Dynamic cultivation of human mesenchymal stem cells in a rotating bed bioreactor system based on the Z®RP platform

Abstract

Because the regeneration of large bone defects is limited by quantitative restrictions and risks of infections, the development of bioartificial bone substitutes is of great importance. To obtain a three-dimensional functional tissue-like graft, static cultivation is inexpedient due to limitations in cell density, nutrition and oxygen support. Dynamic cultivation in a bioreactor system can overcome these restrictions and furthermore provide the possibility to control the environment with regard to pH, oxygen content, and temperature. In this study, a three-dimensional bone construct was engineered by the use of dynamic bioreactor technology. Human adipose tissue derived mesenchymal stem cells were cultivated on a macroporous zirconium dioxide based ceramic disc called Sponceram. Furthermore, hydroxyapatite coated Sponceram was used. The cells were cultivated under dynamic conditions and compared with statically cultivated cells. The differentiation into osteoblasts was initiated by osteogenic supplements. Cellular proliferation during static and dynamic cultivation was compared measuring glucose and lactate concentration. The differentiation process was analysed determining AP-expression and using different specific staining methods. Our results demonstrate much higher proliferation rates during dynamic conditions in the bioreactor system compared to static cultivation measured by glucose consumption and lactate production. Cell densities on the scaffolds indicated higher proliferation on native Sponceram compared to hydroxyapatite coated Sponceram. With this study, we present an excellent method to enhance cellular proliferation and bone lineage specific growth of tissue like structures comprising fibrous (collagen) and globular (mineral) extracellular components.

Bioreactor Systems for Human Bone Tissue Engineering

“…Another study by Diederichs et al demonstrated that adipose tissue-derived hMSCs cultured for 47 days under osteogenic conditions onto macroporous zirconium dioxide-based ceramic discs with a diameter of 65 mm (natural or hydroxyaptatite (HA)-coated) in rotating wall vessels (1 rpm) displayed increased cell proliferation and bone matrix synthesis and mineralization, as evidenced by glucose consumption and positive staining for osteocalcein, Alizarin red and von Kossa respectively [43]. However, no significant improvements were observed on the HA-coated scaffolds, indicating a role played by the biomaterial properties in bone tissue development under the investigated conditions.…”

Proliferation and osteogenic differentiation of human mesenchymal stem cells on zirconia and titanium with different surface topography

“…It was reported that nanostructure surface enhanced the proliferation and osteogenic differentiation of the MSCs on titanium [23][24][25][26] . However, few studies were reported about the hMSCs behavior on the TZP with different surfaces 27,28) . We assumed that the microand nano-topography may have suitable configurations for hMSCs behavior.…”

Approaches for Stimulating Proliferation of Stem Cells in Vitro

“…Recently, various attempts have been reported to expand MSC in three-dimensional (3D) environments, based on suspension culture in the presence of dynamic flow [85,86], on microcarrier beads [87][88][89] or on a rotating bed bioreactor system [90,91]. Papadimitropoulos et al [94] developed a system for the proliferation of MSC which entirely bypasses the use of 2D surfaces by seeding and expanding fresh bone marrow preparations directly within the pores of 3D scaffolds under perfusion flow.…”