Highlights
Osteopontin (OPN) plays an important role in the bone regeneration process.
The plant-produced OPN-Fc increases the protein expression level and facilitates the purification of the recombinant protein.
The plant-produced OPN-Fc can stimulate the expression of osteogenic related genes and the calcium deposition in hPDL cells.
The plant-produced OPN-Fc has potential application in tissue engineering in the future.
Highlights
Recombinant human dentin matrix protein 1 (DMP1) can be produced in
Escherichia coli
.
E. coli
produced DMP1 could induce the expression of osteogenic-related genes and calcium deposition in human PDL cells.
This protein has potential to use for improving tooth repair and regeneration in the future.
Human fibroblast growth factor regulates a broad spectrum of biological functions, including cell proliferation and tissue differentiation, and has a wider application in tissue engineering. Here, we described the production of human basic fibroblast growth factor in plants by using a geminiviral vector system. In this study, we transiently expressed basic fibroblast growth factor containing a C-terminus 8X-Histidine with and without a barley alpha amylase signal peptide in Nicotiana benthamiana. The expression level of basic fibroblast growth factor without the signal peptide was found to be higher than the basic fibroblast growth factor with the signal peptide. Further, the recombinant basic fibroblast growth factor was purified from the plant crude extract by two-step purification viz., ammonium sulfate precipitation and Ni-affinity chromatography. Our results demonstrated that the purified plant-produced basic fibroblast growth factor was biologically active and promotes the proliferation of human periodontal ligament stem cells and human follicle dermal papilla cells in vitro. Moreover, the plant-produced basic fibroblast growth factor also induced collagen production in human dermal fibroblast cells. Our results suggest the potential use of plant-produced basic fibroblast growth factor as an antiaging and hair growth-promoting agent in the cosmetic industry.
Inductive molecules are critical components for successful bone tissue engineering. Dentin matrix protein-1 (DMP1), a non-collagenous protein in the bone matrix, has been shown to play roles in osteogenic differentiation and phosphate homeostasis. This study aimed to produce recombinant human dentin matrix protein-1 (hDMP1) in Nicotiana benthamiana and investigated the ability of this plant-produced DMP1 to induce osteogenesis in human periodontal ligament stem cells (hPDLSCs). The hDMP1 gene was cloned into the geminiviral vector for transient expression in N. benthamiana. We found that hDMP1 was transiently expressed in N. benthamiana leaves and could be purified by ammonium sulphate precipitation followed by nickel affinity chromatography. The effects of hDMP1 on the induction of cell proliferation and osteogenic differentiation were investigated. The results indicated that plant-produced hDMP1 could induce the cell proliferation of hPDLSCs and increase the expression levels of osteogenic genes, including osterix (OSX), type I collagen (COL1), bone morphogenetic protein-2 (BMP2), and Wnt3a. Moreover, the plant-produced hDMP1 promoted calcium deposition in hPDLSCs as determined by alizarin red S staining. In conclusion, our results indicated that plant-produced hDMP1 could induce osteogenic differentiation in hPDLSCs and could potentially be used as a bone inducer in bone tissue engineering.
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