Hydroxyapatite‐hybridized double‐network hydrogel surface enhances differentiation of bone marrow‐derived mesenchymal stem cells to osteogenic cells

jbma37324-sup-0001-FigS1.tifTIFF image, 2.4 MB

FIGURE S1. The Western blotting to evaluate protein expression levels of CD73, CD90, CD105, and CD44 in the cells cultured with Medium ABS at 7 days. These stemness markers were highly expressed in the cells cultured with Medium ABS. In the cells cultured with Medium FBS, CD73, or CD105 was not expressed, and the bands showing expression of CD90 and CD44 appeared to be faint. ABS, autogenous blood serum; FBS, fetal bovine serum.

jbma37324-sup-0004-Tables.docxWord 2007 document , 47.3 KB

TABLE S1. Nucleotide sequences of primers.

TABLE S2. Gene expression of TGF-beta1 in the MSCs cultured on the PS, DN gel, HAp, and HAp/DN gel surfaces at day 7 and 14. The 2-way ANOVA showed a significance between both the materials (p < 0.0001) and the periods (p < 0.0001). In addition, there was a significance on the interaction (p < 0.0001). This table shows the results of multiple comparisons using the post-hoc test. The expression level on the HAp/DN gel disk was significantly higher at both day 7 and day 14 than the levels on the other material surfaces, while the expression level at day 14 was significantly reduced than the level at day 7. The expression levels on the DN gel and HAp disks did not show any significant differences in comparison with the level on the PS dish at both day 7 and day 14.

TABLE S3. Gene expression of BMP-2 in the MSCs cultured on the PS, DN gel, HAp, and HAp/DN gel surfaces at day 7 and 14. The 2-way ANOVA showed a significance between both the materials (p < 0.0001) and the periods (p < 0.0001). In addition, there was a significance on the interaction (p < 0.0001). This table shows the results of multiple comparisons using the post-hoc test. The expression level on the HAp/DN gel disk was significantly higher at both day 7 and day 14 than that the levels on the other material surfaces, while the expression level at day 14 was significantly reduced than the level at day 7. The expression levels on the DN gel and HAp disks did not show any significant differences in comparison with the level on the PS dish at both day 7 and day 14.

TABLE S4. Gene expression of Runx2 in the MSCs cultured on the PS, DN gel, HAp, and HAp/DN gel surfaces at day 7 and 14. The 2-way ANOVA showed a significance between both the materials (p < 0.0001) and the periods (p = 0.0006). In addition, there was a significance on the interaction (p < 0.0001). This table shows the results of multiple comparisons using the post-hoc test. The expression level on the HAp/DN gel disk was significantly higher at only day 7 than the levels on the other material surfaces, while the expression level at day 14 was significantly reduced than the level at day 7. The expression levels on the DN gel and HAp disks were significantly higher in comparison with the level on the PS dish at both day 7 and day 14. Additionally, the expression levels on the PS dish and the DN gel disk at day 14 were significantly higher than those measured at day 7.

TABLE S5. Gene expression of Col-1 in the MSCs cultured on the PS, DN gel, HAp, and HAp/DN gel surfaces at day 7 and 14. The 2-way ANOVA showed a significance between both the materials (p < 0.0001) and the periods (p < 0.0001). In addition, there was a significance on the interaction (p < 0.0001). This table shows the results of multiple comparisons using the post-hoc test. The expression level on the HAp/DN gel disk was significantly higher at both day 7 and day 14 than the levels on the other material surfaces, while the expression level at day 14 was significantly reduced than the level at day 7. The expression levels on the DN gel and HAp disks did not show any significant differences in comparison with the level on the PS dish at both day 7 and day 14.

TABLE S6. Gene expression of ALP in the MSCs cultured on the PS, DN gel, HAp, and HAp/DN gel surfaces at day 7 and 14. The 2-way ANOVA showed a significance between the materials (p < 0.0001), while there was no significance between the periods or on the interaction. This table shows the results of multiple comparisons using the post-hoc test. The expression level on the HAp/DN gel disk was significantly higher at both day 7 and day 14, compared with the levels on the other material surfaces. The expression levels on the DN gel and HAp disks did not show any significant differences in comparison with the level on the PS dish.

TABLE S7. Gene expression of OPN in the MSCs cultured on the PS, DN gel, HAp, and HAp/DN gel surfaces at day 7 and 14. The 2-way ANOVA showed a significance between the materials (p = 0.0003), while there was no significance between the periods or on the interaction. This table shows the results of multiple comparisons using the post-hoc test. The expression level on the HAp/DN gel disk was significantly higher at only day 7, compared with the levels on the other material surfaces. The expression levels on the DN gel and HAp disks did not show any significant differences in comparison with the level on the PS dish.

TABLE S8. Gene expression of BSP in the MSCs cultured on the PS, DN gel, HAp, and HAp/DN gel surfaces at day 7 and 14. The 2-way ANOVA showed a significance between the materials (p < 0.0001), while there was no significance between the periods or on the interaction. This table shows the results of multiple comparisons using the post-hoc test. The expression level on the HAp/DN gel disk was significantly higher at both day 7 and day 14, compared with the levels on the other material surfaces. The expression levels on the DN gel and HAp disks did not show any significant differences in comparison with the level on the PS dish.

TABLE S9. Gene expression of OCN in the MSCs cultured on the PS, DN gel, HAp, and HAp/DN gel surfaces at day 7 and 14. Results of comparisons using the post-hoc test concerning mRNA expression of OCN. The 2-way ANOVA showed a significance between both the materials (p < 0.0001) and the periods (p < 0.0001). In addition, there was a significance on the interaction (p < 0.0001). This table shows the results of multiple comparisons using the post-hoc test. The expression level on the HAp/DN gel disk was significantly higher at both day 7 and day 14 than the levels on the other material surfaces, while the expression level at day 14 was significantly reduced than the level at day 7. The expression levels on the DN gel and HAp disks did not show any significant differences in comparison with the level on the PS dish, although only the expression level on the HAp disk at day 7 was significantly higher than the level on the PS dish.

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