Comparison of Phenotypic Characterization between Differentiated Osteoblasts from Stem Cells and Calvaria Osteoblasts In vitro

Abstract

Background: Characteristics of differentiated osteoblasts from adipose derived stem cells (ADSCs) in compared with isolated osteoblasts from normal bone such as calvaria are unknown. The aim of this study was determination and comparison of phenotypic characterization between differentiated osteoblasts from stem cells and calvaria osteoblasts in vitro.

Methods: In this study, mesenchymal stem cells were isolated from adipose tissue of human by enzymatic digestion and were differentiated into osteoblasts using osteogenic medium. Characteristics of these cells at first, second, third and fourth weeks were comprised with calvaria osteoblasts that were isolated from human calvaria by explanation culture method. To screen the characteristics of both calvaria and the differentiated osteoblasts, we used western blot to identify protein levels, von Kossa staining for mineral matrix detection and alkaline phosphatase (ALP) assay kit (Sigma) for ALP activity measurement. Difference between calvaria and differentiated osteoblast cells were analyzed by one‑way ANOVA and P < 0.05 was considered as statistically significant.

Results: Alkaline phosphatase activity, collagen and mineral material production in differentiated osteoblasts at third week were more significantly than calvaria cells (P < 0.05). Our results indicated that there was no significant different in osteocalcin (OC) production between differentiated osteoblast at first, second and third weeks and calvaria cells but declined at fourth week (P < 0.05).

Conclusions: Our survey showed that cellular traits of differentiated osteoblasts presented better than calvaria osteoblasts in vitro conditions. Therefore, we suggest that ADSCs could be used in next studies for bone tissue engineering.

Keywords: Adipose derived stem cells, bone, calvaria, osteogenesis