Osteogenic Differentiation of ADSCs in Novel Microbeads in Dynamic Environment

Abstract

Feasibility of constructing bone microbeads by culturing encapsulated ADSCs in calcium alginate/bone powder (CABP) microbeads in spinner flask was explored. Experimental group was osteogenic differentiation of encapsulated ADSCs in CABP microbeads in spinner flask, control group I and II were osteogenic differentiation of ADSCs encapsulated in CABP microbeads and in calcium alginate microbeads in T-flasks, respectively. Growth status of encapsulated ADSCs in beads was observed by Calcein-AM/PI Staining. After 14 days of induction, ALP staining was used to qualitatively detect ALP in culture media. After 21 days, alizarin red staining and von-kossa staining were both used to detect mineralized nodules in cells. The growth state and proliferation ability of ADSCs in three groups were quite good. After 4 days of induction, ALP was detected in culture media, and ALP content reached the maximum on day 14, while mineralized nodules began to generate. ALP, alizarin red and von-kossa staining in experimental group showed higher osteogenic capacity than that in control groups. Three-dimensional dynamic environment and bone power could work together to promote osteogenic differentiation of encapsulated ADSCs in CABP microbeads, constructing tissue engineered bone microbeads successfully.