Table 1.
Morphology, matrix accumulation and matrix quality in cartilage-like tissue formed by SZC and DZC overexpressing bone morphogenetic protein (BMP) or Noggin1
| Control | BMP | Noggin | |
|---|---|---|---|
| Cell Diameter (μm)2 | |||
| SZC | 12.8 ± 2.2 | 15.2 ± 0.7 | 10.0 ± 0.3* |
| DZC | 15.2 ± 2.0 | 20.9 ± 1.0 (BMP2)*
17.7 ± 0.6 (BMP7)* |
11.4 ± 0.5* |
| Cellularity (cells per field) | |||
| SZC | 1238 ± 423 | 520 ± 77* | 735 ± 97* |
| DZC | 898 ± 336** | 721 ± 316 | 899 ± 350 |
| Pellet Area (μm2) | |||
| SZC | 2.0 ± 0.7 | 4.6 ± 1.5* | 2.2 ± 0.3 |
| DZC | 3.3 ± 1.0 | 4.0 ± 1.2 | 1.9 ± 1.9 |
| PG:Am1 Ratio (×10−3)3 | |||
| SZC | 0.47 ± 0.17 | 1.67 ± 0.45* | NA |
| DZC | 0.96 ± 0.67 | 1.00 ± 0.52 | NA |
The SZC and DZC were treated with adenovirus vectors encoding no transgene (Control), BMP2, BMP7 or Noggin and maintained as pellet cultures for 7 days as described in Experimental Procedures. Tissues were harvested and fixed sections were analyzed by light microscopy and fourier-transform infra-red imaging spectroscopy (FT-IRIS). Where measurements for BMP2 and -7 overexpressing samples were similar, values were analyzed as a single group (BMP).
Histomorphometric measurements including cell diameter, cell number per field (cellularity) and pellet area were made on sections stained with alcian blue and nuclear fast red as described in Experimental Procedures. Measurements were made in at least 6 sections per pellet in 3 – 6 pellets per group. Mean ± standard deviation are shown. [*] indicates statistically significant differences with p<0.05 compared to its own Control. [**] indicates statistically significant difference with p<0.05 compared to SZC Control.
Matrix quality was assessed by FT-IRIS as described in Experimental Procedures. The ratio of peak area measurements for proteoglycan (PG) and Amide 1 (Am1, mostly collagen) was calculated and normalized to the number of cells per field. Measurements were made in 3 – 6 pellets per group. Mean ± standard deviation are shown. [*] indicates statistically significant differences with p<0.05 compared to Control. [**] indicates statistically significant differences with p<0.02 compared to all other groups.