Electrical Stimulation through Conductive Substrate to Enhance Osteo-Differentiation of Human Dental Pulp-Derived Stem Cells

Human dental pulp-derived stem cells (hDPSCs) are promising cellular sources for bone healing. The acceleration of their differentiation should be beneficial to their clinical application. Therefore, a conductive polypyrrole (PPy)-made electrical stimulation (ES) device was fabricated to provide direct-current electric field (DCEF) treatment, and its effect on osteo-differentiation of hDPSCs was investigated in this study. To determine the optimal treating time, electrical field of 0.33 V/cm was applied to hDPSCs once for 4 h on different days after the osteo-induction. The alizarin red S staining results suggested that ES accelerated the mineralization rates of hDPSCs. The quantification analysis results revealed a nearly threefold enhancement in calcium deposition by ES at day 0, 2, and 4, whereas the promotion effect in later stages was in vain. To determine the ES-mediated signaling pathway, the expression of genes in the bone morphogenetic protein (BMP) family and related receptors were quantified using qPCR. In the early stages of osteo-differentiation, the mRNA levels of BMP2, BMP3, BMP4, and BMP5 were increased significantly in the ES groups, indicating that these genes were involved in the specific signaling routes induced by ES. We are the first using DCEF to improve the osteo-differentiation of hDPSCs, and our results promise the therapeutic applications of hDPSCs on cell-based bone tissue engineering.

[1]  T. Mueller,et al.  Structural insights into BMP receptors: Specificity, activation and inhibition. , 2016, Cytokine & growth factor reviews.

[2]  Y. Seo,et al.  Co-treatment effect of pulsed electromagnetic field (PEMF) with human dental pulp stromal cells and FK506 on the regeneration of crush injured rat sciatic nerve , 2015, The International journal of neuroscience.

[3]  M. H. Fernandes,et al.  Dental stem cells for craniofacial tissue engineering. , 2012, Oral surgery, oral medicine, oral pathology and oral radiology.

[4]  M. Murphy,et al.  Increased Internal Porosity and Surface Area of Hydroxyapatite Accelerates Healing and Compensates for Low Bone Marrow Mesenchymal Stem Cell Concentrations in Critically-Sized Bone Defects , 2018, Applied Sciences.

[5]  M. Sogayar,et al.  Bone Morphogenetic Proteins: structure, biological function and therapeutic applications. , 2014, Archives of biochemistry and biophysics.

[6]  P. Potdar,et al.  Human dental pulp stem cells: Applications in future regenerative medicine. , 2015, World journal of stem cells.

[7]  E. Wolfs,et al.  Ultrastructural and Immunocytochemical Analysis of Multilineage Differentiated Human Dental Pulp- and Umbilical Cord-Derived Mesenchymal Stem Cells , 2010, Cells Tissues Organs.

[8]  K. Lyons,et al.  Bone morphogenetic protein-3 is a negative regulator of bone density , 2001, Nature Genetics.

[9]  Effects of Micro-Electrical Stimulation on Regulation of Behavior of Electro-Active Stem Cells , 2013 .

[10]  L. Duong,et al.  PDGF-BB secreted by preosteoclasts induces CD31hiEmcnhi vessel subtype in coupling osteogenesis , 2014, Nature Medicine.

[11]  Wei-Wen Hu,et al.  The Regulation of Osteogenesis Using Electroactive Polypyrrole Films , 2016, Polymers.

[12]  J. Mao,et al.  Clones of Ectopic Stem Cells in the Regeneration of Muscle Defects In Vivo , 2010, PloS one.

[13]  Qun Wang,et al.  Electroconductive natural polymer-based hydrogels. , 2016, Biomaterials.

[14]  M. Shamos,et al.  Piezoelectric Effect in Bone , 1963, Nature.

[15]  B. Badran,et al.  Molecular mechanisms of mesenchymal stem cell differentiation towards osteoblasts. , 2013, World journal of stem cells.

[16]  M. Rogers,et al.  Turning Bone Morphogenetic Protein 2 (BMP2) on and off in Mesenchymal Cells , 2015, Journal of cellular biochemistry.

[17]  Gordon G Wallace,et al.  Electrical stimulation promotes nerve cell differentiation on polypyrrole/poly (2-methoxy-5 aniline sulfonic acid) composites , 2009, Journal of neural engineering.

[18]  P. Peckham,et al.  Functional electrical stimulation for neuromuscular applications. , 2005, Annual review of biomedical engineering.

[19]  A. Reddi.  Bone morphogenetic proteins (BMPs): from morphogens to metabologens. , 2009, Cytokine & growth factor reviews.

[20]  S Tetè,et al.  In Vitro Stem Cell Cultures from Human Dental Pulp and Periodontal Ligament: New Prospects in Dentistry , 2007, International journal of immunopathology and pharmacology.

[21]  J. Nabekura,et al.  A Novel Stem Cell Source for Vasculogenesis in Ischemia: Subfraction of Side Population Cells from Dental Pulp , 2008, Stem cells.

[22]  Miguel Angel Martin-Piedra,et al.  Human Dental Pulp Stem Cells. A promising epithelial-like cell source. , 2015, Medical hypotheses.

[23]  G. Duruksu,et al.  Human dental pulp stem cells demonstrate better neural and epithelial stem cell properties than bone marrow-derived mesenchymal stem cells , 2011, Histochemistry and Cell Biology.

[24]  C. Tsao,et al.  The effects of substrate-mediated electrical stimulation on the promotion of osteogenic differentiation and its optimization. , 2018, Journal of biomedical materials research. Part B, Applied biomaterials.

[25]  T. Hentunen,et al.  Differentiation of osteoblasts and osteocytes from mesenchymal stem cells. , 2008, Current stem cell research & therapy.

[26]  W. Grayson,et al.  Stromal cells and stem cells in clinical bone regeneration , 2015, Nature Reviews Endocrinology.

[27]  P. Devreotes,et al.  Cell migration directionality and speed are independently regulated by RasG and Gβ in Dictyostelium cells in electrotaxis , 2019, Biology Open.

[28]  C. Chien,et al.  Osteogenic differentiation of placenta-derived multipotent cells in vitro. , 2014, Taiwanese journal of obstetrics & gynecology.

[29]  Fan Wang,et al.  Human dental pulp stem cell is a promising autologous seed cell for bone tissue engineering. , 2011, Chinese medical journal.

[30]  Kozo Nakamura,et al.  Physiological role of bone morphogenetic proteins in osteogenesis , 2005, Journal of Bone and Mineral Metabolism.

[31]  D. Greenspan,et al.  Developmental roles of the BMP1/TLD metalloproteinases. , 2006, Birth defects research. Part C, Embryo today : reviews.

[32]  S. Cartmell,et al.  Direct electrical stimulation enhances osteogenesis by inducing Bmp2 and Spp1 expressions from macrophages and preosteoblasts , 2019, Biotechnology and bioengineering.

[33]  A. James,et al.  BMP-2-induced bone formation and neural inflammation. , 2017, Journal of orthopaedics.

[34]  M. Zayzafoon.  Calcium/calmodulin signaling controls osteoblast growth and differentiation , 2006, Journal of cellular biochemistry.

[35]  E. Lo,et al.  Current Advance and Future Prospects of Tissue Engineering Approach to Dentin/Pulp Regenerative Therapy , 2016, Stem cells international.

[36]  O. Forlenza,et al.  The neuroprotective effect of dental pulp cells in models of Alzheimer’s and Parkinson’s disease , 2008, Journal of Neural Transmission.

[37]  V. Rosen,et al.  Expression and function of BMP3 during chick limb development , 2008, Developmental dynamics : an official publication of the American Association of Anatomists.

[38]  R. Haydon,et al.  Bone Morphogenetic Protein (BMP) signaling in development and human diseases , 2014, Genes & diseases.

[39]  Ze Zhang,et al.  Polypyrrole as Electrically Conductive Biomaterials: Synthesis, Biofunctionalization, Potential Applications and Challenges. , 2018, Advances in experimental medicine and biology.

[40]  H. M. Jamil,et al.  TGF-β/BMP signaling and other molecular events: regulation of osteoblastogenesis and bone formation , 2015, Bone Research.

[41]  C. McCaig,et al.  Electrical dimensions in cell science , 2009, Journal of Cell Science.

[42]  Han Lu,et al.  Electroconductive hydrogels for biomedical applications. , 2019, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[43]  K. Lyons,et al.  A requirement for bone morphogenetic protein-7 during development of the mammalian kidney and eye. , 1995, Genes & development.

[44]  C. Wildsoet,et al.  Differential gene expression of BMP2 and BMP receptors in chick retina & choroid induced by imposed optical defocus , 2016, Visual Neuroscience.

[45]  Rohaya Megat Abdul Wahab,et al.  A Perspective on Stem Cells as Biological Systems that Produce Differentiated Osteoblasts and Odontoblasts. , 2017, Current Stem Cell Research & Therapy.

[46]  S. Gronthos,et al.  Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[47]  Sahba Mobini,et al.  In vitro effect of direct current electrical stimulation on rat mesenchymal stem cells , 2017, PeerJ.

[48]  C. Stanford,et al.  Calcium and phosphate supplementation promotes bone cell mineralization: implications for hydroxyapatite (HA)-enhanced bone formation. , 2000, Journal of biomedical materials research.

[49]  Yu-Che Cheng,et al.  Electrical stimulation to promote osteogenesis using conductive polypyrrole films. , 2014, Materials science & engineering. C, Materials for biological applications.

[50]  P. Gervois,et al.  Effect of isolation methodology on stem cell properties and multilineage differentiation potential of human dental pulp stem cells , 2013, Cell and Tissue Research.

[51]  V. Campanella.  Dental Stem Cells: Current research and future applications. , 2018, European Journal of Paediatric Dentistry.

[52]  P H Krebsbach,et al.  Craniofacial Tissue Engineering by Stem Cells , 2006, Journal of dental research.

[53]  Yi-Ping Li,et al.  TGF-β and BMP signaling in osteoblast, skeletal development, and bone formation, homeostasis and disease , 2016, Bone Research.

[54]  Joyce Doorn,et al.  cAMP/PKA pathway activation in human mesenchymal stem cells in vitro results in robust bone formation in vivo , 2008, Proceedings of the National Academy of Sciences.

[55]  B. Kramer,et al.  Electrical stimulation promotes the angiogenic potential of adipose-derived stem cells , 2019, Scientific Reports.