ABSTRACT
Osteoporosis is a common skeletal disease that results in an increased risk of fractures. However, there is no definitive cure, warranting the development of potential therapeutic agents. 3'-Sialyllactose (3'-SL) in human milk regulates many biological functions. However, its effect on bone metabolism remains unknown. This study aimed to investigate the molecular mechanisms underlying the effect of 3'-SL on bone homeostasis. Treatment of human bone marrow stromal cells (hBMSCs) with 3'-SL enhanced osteogenic differentiation and inhibited adipogenic differentiation of hBMSCs. RNA sequencing showed that 3'-SL enhanced laminin subunit gamma-2 expression and promoted osteogenic differentiation via the phosphatidylinositol 3kinase/protein kinase B signaling pathway. Furthermore, 3'-SL inhibited the receptor activator of nuclear factor κB ligand-induced osteoclast differentiation of bone marrow-derived macrophages through the nuclear factor κB and mitogenactivated protein kinase signaling pathway, ameliorated osteoporosis in ovariectomized mice, and positively regulated bone remodeling. Our findings suggest 3'-SL as a potential drug for osteoporosis.
Subject(s)
Oligosaccharides , Osteogenesis , Osteoporosis , Mice , Humans , Animals , Osteogenesis/genetics , Cell Differentiation/genetics , Osteoporosis/drug therapy , HomeostasisABSTRACT
An accurate microscopical analysis of blood smears requires a reproducible and convenient method of staining. Solution-based staining procedures can be cumbersome. Especially in low- and middle-income countries, the lack of skilled technicians and adequate laboratory facilities, as well as insufficient water and reagent quality, often become confounding factors. To overcome these obstacles, we developed a new cell staining method based on sequential stamping of agarose gel patches that contain eosin, methylene blue/oxidized methylene blue, Azure B, and buffer, respectively. Our method, termed "hydrogel staining", provides a simple, reproducible, solution-free, and inexpensive approach to stain blood cells. We have optimized incubation times to achieve the optimal transfer of dyes to fixed blood cells on a glass slide, with outcomes comparable to conventional solution-based methods for white blood cells and malaria-infected red blood cells. This hydrogel staining method does not require special skills to produce excellent quality stained blood film slides. The new method could enhance the accuracy of microscopical examination of blood smears, especially in resource-limited settings.