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VOLUME 21 , ISSUE 7 ( July, 2020 ) > List of Articles

ORIGINAL RESEARCH

Osteogenic Potential of Periodontal Ligament Stem Cells Cultured in Osteogenic and Regular Growth Media: Confocal and Scanning Electron Microscope Study

Areej H Mukhtar, Montaser N Alqutub

Citation Information : Mukhtar AH, Alqutub MN. Osteogenic Potential of Periodontal Ligament Stem Cells Cultured in Osteogenic and Regular Growth Media: Confocal and Scanning Electron Microscope Study. J Contemp Dent Pract 2020; 21 (7):776-780.

DOI: 10.5005/jp-journals-10024-2822

License: CC BY-NC 4.0

Published Online: 19-08-2020

Copyright Statement:  Copyright © 2020; Jaypee Brothers Medical Publishers (P) Ltd.


Abstract

Aim: To evaluate the ability of osteogenic culture media in comparison with regular growth culture media in enhancing the osteoblastic cell differentiation of human periodontal ligament stem cells (hPDLSCs). Materials and methods: In vitro cultures of commercially obtained hPDLSCs were seeded onto xenograft bone blocks in both regular and osteogenic media. Confocal laser microscope images were obtained for cellular differentiation and adhesion, and scanning electron microscopy (SEM) images were obtained to validate the osteogenic differentiation by showing the morphological characteristics of the newly formed cells. Results: Confocal laser microscope analysis showed positive staining for new bone cells with an increased signal intensity when samples were cultured in osteogenic culture media compared with regular culture media. These findings indicate the effect of the active ingredients of the osteogenic culture media in enhancing the osteogenic differentiation hPDLSC. Scanning electron microscopy images validated the osteogenic differentiation showing a flattened, polygonal morphology with multiple extending cytoplasmic processes of new cells. Conclusion: Xenograft bone blocks are biocompatible scaffold for the osteogenic differentiation of seeded hPDLSCs. Osteogenic culture media enhances and increases the osteogenic differentiation of hPDLSCs into new bone cells more than regular growth culture media. Periodontal ligament stem cells are a predictable biological input as a cell-based tissue-engineered construct and biologically acceptable when it is cultured in a suitable growth media that mimics the intended environment. Clinical significance: Consideration of the clinical use of equine bone blocks and periodontal ligament stem cells in a suitable biological environment as a potential new option for bone regeneration techniques.


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  1. Bartold PM, Mcculloch CA, Narayanan AS, et al. Tissue engineering: a new paradigm for periodontal regeneration based on molecular and cell biology. periodontology 2000. 2000;24:253–269. DOI: 10.1034/j.1600-0757.2000.2240113.x.
  2. Pagni G, Kaigler D, Rasperini G, et al. Bone repair cells for craniofacial regeneration. Adv Drug Deliv Rev 2012;64(12):1310–1319. DOI: 10.1016/j.addr.2012.03.005.
  3. Moore WR, Graves SE, Bain GI. Synthetic bone graft substitutes. ANZ J Surgery 2001;71(6):354–361. DOI: 10.1046/j.1440-1622.2001.02128.x.
  4. Scaglione M, Fabbri L, Dell'Omo D, et al. Long bone nonunions treated with autologous concentrated bone marrow-derived cells combined with dried bone allograft. Musculoskelet Surg 2014;98(2):101–106. DOI: 10.1007/s12306-013-0271-2.
  5. Han J, Menicanin D, Marino V, et al. Assessment of the regenerative potential of allogeneic periodontal ligament stem cells in a rodent periodontal defect model. J Periodontal Res 2014;49(3):333–345. DOI: 10.1111/jre.12111.
  6. Murabayashi D, Mochizuki M, Tamaki Y, et al. Practical methods for handling human periodontal ligament stem cells in serum-free and serum-containing culture conditions under hypoxia: implications for regenerative medicine. Hum Cell 2017;30(3):169–180. DOI: 10.1007/s13577-017-0161-2.
  7. Zhang Y, Li X, Qian S, et al. Down-regulation of type I Runx2 mediated by dexamethasone is required for 3T3-L1 adipogenesis. Mol Endocrinol 2012;26(5):798–808. DOI: 10.1210/me.2011-1287.
  8. El-Sabban ME, El-Khoury H, Hamdan-Khalil R, et al. Xenogenic bone matrix extracts induce osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells. Regen Med 2007;2(4): 383–390. DOI: 10.2217/17460751.2.4.383.
  9. Boyde A. Stereoscopic images in confocal (tandem scanning) microscopy. Science 1985;230(4731):1270–1272. DOI: 10.1126/science.4071051.
  10. Burridge K, Nuckolls G, Otey C, et al. Actin-membrane interaction in focal adhesions. Cell Differ Dev 1990;32(3):337–342. DOI: 10.1016/0922-3371(90)90048-2.
  11. Born A-K, Rottmar M, Lischer S, et al. Correlating cell architecture with osteogenesis: first steps towards live single cell monitoring. Eur Cell Mater 2009;18:49–62. DOI: 10.22203/ecm.v018a05.
  12. Manescu A, Giuliani A, Mohammadi S, et al. Osteogenic potential of dualblocks cultured with human periodontal ligament stem cells: in vitro and synchrotron microtomography study. J Periodontal Res 2016;51(1):112–124. DOI: 10.1111/jre.12289.
  13. Schroeder TM, Jensen ED, Westendorf JJ. Runx2: a master organizer of gene transcription in developing and maturing osteoblasts. Birth Defects Res C Embryo Today 2005;75(3):213–225. DOI: 10.1002/bdrc.20043.
  14. Matsuoka F, Takeuchi I, Agata H, et al. Morphology-based prediction of osteogenic differentiation potential of human mesenchymal stem cells. PLoS ONE 2013;8(2):e55082. DOI: 10.1371/journal.pone. 0055082.
  15. Annaz B, Hing KA, Kayser M, et al. Porosity variation in hydroxyapatite and osteoblast morphology: a scanning electron microscopy study. J Microsc 2004;215(1):100–110. DOI: 10.1111/j.0022-2720.2004.01354.x.
  16. Trubiani O, Orsini G, Zini N, et al. Regenerative potential of human periodontal ligament derived stem cells on three-dimensional biomaterials: a morphological report. J Biomed Mater Res A 2008;87(4):986–993. DOI: 10.1002/jbm.a.31837.
  17. Beloti MM, Rosa AL. Osteoblast differentiation of human bone marrow cells under continuous and discontinuous treatment with dexamethasone. Braz Dent 2005;16(2):156–161. DOI: 10.1590/s0103-64402005000200013.
  18. Atmani H, Audrain C, Mercier L, et al. Phenotypic effects of continuous or discontinuous treatment with dexamethasone and/or calcitriol on osteoblasts differentiated from rat bone marrow stromal cells. J Cell Biochem 2002;85(3):640–650. DOI: 10.1002/jcb.10165.
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