resembling the inflammatory and fibrotic microenvironment observed in RDEB patients’ chronic wounds. This work describes a novel, non-invasive and painless strategy to obtain human fibroblasts chronically subjected to an inflammatory and fibrotic environment, supporting their use as an accessible model for in vitro studies of RDEB wound healing pathogenesis. As such, this approach is well suited to testing new therapeutic strategies under controlled laboratory conditions.

Los primates son uno de los grupos de mamíferos más decomisados por la autoridades ambientales (SDA) en Bogotá, Colombia. Un total de 133 primates fueron confiscados en Bogotá durante el año 2008 y mantenidos en las instalaciones de la SDA. De ellos, 115 fueron secuenciados para el gen mitocondrial citocromo oxidasa II (mtCOII) y en 112 ejemplares, las secuencias obtenidas fueron de alta calidad. Esas secuencias se compararon con las obtenidas para ejemplares muestreados directamente en campo por nuestro grupo de investigación y con origen geográfico conocido. De ese modo, se pudo determinar las áreas geográficas, en el territorio colombiano, donde pueden liberarse esos ejemplares después del tratamiento de rehabilitación oportuno. Los resultados principales para las cinco especies de primates fueron como siguen: 1- Para Saguinus leucopus, los animales analizados pueden ser liberados en cualquier área geográfica dentro del rango de distribución de la especie, ya que solo se detectó un acervo genético sin estructura espacial. 2- Para los 14 Aotus sp. secuenciados procedentes de la SDA, se determinó que: uno de ellos pertenecía a A. vociferans, propio de la Amazonía; siete ejemplares pertenecieron a A. griseimembra, propio del valle del Magdalena hasta la costa Caribe colombiana; cuatro ejemplares representaron a A. brumbacki, de los Llanos Orientales de Colombia; y dos ejemplares se asociaron con A. azarae azarae del norte de Argentina y Paraguay, con lo cual se muestra que en Colombia se está recibiendo fauna ilegal procedente de otros países. 3- De los 14 Cebus albifrons secuenciados, dos pertenecieron al área geográfica de distribución de C. a. versicolor; uno al de C. a. pleei, 10 al de C. a. leucocephalus, y uno no pudo ser asignado ya que su secuencia mostraba gran divergencia respecto a los otros ejemplares secuenciados de esta especie. 4- Los dos Cebus capucinus secuenciados mostraron estar asociados a un acervo genético encontrado en el norte del Chocó, Sucre y Córdoba. 5- De 11 Cebus apella secuenciados, 10 mostraron pertenecer al acervo genético que se encuentra en los Llanos Orientales de Colombia y altamente relacionado a Cebus apella apella de la Guyana Francesa, aunque podrían representar un acervo propio de Colombia, C. a. fatuellus sensu Groves (2001). Un individuo no pudo ser relacionado con ningún grupo de los otros C. apella estudiados, ni con los taxones relacionados a la especie mencionada, pero, probablemente, con su propio estatus taxonómico (C. a. paraguayanus = C. cay, C. xanthosternos, C. nigritus).

Genetic methods for the reintroduction of primates Saguinus, Aotus and Cebus (Primates: Cebidae) seized in Bogota, Colombia. Primates are one of more confiscated taxa by the environmental authorities in Bogota, Colombia. During 2008, 133 monkeys were confiscated; samples from 115 of them were sequenced by the mitochondrial cythocrome oxidase II gene (mtCOII) and 112 sequences obtained were of high quality. These sequences were compared with those obtained by our research group from individuals directly sampled in the field, with precise geographic origin. So, a more specific geographic area of the Colombian territory could be considered for a correct rehabilitation treatment during the reintroduction of these confiscated animals. The main results with five primate species were: 1- For all the specimens analyzed of Saguinus leucopus, they could be liberated in any geographical area of its distribution range, since only one gene pool was found. 2- For the 14 Aotus sp. individuals sequenced from the SDA (Environmental District Secretariat), one of them (A. vociferans) was coming from the Amazon, seven exemplars belonged to A. griseimembra from the Magdalena Valley and the Colombian Caribbean coasts, four individuals represented to A. brumbacki from the Colombian Eastern Llanos, and two were associated to A. azarae azarae from Northern Argentina and Paraguay (which means that illegal traffic of animals is arriving to Colombia from other South-American countries). 3-Out 14 Cebus albifrons sequenced, two belonged to the geographical area of C. a. versicolor, one to C. a. pleei, 10 to C. a. leucocephalus and one could be not assigned because its sequence yielded a great genetic divergence with respect to the other specimens sequenced of this species. 4- The two Cebus capucinus sequenced showed to be associated to a gene pool found in the Northern of Chocó, Sucre and Córdoba Departments. 5- Out 11 Cebus apella sequenced, 10 showed to belong to the gene pool presented in the Colombian Eastern Llanos and highly related (but differentiable) to Cebus apella apella from the French Guyana. It could be named C. a. fatuellus sensu Groves (2001). One exemplar sequenced could be not related with the other C. apella analyzed, nor the related taxa to the aforementioned species (C. a. paraguayanus =C. cay; C. xanthosternos; C. nigritus). Rev. Biol. Trop. 58 (3): 1049-1067. Epub 2010 September 01.