HA-Col scaffolds selleck kinase inhibitor prepared by self-assembly method showed an irregular porous and fibrous structure (Fig. 1). The scaffold porosity percentage and pore width were measured in order to verify the ability of these scaffolds to be colonised by cells. A homogenous porosity of 65�C70% in the entire scaffold was measured from histological transversal cross-sections (Fig. 1A-B) and confirmed by liquid displacement measurements that gave 62% (v/v) of scaffold permeability. By image treatment, average pore diameters presented a heterogeneous distribution with a mean of 280 �� 80 ��m (Fig. 1D). Figure 1. (A) A schematic diagram of the image treatment method to calculate the scaffold porosity on transversal sections; (B) Scaffold porosity on ten successive transversal sections; (C) SEM micrograph of HA-Col scaffold surface; (D) Histological .
.. HA-Col scaffold stability The scaffold stability in static culture conditions was observed by SEM for up to 21 d. No morphological changes in structural parameters such as collagen fibers density, mineralization and scaffold global shape (Fig. 2A, B and C) were observed. The scaffold porosity measured on longitudinal sections was relatively stable even with the application of mechanical forces due to fluid flow (Fig. 2D). Higher porosity could be observed in scaffolds submitted to high flow-rate (except at 7 d). Figure 2. (A, B and C) SEM micrographs of scaffolds immersed under static culture condition up to 21 d and (D) total porosity of scaffolds maintained under different culture conditions up to 21 d.
The dotted line indicates the initial porosity … HA-Col scaffold colonization Cell colonization First, histological sections were analyzed in order to determine the total number of STRO-1A cell nuclei inside each sample. No significant differences were found in the number of cell nuclei per section after 1 and 3 d of culture in all culture conditions (Fig. 3). A lower quantity of cell nuclei was observed in scaffolds cultured for 24 h under high flow-rate. It seems that the dynamic high flow-rate was able to induce a cell detachment after 24 h in spite of the maintenance of the samples during 24 h in static conditions after inoculation to permit cell adhesion. After the first week, significant differences appeared between the conditions.
Cell nuclei number was significantly higher in scaffolds cultured under high flow-rate conditions for 7, 14 and 21 d compared with scaffolds cultured in the two other conditions. At 21 d, a significantly higher number of nuclei were observed in scaffolds cultured in static conditions than in low Anacetrapib flow-rate conditions (Fig. 3). Figure 3. STRO-1A cell nuclei number stained by DAPI per longitudinal cross-section of HA-Col scaffolds cultured under different conditions up to 21 d. Error bars represent mean �� SD with n = 4. The symbols (*; **) indicate statistical …