Analysis of Spontaneous and Induced Osteogenic Differentiation in 3D-micromasses of Human Multipotent Stem Cells.

Background/aim: Craniofacial reconstruction of extensive bone defects causes high morbidity to patients. Contemporary reconstructive surgery aims at restoring lost bone with either autogenous bone or substitutes. Multipotent unrestricted somatic stem cells (USSC) show a potential for osteoblast differentiation and are increasingly used in tissue engineering.

Effects of dexamethasone, ascorbic acid and β-glycerophosphate on the osteogenic differentiation of stem cells in vitro.

The standard procedure for the osteogenic differentiation of multipotent stem cells is treatment of a confluent monolayer with a cocktail of dexamethasone (Dex), ascorbic acid (Asc) and β-glycerophosphate (β-Gly). This review describes the effects of these substances on intracellular signaling cascades that lead to osteogenic differentiation of bone marrow stroma-derived stem cells. We conclude that Dex induces Runx2 expression by FHL2/β-catenin-mediated transcriptional activation and that Dex enhances Runx2 activity by upregulation of TAZ and MKP1.

Biocompatibility of membranes with unrestricted somatic stem cells.

Aim: The biocompatibility of human osteoblasts (HOB) and human unrestricted somatic stem cells (USSCs) with membranes (BioGide®, GORE-TEX®, GENTA-FOIL resorb®, RESODONT®, BioMend®, BioMend® Extend™) was evaluated.

Materials And Methods: After osteogenic differentiation (dexamethasone, ascorbic acid and β-glycerolphosphate) cells were seeded on membranes. On days 1, 3 and 7, attachment, proliferation, cell vitality, cytotoxicty and cell morphology were analyzed.

Scaffold-free microtissues: differences from monolayer cultures and their potential in bone tissue engineering.

Objectives: Cell-based therapies for bone augmentation after tooth loss and for the treatment of periodontal defects improve healing defects. Usually, osteogenic cells or stem cells are cultivated in 2D primary cultures, before they are combined with scaffold materials, even though this means a loss of the endogenous 3D microenvironment for the cells. Moreover, the use of single-cell suspensions for the inoculation of scaffolds or for the direct application into an area of interest has the disadvantages of low initial cell numbers and susceptibility to unwanted cellular distribution, respectively.

Impact of DAG stimulation on mineral synthesis, mineral structure and osteogenic differentiation of human cord blood stem cells.

It remains unexplored in what way osteogenic stimulation with dexamethasone, ascorbic acid and β-glycerol phosphate (DAG) influences the process of mineralization, the composition and structure of the assembled mineral. Therefore, we analyzed and characterized biomineralization in DAG-stimulated and unstimulated 3D human unrestricted somatic stem cell (USSC) cultures. Microspheres were analyzed by histological staining, scanning electron microscopy (SEM), semi-quantitative energy-dispersive X-ray spectroscopy (EDX), quantitative wavelength-dispersive X-ray spectroscopy (WDX), transmission electron microscopy (TEM), selected area electron diffraction (SAED) and Raman spectroscopy.

Generation and differentiation of microtissues from multipotent precursor cells for use in tissue engineering.

This protocol describes an effective method for the production of spherical microtissues (microspheres), which can be used for a variety of tissue-engineering purposes. The obtained microtissues are well suited for the study of osteogenesis in vitro when multipotent stem cells are used. The dimensions of the microspheres can easily be adjusted according to the cell numbers applied in an individual experiment.

Improvement of the cell-loading efficiency of biomaterials by inoculation with stem cell-based microspheres, in osteogenesis.

In critical-size bone defects, autologous or allogenic cells are required in addition to compatible biomaterials for the successful defect healing. State of the art inoculation methods of biomaterials are based on the application of cell suspensions to the biomaterial. However, only less amounts of cells can be applied and sufficient adhesion to the material is required.

Comparison of ectopic bone formation of embryonic stem cells and cord blood stem cells in vivo.

Cell-based reconstruction therapies promise new therapeutic opportunities for bone regeneration. Unrestricted somatic stem cells (USSC) from cord blood and embryonic stem cells (ESCs) can be differentiated into osteogenic cells. The purpose of this in vivo study was to compare their ability to induce ectopic bone formation in vivo.

Biocompatibility of osteogenic predifferentiated human cord blood stem cells with biomaterials and the influence of the biomaterial on the process of differentiation.

Modern cell-based bone reconstruction therapies offer new therapeutic opportunities and tissue engineering represents a more biological-oriented approach to heal bone defects of the skeleton. Human unrestricted somatic stem cells (USSCs) derived form umbilical cord blood offer new promising aspects e.g.

Osteogenic differentiation influences stem cell migration out of scaffold-free microspheres.

Complete bone regeneration of critical-size defects frequently fail because of the use of acellular bone substitutes and because of partially negative influences of artificial scaffolds. However, the supply of cells to critical-size defects is essential for the regeneration. Therefore, engineered scaffold-free tissues, with outgrowing cells that fill up spaces in the bony defect, are promising candidates for bone regeneration approaches.