Title : Adhesion, proliferation and osteogenic differentiation of hSCAPs on HAP and PDA-HAP
hSCAPs were extracted and identified and inoculated on HAP and PDA-HAP so that we can compare cell adhesion, proliferation and osteogenic differentiation of the two scaffolds. Methods: 1.hSCAPs were extracted by improved tissue block method, and cell surface antigens were identified by flow cytometry: STRO-1, CD90, CD146, CD45 and CD34. 2.hSCAPs were inoculated into HAP group and PDA-HAP group respectively. DAPI and rhodamine phalloidin stained the nucleus and cytoskeleton respectively so that we can observe cell morphology and counted for statistical analysis. 3.hSCAPs were inoculated into HAP group and PDA-HAP group respectively and cultured for 1, 3, 5 and 7 days. The OD values were measured by CCK8 to detect the cell proliferation. 4.hSCAPs were inoculated into HAP group and PDA-HAP group for 1, 7 and 14 days. ALP activity was detected. Real-time PCR was used to detect the expression of ALP, OCN and Runx2. Results: 1.The hSCAPs expressed STRO-1 (+), CD90 (+), CD146 (+), CD45 (-), CD34 (-). 2.The number of hSCAPs attached to PDA-HAP group was more than that on HAP group (P < 0.001), and the cell area on PDA-HAP was larger and more fully extended than that on HAP group, we could observe cell antennae. 3.There was no significant difference in toxicity between HAP and P DA-HAP cultured hSCAPs 1, 3, 5 and 7 days (P > 0.05). 4.ALP activity: There was no significant difference between the two groups on the 1st and 14th day of induction (P > 0.05), but the activity of PDA-HAP group was significantly higher than that of HAP group on the 7th day (P < 0.001). Real-time PCR: The expression of ALP was the same as that of ALP activity; the expressions of Run x2 in PDA-HAP group were higher than that in HAP group on 1, 7 and 14 days (P < 0.05); there was no significant difference between the two groups on the 1st day of OCN (P > 0.05), but the expressions of OCN in PDA-HAP group were higher than that in HAP group on the 7th and 14th day (P < 0.05). Conclusion:PDA-HAP can significantly promote the expression of ALP, Runx2 and OCN, and promote the osteogenic differentiation of hSCAPs.