title: Knockdown of SVCT2 impairs in-vitro cell attachment, migration and wound healing in bone marrow stromal cells.
abstract: Bone marrow stromal cell (BMSC) adhesion and migration are fundamental to a number of pathophysiologic processes, including fracture and wound healing. Vitamin C is beneficial for bone formation, fracture repair and wound healing. However, the role of the vitamin C transporter in BMSC adhesion, migration and wound healing is not known. In this study, we knocked-down the sodium-dependent vitamin C transporter, SVCT2, the only known transporter of vitamin C in BMSCs, and performed cell adhesion, migration, in-vitro scratch wound healing and F-actin re-arrangement studies. We also investigated the role of oxidative stress on the above processes. Our results demonstrate that both oxidative stress and down-regulation of SVCT2 decreased cell attachment and spreading. A trans-well cell migration assay showed that vitamin C helped in BMSC migration and that knockdown of SVCT2 decreased cell migration. In the in-vitro scratch wound healing studies, we established that oxidative stress dose-dependently impairs wound healing. Furthermore, the supplementation of vitamin C significantly rescued the BMSCs from oxidative stress and increased wound closing. The knockdown of SVCT2 in BMSCs strikingly decreased wound healing, and supplementing with vitamin C failed to rescue cells efficiently. The knockdown of SVCT2 and induction of oxidative stress in cells produced an alteration in cytoskeletal dynamics. Signaling studies showed that oxidative stress phosphorylated members of the MAP kinase family (p38) and that vitamin C inhibited their phosphorylation. Taken together, these results indicate that both the SVCT2 transporter and oxidative stress play a vital role in BMSC attachment, migration and cytoskeletal re-arrangement. BMSC-based cell therapy and modulation of SVCT2 could lead to a novel therapeutic approach that enhances bone remodeling, fracture repair and wound healing in chronic disease conditions.
authors.affiliation: Department of Orthopaedic Surgery, Georgia Regents University, Augusta, GA 30912, USA.
authors.forename: Rajnikumar
authors.initials: R
authors.lastname: Sangani
authors.affiliation: Department of Psychiatry and Health Behavior, Georgia Regents University, Augusta, GA 30912, USA.
authors.forename: Chirayu D
authors.initials: CD
authors.lastname: Pandya
authors.affiliation: Department of Orthopaedic Surgery, Georgia Regents University, Augusta, GA 30912, USA.
authors.forename: Maryka H
authors.initials: MH
authors.lastname: Bhattacharyya
authors.affiliation: Cellular Biology and Anatomy, Georgia Regents University, Augusta, GA 30912, USA.
authors.forename: Sudharsan
authors.initials: S
authors.lastname: Periyasamy-Thandavan
authors.affiliation: Department of Orthopaedic Surgery, Georgia Regents University, Augusta, GA 30912, USA.
authors.forename: Norman
authors.initials: N
authors.lastname: Chutkan
authors.affiliation: Cellular Biology and Anatomy, Georgia Regents University, Augusta, GA 30912, USA.
authors.forename: Shanu
authors.initials: S
authors.lastname: Markand
authors.affiliation: Cellular Biology and Anatomy, Georgia Regents University, Augusta, GA 30912, USA; Institute of Regenerative and Reparative Medicine, Georgia Regents University, Augusta, GA 30912, USA.
authors.forename: William D
authors.initials: WD
authors.lastname: Hill
authors.affiliation: Cellular Biology and Anatomy, Georgia Regents University, Augusta, GA 30912, USA; Institute of Regenerative and Reparative Medicine, Georgia Regents University, Augusta, GA 30912, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Hamrick
authors.affiliation: Department of Orthopaedic Surgery, Georgia Regents University, Augusta, GA 30912, USA; Institute of Regenerative and Reparative Medicine, Georgia Regents University, Augusta, GA 30912, USA.
authors.forename: Carlos
authors.initials: C
authors.lastname: Isales
authors.affiliation: Department of Orthopaedic Surgery, Georgia Regents University, Augusta, GA 30912, USA; Institute of Regenerative and Reparative Medicine, Georgia Regents University, Augusta, GA 30912, USA. Electronic address: sfulzele@gru.edu.
authors.forename: Sadanand
authors.initials: S
authors.lastname: Fulzele
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D015536:Down-Regulation; D055785:Gene Knockdown Techniques; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018384:Oxidative Stress / Q000502:physiology; D010766:Phosphorylation; D015398:Signal Transduction; D060347:Sodium-Coupled Vitamin C Transporters / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation; D014945:Wound Healing / Q000502:physiology*
chemical_list: C555165:Slc23a2 protein, mouse; D060347:Sodium-Coupled Vitamin C Transporters