Collagen I-matrigel scaffolds for enhanced Schwann cell survival and control of three-dimensional cell morphology.
We report on the ability to control three-dimensional Schwann cell (SC) morphology using collagen I-Matrigel composite scaffolds for neural engineering applications. SCs are supportive of nerve regeneration after injury, and it has recently been reported that SCs embedded in collagen I, a material frequently used in guidance channel studies, do not readily extend processes, instead adopting a spherical morphology indicative of little interaction with the matrix. We have modified collagen I matrices by adding Matrigel to make them more supportive of SCs and characterized these matrices and SC morphology in vitro. Incorporation of 10%, 20%, 35%, and 50% Matrigel by volume resulted in 2.4, 3.5, 3.7, and 4.2 times longer average SC process length after 14 days in culture than with collagen I-only controls. Additionally, only 35% and 50% Matrigel constructs were able to maintain SC number over 14 days, whereas an 88% decrease in cells from initial seeding density was observed in collagen-only constructs over the same time period. Mechanical testing revealed that the addition of 50% Matrigel increased matrix stiffness from 6.4 kPa in collagen I-only constructs to 9.8 kPa. Furthermore, second harmonic generation imaging showed that the addition of Matrigel resulted in non-uniform distribution of collagen I, and scanning electron microscope imaging illustrated distinct differences in the fibrillar structure of the different constructs. Collectively, this work lays a foundation for developing scaffolding materials that are concurrently supportive of neurons and SCs for future neural engineering applications.
Dewitt
Daniel D
DD
Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Troy, New York, USA.
Kaszuba
Stephanie N
SN
Thompson
Deanna M
DM
Stegemann
Jan P
JP
D000818:Animals; D000831:Animals, Newborn; D003094:Collagen / Q000737:chemistry*; D004338:Drug Combinations; D007797:Laminin / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D011509:Proteoglycans / Q000737:chemistry*; D051381:Rats; D012583:Schwann Cells / Q000166:cytology* / Q000648:ultrastructure; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen