Knockdown of SVCT2 impairs in-vitro cell attachment, migration and wound healing in bone marrow stromal cells

Rajnikumar Sangani, Chirayukumar D Pandya, Maryka H. Bhattacharyya, Sudharsan Periyasamy-Thandavan, Norman Chutkan, Shanu Markand, William D. Hill, Mark W Hamrick, Carlos M Isales, Sadanand T Fulzele

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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.

Original languageEnglish (US)
Pages (from-to)354-363
Number of pages10
JournalStem Cell Research
Volume12
Issue number2
DOIs
StatePublished - Jan 1 2014

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Mesenchymal Stromal Cells
Wound Healing
Cell Movement
Ascorbic Acid
Oxidative Stress
Fracture Healing
Cell Adhesion
Bone Fractures
Sodium-Coupled Vitamin C Transporters
Cell Migration Assays
In Vitro Techniques
Bone Remodeling
p38 Mitogen-Activated Protein Kinases
Cell- and Tissue-Based Therapy
Osteogenesis
Actins
Chronic Disease
Down-Regulation
Phosphorylation
Wounds and Injuries

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Medicine(all)

Cite this

Sangani, R., Pandya, C. D., Bhattacharyya, M. H., Periyasamy-Thandavan, S., Chutkan, N., Markand, S., ... Fulzele, S. T. (2014). Knockdown of SVCT2 impairs in-vitro cell attachment, migration and wound healing in bone marrow stromal cells. Stem Cell Research, 12(2), 354-363. https://doi.org/10.1016/j.scr.2013.11.002

Knockdown of SVCT2 impairs in-vitro cell attachment, migration and wound healing in bone marrow stromal cells. / Sangani, Rajnikumar; Pandya, Chirayukumar D; Bhattacharyya, Maryka H.; Periyasamy-Thandavan, Sudharsan; Chutkan, Norman; Markand, Shanu; Hill, William D.; Hamrick, Mark W; Isales, Carlos M; Fulzele, Sadanand T.

In: Stem Cell Research, Vol. 12, No. 2, 01.01.2014, p. 354-363.

Research output: Contribution to journalArticle

Sangani, R, Pandya, CD, Bhattacharyya, MH, Periyasamy-Thandavan, S, Chutkan, N, Markand, S, Hill, WD, Hamrick, MW, Isales, CM & Fulzele, ST 2014, 'Knockdown of SVCT2 impairs in-vitro cell attachment, migration and wound healing in bone marrow stromal cells', Stem Cell Research, vol. 12, no. 2, pp. 354-363. https://doi.org/10.1016/j.scr.2013.11.002
Sangani R, Pandya CD, Bhattacharyya MH, Periyasamy-Thandavan S, Chutkan N, Markand S et al. Knockdown of SVCT2 impairs in-vitro cell attachment, migration and wound healing in bone marrow stromal cells. Stem Cell Research. 2014 Jan 1;12(2):354-363. https://doi.org/10.1016/j.scr.2013.11.002
Sangani, Rajnikumar ; Pandya, Chirayukumar D ; Bhattacharyya, Maryka H. ; Periyasamy-Thandavan, Sudharsan ; Chutkan, Norman ; Markand, Shanu ; Hill, William D. ; Hamrick, Mark W ; Isales, Carlos M ; Fulzele, Sadanand T. / Knockdown of SVCT2 impairs in-vitro cell attachment, migration and wound healing in bone marrow stromal cells. In: Stem Cell Research. 2014 ; Vol. 12, No. 2. pp. 354-363.
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