In vitro construction and in vivo regeneration of esophageal bilamellar muscle tissue

Abstract

In order to induce esophageal muscle cells' orientation, the silicon wafer with prototype 1 and prototype 2 was designed. Prototype 1 has micro-channels of 200 µm width and 30 µm depth with 30 µm wide wall as the interval. Prototype 2 has channels of 100 µm width and 30 µm depth with a discontinuous wall which has 30 µm gap for each 100 µm channel. The poly(ester urethane) scaffolds with pattern prototype 1 and prototype 2 were fabricated using solution casting method and abbreviated as PU1 and PU2, respectively. Silk fibroin was grafted individually on PU1 and PU2 surface (PU1-SF, PU2-SF) using our previous protocol, aiming at improving scaffolds' biocompatibility. The primary esophageal smooth muscle cell was seeded to evaluate the scaffolds' cytocompatibility in vitro. Characterizations like MTT assay, immunocytochemistry, scanning electron microscope, and Western blotting were applied. After that, poly(ester urethane) scaffolds with double patterns, prototype 1 on the exterior, and prototype 2 in the lumen were implanted into the rabbit esophagous to test the regeneration of the muscle tissue. Results from these preliminary tests showed that the growth and differentiation of primary smooth muscle cells were promoted, but also the muscle tissue with endocircular and exolongitudinal architecture was in regenerating, against non-constitution in the animals without the patterned scaffold or with poly(ester urethane) plane membrane at the defaulted sites. This micro-channel pattern together with silk fibroin grafting and vascular endothelial growth factor coating greatly promoted the regeneration of esophageal muscle with normal histological structure.

Recovery from volumetric muscle loss injury: A comparison between young and aged rats

“…The poor clinical outcome following VML injury combined with a lack of effective treatment options motivates our exploration of VML regeneration. To better understand the sequence of events that follow VML injury and aid in the development of effective treatment strategies, various pre-clinical animal models have been developed (Chen and Walters 2013; Hou and others 2016; Turner and others 2012; Wu and others 2012; Zhang and others 2016). Among these, the hind limb muscles of the rat are a reliable pre-clinical model that recapitulates healing with non-contractile scar tissue observed in humans following VML injury (Wu and others 2012).…”

Tissue Engineering of Esophagus

“…Further, the scaffold surface was grafted with silk fibroin using our method of diamine aminolysis and GA crosslinking. The primary esophageal SMC was cultured in these 3D protein-grafted channels in order to achieve SMC phenotype regulation and in situ muscle formation [63]. The results confirmed that primary esophageal smooth muscle cells exhibited fine alignment in all types of microchannels while SMCs in the interval channels communicated well through the gaps (Figure 5).…”

Development and Prospect of Esophageal Tissue Engineering

“…From the perspective of single-layer scaffold structure, the classification mainly includes acellular matrix, membranes grafted with biomolecules, electrospinning scaffolds, micro-pattern scaffolds, etc. ( Hou et al, 2016 ; Wei et al, 2018 ; Zhuravleva et al, 2019 ; Chaitin et al, 2021 ; Levenson et al, 2021 ) ( Table 2 ). In order to accurately simulate the inner ring and outer longitudinal structure of the muscle layer, electrospinning scaffolds and micro-pattern scaffolds are mainly used ( Gong et al, 2013 ; Kuppan et al, 2017 ).…”