Dynamic behavior of different quantities of osteoblasts during formation of micromass cultures.

Implantation of micromass cultures of osteoblastic cells offers the possibility of scaffold free tissue engineering for example, regeneration of bone defects. However, the details of cell dynamics during the formation of these micromasses are still not well understood. This study aims to investigate and clarify the extent to which cell quantity influences the dynamics of micromass formation of osteoblastic cell cultures. For this purpose, the migration and aggregation during this process are investigated by optical inspection employing image processing software that allows for automated tracking of cell groups using digital image correlation. An exponential time behavior is observed with respect to the velocity of the cells and the distance of the cells to their common center of gravity. Characteristic time constants are derived as quantitative measures of the cell dynamics. The results indicate that the time constants strongly depend on the quantity of cells, that is, will decrease with increasing cell quantity. © 2018 International Society for Advancement of Cytometry.

Quantitative analysis of dynamic behavior of osteoblasts during in vitro formation of micro-mass cell cultures.

Improvements in bone tissue engineering require an understanding of cellular and tissue level behavior of osteoblast-like cells. Experiments indicate that in the absence of an anchoring material, intercellular adhesion may be based on signals that promote cell activity resulting in the formation of a spheroid cell-matrix. The aim of the present study is to investigate the formation of scaffold-free three-dimensional micro-mass cell spheroids in vitro, and to characterize quantitatively the cell movement. A new correlation based automated tracking method is evaluated in order to optimize the processing parameters and to identify statistical parameters that characterize the cell behavior. Results suggest that the temporal development of the mean distance of the cells to the center of gravity may be described by an exponential function, thus providing a characteristic time constant as a quantitative measure of cell dynamics. (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).

An in vitro study of osteoblast vitality influenced by the vitamins C and E.

Vitamin C and vitamin E are known as important cellular antioxidants and are involved in several other non-antioxidant processes. Generally vitamin C and vitamin E are not synthesized by humans and therefore have to be applied by nutrition. The absence or deficiency of the vitamins can lead to several dysfunctions and even diseases (e.g. scurvy). The main interest in this study is that vitamin C and E are known to influence bone formation, e.g. vitamin C plays the key role in the synthesis of collagen, the major component of the extracellular bone matrix.In the present study we evaluate the effect of ascorbic acid (vitamin C) and α-tocopherol (vitamin E) on the proliferation and differentiation of primary bovine osteoblasts in vitro. Starting from standard growth medium we minimized the foetal calf serum to reduce their stimulatory effect on proliferation.An improved growth and an increased synthesis of the extracellular matrix proteins collagen type I, osteonectin and osteocalcin was observed while increasing the ascorbic acid concentration up to 200 µg/ml. Furthermore the effects of α-tocopherol on cell growth and cell differentiation were examined, whereby neither improved growth nor increased synthesis of the extracellular matrix proteins collagen type I, osteonectin and osteocalcin were detected.Further investigations are necessary to target at better supportive effect of vitamins on bone regeneration, and healing.

Kinetic oxygen measurements by CVC96 in L-929 cell cultures.

Generally animal and human cells use oxygen during their whole life. Consequently the oxygen use is a simple indicator to test the vitality of cells. When the vitality decreases by the delivery of toxic substances the decrease can be observed directly by the oxygen-use of the cells. To get fast information of the vitality of cells we have measured the O2-tension by testing a new model of a bioreactor, the Cell Vitality Checker 96 (CVC96), in practical application. With this CVC96, soon a simple test will exist for the measurement of the oxygen use. In this respect the question had to be answered whether the use in the laboratory is easy and whether oxygen as a parameter in the vitality test can also be applied in future for problems in the field of material testing.