Decellularized small intestine submucosa/polylactic-co-glycolic acid composite scaffold for potential application in hypopharyngeal and cervical esophageal tissue repair

Abstract

There has been an increase in the incidence of hypopharyngeal and cervical esophageal cancer worldwide, and hence growing needs for hypopharyngeal and cervical esophageal tissue repair. This work produced a bi-layer composite scaffold with decellularized small intestine submucosa and polylactic-co-glycolic acid, which resembled the layered architectures of its intended tissues. The decellularized small intestine submucosa contained minimal residual DNA (52.5 ± 1.2 ng/mg) and the composite scaffold exhibited satisfactory mechanical properties (a tensile modulus of 21.1 ± 4.8 MPa, an ultimate tensile strength of 14.0 ± 2.9 MPa and a failure strain of 26.9 ± 5.1%). The interactions between cells and the respective layers of the scaffold were characterized by CCK-8 assays, immunostaining and Western blotting. Desirable cell proliferation and phenotypic behaviors were observed. These results have provided an important basis for the next-step in vivo studies of the scaffold, and bode well for its future clinical applications.

Keywords: composite scaffold; hypopharyngeal and cervical esophageal cancer; polylactic-co-glycolic acid; small intestine submucosa; tissue repair.

Figure 1.

H&E staining images of (a) SIS and (b) dSIS

Figure 2.

SEM micrographs of dSIS/PLGA composite scaffold: (a) principal view of the dSIS side; (b) the PLGA side with an angled view of the cross-section.

Figure 3.

Cell proliferation measured by CCK-8 assays on Day 1, 3 and 7: (a) ESMCs on the dSIS side of scaffold; (b) EECs on the PLGA side of scaffold. *P < 0.05.

Figure 4.

Immunofluorescent images of ESMCs and ECs on composite scaffolds: (a) ESMCs with anti-actin antibody on Day 1, 3 and 7. α-actin displayed green and nuclei displayed blue. (b) EECs with anti-CK-14 antibody on Day 1, 3 and 7. CK-14 displayed green and nuclei displayed blue.

Figure 5.

Western blotting analysis of ESMCs and ECs on the scaffolds: (a) Western blotting bands of α-actin and GAPDH of ESMCs on the dSIS side of scaffold and in control group. (b) Normalized intensities of α-actin bands. (c) Western blotting bands of CK-14 and GAPDH of EECs on the PLGA side of scaffold and in control group. (d) Normalized intensities of CK-14 bands. *P < 0.05.