Tissue Engineered Esophageal Patch by Mesenchymal Stromal Cells: Optimization of Electrospun Patch Engineering

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.

Graphene Nanoplatelets for the Development of Reinforced PLA–PCL Electrospun Fibers as the Next-Generation of Biomedical Mats

“…Electrospun PLA–PCL-based fibrous mats cellularization was deeply investigated in our previous work [ 7 ] demonstrating that the cells adhered, proliferated and colonized patches, thus confirming the biocompatibility of the PLA–PCL-based fibrous mats.…”

“…Electrospinning has been revealed a relatively simple, robust and up-scalable technique at the forefront of micro-nanofiber mats fabrication for industrial (e.g., water/air filtration, textile and packaging materials, optical electronics and biosensors) and biomedical/pharmaceutical applications (e.g., tissue engineering and regenerative medicine, wound dressing, implant coating films and in-product labeling of tablets, drug delivery) [ 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 ]. This process offers unique capabilities for producing nanofibrous fabrics with wide variety of sizes and shapes, extremely high surface-to-volume ratio and tunable porosity [ 1 , 13 ].…”

Standardized User-Independent Confocal Microscopy Image Acquisition and Analysis for Thickness Measurements of Microscale Collagen Scaffolds

“…EM is one of the most common thickness determination approaches in the tissue engineering literature (Versteegden et al, 2017;Pisani et al, 2020). Although cryo-EM permits measurements in hydrated samples, including collagen structures (Quan & Sone, 2013), it has not been widely employed for the determination of tissue engineering scaffold thickness.…”

“…Accurate and precise thickness measurements of biomaterial scaffolds are essential for many tissue engineering applications (Soletti et al, 2010;Fotticchia et al, 2018;Pisani et al, 2020). Thickness values play a key role in determining dimensional and mechanical properties of tissue-engineered constructs, such as elastic modulus, ultimate tensile strength, and compliance (Dahl et al, 2011;Wise et al, 2011;Kumar et al, 2013).…”

“…Electron microscopy (EM; Zhang et al, 2012) and optical approaches, such as ellipsometry (Sileika et al, 2013) and white light interferometry (Dzhoyashvili et al, 2016), have been used for the determination of nanometer scale film coatings. When determining tissue engineering-relevant microscale features (100 nm-100 μm), preferred measurement strategies have largely included optical (Dahl et al, 2007;Konig et al, 2009), confocal (Lawrence et al, 2010;Jiang et al, 2017), and most commonly, electron microscopy (Versteegden et al, 2017;Pisani et al, 2020).…”