Immature Bone

Abstract: A conceptual framework is presented for understanding and investigating structural adaptation of cortical bone. The magnitudes, orientations, and sense (tension or compression) of the physiologically incurred cyclic principal strains vary markedly throughout the skeleton. It is probable, therefore, that the strain/remodeling response of bone is site specific. Furthermore, there is some indication that immature bone is more responsive to alterations of cyclic strains than mature bone. Animal experimental studies and complementary stress and strain analyses suggest that the structural adaptation due to changes in cyclic strain fields may be a very nonlinear response. Bone loss in mature animals due to immobilization is sensitive to even small changes in the cyclic bone strains. Under normal conditions, however, there appears to be a broad range of physical activity in which bone is relatively unresponsive to changes in loading history. With severe repeated loading, bone hypertrophy can be pronounced. These observations open the possibility that bone atrophy and hypertrophy are controlled by different mechanisms. Therefore, two (or more) complementary control systems may be involved in the regulation of bone mass by bone cyclic strain histories. It is probable that bone mechanical microdamage is one control stimulus for affecting an increase in bone mass.

Abstract: Runx2 and Sp7/Osterix determine the osteoblast lineage from mesenchymal stem cells with canonical Wnt signaling. In the process of osteoblast differentiation, these factors and canonical Wnt signaling molecules inhibit mesenchymal cells from differentiating into chondrocytes and adipocytes. After the commitment to osteoblast lineage, Runx2 maintains the osteoblasts in an immature stage, during which immature bone forms with randomly and loosely packed collagen fibrils and low mineralization. Runx2 must be suppressed for immature osteoblasts to become fully mature osteoblasts, which form mature bone with regularly and densely packed collagen fibrils and high mineralization. During the early stage of osteoblast differentiation, Runx2 regulates the expression of major bone matrix protein genes. However, Runx2 is not essential for the maintenance of the expression of the major bone matrix protein genes in mature osteoblasts. Estrogen and parathyroid hormone (PTH) enhance Runx2 expression and activity through anabolic effects, however, estrogen negatively regulates Runx2 in osteoclastogenesis. Runx2 is also involved in the catabolic effect of PTH through the induction of Tnfsf11. Thus, Runx2 regulates bone development, bone maturation, and bone maintenance through the regulation of osteoblast differentiation and function.

Abstract: Apatite formation on the surface of three kinds of bioactive material at an early stage after implantation in bone was studied using transmission electron microscopy (TEM). The materials were apatite- and wollastonite-containing glass-ceramic (A-W GC) as a surface-active glass-ceramic, dense sintered hydroxyapatite (HA) as a surface-active ceramic, and dense sintered beta-tricalcium phosphate (beta-TCP) as a resorbable ceramic. Particles of these materials, ranging from 100-300 microns in diameter, were implanted into rat tibiae, and specimens were prepared at 3, 7, 10, and 14 days after implantation. For A-W GC, dissolution of the glassy and probably wollastonite phase was observed in the surface region on and after the third day, and a collagen-free thin apatite layer on the surface of the material was evident on and after the seventh day. This apatite layer was observed before the mineralization of the surrounding bone matrix and was sometimes evident even where the material bordered on the bone marrow. On and after the tenth day, the surrounding bone matrix calcified and A-W GC-bone bonding through an apatite layer was completed. For HA, a mineralized collagen-free layer was observed on the surface of the ceramic on and after the tenth day. This layer was always present near calcifying bone and it was difficult to distinguish from immature bone. For beta-TCP, such a surface mineralized layer was rarely evident, even just before bone-ceramic contact, and finally the bone bonded to beta-TCP directly. Cell-mediated degradation of beta-TCP was frequently observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: Facial deformities require precise reconstruction of the appearance and function of the original tissue. The current standard of care-the use of bone harvested from another region in the body-has major limitations, including pain and comorbidities associated with surgery. We have engineered one of the most geometrically complex facial bones by using autologous stromal/stem cells, native bovine bone matrix, and a perfusion bioreactor for the growth and transport of living grafts, without bone morphogenetic proteins. The ramus-condyle unit, the most eminent load-bearing bone in the skull, was reconstructed using an image-guided personalized approach in skeletally mature Yucatan minipigs (human-scale preclinical model). We used clinically approved decellularized bovine trabecular bone as a scaffolding material and crafted it into an anatomically correct shape using image-guided micromilling to fit the defect. Autologous adipose-derived stromal/stem cells were seeded into the scaffold and cultured in perfusion for 3 weeks in a specialized bioreactor to form immature bone tissue. Six months after implantation, the engineered grafts maintained their anatomical structure, integrated with native tissues, and generated greater volume of new bone and greater vascular infiltration than either nonseeded anatomical scaffolds or untreated defects. This translational study demonstrates feasibility of facial bone reconstruction using autologous, anatomically shaped, living grafts formed in vitro, and presents a platform for personalized bone tissue engineering.