Project description:The production of skeletal muscle constructs useful for in vivo large defects replacement, such as congenital diaphragmatic hernia (CDH), is still considered a challenge. The standard application of prosthetic material presents strong limitations, like hernia recurrences in a remarkable number of CDH patients. With this work, we developed a tissue engineering approach based on decellularized diaphragmatic muscle and human cells for the generation in vitro of diaphragmatic-like tissues as proof-of-concept of a new option for the surgical treatment of large diaphragm defects. A specific bioreactor for diaphragmatic muscle was achieved to control mechanical stimulation during the construct generation. In vitro tests demonstrated the increased maturation of mechanically stimulated constructs, with the formation of new oriented and aligned muscle fibres. Moreover, after in vivo orthotopic implantation in a CDH mouse model, mechanically stimulated muscles maintained the presence of human cells within myofibers, supporting the idea of a new therapeutic option for this dramatic congenital malformation. Production of diaphragmatic-like tissues using mouse decellularized extracellular matrix, human cells and mechanical stimulation with bioreactor
Project description:Investigation into the effects of Congenital Diaphragmatic Hernia (CDH) and subsequent treatment with tracheal occlusion (TO) on the pulmonary transcriptome. A diaphragm defect was created by surgical means in fetal rabbits. The surgical creation of diaphragmatic hernia (DH) allows for direct analysis of changes in pulmonary gene expression due to pulmonary hypoplasia, without the need for gene knockdown (as for KO mice) or use of teratogens (such as nitrofen). The subsequent treatment with tracheal occlusion (TO) was also investigated to determine the changes in gene expression due to forced lung growth in the prenatal phase. RNA-Seq analysis was performed on left lung samples from fetal rabbits. Samples were generated and analysed for DH (n=4), TO (n=6), and control lungs (n=4)
Project description:Congenital diaphragmatic hernia (CDH) is a life-threatening anomaly with high morbidity and mortality. To investigate the pathogenesis of CDH, miRNA sequencing was performed using amniotic fluid-derived extracellular vesicles (AF-EVs) of CDH patients.
Project description:Congenital diaphragmatic hernia (CDH) is a life-threatening anomaly with high morbidity and mortality. To investigate the pathogenesis of CDH, miRNA sequencing was performed using amniotic fluid-derived extracellular vesicles (AF-EVs) and fetal lung tissue of nitrofen-induced CDH rat model.
Project description:Lung organoids made from pluripotent stem cells have the potential to enhance our understanding of disease mechanisms in pediatric lung disorders. As proof of concept, we have established a reproducible ex vivo model of lung organoid development derived from human induced pluripotent stem cells generated from fetuses and infants with Bockdalek congenital diaphragmatic hernia (CDH), a polygenic disorder associated with fetal lung compression and often lethal pulmonary hypoplasia at birth. We used microarrays to compare transcriptomes among the different cell types focusing on genes associated with lung development and extracellular matrix. We seek to identity anomalous gene expression during lung development, using lung organoids generated from hiPS of patients with congenital diafragmatic hernia.
