Download PDF
CC BY-NC-ND 4.0 · Eur J Dent 2019; 13(02): 131-136
DOI: 10.1055/s-0039-1694904
Original Article
Dental Investigation Society

Dimethyl Sulfoxide Leads to Decreased Osteogenic Differentiation of Stem Cells Derived from Gingiva via Runx2 and Collagen I Expression

Hyunjin Lee
1   Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea
,
Jun-Beom Park
1   Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea
› Author Affiliations Funding This research was supported by Research Fund of Seoul St. Mary's Hospital, The Catholic University of Korea.
Further Information

Publication History

Publication Date:
01 October 2019 (online)

   Permissions and Reprints

Abstract

Objectives Dimethyl sulfoxide (DMSO) plays various functions, including cellular functions such as cellular growth. The aim of this study was to evaluate the effects of DMSO on the proliferation and osteogenic differentiation of human gingiva-derived stem cells.

Materials and Methods Stem cells derived from gingiva were cultured in the presence of DMSO at concentrations ranging from 0.01 to 10%.

Statistical Analysis We performed a one-way analysis of variance (ANOVA) with post hoc test to determine the differences between the groups using a commercially available program and the level of significance was 0.05.

Results The cells in the control group showed normal fibroblast morphology. The cells treated with 0.01%, 0.01%, 0.1%, and 1% DMSO were morphologically similar to those from the control group on each day. Statistically significant decreases in cell counting kit-8 (CCK-8) values were seen in the 3% and 10% DMSO groups (p < 0.05). A statistically significant decrease in alkaline phosphatase activity was seen in the 3% DMSO group. (p < 0.05). The application of DMSO produced a decrease in alizarin red S staining. The expression of Runx2 and collagen I by immunofluorescence decreased as the dose of lovastatin increased.

Conclusion The effects of DMSO on the viability of osteogenic differentiation among stem cells derived from human gingiva were evaluated. Applying DMSO produced decreased cell viability and decreased osteogenic differentiation in this experimental setting. This should be considered when designing and interpreting the data, and a DMSO-free method may be considered for bone regeneration applications.

Keywords

osteogenic differentiation - proliferation - dimethyl sulfoxide - gingiva - stem cells

Supplementary Material

 

  • References

  • 1 Santos NC, Figueira-Coelho J, Martins-Silva J, Saldanha C. Multidisciplinary utilization of dimethyl sulfoxide: pharmacological, cellular, and molecular aspects. Biochem Pharmacol 2003; 65 (07) 1035-1041
    CrossrefPubMedGoogle Scholar
  • 2 Pal R, Mamidi MK, Das AK, Bhonde R. Diverse effects of dimethyl sulfoxide (DMSO) on the differentiation potential of human embryonic stem cells. Arch Toxicol 2012; 86 (04) 651-661
    CrossrefPubMedGoogle Scholar
  • 3 Yuan C, Gao J, Guo J. et al. Dimethyl sulfoxide damages mitochondrial integrity and membrane potential in cultured astrocytes. PLoS One 2014; 9 (09) e107447
    CrossrefPubMedGoogle Scholar
  • 4 Hanslick JL, Lau K, Noguchi KK. et al. Dimethyl sulfoxide (DMSO) produces widespread apoptosis in the developing central nervous system. Neurobiol Dis 2009; 34 (01) 1-10
    CrossrefPubMedGoogle Scholar
  • 5 Li YM, Wang HB, Zheng JG. et al. Dimethyl sulfoxide inhibits zymosan-induced intestinal inflammation and barrier dysfunction. World J Gastroenterol 2015; 21 (38) 10853-10865
    CrossrefPubMedGoogle Scholar
  • 6 Morris C, de Wreede L, Scholten M. et al; Chronic Malignancies and Lymphoma Working Parties of EBMT. Should the standard dimethyl sulfoxide concentration be reduced? Results of a European Group for Blood and Marrow Transplantation prospective noninterventional study on usage and side effects of dimethyl sulfoxide. Transfusion 2014; 54 (10) 2514-2522
    CrossrefPubMedGoogle Scholar
  • 7 Windrum P, Morris TC, Drake MB, Niederwieser D, Ruutu T. EBMT Chronic Leukaemia Working Party Complications Subcommittee. Variation in dimethyl sulfoxide use in stem cell transplantation: a survey of EBMT centres. Bone Marrow Transplant 2005; 36 (07) 601-603
    CrossrefPubMedGoogle Scholar
  • 8 Jin SH, Lee JE, Yun JH, Kim I, Ko Y, Park JB. Isolation and characterization of human mesenchymal stem cells from gingival connective tissue. J Periodontal Res 2015; 50 (04) 461-467
    CrossrefPubMedGoogle Scholar
  • 9 Ha DH, Pathak S, Yong CS, Kim JO, Jeong JH, Park JB. Potential differentiation ability of gingiva originated human mesenchymal stem cell in the presence of tacrolimus. Sci Rep 2016; 6: 34910
    CrossrefPubMedGoogle Scholar
  • 10 Qiu Z, Mishra A, Li M. et al. Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide. Stem Cell Res (Amst) 2015; 15 (01) 141-150
    CrossrefPubMedGoogle Scholar
  • 11 Lee SI, Yeo SI, Kim BB, Ko Y, Park JB. Formation of size-controllable spheroids using gingiva-derived stem cells and concave microwells: Morphology and viability tests. Biomed Rep 2016; 4 (01) 97-101
    CrossrefPubMedGoogle Scholar
  • 12 Yook S, Jeong JH, Jung YS. et al. Molecularly engineered islet cell clusters for diabetes mellitus treatment. Cell Transplant 2012; 21 (08) 1775-1789
    CrossrefPubMedGoogle Scholar
  • 13 Park JB, Zhang H, Lin CY. et al. Simvastatin maintains osteoblastic viability while promoting differentiation by partially regulating the expressions of estrogen receptors α. J Surg Res 2012; 174 (02) 278-283
    CrossrefPubMedGoogle Scholar
  • 14 Jeong SH, Lee JE, Kim BB, Ko Y, Park JB. Evaluation of the effects of Cimicifugae Rhizoma on the morphology and viability of mesenchymal stem cells. Exp Ther Med 2015; 10 (02) 629-634
    CrossrefPubMedGoogle Scholar
  • 15 Jin SH, Kweon H, Park JB, Kim CH. The effects of tetracycline-loaded silk fibroin membrane on proliferation and osteogenic potential of mesenchymal stem cells. J Surg Res 2014; 192 (02) e1-e9
    CrossrefPubMedGoogle Scholar
  • 16 Komori T. Regulation of osteoblast differentiation by Runx2. Adv Exp Med Biol 2010; 658: 43-49
    CrossrefPubMedGoogle Scholar
  • 17 Shi S, Kirk M, Kahn AJ. The role of type I collagen in the regulation of the osteoblast phenotype. J Bone Miner Res 1996; 11 (08) 1139-1145
    PubMedGoogle Scholar
  • 18 González-López TJ, Sánchez-Guijo FM, Ortín A. et al. Ischemic stroke associated with the infusion of DMSO-cryopreserved auto-PBSCs. Bone Marrow Transplant 2011; 46 (07) 1035-1036
    CrossrefPubMedGoogle Scholar
  • 19 Akkök CA, Holte MR, Tangen JM, Ostenstad B, Bruserud O. Hematopoietic engraftment of dimethyl sulfoxide-depleted autologous peripheral blood progenitor cells. Transfusion 2009; 49 (02) 354-361
    CrossrefPubMedGoogle Scholar
  • 20 Sánchez-Salinas A, Cabañas-Perianes V, Blanquer M. et al. An automatic wash method for dimethyl sulfoxide removal in autologous hematopoietic stem cell transplantation decreases the adverse effects related to infusion. Transfusion 2012; 52 (11) 2382-2386
    CrossrefPubMedGoogle Scholar
  • 21 Rogulska O, Petrenko Y, Petrenko A. DMSO-free cryopreservation of adipose-derived mesenchymal stromal cells: expansion medium affects post-thaw survival. Cytotechnology 2016; 69 (02) 265-276
    PubMedGoogle Scholar
  • 22 Kim JH, Ko SY, Lee JH, Kim DH, Yun JH. Evaluation of the periodontal regenerative properties of patterned human periodontal ligament stem cell sheets. J Periodontal Implant Sci 2017; 47 (06) 402-415
    CrossrefPubMedGoogle Scholar