Recombinant human osteopontin expressed in Nicotiana benthamiana stimulates osteogenesis related genes in human periodontal ligament cells

Kaewta Rattanapisit, Supaniga Abdulheem, Daneeya Chaikeawkaew, Anchanee Kubera, Hugh Mason, Julian K.C. Ma, Prasit Pavasant, Waranyoo Phoolcharoen

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Tissue engineering aims to utilise biologic mediators to facilitate tissue regeneration. Several recombinant proteins have potential to mediate induction of bone production, however, the high production cost of mammalian cell expression impedes patient access to such treatments. The aim of this study is to produce recombinant human osteopontin (hOPN) in plants for inducing dental bone regeneration. The expression host was Nicotiana benthamiana using a geminiviral vector for transient expression. OPN expression was confirmed by Western blot and ELISA, and OPN was purified using Ni affinity chromatography. Structural analysis indicated that plant-produced hOPN had a structure similar to commercial HEK cell-produced hOPN. Biological function of the plant-produced hOPN was also examined. Human periodontal ligament stem cells were seeded on an OPN-coated surface. The results indicated that cells could grow normally on plant-produced hOPN as compared to commercial HEK cell-produced hOPN determined by MTT assay. Interestingly, increased expression of osteogenic differentiation-related genes, including OSX, DMP1, and Wnt3a, was observed by realtime PCR. These results show the potential of plant-produced OPN to induce osteogenic differentiation of stem cells from periodontal ligament in vitro, and suggest a therapeutic strategy for bone regeneration in the future.

Original languageEnglish (US)
Article number17358
JournalScientific Reports
Volume7
Issue number1
DOIs
StatePublished - Jan 1 2017

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Periodontal Ligament
Osteopontin
Osteogenesis
Tobacco
Genes
Bone Regeneration
Stem Cells
Tissue Engineering
Affinity Chromatography
Recombinant Proteins
Regeneration
Tooth
Western Blotting
Enzyme-Linked Immunosorbent Assay
Costs and Cost Analysis
Bone and Bones
Polymerase Chain Reaction
Therapeutics

ASJC Scopus subject areas

  • General

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Recombinant human osteopontin expressed in Nicotiana benthamiana stimulates osteogenesis related genes in human periodontal ligament cells. / Rattanapisit, Kaewta; Abdulheem, Supaniga; Chaikeawkaew, Daneeya; Kubera, Anchanee; Mason, Hugh; Ma, Julian K.C.; Pavasant, Prasit; Phoolcharoen, Waranyoo.

In: Scientific Reports, Vol. 7, No. 1, 17358, 01.01.2017.

Research output: Contribution to journalArticle

Rattanapisit, Kaewta ; Abdulheem, Supaniga ; Chaikeawkaew, Daneeya ; Kubera, Anchanee ; Mason, Hugh ; Ma, Julian K.C. ; Pavasant, Prasit ; Phoolcharoen, Waranyoo. / Recombinant human osteopontin expressed in Nicotiana benthamiana stimulates osteogenesis related genes in human periodontal ligament cells. In: Scientific Reports. 2017 ; Vol. 7, No. 1.
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