Abstract
Aim of work was to locate a simple, reproducible protocol for uniform seeding and optimal cellularization of biodegradable patch minimizing the risk of structural damages of patch and its contamination in long-term culture. Two seeding procedures are exploited, namely static seeding procedures on biodegradable and biocompatible patches incubated as free floating (floating conditions) or supported by CellCrownTM insert (fixed conditions) and engineered by porcine bone marrow MSCs (p-MSCs). Scaffold prototypes having specific structural features with regard to pore size, pore orientation, porosity, and pore distribution were produced using two different techniques, such as temperature-induced precipitation method and electrospinning technology. The investigation on different prototypes allowed achieving several implementations in terms of cell distribution uniformity, seeding efficiency, and cellularization timing. The cell seeding protocol in stating conditions demonstrated to be the most suitable method, as these conditions successfully improved the cellularization of polymeric patches. Furthermore, the investigation provided interesting information on patches’ stability in physiological simulating experimental conditions. Considering the in vitro results, it can be stated that the in vitro protocol proposed for patches cellularization is suitable to achieve homogeneous and complete cellularizations of patch. Moreover, the protocol turned out to be simple, repeatable, and reproducible.
Highlights
There are several conditions, both congenital and acquired, for which esophageal replacement is needed [1]
The results demonstrated the fabrication process and their resulting structure significantly affected patches wettability, since temperature-induced precipitation (TIP)-Fs and electrospun matrices (EL-Ms) prototypes are based on the same polymer combination
Our results demonstrated that porcine bone marrow Mesenchymal stem cells (MSCs) (p-MSCs) were able to adhere, proliferate, and colonize both type of patches, confirming their biocompatibility
Summary
There are several conditions, both congenital and acquired (malignant or benign tumor, in some forms of atresia, caustic lesion, and severe reflux), for which esophageal replacement is needed [1]. The surgical intervention remains the preferred treatment, alternative options are essential for restoring esophageal continuity and functionality [2,3,4,5] These alternative approaches may be associated with several complications, such as inflammatory response, scar deposition, stricture formation, and in the most serious cases, they could cause the morbidity of surrounding tissues [6]. In the last few years, tissue engineering (TE) has emerged as a promising solution for the replacement of physiological esophageal functions [7,8,9] This field of science has deeply benefited from meaningful progresses in biomaterial science, nanotechnologies, bioreactor technology, molecular biology, and stem cells discovery [10,11]. The scaffolds are turning into sophisticated structure for local delivering of active agents (such as proteins, genes, growth factors, antibiotics, and anti-inflammatory drugs) in their free form or loaded in micro- and nano-carriers [12]
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