Medium flow rate regulates viability and barrier function of engineered skin substitutes in perfusion culture

Tissue Engineering. Part a
Balaji KalyanaramanSteven Boyce

Abstract

Perfusion culture of engineered tissues improves mass transfer of nutrients and provides flow-mediated mechanical stimulation to the developing constructs, thereby improving their anatomy and physiology in vitro. In this study, the responses to medium flow rate of engineered skin substitutes (ESS) incubated in perfusion at the air-liquid interface were investigated. ESS fabricated with autologous keratinocytes, fibroblasts, and collagen-glycosaminoglycan (GAG) sponges were incubated for 21 days at the air-liquid interface in a custom-built recirculating bioreactor system at flow rates of 5, 15, and 50 mL/min (n = 8 per condition). ESS were evaluated in vitro using histology, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, bromodeoxyuridine (BrdU) incorporation, and surface hydration. ESS incubated at 5 and 15 mL/min had histological organization comparable with that of control ESS incubated in static conditions. ESS incubated at 50 mL/min displayed a disorganized epidermal substitute and, at later time points in culture, showed greater degradation of the dermal scaffold. Cell viability measured using MTT assay was significantly higher in ESS incubated at 5 mL/min than in static controls at day 14 (mean...Continue Reading

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