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Published in final edited form as: Biomaterials. 2011 Nov 8;33(4):1065–1078. doi: 10.1016/j.biomaterials.2011.10.046

Co-cultures of free floating calvarial bones with cancer cells in suspension using bone formation model and the roller tube system under hypoxic conditions. (a)–(c) Control calvaria: (a) H & E sections of the parietal bone region. (b) NR stained calvaria showing no multinucleated osteoclasts. (c) NR + silver nitrate counter stained calvaria showing lack of bone resorption. (d)–(f) Co-cultures of MCF-7 cells and calvarial bone: (d) H & E sections of the parietal bone region. (e) NR stained calvaria showing numerous active multinucleated osteoclasts. (f) NR + silver nitrate counter stained calvaria with light transparency reflecting bone resorption. (g)-(i) Co-cultures of MDA-231 cells and calvarial bone: (g) H & E sections of the parietal bone region. (h) NR stained calvaria showing numerous active multinucleated osteoclasts. (i) NR stained calvaria counter stained with silver nitrate showing substantial bone resorption. (k)-(m) Co-cultures of LnCap cancer cells and calvarial bone: (k) H & E sections of the parietal bone region. (l) NR stained calvaria showing no multinucleated osteoclasts. (m) NR + silver nitrate counter stained calvaria. (n)-(q) Co-cultures of PC3 cancer cells and calvarial bone: (n) H & E sections of the parietal bone region showing PC3 cells colonizing the endosteal side with visible new osteoid and bone formation. (o) A higher magnification, 400 x, of a section of bone grown in the presence of PC3 cells showing clear new osteoid formation, active osteoblasts and new bone. (p) NR stained calvaria showing no multinucleated osteoclasts. (q) NR + silver nitrate counter stained calvaria. (r) Cummulative calcium release and/or uptake in control and those with different cancer cells. OB, old bone, NB, new bone, OSC-RA, osteoclastic resorption areas; TC, tumor cells; MNOC, multinucleated osteoclasts; RA, resorption areas. All magnifications unless otherwise stated were: H & E 100x, NR 200x and silver nitrate counter stain 30x.

Co-cultures of free floating calvarial bones with cancer cells in suspension using bone formation model and the roller tube system under hypoxic conditions. (a)–(c) Control calvaria: (a) H & E sections of the parietal bone region. (b) NR stained calvaria showing no multinucleated osteoclasts. (c) NR + silver nitrate counter stained calvaria showing lack of bone resorption. (d)–(f) Co-cultures of MCF-7 cells and calvarial bone: (d) H & E sections of the parietal bone region. (e) NR stained calvaria showing numerous active multinucleated osteoclasts. (f) NR + silver nitrate counter stained calvaria with light transparency reflecting bone resorption. (g)-(i) Co-cultures of MDA-231 cells and calvarial bone: (g) H & E sections of the parietal bone region. (h) NR stained calvaria showing numerous active multinucleated osteoclasts. (i) NR stained calvaria counter stained with silver nitrate showing substantial bone resorption. (k)-(m) Co-cultures of LnCap cancer cells and calvarial bone: (k) H & E sections of the parietal bone region. (l) NR stained calvaria showing no multinucleated osteoclasts. (m) NR + silver nitrate counter stained calvaria. (n)-(q) Co-cultures of PC3 cancer cells and calvarial bone: (n) H & E sections of the parietal bone region showing PC3 cells colonizing the endosteal side with visible new osteoid and bone formation. (o) A higher magnification, 400 x, of a section of bone grown in the presence of PC3 cells showing clear new osteoid formation, active osteoblasts and new bone. (p) NR stained calvaria showing no multinucleated osteoclasts. (q) NR + silver nitrate counter stained calvaria. (r) Cummulative calcium release and/or uptake in control and those with different cancer cells. OB, old bone, NB, new bone, OSC-RA, osteoclastic resorption areas; TC, tumor cells; MNOC, multinucleated osteoclasts; RA, resorption areas. All magnifications unless otherwise stated were: H & E 100x, NR 200x and silver nitrate counter stain 30x.