![]() FITC-conjugated AffiniPure goat anti-mouse IgG (H+L) was purchased from Jackson ImmunoResearch. Anti-MyoD1 antibody was supplied by Abcam (Hong Kong) Ltd. Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), antibiotic antimycotic solution (AAS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 100 μg/mL amphotericin B), and Trypsin-EDTA solution were purchased from Gibco (Invitrogen, Co., New York). Poly(ester urethane) (PU, 58213 NAT 022) was purchased from Estane Co., China. Furthermore, we will offer a new perspective on the implications of ex vivo niche system to advance the scientific understanding of stem cell functions as well as supporting clinical applications. Protein grafted surfaces promote human hypopharyngeal cell proliferation and differentiation. In our experiment, scaffolds with linearly aligned microchannels were fabricated to align the seeded myoblasts through contact guidance. These results suggest that 3D muscle grafts created in vitro from collagen composite scaffolds seeded with myoblasts can be used for defect muscle tissue repair in vivo. The scaffolds were found to degrade slowly over time, and muscle healing was improved as shown by an increased quantity of innervated and vascularized muscle fibers. The 3D graft was then used for in vivo muscle tissue repair by implanting it into defect sites created in mice models. In another study by Ma and coworkers, ATCC were seeded onto a collagen composite scaffold and cultured in a roller bottle cell culture system to create a 3-dimensional (3D) tissue graft in vitro. The myoblasts formed long and unbranched myotubes that had uniform diameter and were aligned parallel to the microgroove direction to suggest that microgrooves promote end to end fusion of myoblasts. Huang and colleagues cultured murine myoblasts (ATCC C2C12) on elastic poly(dimethylsiloxane) (PDMS) films topographically micropatterned with 10 μm wide microgrooves. Muscle fiber consists of a longitudinal arrangement of myofilaments with actin and myosin as major components. Skeletal muscle is the most abundant tissue type in the human body. However, these grafts cannot fully restore the function of hypopharynx tissue, in particular the reconstruction of the complex structures surrounding the larynx and hypopharynx, while postoperative dyspnea may be inevitable when hypopharynx pharyngeal constrictors were sectioned. For the severe and large area hypopharynx defects, visceral or myocutaneous flaps, such as an ileocolic flap, a radial forearm flap, and an anterolateral thigh flap, a submental island flap, and an infrahyoid myocutaneous flap, are common methods used for surgical repair. Also, physical deformities and emotional trauma persist long after the conclusion of surgical intervention. However, surgical interventions are inevitably associated with quality-of-life impairments including severe speech and swallowing disability. Laryngopharyngectomy and reconstruction followed by chemoradiotherapy has been the traditional and relatively practicable treatment presently. Hypopharynx carcinoma is one of the common head and neck cancers approximately 2500 new cases are diagnosed in the United States each year with the peak incidence in males and females aged 50 to 60 years. Our results demonstrate that polyurethane scaffolds seeded with myoblasts hold promise to guide hypopharynx muscle regeneration. Also, the myoblasts seeded on PU-SF had the higher proliferative rate and better differentiation compared with those on the control or PU-Gel. Scaffolds were observed to degrade slowly over time, and their mechanical properties and hydrophilicities were improved through the surface grafting. Due to the low hydrophilicity of PU, the scaffold was grafted with silk fibroin (PU-SF) or gelatin (PU-Gel) to improve its cell adhesion properties. In this study, a polyurethane (PU) scaffold with microchannel patterns was used to provide aligning guidance for the seeded human myoblasts. For hypopharynx tissue engineering, skeletal muscle is one important functional part of the whole organ, which assembles the unidirectionally aligned myotubes. In the field of tissue engineering, polymeric materials with high biocompatibility like polylactic acid and polyglycolic acid have been widely used for fabricating living constructs.
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