Comparison of Fetal and Adult Marrow Stromal Cells in Osteogenesis with and without Glucocorticoids

Abstract

To better understand the potential use of fetal marrow stromal cells (MSCs) in bone tissue engineering, we compared the ability of these cells with those of adult MSCs with respect to osteoblasts differentiation in the presence or absence of glucocorticoids. Cells were grown for 3-4 weeks in basal medium or supplemented with 100 nM dexamethasone (DEX, a synthetic glucocorticoid analog) or with 50 microM L-ascorbate and 10 mM glycerol-2-phosphate (AS+GP) or with AS+GP+DEX. At various time points in culture, the following parameters were compared between fetal and adult MSCs: cell morphology, cell proliferation, alkaline phosphatase activity, calcium (45Ca) uptake, von Kossa staining, and glucocorticoids receptor expression were analyzed. Compared with adult MSCs, fetal cells showed a less dramatic change to cuboidal morphology in DEX-containing media. Fetal MSCs in all media conditions showed higher proliferation rates and lower alkaline phosphatase activities (p < 0.001) than adult cells. Both fetal and adult MSCs responded similarly in DEX-containing media with respect to suppressing cell proliferation, stimulating alkaline phosphatase activity, and consistently accumulating calcium (usually higher in fetal cells) with subsequent formation of mineralized matrix when compared with cells cultured in AS+GP. Our findings further implicate the requirement of glucocorticoids in osteogenesis. In conclusion, compared with adult MSCs, fetal cells showed greater ability in sustaining cell proliferation and calcium uptake suggesting that they may be useful for bone tissue repair.

Glucocorticoid-induced gene tripartite motif-containing 63 (TRIM63) promotes differentiation of osteoblastic cells

“…The effect of TRIM63 overexpression is consistent with previous reports on the global effects of glucocorticoids on human bone marrow stromal cells and SaOS2 cells [7,[10][11]. According to the previous report, glucocorticoids suppressed the proliferation and promoted osteoblastic differentiation.…”

“…The synthetic glucocorticoid dexamethasone induces osteoblastic differentiation and is often included in the osteogenic medium [7]. However, glucocorticoids are also known to induce apoptosis of osteoblasts and osteocytes, which is consistent with clinical glucocorticoid-induced osteoporosis [8].…”

Effects of silica&ndash;gentamicin nanohybrids on osteogenic differentiation of human osteoblast-like SaOS-2 cells

“…In another two studies, ECM mineralization was observed in human BMSCs that achieved a minimal ALP activity (~0.25 nmol/min/μg protein or 1.2 nmol/min/10,000 cells) during the culture period of 2-3 weeks. 46,47 From these aforementioned studies, it was observed that the levels of ALP activity were not in proportion to the observed mineralization levels. In the present study, the cells can still express low levels of ALP after exposure to a high concentration of SiO 2 -gentamicin nanohybrids or native SiO 2 NPs (Figure 6).…”

Pre-receptorial regulation of steroid hormones in bone cells: Insights on glucocorticoid-induced osteoporosis

“…In cultured MSC, glucocorticoids decrease the proliferation and trigger osteogenesis through the expression of osteogenic markers [36][37][38][39][40][41]. The presence of dexamethasone does not appear to be required for the full development of the osteoblast phenotype, but it appears to be necessary in the earliest stages of commitment to allow pre-osteoblast recruitment (Fig.…”