Comparative Analysis of Dental Stem Cells from a Single Donor: Differences in Adipogenic Differentiation and Protein Profile
DOI:
https://doi.org/10.15517/ijds.2024.58892Keywords:
Stem cells; Dental stem cells; Dental pulp; Periodontal ligament; Adipogenesis.Abstract
Dental stem cells (DSCs) are multipotent cells with high proliferation capacity and multilineage differentiation. Few studies have compared the cellular characteristics and adipogenic differentiation potential of DSCs derived from tissues of the same individual. The objective of this work was to evaluate the differences in growth characteristics, expression of mesenchymal stem cells (MSCs)-specific markers,
and the proteomic profile in response to adipogenic differentiation, of dental pulp and periodontal ligament cells obtained from a single donor. Dental cells were isolated from the third molar of a single donor using the outgrowth method. To obtain the proliferation curve of the cells was evaluated by trypan blue analysis. After the cells were cultured in adipogenic medium, morphological changes were monitored by oil red O staining, as well as adipogenic markers PPARγ and adiponectin by RT-qPCR. Finally, a two-dimensional electrophoresis of the proteins isolated from these cells was performed to analyze the proteomic profile. The two types of DSCs share similar cellular characteristics; however, their capacity for adipogenic differentiation is different. Based on the protein profiling results, we identified five differentially expressed proteins between both types of stem cells. Our findings showed that dental pulp and periodontal ligament stem cells from a single donor have similar cellular characteristics but a different response to adipogenesis, which would explain the differences in the expression of their proteins.
Downloads
References
Rodas-Junco B.A., Canul-Chan M., Rojas-Herrera R.A., De-la-Peña C., Nic-Can G.I. Stem cells from dental pulp: what epigenetics can do with your tooth. Front Physiol. 2017; 8: 999. DOI: https://doi.org/10.3389/fphys.2017.00999
Bansal R., Jain A. Current overview on dental stem cells applications in regenerative dentistry. J Nat Sci Biol Med. 2015; 6 (1): 29. DOI: https://doi.org/10.4103/0976-9668.149074
Sarjeant K., Stephens J.M. Adipogenesis. Cold Spring Harb Perspect Biol. 2012; 4 (9): a008417. DOI: https://doi.org/10.1101/cshperspect.a008417
Lee H., Lee B., Park S., Kim C. The proteomic analysis of an adipocyte differentiated from human mesenchymal stem cells using two-dimensional gel electrophoresis. Proteomics. 2006; 6 (4): 1223-9. DOI: https://doi.org/10.1002/pmic.200500385
Zhuang W., Ge X., Yang S., Huang M., Zhuang W., Chen P., et al. Upregulation of lncRNA MEG3 promotes osteogenic differentiation of mesenchymal stem cells from multiple myeloma patients by targeting BMP4 transcription. Stem Cells. 2015; 33 (6): 1985-97. DOI: https://doi.org/10.1002/stem.1989
Khurshid Z., Zohaib S., Najeeb S., Zafar M.S., Rehman R., Rehman I.U. Advances of proteomic sciences in dentistry. Int J Mol Sci. 2016;17 (5): 728. DOI: https://doi.org/10.3390/ijms17050728
Li L., Zuo Y., Zou Q., Yang B., Lin L., Li J., et al. Hierarchical Structure and Mechanical Improvement of an n-HA/GCO–PU Composite Scaffold for Bone Regeneration. ACS Appl Mater Interfaces [Internet]. 2015;151002103911000. Available from: http://pubs.acs.org/doi/10.1021/acsami.5b07327 DOI: https://doi.org/10.1021/acsami.5b07327
DeLany J.P., Floyd Z.E., Zvonic S., Smith A., Gravois A., Reiners E., et al. Proteomic Analysis of Primary Cultures of Human Adipose-derived Stem Cells: Modulation by Adipogenesis* S. Mol Cell Proteomics. 2005; 4 (6): 731-40. DOI: https://doi.org/10.1074/mcp.M400198-MCP200
Lo T., Tsai C.-F, Shih Y.-R.V., Wang Y.-T., Lu S.-C., Sung T.-Y., et al. Phosphoproteomic analysis of human mesenchymal stromal cells during osteogenic differentiation. J Proteome Res. 2012; 11 (2): 586-98. DOI: https://doi.org/10.1021/pr200868p
Jeong J.A., Ko K., Park H.S., Lee J., Jang C., Jeon C., et al. Membrane proteomic analysis of human mesenchymal stromal cells during adipogenesis. Proteomics. 2007; 7 (22): 4181-91. DOI: https://doi.org/10.1002/pmic.200700502
Pelaez-Garcia A., Barderas R., Batlle R., Vinas-Castells R., Bartolome R.A., Torres S., et al. A proteomic analysis reveals that Snail regulates the expression of the nuclear orphan receptor Nuclear Receptor Subfamily 2 Group F Member 6 (Nr2f6) and interleukin 17 (IL-17) to inhibit adipocyte differentiation. Mol Cell Proteomics. 2015; 14 (2): 303-15. DOI: https://doi.org/10.1074/mcp.M114.045328
Guerrero-Jiménez M., Nic-Can G.I., Castro-Linares N., Aguilar-Ayala F.J., Canul-Chan M., Rojas-Herrera R.A., et al. In vitro histomorphometric comparison of dental pulp tissue in different teeth. PeerJ. 2019; 7: e8212. DOI: https://doi.org/10.7717/peerj.8212
Peterson G.L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977; 83 (2): 346-56. DOI: https://doi.org/10.1016/0003-2697(77)90043-4
Zuk P.A. The adipose-derived stem cell: looking back and looking ahead. Mol Biol Cell. 2010; 21 (11): 1783-7. DOI: https://doi.org/10.1091/mbc.e09-07-0589
Zhang X., Liu J., Liang X., Chen J., Hong J., Li L., et al. History and progression of Fat cadherins in health and disease. Onco Targets Ther. 2016; 9: 7337. DOI: https://doi.org/10.2147/OTT.S111176
Chakrabarty R.P., Chandel N.S. Mitochondria as signaling organelles control mammalian stem cell fate. Cell Stem Cell. 2021; 28 (3): 394-408. DOI: https://doi.org/10.1016/j.stem.2021.02.011
Zhou D., Gan L., Peng Y., Zhou Y., Zhou X., Wan M., et al. Epigenetic regulation of dental pulp stem cell fate. Stem Cells Int. 2020; 2020: 8876265 DOI: https://doi.org/10.1155/2020/8876265
Li B., Ouchi T., Cao Y., Zhao Z., Men Y. Dental-derived mesenchymal stem cells: state of the art. Front Cell Dev Biol. 2021; 9: 654559. DOI: https://doi.org/10.3389/fcell.2021.654559
Drela K., Stanaszek L., Nowakowski A., Kuczynska Z, Lukomska B. Experimental strategies of mesenchymal stem cell propagation: adverse events and potential risk of functional changes. Stem Cells Int. 2019; 6: 7012692 DOI: https://doi.org/10.1155/2019/7012692
Mercado-Rubio M.D., Pérez-Argueta E., Zepeda-Pedreguera A., Aguilar-Ayala F.J., Peñaloza-Cuevas R., Kú-González A., et al. Similar Features, Different Behaviors: A Comparative In Vitro Study of the Adipogenic Potential of Stem Cells from Human Follicle, Dental Pulp, and Periodontal Ligament. J Pers Med. 2021; 11 (8): 738. DOI: https://doi.org/10.3390/jpm11080738
Kotova A.V., Lobov A.A., Dombrovskaya J.A., Sannikova V.Y., Ryumina N.A., Klausen P., et al. Comparative Analysis of Dental Pulp and Periodontal Stem Cells: Differences in Morphology, Functionality, Osteogenic Differentiation and Proteome. Biomedicines. 2021; 9 (11): 1606. DOI: https://doi.org/10.3390/biomedicines9111606
Navabazam A.R., Sadeghian Nodoshan F., Sheikhha M.H., Miresmaeili S.M., Soleimani M., Fesahat F. Characterization of mesenchymal stem cells from human dental pulp, preapical follicle and periodontal ligament. Iran J Reprod Med. 2013 Mar; 11 (3): 235-42.
Frank D., Cser A., Kolarovszki B., Farkas N., Miseta A., Nagy T. Mechanical stress alters protein O-GlcNAc in human periodontal ligament cells. J Cell Mol Med. 2019; 23 (9): 6251-9. DOI: https://doi.org/10.1111/jcmm.14509
Chukkapalli S.S., Lele T.P. Periodontal cell mechanotransduction. Open Biol. 2018; 8 (9): 180053. DOI: https://doi.org/10.1098/rsob.180053
Trubiani O., Zalzal S.F., Paganelli R., Marchisio M., Giancola R., Pizzicannella J., et al. Expression profile of the embryonic markers nanog, OCT-4, SSEA-1, SSEA-4, and frizzled-9 receptor in human periodontal ligament mesenchymal stem cells. J Cell Physiol. 2010; 225 (1): 123-31. DOI: https://doi.org/10.1002/jcp.22203
Tatullo M., Marrelli M., Shakesheff K.M., White L.J. Dental pulp stem cells: function, isolation and applications in regenerative medicine. J Tissue Eng Regen Med. 2015; 9 (11): 1205-16. DOI: https://doi.org/10.1002/term.1899
Silvério K.G., Rodrigues T.L., Coletta R. Dela, Benevides L., Da Silva J.S., Casati M.Z., et al. Mesenchymal stem cell properties of periodontal ligament cells from deciduous and permanent teeth. J Periodontol. 2010; 81 (8): 1207-15. DOI: https://doi.org/10.1902/jop.2010.090729
Jang J.Y., Park S.H., Park J.H., Lee B.K., Yun J.H., Lee B., et al. In Vivo Osteogenic Differentiation of Human Dental Pulp Stem Cells Embedded in an Injectable In Vivo-Forming Hydrogel. Macromol Biosci. 2016; 1158-69. DOI: https://doi.org/10.1002/mabi.201600001
Miletić M., Mojsilović S., Okić-Đorđević I., Kukolj T., Jauković A., Santibanez J., et al. Mesenchymal stem cells isolated from human periodontal ligament. Arch Biol Sci. 2014; 66 (1): 261-71. DOI: https://doi.org/10.2298/ABS1401261M
Piva E., Tarlé S.A., Nör J.E., Zou D., Hatfield E., Guinn T., et al. Dental pulp tissue regeneration using dental pulp stem cells isolated and expanded in human serum. J Endod. 2017; 43 (4): 568-74. DOI: https://doi.org/10.1016/j.joen.2016.11.018
Diomede F., Rajan T.S., Gatta V., D’Aurora M., Merciaro I., Marchisio M., et al. Stemness maintenance properties in human oral stem cells after long-term passage. Stem Cells Int. 2017: 5651287. DOI: https://doi.org/10.1155/2017/5651287
Duff S.E., Li C., Garland J.M., Kumar S. CD105 is important for angiogenesis: evidence and potential applications. FASEB J. 2003; 17 (9): 984-92. DOI: https://doi.org/10.1096/fj.02-0634rev
Saghiri M.A., Asatourian A., Sorenson C.M., Sheibani N. Mice dental pulp and periodontal ligament endothelial cells exhibit different proangiogenic properties. Tissue Cell. 2018; 50: 31-6. DOI: https://doi.org/10.1016/j.tice.2017.11.004
Tsai C.-C., Su P.-F., Huang Y.-F., Yew T.-L., Hung S.-C. Oct4 and Nanog directly regulate Dnmt1 to maintain self-renewal and undifferentiated state in mesenchymal stem cells. Mol Cell. 2012; 47 (2): 169-82. DOI: https://doi.org/10.1016/j.molcel.2012.06.020
Kawanabe N., Murata S., Murakami K., Ishihara Y., Hayano S., Kurosaka H., et al. Isolation of multipotent stem cells in human periodontal ligament using stage-specific embryonic antigen-4. Differentiation. 2010; 79 (2): 74-83. DOI: https://doi.org/10.1016/j.diff.2009.10.005
Ponnaiyan D., Jegadeesan V. Comparison of phenotype and differentiation marker gene expression profiles in human dental pulp and bone marrow mesenchymal stem cells. Eur J Dent. 2014; 8 (03): 307-13. DOI: https://doi.org/10.4103/1305-7456.137631
Pierantozzi E., Gava B., Manini I., Roviello F., Marotta G., Chiavarelli M., et al. Pluripotency regulators in human mesenchymal stem cells: expression of NANOG but not of OCT-4 and SOX-2. Stem Cells Dev. 2011; 20 (5): 915-23. DOI: https://doi.org/10.1089/scd.2010.0353
Greco S.J., Liu K., Rameshwar P. Functional similarities among genes regulated by OCT4 in human mesenchymal and embryonic stem cells. Stem Cells. 2007; 25 (12): 3143-54. DOI: https://doi.org/10.1634/stemcells.2007-0351
Trivanović D., Jauković A., Popović B., Krstić J., Mojsilović S., Okić-Djordjević I., et al. Mesenchymal stem cells of different origin: comparative evaluation of proliferative capacity, telomere length and pluripotency marker expression. Life Sci. 2015; 141: 61-73. DOI: https://doi.org/10.1016/j.lfs.2015.09.019
Volponi A.A., Gentleman E., Fatscher R., Pang Y.W.Y., Gentleman M.M., Sharpe P.T. Composition of mineral produced by dental mesenchymal stem cells. J Dent Res. 2015; 94 (11): 1568-74. DOI: https://doi.org/10.1177/0022034515599765
Okajcekova T., Strnadel J., Pokusa M., Zahumenska R., Janickova M., Halasova E., et al. A comparative in vitro analysis of the osteogenic potential of human dental pulp stem cells using various differentiation conditions. Int J Mol Sci. 2020; 21 (7): 2280. DOI: https://doi.org/10.3390/ijms21072280
Korkmaz Y., Imhof T., Kaemmerer P.W., Bloch W., Rink-Notzon S., Moest T., et al. The colocalizations of pulp neural stem cells markers with dentin matrix protein-1, dentin sialoprotein and dentin phosphoprotein in human denticle (pulp stone) lining cells. Ann Anatomy-Anatomischer Anzeiger. 2022; 239: 151815. DOI: https://doi.org/10.1016/j.aanat.2021.151815
James A.W. Review of signaling pathways governing MSC osteogenic and adipogenic differentiation. Scientifica (Cairo). 2013: 684736. DOI: https://doi.org/10.1155/2013/684736
Kolar M.K., Itte V.N., Kingham P.J., Novikov L.N., Wiberg M., Kelk P. The neurotrophic effects of different human dental mesenchymal stem cells. Sci Rep. 2017; 7 (1): 1-12. DOI: https://doi.org/10.1038/s41598-017-12969-1
Monterubbianesi R., Bencun M., Pagella P., Woloszyk A., Orsini G., Mitsiadis T.A. A comparative in vitro study of the osteogenic and adipogenic potential of human dental pulp stem cells, gingival fibroblasts and foreskin fibroblasts. Sci Rep. 2019; 9 (1):1-13. DOI: https://doi.org/10.1038/s41598-018-37981-x
Shen W.-C., Lai Y.-C., Li L.-H., Liao K., Lai H.-C., Kao S.-Y., et al. Methylation and PTEN activation in dental pulp mesenchymal stem cells promotes osteogenesis and reduces oncogenesis. Nat Commun. 2019; 10 (1): 1-13. DOI: https://doi.org/10.1038/s41467-019-10197-x
Fracaro L., Senegaglia A.C., Herai R.H., Leitolis A., Boldrini-Leite L.M., Rebelatto C.L.K., et al. The expression profile of dental pulp-derived stromal cells supports their limited capacity to differentiate into adipogenic cells. Int J Mol Sci. 2020; 21 (8): 2753. DOI: https://doi.org/10.3390/ijms21082753
Xing Y., Zhang Y., Wu X., Zhao B., Ji Y., Xu X. A comprehensive study on donor-matched comparisons of three types of mesenchymal stem cells-containing cells from human dental tissue. J Periodontal Res. 2019; 54 (3): 286-99. DOI: https://doi.org/10.1111/jre.12630
Um S., Choi J., Lee J., Zhang Q., Seo B.M. Effect of leptin on differentiation of human dental stem cells. Oral Dis. 2011; 17 (7): 662-9. DOI: https://doi.org/10.1111/j.1601-0825.2011.01820.x
Argaez-Sosa A.A., Rodas-Junco B.A., Carrillo-Cocom L.M., Rojas-Herrera R.A., Coral-Sosa A., Aguilar-Ayala F.J., et al. Higher Expression of DNA (de) methylation-Related Genes Reduces Adipogenicity in Dental Pulp Stem Cells. Front cell Dev Biol. 2022; 10: 791667 DOI: https://doi.org/10.3389/fcell.2022.791667
Li Y.-D., Lv Z., Zhu W-F. RBBP4 promotes colon cancer malignant progression via regulating Wnt/β-catenin pathway. World J Gastroenterol. 2020; 26 (35): 5328. DOI: https://doi.org/10.3748/wjg.v26.i35.5328
Christodoulides C., Lagathu C,. Sethi J.K., Vidal-Puig A. Adipogenesis and WNT signalling. Trends Endocrinol Metab. 2009; 20 (1): 16-24. DOI: https://doi.org/10.1016/j.tem.2008.09.002
Prestwich T.C., MacDougald O.A. Wnt/β-catenin signaling in adipogenesis and metabolism. Curr Opin Cell Biol. 2007; 19 (6): 612-7. DOI: https://doi.org/10.1016/j.ceb.2007.09.014
De Winter T.J.J., Nusse R. Running against the Wnt: How Wnt/β-catenin suppresses adipogenesis. Front Cell Dev Biol. 2021; 9: 627429. DOI: https://doi.org/10.3389/fcell.2021.627429
Shi H., DiRienzo D., Zemel M.B. Effects of dietary calcium on adipocyte lipid metabolism and body weight regulation in energy-restricted aP2-agouti transgenic mice. FASEB J. 2001; 15 (2): 291-3. DOI: https://doi.org/10.1096/fj.00-0584fje
Gherardi G., Monticelli H., Rizzuto R., Mammucari C. The mitochondrial Ca2+ uptake and the fine-tuning of aerobic metabolism. Front Physiol. 2020; 11: 554904. DOI: https://doi.org/10.3389/fphys.2020.554904
Zhao J., Zhou A., Qi W. The Potential to Fight Obesity with Adipogenesis Modulating Compounds. Int J Mol Sci. 2022; 23 (4): 2299. DOI: https://doi.org/10.3390/ijms23042299
Published
Issue
Section
License
Copyright (c) 2024 José A. Marín-Uc, Víctor Aguilar-Hernández, Teresa Hernández-Sotomayor, Ligia Brito Argáez, Geovanny I. Nic-Can, Martha Gabriela Chuc-Gamboa, Fernando Aguilar-Ayala, Fernando Aguilar-Pérez, Beatriz A. Rodas-Junco

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
ODOVTOS - Int. J. Dent. Sc. endorses CC BY-NC-SA
This license enables reusers to distribute, remix, adapt, and build upon the material in any medium or format for noncommercial purposes only, and only so long as attribution is given to the creator. If you remix, adapt, or build upon the material, you must license the modified material under identical terms. CC BY-NC-SA includes the following elements:
BY: credit must be given to the creator.
NC: Only noncommercial uses of the work are permitted.
SA: Adaptations must be shared under the same terms.





