Supplementary MaterialsFigure?S1 Assessment of 3d microCT pictures of PCSD1-injected femurs demonstrated osteolytic and osteoblastic shifts in comparison to Control (Matrigel alone)-injected femurs. hole was used to confirm successful intra-femoral direct injection onto the endosteal space of the right femur. Success rate: Needle hole?+?tumor formation: 13/14 PCSD1-injected mice?=?92.9%. The Control media-injected femurs had smooth bone surfaces and overall bone structure was similar to the un-injected contra-lateral femurs. In contrast, the PCSD1-injected femurs showed significant malformations in the 3D microCT images. The bone surface of the PCSD1 group was irregular and bumpy as seen in the 3D whole bone images of PCSD1-injected femurs (Supplemental fig. 1A, B and C). mmc1.pdf (1.9M) GUID:?78CE43A0-5BCF-416D-AAAD-929FC3A55BA6 Figure?S2 Comparison of hematoxylin and eosin (H&E) stained cortical periosteal regions sagittal sections of PCSD1-injected femurs showing periosteal reaction along AG-1478 kinase inhibitor femur shaft. A-A. PCSD1 tumor cells (yellow arrow) were found adjacent to osteoblastic bone reaction in cortical femur and in regions of periosteal reaction in cortical bone and third trochanter. A-B. Enlarged image of hatched, AG-1478 kinase inhibitor black box in A. shows irregular, outer edge of cortical femur bone (black bracketed line, P?=?periosteal region), multi-cell periosteal layer indicating periosteal reaction adjacent to skeletal muscle layer and PCSD1 tumor. A-C. H&E staining of Control-injected femur bone sagittal sections at 50 magnification. Track of intra-femorally injected Matrigel as shown by the yellow arrow can be seen in the endosteum where it has displaced the bone marrow (small, dark purple nuclei); A-D. Enlarged image (200) of Control cortical bone region (black, hatched box in C.) showing single layer of periosteal cells and smooth edge of cortical femur bone (black bracketed line, P?=?periosteal region). Yellow arrow?=?PCSD1 cells, Dark blue arrow?=?femur bone, Green arrow?=?periosteum, Light blue arrow?=?skeletal muscle fibers, Black arrow?=?Matrigel track that was injected intra-femorally, Red arrow?=?bone marrow. B. PCSD1 cells were present within osteoblastic lesions in cortical bone. Histochemical H&E stained images of the sagittal sections from the mice analyzed using microCT scanning were examined in the femur shaft regions with osteoblastic lesions. Examination of H&E stained sections in the third trochanter region of the PCSD1 group revealed that it was encased in PCSD1 tumor cells in these mice (Supplemental fig. 3A, yellow arrow) and showed aberrant bone structures as detected in microCT analysis (Supplemental fig. 3A, center). The higher magnification image shows from left to right: PCSD1 tumor cells (yellow arrow), PCSD1 tumor cells attached to muscle fiber near the top of the image (light blue arrow), the porous structure of new bone formation (pink arrow), the multi-cellular periosteal layer (green arrow), bone (dark blue arrow), and interspersed bone marrow cells. mmc2.pdf (20M) GUID:?1B39DA4C-F48B-47F0-8FB9-DB81A967BD9A Figure?S3 Shortened femur length in right femur compared to ipsi-lateral, un-injected femur due to intra-femoral injection in both Control Stx2 and PCSD1-injected groups. Femur length was measured from the greater trochanter to the medial condyle. Femur length between right side and left side was compared in both Control AG-1478 kinase inhibitor group and PCSD1 group to reveal the influence of tumor injection. The right femurs were significantly shortened compared to the left femurs in Control group as well as PCSD1 group. mmc3.pdf (117K) GUID:?D34EE69A-51B8-4791-8C15-EBFF1CB90012 Figure?S4 Proximal bone at femur head had lower bone volume in PCSD1-injected femurs than in Control femurs. The maximum diameter in the bone shaft was measured on Mimics? in which we can visualize the maximum diameter on both 2D and 3D. The longitudinal axis of the femur was set up in advance to be paralleled with the bone tissue shaft on Data Audience?. BV and BV/Television of PCSD1 group was less than Control group (p? ?0.01**). BS/BV of PCSD1 group was more than Control group (p? ?0.01**). mmc4.pdf (94K) GUID:?D243C84E-8AB0-4C92-9359-6BF9D7B9A870 Figure?S5 Trabecular bone lesions were in the osteolytic or osteoblastic lesion predominantly. Bone tissue parameters were re-evaluated between high BV/TV PCSD1 group and.