Expression of human dentin matrix protein 1 in escherichia coli and nicotiana benthamiana and their effect on proliferation and osteogenic differentiation of human periodontal ligament stem cells

Abstract

This study aimed to express recombinant protein human Dentin Matrix Protein 1 (hDMP1) in Escherichia coli and Nicotiana benthamiana and investigated the osteogenic differentiation effects in human Periodontal Ligament Stem Cells (hPDLSCs). The gene encoding hDMP1 was ligated into the pET22b and pBY2R2e expression vectors. Recombinant hDMP1 protein has been productively expressed in E. coli and N. benthamiana. E. coli-produced hDMP1 resulted in 60 kDa and 100, and the optimal condition to produce highly expression was demonstrated at 0.5 mM IPTG and incubation at 37oC for 6 hr. N. benthamiana-produced hDMP1 protein showed in a 100 kDa that indicated the presence of post-translational modification, and the highest-level expression was obtained at 2 dpi and OD600 0.4. Recombinant hDMP1 could induce ALP, BMP2, CBFA1, OSX, OPN, and WNT3a osteogenic marker genes and calcium deposition in hPDLSCc. In conclusion, the results indicated that recombinant hDMP1 could induce osteogenic differentiation in hPDLSCs. However, N. benthamiana-produced hDMP1 could induce osteogenic differentiation and calcium deposition better than E. coli-produced hDMP1. Therefore, N. benthamiana-produced hDMP1 has potential to use as an inductive molecule in bone tissue engineering.