Immunocytochemistry and western blot test for the in-situ detection of biomarkers of osteogenesis

Abstract

There are two ways that bones can form: intramembranous and endochondral ossification. Specific osteogenic markers, such as insulin-like growth factor (IGF-1), osteocalcin (OCN), Runt-related transcription factor 2 (RUNX2), and osterix (OSX), accompany osteoblast differentiation from an undifferentiated state to a functional state. IGF-1 hormones are the main regulators in growth, differentiation, and apoptosis in cells and tissues mediated by IGF-1 receptors (IGF-1R). Biomolecular technology aims to study nucleic acids and their regulation and expression of proteins. Techniques that can be used when analyzing proteins include the Bradford protein assay, immunocytochemistry, immunohistochemistry, sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and the western blot test. To better understand the biomarkers of osteogenesis, the use of in-situ detection is suggested, such as immunocytochemistry and the western blot test. For this review, the author adhered to the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) standards. The research examined 50 articles; only 4 articles were selected for this review. In the immunocytochemical test, IGF-1 expression was found in the nucleus and IGF-1R expression in the cell membrane, and it was found that RUNX2, OSN, and OCN are important for osteogenesis. During the western blot test, it was found that the addition of IGF-1 in dental stem cells could increase the expression of RUNX2, OSX, and OCN proteins. Based on this information, it appears that both immunocytochemistry and western blot tests can detect the main biomarkers of osteogenesis.