Inhibition of osteoblast differentiation but not adipocyte differentiation of mesenchymal stem cells by sera obtained from aged females

Abstract

Aging is associated with decreased osteoblast-mediated bone formation leading to bone loss and increased risk for osteoporotic fractures. However, the cellular mechanisms responsible for impaired osteoblast functions are poorly understood. In the present study, we hypothesized that changes in bone microenvironment composition with aging are responsible for impaired osteoprogenitor cell recruitment and differentiation. As a model for bone microenvironment, we examined the effects of sera obtained from young (age 20-30 year old [yo], n = 20) and old (70-84 yo, n = 19) healthy female donors on cell proliferation and differentiation capacity into osteoblasts and adipocytes of human mesenchymal stem cells (hMSC). Cell proliferation rate determined by counting cell number was similar when the cells were cultured in the presence of media containing 5% sera from old or from young donors. Similarly, the number of adipocytes and levels of adipocytic gene expression was similar in cultures incubated with sera from young or old donors. We observed decreased osteoblastic gene expression in hMSC cultured either in pooled or individual sera of old donors compared to sera from young donors: core binding factor/runt-related binding factor 2 (Cbfa1/Runx2) 46% ± 2% (P < 0.05), alkaline phosphatase (ALP) 45% ± 2% (P < 0.05), collagen type I (Col-I) 50% ± 1% (P < 0.05), and osteocalcin 65% ± 3% (P < 0.05). This down-regulation of the mRNA was accompanied by reduced ALP enzyme activity by 25% ± 1% (P < 0.01), immunocytochemical staining for osteoblastic markers: ALP, Col-I, and bone sialoprotein (BSP) as well as reduced in vitro mineralization as determined by Alizarin red staining. In conclusion, age-related changes in the serum composition and possibly hMSC microenvironment may contribute to the impaired osteoblast functions with aging. The factors mediating these changes remain to be determined. © 2006 Elsevier Inc. All rights reserved.