Bioengineered human bone tissue using autogenous osteoblasts cultured on different biomatrices

Abstract

Surgical treatment of critical-size posttraumatic bone defects is still a challenging problem, even in modern bone and joint surgery. Progress in cellular and molecular biology during the last decade now permits novel approaches in bone engineering. Recent conceptual and technical advances have enabled the use of mitotically expanded, bone-derived cells as a therapeutic approach for tissue repair. Using three different tissue carrier systems, we successfully cultivated human osteoblasts in a newly developed perfusion chamber. We studied cell proliferation and the expression of osteocalcin, osteopontin, bone morphogenetic protein-2A, alkaline phosphatase, and vascular endothelial growth factor as parameters for osteoblast function and viability. Adherence of highly enriched human osteoblasts had already started after 1 h and resulted in completely overgrown human bone pieces after 10 days. Expression analysis of bone-specific alkaline phosphatase indicated differentiating osteoblasts, whereas the high mRNA expression of osteocalcin and osteopontin revealed terminally differentiated osteoblasts and the process of mineralization. Additionally, gene expression was significantly higher when demineralized bone was used as biomatrix, compared to autoclaved bone and hydroxyapatite ceramics. We conclude that with our newly developed perfusion culture system, vital autogenous bone implants of clinically applicable size can be generated within 17 days in order to manage critical-size bone defects.

Bioreactor Systems for Human Bone Tissue Engineering

“…2013 [25] hiPSC-MPs A relatively large amount of these studies have also explored the bone formation potential of these substitutes in vivo using ectopic and orthotopic animal models (Table 2). In a study by Hofmann et al, human osteoblasts were cultured in a perfusion bioreactor (flow rate: 1 mL/h) for ten days after seeding onto demineralized and non-demineralized human spongiosa, and HA cylinder scaffolds (7.5 mm diameter and 10 mm height) [63]. The results demonstrated significant increased expression of bone-specific genes when cells where seeded onto demineralized matrices compared to the other scaffolds.…”

Macrophage-mediated angiogenic activation of outgrowth endothelial cells in co-culture with primary osteoblasts

“…Human pOBs were isolated from cancellous bone fragments from healthy donors according to a previously published method (Hofmann et al, 2003) and cultivated in Dulbecco´s Modified Eagle´s Medium Nutrient Mixture F-12 (DMEM/F-12, Sigma-Aldrich, St. Louis, MO, USA), supplemented with 10 % FCS (Gibco) + 1 % Penicillin/ Streptomycin at 37 °C in an atmosphere of 95 % air and 5 % CO 2 . Cells were passaged in a ratio of 1:2.…”

MC3T3-E1 cell adhesion to hydroxyapatite with adsorbed bone sialoprotein, bone osteopontin, and bovine serum albumin

“…1 A number of investigators are currently examining the influence of various calcium phosphates on the adhesion, proliferation, and differentiation of bone-related cells. [2][3][4][5][6] These studies are primarily focused on the influence that characteristics of the calcium phosphates, such as the surface coverage, structure, and the exact composition, have on the cells and their function. In one of these studies, Chou et al examined four different forms of HAP and determined that while the HAP samples induced osteoblast differentiation, there were more cells adherent on the tissue culture polystyrene controls at all of the time points examined.…”