Bone formation by isolated calvarial osteoblasts in syngeneic and allogeneic transplants: Light microscopic observations

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Abstract
Taking advantage of recently developed methods for osteoblast isolation, we used these cells to study bone morphogenesis in syngeneic and allogeneic intramuscular transplants. Syngeneic osteoblasts from fetal rat calvaria produced small islands of bone by the third day after transplantation. These islands increased in size and began to fuse after about 14 days. At the surface of the woven bone laid down first, lamellar bone developed. The amount of this bone increased, and in 56-day-old transplants solid blocks of bone were present. Osteoclasts were scarce, and the woven bone remained unresorbed. Bone marrow was absent. The structure of bone in transplants differed from that of mature calvarial bones in which only remnants of woven bone remained and bone marrow was well developed. The scarcity of osteoclasts in transplants could be caused by their relative paucity among the injected cells, since these cells responded strongly to added parathyroid hormone by increased production of cyclic adenosine monophosphate (cAMP) but only weakly to calcitonin. Osteoblasts isolated from the surface of calvarial lamellar bone of 28-day-old rats formed woven bone similar to the bone formed by fetal cells. This suggests that the type of bone produced does not depend on the intrinsic properties of the osteoblasts. Bone formed in an allogeneic system was surrounded by infiltrations containing lymphocytes, macrophages, and osteoclastlike cells in 14-day-old transplants. Osteoblasts at the bone periphery were destroyed and bone matrix was resorbed by infiltrating cells. Numerous bone lacunae were enlarged, suggesting the occurrence of osteocytic osteolysis. Isolated osteoblasts cultured for three population doublings or longer did not form bone after transplantation, although they retained some reactivity toward parathyroid hormone.