A 3D in vitro bone organ model using human progenitor cells

Abstract

A Papadimitropoulos et al.A 3D human bone organ model European Cells and Materials Vol. 21 2011 (pages 445-458) DOI: 10.22203/eCM.v021a33 ISSN 1473-2262 AbstractThree-dimensional (3D) organotypic culture models based on human cells may reduce the use of complex and costly animal models, while gaining clinical relevance. This study aimed at developing a 3D osteoblastic-osteoclasticendothelial cell co-culture system, as an in vitro model to mimic the process of bone turnover. Osteoprogenitor and endothelial lineage cells were isolated from the stromal vascular fraction (SVF) of human adipose tissue, whereas CD14+ osteoclast progenitors were derived from human peripheral blood. Cells were co-cultured within 3D porous ceramic scaffolds using a perfusion-based bioreactor device, in the presence of typical osteoclastogenic factors. After 3 weeks, the scaffolds contained cells with endothelial (2.0 ±0.3%), pre/osteoclastic (14.0 ±1.4%) and mesenchymal/ osteoblastic (44.0 ±8.4%) phenotypes, along with tartrateresistant acid phosphatase-positive (TRAP + ) osteoclastic cells in contact with deposited bone-like matrix. Supernatant analysis demonstrated sustained matrix deposition (by C-terminus procollagen-I propeptides), resorption (by N-terminus collagen-I telopeptides and phosphate levels) and osteoclastic activity (by TRAP-5b) only when SVF and CD14+ cells were co-cultured. Scanning electron microscopy and magnetic resonance imaging confi rmed the pattern of matrix deposition and resorption. The effectiveness of Vitamin D in replacing osteoclastogenic factors indicated a functional osteoblast-osteoclast coupling in the system. The formation of human-origin bonelike tissue, blood vessels and osteoclasts upon ectopic implantation validated the functionality of the developed cell types. The 3D co-culture system and the associated non-invasive analytical tools can be used as an advanced model to capture some aspects of the functional coupling of bone-like matrix deposition and resorption and could be exploited toward the engineering of multi-functional bone substitute implants.

From mechanical stimulus to bone formation: A review

“…This approach overlooks many parameters known to be essential for accurately reproducing the mechanotransduction process. Three-dimensional cell-growth environments have been developed in an at-tempt to overcome some cell culture disadvantages, mimic the physiological complexity of real tissue, and avoid the use of bone organ cultures [121]. Several materials have been tested as scaffolds, such as collagen glycosaminoglycan [131,132] and porous chitosan [133].…”

“…Although tremendous advances have been made in the development of threedimensional scaffold-based systems for bone cells studies that could reproduce both the formation of an organized bone-like matrix and cell-mediated substrate degradation, this methodology is also associated with limitations depending on the aim of a study [121,136].…”

“…In recent years, the evolution of bioreactor devices for the application of specific mechanical stimuli and controlled medium recirculation has favoured cell viability and prolonged culture time of bone explants [121]. A system that shows a successful applicability of these strategy was developed by Davies et al [153].…”

Bone remodeling markers and bone metastases: From cancer research to clinical implications

“…[42][43][44][45][46] Biomarkers of bone remodeling can also be used in in vitro models. 47 These cellular models can be reliably used for enhancing preclinical evaluation of pharmacological agents in metastatic bone disease. Nevertheless, in the vast majority of in vitro studies, the determination of osteoclast formation and activity is usually performed by TRAP staining, TRAP activity, pit resorption assays and expression of osteoclast-related markers such as cathepsin K and MMP-9.…”

Adipose-derived stromal/stem cells

“…51,52 There is equally exciting work that's underway in a project that is in the Orthopedic and Trauma department in Basel, Switzerland under Dr Ivan Martin. 65,66 He is using these cells to do a clinical long bone fracture model where they are going to conducting a very nicely controlled study comparing large cohorts of ASC treated and untreated patients side by side based on recovery of bone formation. There are a number of studies that are taking place in places like Korea and China in conjunction with Korean companies.…”