The design of in vitro liver sinusoid mimics using chitosan-hyaluronic acid polyelectrolyte multilayers.
Interactions between hepatocytes and liver sinusoidal endothelial cells (LSECs) are essential for the development and maintenance of hepatic phenotypic functions. We report the assembly of three-dimensional liver sinusoidal mimics comprised of primary rat hepatocytes, LSECs, and an intermediate chitosan-hyaluronic acid polyelectrolyte multilayer (PEM). The height of the PEMs ranged from 30 to 55 nm and exhibited a shear modulus of approximately 100 kPa. Hepatocyte-PEM cellular constructs exhibited stable urea and albumin production over a 7-day period, and these values were either higher or similar to cells cultured in a collagen sandwich. This is of significance because the thickness of a collagen gel is approximately 1000-fold higher than the height of the chitosan-hyaluronic acid PEM. In the hepatocyte-PEM-LSEC liver-mimetic cellular constructs, LSEC phenotype was maintained, and these cultures exhibited stable urea and albumin production. CYP1A1/2 activity measured over a 7-day period was significantly higher in the hepatocyte-PEM-LSEC constructs than in collagen sandwich cultures. A 16-fold increase in CYP1A1/2 activity was observed for hepatocyte-PEM-10,000 LSEC samples, thereby suggesting that interactions between hepatocytes and LSECs are critical in enhancing the detoxification capability in hepatic cultures in vitro.
Kim
Yeonhee
Y
Department of Chemical Engineering, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061, USA.
Larkin
Adam L
AL
Davis
Richey M
RM
Rajagopalan
Padmavathy
P
D000818:Animals; D048271:Chitosan / Q000737:chemistry*; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D051381:Rats; D046509:Tissue Culture Techniques
D011108:Polymers; D006820:Hyaluronic Acid; D048271:Chitosan