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. 2021 Oct 25;9:672959. doi: 10.3389/fbioe.2021.672959

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Copyright © 2021 Delaine-Smith, Hann, Green and Reilly.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Aligned fibers promote mature osteoblasts (MLO-A5) cell mineralization and organization. MLO-A5-polycaprolactone (PCL) fibrous constructs cultured for (A) 3 days and cell nucleus 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI in blue) and cytoskeleton (Phalloidin-TRITC in red) were visualized via IF at two depths. Constructs were cultured for 10 days and assayed for (B) total DNA, (C) Alkaline phosphatase (ALP) activity, (D,E) matrix deposition (total and normalized to DNA) measured via (D) sirius red (SR) staining (collagen) and (E) alizarin red (AR) staining (calcium; inserts show stained constructs). (F) Representative scanning electron microscope (SEM) images of 10-day-old constructs. Data are mean ± 95% CI for n = 9. ^*p < 0.05, and ^***p < 0.001 (two-tailed unpaired t-test: random vs. aligned).