Table 2.
Fold Change (2−ΔΔCt) in ALPL, COL1A1, BGLAP, CNR1, CNR2, TNFRSF11B/TNFSF11 (Mean ± SEM) Gene Expression in C, HT, and CT ASC-Scaffold Constructs Cultured in Stromal (S), Osteogenic for 48 h Followed by Stromal (OS) or Osteogenic (O) Medium for 7, 14, and 28 Days
| S | OS | O | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Gene | Scaffold | Day 7 | Day 14 | Day 28 | Day 7 | Day 14 | Day 28 | Day 7 | Day 14 | Day 28 |
| ALPL | C | 1.4 ± 1.7A | 6.7 ± 1.7B | 6.0 ± 1.7B | 3.0 ± 1.7A | 7.9 ± 1.7B | 9.9 ± 1.7B | 3.7 ± 1.7A | 18.1 ± 1.7B | 15.1 ± 1.7B |
| HT | 3.4 ± 1.7A | 14.7 ± 1.7B | 10.2 ± 1.7B | 16.7 ± 1.7A | 11.4 ± 1.7B | 12.7 ± 1.7A/B | 11.8 ± 1.7A/B | 18.4 ± 1.7A | 9.1 ± 1.7B | |
| CT | 4.6 ± 1.7 | 4.0 ± 1.7 | 4.5 ± 1.7 | 3.2 ± 1.7 | 4.6 ± 1.7 | 2.2 ± 1.7 | 4.6 ± 1.7 | 5.1 ± 1.7 | 2.8 ± 1.7 | |
| COL1A1 | C | 1.0 ± 0.8A | 3.6 ± 0.8B | 4.0 ± 0.8B | 2.2 ± 0.8 | 1.2 ± 0.8 | 1.1 ± 0.8 | 2.2 ± 0.8 | 1.3 ± 0.8 | 0.6 ± 0.8 |
| HT | 1.4 ± 0.8A | 5.3 ± 0.8B | 6.6 ± 0.8B | 2.0 ± 0.8 | 0.5 ± 0.8 | 0.5 ± 0.8 | 2.5 ± 0.8A | 0.5 ± 0.8A/B | 0.2 ± 0.8B | |
| CT | 0.08 ± 0.8 | 0.9 ± 0.8 | 1.1 ± 0.8 | 0.8 ± 0.8 | 0.2 ± 0.8 | 0.4 ± 0.8 | 0.1 ± 0.8 | 0.2 ± 0.8 | 0.2 ± 0.8 | |
| BGLAP | C | 1.1 ± 0.6A | 3.6 ± 0.6B | 5.1 ± 0.6C | 1.5 ± 0.6A | 4.1 ± 0.6B | 4.7 ± 0.6B | 1.9 ± 0.6A | 4.9 ± 0.6B | 6.8 ± 0.6B |
| HT | 1.3 ± 0.6 | 1.9 ± 0.6 | 2.7 ± 0.6 | 1.1 ± 0.6 | 1.8 ± 0.6 | 2.2 ± 0.6 | 1.3 ± 0.6A | 2.3 ± 0.6A | 4.8 ± 0.6B | |
| CT | 0.9 ± 0.6 | 1.4 ± 0.6 | 1.9 ± 0.6 | 1.1 ± 0.6 | 1.9 ± 0.6 | 2.5 ± 0.6 | 0.9 ± 0.6 | 1.7 ± 0.6 | 2.3 ± 0.6 | |
| CNR1 ( × 104) | C | 0.0006 ± 0.3 | 0.0003 ± 0.3 | 0.01 ± 0.3 | 0.002 ± 0.3 | 0.007 ± 0.3 | 0.03 ± 0.3 | 4 ± 0.3A | 4.6 ± 0.3B | 5.3 ± 0.3B |
| HT | 0.0002 ± 0.3 | 0.0004 ± 0.3 | 0.03 ± 0.3 | 0.1 ± 0.3 | 0.03 ± 0.3 | 0.02 ± 0.3 | 0.3 ± 0.3A | 2.3 ± 0.3B | 1.1 ± 0.3C | |
| CT | 0.0001 ± 0.3 | 0.0002 ± 0.3 | 0.0003 ± 0.3 | 0.02 ± 0.3 | 0.01 ± 0.3 | 0.002 ± 0.3 | 0.02 ± 0.3A | 0.6 ± 0.3A/B | 1.2 ± 0.3B | |
| CNR2 | C | 13.5 ± 4.8A | 39.9 ± 4.8B | 16.9 ± 4.8A | 8.1 ± 4.8 | 15.6 ± 4.8 | 12.8 ± 4.8 | 7.7 ± 4.8A | 40.3 ± 4.8B | 6.0 ± 4.8A |
| HT | 3.0 ± 4.8 | 9.0 ± 4.8 | 5.1 ± 4.8 | 8.5 ± 4.8 | 7.8 ± 4.8 | 8.1 ± 4.8 | 9.3 ± 4.8 | 16.5 ± 4.8 | 18.1 ± 4.8 | |
| CT | 6.1 ± 4.8A | 36.1 ± 4.8B | 21.5 ± 4.8C | 6.6 ± 4.8A | 32.7 ± 4.8B | 16.9 ± 4.8A | 3.4 ± 4.8A | 22.2 ± 4.8B | 7.7 ± 4.8A | |
| TNFRSF11B/ TNFSF11 | C | 1.1 ± 2.5 | 2.6 ± 2.5 | 3.7 ± 2.5 | 2.1 ± 2.5 | 3.0 ± 2.5 | 7.8 ± 2.5 | 7.5 ± 2.5A | 6.2 ± 2.5A | 67.8 ± 2.5B |
| HT | 4.1 ± 2.5A | 13.7 ± 2.5B | 15.7 ± 2.5B | 4.4 ± 2.5A | 16.1 ± 2.5B | 14.5 ± 2.5B | 2.7 ± 2.5A | 49.1 ± 2.5B | 90.4 ± 2.5C | |
| CT | 0.3 ± 2.5 | 0.8 ± 2.5 | 3.8 ± 2.5 | 0.2 ± 2.5 | 1.0 ± 2.5 | 2.4 ± 2.5 | 0.7 ± 2.5 | 3.9 ± 2.5 | 6.2 ± 2.5 | |
Fold change was calculated relative to the reference gene GAPDH and target gene expression in constructs immediately after cell loading. Columns with different superscripts within constructs and media are significantly different among time points.
ASC, adipose-derived multipotent stromal cell; SEM, standard error of the mean.