Open AccessBone morphogenetic protein 2: heterologous expression and potential in bone regeneration
Author(s) -
Andrea Mesa Restrepo,
Juan Fernando Alzate,
Edwin Bairon Patiño González
Publication year - 2021
Language(s) - English
Resource type - Journals
ISSN - 2145-7166
DOI - 10.17533/udea.acbi.v43n114a01
Subject(s) - heterologous , bone morphogenetic protein 2 , chemistry , bone morphogenetic protein , heterologous expression , in vitro , recombinant dna , microbiology and biotechnology , biochemistry , biology , gene
Currently, bone morphogenetic protein 2 (BMP-2) is one of the two osteoinductive growth factors used in medical devices to promote bone formation. Typically, this protein is bought from commercial houses at high rates and in small quantities that are not enough to cover clinical needs. Because of this, it has been proposed that research centers use their own heterologous expression systems to have a constant supply of BMP-2. The aim of this study was to standardize the heterologous expression of BMP-2 and evaluate its osteoinductive activity in vitro. Our procedure for expression and purification was based on recombinant DNA technology using the plasmid pET-28 and IPTG as inductor. After extracting the protein from inclusion bodies, folding it and modifying it via a redox system, we observed via electrophoresis a 26 kDa dimer. We evaluated its osteoinductive activity in myoblastic C2C12 by quantifying enzymatically the activity of alkaline phosphate (ALP) and staining mineralization nodules. ALP activity is proportional to BMP-2 concentration, increasing 90% at 3 µg/mL. These cells form calcium nodules, mineralizing 50% of the area.