The Effects of Plasma Treated Electrospun Nanofibrous Poly (ε-caprolactone) Scaffolds with Different Orientations on Mouse Embryonic Stem Cell Proliferation.

OBJECTIVE: Assessments of cell reactions such as motility, orientation and activation to the topography of the substratum will assist with the fabrication of a proper implantable scaffold for future tissue engineering applications.The current challenge is to analyze the orientation effect of elecrospun nanofibers of poly (ε-caprolactone) (PCL) on viability and proliferation of mouse embryonic stem cells (mESCs).

MATERIALS AND METHODS: In this experimental study, we used the electrospinning method to fabricate nanofibrous PCL scaffolds. Chemical and mechanical characterizations were specified by the contact angle and tensile test. O2plasma treatment was used to improve surface hydrophilicity. We used the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to evaluate mESCs adhesion and proliferation before and after surface modification. The influence of the orientation of the nanofibers on mESCs growth was evaluated by scanning electron microscopy (SEM). Statistical analysis was performed using one-way analysis of variance (ANOVA) With differences considered statistically significant at p≤0.05.

RESULTS: The results showed that plasma treatment improved the hydrophilic property of PCL scaffolds. MTT assay showed a significant increase in proliferation of mESCs on plasma treated PCL (p-PCL) scaffolds compared to non-treated PCL (p=0.05). However gelatin coated tissue culture plate (TCP) had a better effect in initial cell attachment after one day of cell seeding. There was more cell proliferation on day 3 in aligned plasma treated (AP) nanofibers compared to the TCP. SEM showed optical density of the cell colonies. Aligned nanofibrous scaffolds had larger colony sizes and spread more than random nanofibrous scaffolds.

CONCLUSION: This study showed that plasma treating of scaffolds was a more suitable substrate for growth and cell attachment. In addition, aligned nanofibrous scaffolds highly supported the proliferation and spreading of mESCs when compared to random nanofibrous scaffolds and TCP.