safranine-t and ascorbate-2-phosphate

An unsolved problem with stem cell-based engineering of bone tissue is how to provide a microenvironment that promotes the osteogenic differentiation of multipotent stem cells. Previously, we fabricated porous poly(D,L-lactide-co-glycolide) (PLGA) scaffolds that released biologically active dexamethasone (Dex) and ascorbate-2-phosphate (AsP), and that acted as osteogenic scaffolds. To determine whether these osteogenic scaffolds can be used for bone formation in vivo, we seeded multipotent human marrow stromal cells (hMSCs) onto the scaffolds and implanted them subcutaneously into athymic mice. Higher alkaline phosphatase expression was observed in hMSCs in the osteogenic scaffolds compared with that of hMSCs in control scaffolds. Furthermore, there was more calcium deposition and stronger von Kossa staining in the osteogenic scaffolds, which suggested that there was enhanced mineralized bone formation. We failed to detect cartilage in the osteogenic scaffolds (negative Safranin O staining), which implied that there was intramembranous ossification. This is the first study to demonstrate the successful formation of mineralized bone tissue in vivo by hMSCs in PLGA scaffolds that release Dex and AsP.

Topics: Alkaline Phosphatase; Animals; Anti-Inflammatory Agents; Ascorbic Acid; Biocompatible Materials; Bone and Bones; Bone Development; Bone Marrow Cells; Calcium; Cartilage; Cells, Cultured; Coloring Agents; Dexamethasone; Guided Tissue Regeneration; Humans; In Situ Hybridization; Lactic Acid; Mice; Mice, Inbred BALB C; Osteogenesis; Phenazines; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Polymers; Stromal Cells; Time Factors; Tissue Engineering; Up-Regulation