Pediatric Short Bowel Syndrome is a rare malabsorption disease occurring because of massive surgical resections of the small intestine. To date, the issues related to current strategies including intestinal transplantation prompted the attention towards tissue engineering (TE). Tis work aimed to develop and compare two composite scaffolds for intestinal TE consisting of a novel hydrogel, that is, oxidized polyvinyl alcohol (OxPVA), cross-linked with decellularized intestinal wall as a whole (wW/OxPVA) or homogenized (hW/OxPVA). A characterization of the supports was performed by histology and Scanning Electron Microscopy and their interaction with adipose mesenchymal stem cells occurred by MTT assay. Finally, the scaffolds were implanted in the omentum of Sprague Dawley rats for 4 weeks prior to being processed by histology and immunohistochemistry (CD3; F4/80; Ki-67; desmin; alpha-SMA; MNF116). In vitro studies proved the effectiveness of the decellularization, highlighting the features of the matrices; moreover, both supports promoted cell adhesion/proliferation even if the wW/OxPVA ones were more effective ( < 0.01). Analysis of explants showed a continuous and relatively organized tissue wall around the supports with a connective appearance, such as myofibroblastic features, smooth muscle, and epithelial cells. Both scaffolds, albeit with some difference, were promising; nevertheless, further analysis will be necessary.
La sindrome dell’intestino corto in età pediatrica è una condizione severa rara che si associa a malassorbimento e nei casi più gravi porta ad insufficienza intestinale. Attualmente, non esistono degli approcci chirurgici/ricostruttivi efficaci, ed il trapianto di intestino non è una strategia facilmente percorribile; in tale contesto, l’ingegneria tissutale si propone come uno strumento utile per poter identificare dei device d’avanguardia. Lo scopo di questo studio è stato quello di allestire e comparare (istologia, microscopia elettronica a scansione, test in vitro) due scaffold compositi costituiti da polivinil alcol ossidato (OxPVA) e matrice intestinale acellulare, intera o omogeneizzata. In seguito, i device sono stati impiantati nell’omento di ratti Sprague Dawley per 4 settimane per poi essere analizzati sia tramite istologia che immunoistochimica. Gli studi in vitro hanno mostrato l’efficienza della decellularizzazione e la capacità degli scaffold compositi di promuovere l’adesione e la proliferazione cellulare. Analisi sugli espianti hanno mostrato la presenza, attorno ai supporti, di uno strato tissutale organizzato dove si potevano riconoscere componenti connettivali, muscolari lisce ed epiteliali. Entrambi i supporti sono promettenti, tuttavia ulteriori analisi sono necessarie per una più specifica caratterizzazione degli stessi, oltre che del potenziale intrinseco in vivo.
Validazione in vitro e in vivo di scaffold compositi formati da matrice extracellulare intestinale e polivinil alcol ossidato per un nuovo approccio rigenerativo nella sindrome dell’intestino corto
DAL ZOVO, ERICA
2021/2022
Abstract
Pediatric Short Bowel Syndrome is a rare malabsorption disease occurring because of massive surgical resections of the small intestine. To date, the issues related to current strategies including intestinal transplantation prompted the attention towards tissue engineering (TE). Tis work aimed to develop and compare two composite scaffolds for intestinal TE consisting of a novel hydrogel, that is, oxidized polyvinyl alcohol (OxPVA), cross-linked with decellularized intestinal wall as a whole (wW/OxPVA) or homogenized (hW/OxPVA). A characterization of the supports was performed by histology and Scanning Electron Microscopy and their interaction with adipose mesenchymal stem cells occurred by MTT assay. Finally, the scaffolds were implanted in the omentum of Sprague Dawley rats for 4 weeks prior to being processed by histology and immunohistochemistry (CD3; F4/80; Ki-67; desmin; alpha-SMA; MNF116). In vitro studies proved the effectiveness of the decellularization, highlighting the features of the matrices; moreover, both supports promoted cell adhesion/proliferation even if the wW/OxPVA ones were more effective ( < 0.01). Analysis of explants showed a continuous and relatively organized tissue wall around the supports with a connective appearance, such as myofibroblastic features, smooth muscle, and epithelial cells. Both scaffolds, albeit with some difference, were promising; nevertheless, further analysis will be necessary.File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.12608/34397