engleski

Synthetic vs natural scaffolds for human limbal stem cell cultivation

Aim was to investigate the impact of synthetic electrospun polyurethane and polycaprolactone nanoscaffolds, before and after hydrolytic surface modification, on viability and differentiation of cultured human eye epithelial cells, in comparison with natural scaffolds: fibrin and human amniotic membrane. Human placenta was taken at elective caesarean delivery. Fibrin scaffolds were prepared from commercial fibrin glue kits. Nanoscaffolds were fabricated by electrospinning. Limbal cells were isolated from surpluses of human cadaveric cornea and seeded on feeder 3T3 cells. The scaffolds used for viability testing and immunofluorescence analysis were: amniotic membrane, fibrin, polyurethane (PU) and polycaprolactone (PCL) nanoscaffolds, with or without prior NaOH treatment. SEM photographs of all tested scaffolds showed good colony spreading of seeded limbal cells. In regard to viability performance there was statistically significant difference between cells with highest viability cultured on tissue culture plastic and cells cultured on all other scaffolds. On the other hand, electrospun PU, PCL and electrospun PCL treated with NaOH had more than 80% of limbal cells positive for stem cell marker p63 compared to only 27%of p63 positive cells on fibrin. Natural scaffolds, fibrin and amniotic membrane, showed better cell viability compared to electrospun scaffolds. On contrary, high percentages of p63 positive cells on these scaffolds still makes them good candidates as efficient delivery systems for therapeutic purposes.

Scaffold ; Human ; Limbal stem cells

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