Significant downregulation of BSP and ALP expression due to the presence of coating was observed for both sets of coated scaffolds in basic medium. Effect of electrospun fibers On day 7, the presence of electrospun fibers led to a significant downregulation in www.selleckchem.com/products/Lenalidomide.html the expression of BSP, ALP and OC in mineralization medium while no significant differences in gene expression of osteogenic markers was observed in the basic medium. On day 21, statistically significant differences in gene expression were observed only for BMP-2 and OP in basic medium. In the presence of Ca-P coating, a positive effect of the electrospun mesh was observed in the case of BMP-2 whereas the addition of an electrospun mesh led to a significantly lower expression of OP. No differences due to the presence of an electrospun mesh were observed in mineralization medium.

Discussion RP in the form of 3DF deposition was combined with ESP and Ca-P coatings to successfully fabricate hybrid scaffolds for use in bone tissue engineering. Although 3DF and ESP based scaffolds have been separately used for tissue engineering applications, we hypothesized that a combination or merger of these two technologies would lead to improved scaffold properties at different scales. While 3DF scaffolds can provide the necessary mechanical stability6,37 and support in compression, the ESP scaffolds serve to mimic the fibrillar nature of ECM and they generally provide a suitable surface for cell attachment. The combination of 3DF and ESP has already shown enhanced cellular response in terms of cell numbers and increased ALP or GAG production when used as a scaffold for bone19 and cartilage20 tissue engineering respectively.

In an earlier study,38 we demonstrated the in vivo bone forming ability of Ca-P coated electrospun scaffolds in combination with goat MSCs in a subcutaneous nude mouse model and hence decided to add another layer of functionality to the scaffolds by coating them with Ca-P, thereby providing chemical cues for differentiation. By using biomimetic method based on immersion in an aqueous solution of inorganic salts at near-physiological conditions, we succeeded in homogenously coating both 3DF and 3DF + ESP scaffolds with a layer of Ca-P, that was a mixture of OCP and CA, which are considered precursor and end phase of mineral part of bone24,39,40 respectively.

All scaffolds allowed the adhesion and proliferation of hMSCs, and at 21 d, ECM production was observed inside the scaffolds. No effect of either ESP mesh or Ca-P coating was found on cell proliferation; however, cell number at 21 d, expressed in terms of total DNA amount, Dacomitinib was lower in coated scaffolds containing ESP mesh than in coated 3DF scaffolds. Similarly, in mineralization medium, uncoated 3DF + ESP scaffolds had the highest amount of DNA after 21 d and this amount was significantly higher than that observed in the Ca-P coated 3DF + ESP scaffolds.