Dynamic culture enhances stem cell infiltration and modulates extracellular matrix production on aligned electrospun nanofibrous scaffolds.
Electrospun nanofibrous scaffolds have become widely investigated for tissue engineering applications, owing to their ability to replicate the scale and organization of many fiber-reinforced soft tissues such as the knee meniscus, the annulus fibrosus of the intervertebral disc, tendon, and cartilage. However, due to their small pore size and dense packing of fibers, cellular ingress into electrospun scaffolds is limited. Progress in the application of electrospun scaffolds has therefore been hampered, as limited cell infiltration results in heterogeneous deposition of extracellular matrix and mechanical properties that remain below native benchmarks. In the present study, dynamic culture conditions dramatically improved the infiltration of mesenchymal stem cells into aligned nanofibrous scaffolds. While dynamic culture resulted in a reduction of glycosaminoglycan content, removal from dynamic culture to free-swelling conditions after 6 weeks resulted recovery of glycosaminoglycan content. Dynamic culture significantly increased collagen content, and collagen was more uniformly distributed throughout the scaffold thickness. While mechanical function was assessed and tensile modulus increased with culture duration, dynamic culture did not result in any additional improvement beyond free-swelling culture. Transient dynamic (6 weeks dynamic followed by 6 weeks free-swelling) culture significantly enhanced cell infiltration while permitting GAG accumulation. In this study, we demonstrated that a simple modification to standard in vitro culture conditions effectively improves cellular ingress into electrospun scaffolds, resolving a challenge which has until now limited the utility of these materials for various tissue engineering applications.
Nerurkar
Nandan L
NL
Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, Philadelphia, PA 19104, USA.
Sen
Sounok
S
Baker
Brendon M
BM
Elliott
Dawn M
DM
Mauck
Robert L
RL
D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002465:Cell Movement; D002467:Cell Nucleus / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000378:metabolism; D007211:Indoles / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D057139:Nanofibers / Q000737:chemistry*; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
D006025:Glycosaminoglycans; D007211:Indoles; C007293:DAPI; D003094:Collagen