Osteoblast differentiation of human bone marrow cells under continuous and discontinuous treatment with dexamethasone

Abstract

Dexamethasone (Dex) has been shown to induce osteoblast differentiation in several cell culture systems. This study investigated the effect of continuous and discontinuous treatment with Dex on osteoblast differentiation of human bone marrow stromal cells (BMSC). Primary culture and first passage were cultured in media with or without Dex 10(-7) M. During the culture period, cells were incubated at 37 degrees C in humidified atmosphere of 5% CO2 and 95% air. At 7, 14, and 21 days, cell proliferation, cell viability, total protein content, alkaline phosphatase (ALP) activity and bone-like formation were evaluated. Data were compared by two-way analysis of variance. Dex did not affect cell viability and total protein content, but reduced cell number. ALP activity and bone-like formation increased when only first passage or both primary culture and first passage were treated with Dex, in comparison to the groups that did not have contact with Dex after first passage. The results of this study indicate that, for human BMSC, continuous presence of Dex did not appear to be required for development of the osteoblast phenotype, but Dex must be present after first passage to allow osteoblast differentiation expressed by reduced cell proliferation and increased ALP activity and bone-like formation.

Long-term effects of tripeptide Ile-Pro-Pro on osteoblast differentiation in vitro

“…bALP serves as a useful marker of the transition from the proliferative period to matrix maturation in osteogenesis [16,17]. In our study, bALP activity started to rise in control and IPP-treated cells after 1 week of culture.…”

TGF-β1 on induced osteogenic differentiation of human dermal fibroblast

“…The amount of OC in the supernatant was measured by ELISA, which revealed the presence of protein in the supernatant, the OC was detected from the seventh reaching its maximum peak in the twenty-first day and soon after we were able to observe its Although bone marrow cells have been used for this purpose, in tissue engineering these cells require a specialized culture and a purification procedure that may result in a cell population that is smaller than desired 23 . Non-osteogenic cells, such as dermal fibroblasts, are a potential cell source because they are easily obtainable from autologous tissues and can rapidly expand in vitro 13,[20][21][22]24 .…”

“…Slides with 0.5-µm-thick section cuts of collagen sponge were prepared 21 . First, a dewaxing process with xylol was performed, followed by hydration with distilled water, which was exchanged at every rinse (using at least two rinses).…”

“…This alternative cell population should be nonimmunogenic, easily expanded in vitro, and obtainable by minimally invasive tissue sampling 21 . Dermal fibroblasts can be easily isolated by biopsy of small fragments of the patient's own skin and utilized for up to 14 subcultures without any decrease in collagen biosynthesis or growth level 5,6 .…”

Porous titanium scaffolds fabricated using a rapid prototyping and powder metallurgy technique

“…The medium was changed every 2-3 days. After 14 days of culture, the medium was changed to complete McCoy's medium supplemented with ascorbic acid (50 mg/L; SigmaAldrich, Ireland), dexamethasone (10 ng/L; Sigma-Aldrich, Ireland) and b-glycerophosphate (10 mM; Sigma-Aldrich, Ireland) [40][41][42]. All assays were performed after 1, 7, 14 and 21 days in culture.…”