Andrea Cristina Baptista Coelho de Faria*, Antonio Carlos Aloise and Lidya Masako Ferreira

Surgical Translational Graduate Program at Unifesp, Rua Botucatu, São Paulo- SP, Brazil

*Corresponding Author: Andrea Cristina Baptista Coelho de Faria, Surgical Translational Graduate Program at Unifesp, Rua Botucatu, São Paulo- SP, Brazil.

Received: August 20, 2021; Published: September 25, 2021

Abstract

Introduction: The cultivation of periodontal ligament fibroblasts, associated with the simulation of compression or tension on the cell, enables the analysis of changes in morphology and identification of molecules and protein expression. The system formed by the receptor activator of nuclear factor κB (RANK), receptor activator of nuclear factor κB ligand (RANKL), and osteoprotegerin (OPG) is directly linked to bone remodeling, in the formation and reabsorption of bone tissue.

Objective: To evaluate gene expression of RANK \ RANKL and OPG in a three-dimensional culture of human periodontal fibroblasts in chitosan and xanthan membranes.

Method: Immortalized human periodontal fibroblasts were used. Cells were randomized into two groups: control group (CG), with three-dimensional cultivation without application of load for 6h, and experimental group (EG1, EG2, EG3 and EG4), with three-dimensional cultivation with application of 4, 12, 24, and 48 g/cm² loads for 6h, respectively. Cell mRNA was extracted from all groups and evaluated through real-time polymerase chain reaction (RT-PCR) and TaqMan probe. Results: No RANK / RANKL expression was detected, however OPG expression occurred, which was significantly higher in EG2 compared to CG, EG1, EG3 and EG4.

Conclusion: The 12g/cm² load applied onto a three-dimensional culture of periodontal fibroblasts for 6 hours displayed greater OPG expression.

Keywords: RANK; RANKL; OPG; RT-PCR

References

1. Alves LB., et al. “Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel”. Journal of Applied Oral Science – SciELO2 (2015): 206-214.
2. Bellini MZ., et al. “Comparison of the properties of compacted and porous lamelar chitosan-xanthan membranes as dressings and scaffolds for the treatment of skin lesions”. Journal of Applied Polymer Science 125 (2012): 421-431.
3. Bellini MZ., et al. “Combining Xanthan and chitosan membranes to multipotent mesenchimal stromal cells as bioactive dressings for dermo-epidermal wounds”. Journal of Biomaterials Applications 8 (2015): 1155-1166.
4. Chen FM and Jin Y. “Periodontal tissue engineering and regeneration: current approaches and expanding opportunities”. Tissue Engineering. Part B, Reviews 16 (2010): 219-255.
5. Chevallay B and Herbage D. “Collagen-based biomaterials as 3D scaffold for cell cultures: applications for tissue engineering and gene therapy”. Medical and Biological Engineering and Computing 38 (2000): 211-218.
6. Daley WP., et al. “Extracellular matrix dynamics in development and regenerative medicine”. Journal of Cell Science 121 (2008): 255-264.
7. Griffith LG and Naughton G. “Tissue engineering-current challenges and expanding opportunities”. Science 295 (2002): 1009-1014.
8. Heckler AF., et al. “Development of a three-dimensional in vitro model system to study orthodontic tooth movement”. Archives of Oral Biology 58 (2013): 498-410.
9. Jianru YI., et al. “Static compression regulates OPG expression in periodontal ligament cells via the CAMK II pathway”. Journal of Applied Oral Science – SciELO 6 (2015): 549-554.
10. Kanzaki H., et al. “Periodontal ligament cells under stress induce osteoclastogenesis by Receptor Activator of Nuclear Factor κB Ligand up-regulation via prostaglandin E2 synthesis”. Journal of Bone and Mineral Research 17 (2002): 210-220.
11. Kanzaki H., et al. “Compression and tension variably alter Osteoprotegerin expression via miR-3198 in periodontal ligament cells”. BMC Molecular and Cell Biology6 (2019): 187.
12. Kang KL., et al. “Bioinformatic analysis of responsive genes in two-dimension and three-dimension cultured human periodontal ligament cells subjected to compressive stress”. Journal of Periodontal Research 48 (2013): 87-97.
13. Kearns AE., et al. “Receptor activator of nuclear factor kappaB ligand and osteoprotegerin regulation of bone remodeling in health and disease”. Nature Reviews Endocrinology 29 (2008): 155-192.
14. Kim JW., et al. “Effects of compressive stress on the expression of IL-1β, RANKL and OPG mRNA in periodontal ligament cells”. The Korean Journal of Orthodontics4 (2009): 248-256.
15. Kook SH., et al. “Mechanical force inhibits osteoclastogenic potential of human periodontal ligament fibroblasts through OPG production and ERK-mediated signaling”. Journal of Cellular Biochemistry 106 (2009): 101-119.
16. Kook SH., et al. “Human periodontal ligament fibroblasts stimulate osteoclastogenesis in response to compression force through TNF-α-mediated activation of CD 4+ T cells”. Journal of Cellular Biochemistry 112 (2011): 2891-2801.
17. Lee YH., et al. “Differential gene expression of periodontal ligament cells after loading of static compressive force”. Journal of Periodontology 41 (2007): 446-452.
18. Li Y., et al. “Expression of osteoclastogenesis inducers in a tissue model of peri- odontal ligament under compression”. Journal of Dental Research 90 (2011): 115-120.
19. Li M., et al. “Effects of mechanical forces on osteogenesis and osteoclastogenesis in human periodontal ligament fibroblasts”. Bone and Joint Research 8 (2019): 19-31.
20. Lossdorfer S., et al. “Induced changes in RANKL and OPG expression by human PDL cells modify osteoclast biology in a co-culture model with RAW 264.7 cells”. Clinical Oral Investigations 15 (2011): 941-952.
21. Meikle MC. “The tissue, cellular, and molecular regulation of orthodontic tooth movement: 100 years after Carl Sandstedt”. European Journal of Orthodontics 28 (2006): 221-240.
22. Minuth WW., et al. “Tissue engineering: generation of differentiated artificial tissues for biomedical applications”. Cell and Tissue Research 291 (1998): 1-11.
23. Mueller-Klieser W. “Three-dimensional cell cultures: from molecular mechanisms to clinical applications”. American Journal of Physiology 273 (1997): 1109-1123.
24. Mitsuhashi M., et al. “Effects of HSP70 on the compression force-induced TNF and RANKL expression. In human periodontal ligament cells”. Inflammation Research 60 (2011): 187-194.
25. Nakajima R., et al. “Effects of compression force on fibroblast growth fator-2 and receptor activator of nuclear fator kappa B ligand production by periodontal ligament cells in vitro”. Journal of Periodontal Research 43 (2008): 168-173.
26. Nishijima Y., et al. “Levels of RANKL and OPG in gingival crevicular fluid during orthodontictooth movement and effect of compression force on releases from periodontal ligament cells in vitro”. Orthodontics and Craniofacial Research 9 (2006): 63-70.
27. Romer P. “The molecular mechanism behind bone remodelling: A review”. Clinical Oral Investigations 13 (2009): 355-362.
28. Sant’ana ACP., et al. “Culture and caractherization of human derived periodontal ligament cells”. Revista da Faculdade de Odontologia de Bauru 10 (2002): 134-140.
29. Souza RFB., et al. “Mechanically-enhanced polysaccharide-based scaffolds for tissue engineering of soft tissues”. Materials Science and Engineering C 94 (2019): 364-375.
30. Uygun BE., et al. “Membrane thickness in an important variable in membrane scaffolds: influence of chitosan membrane structure on the behavior of cells”. Acta Biomater 6 (2010): 2126-2131.
31. Varoni EM., et al. “Chitosan-Based Trilayer Scaffold for Multitissue Periodontal Regeneration”. Journal of Dental Research3 (2018): 303-311.
32. Wescott DC., et al. “Osteogenic gene expression by human periodontal ligament cells under cyclic tension”. Journal of Dental Research 86 (2007): 1212-1216.
33. Xu T., et al. “Electrophysiological characterization of embryonic hippocampal neurons cultured in a 3D collagen hydrogel”. Biomaterials 30 (2009): 4377-4383.
34. Yasuda H., et al. “Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis inhibitory factor and is identical to TRANCE/RANKL”. Proceedings of the National Academy of Sciences of the United States of America 95 (2009): 3597-3502.

Citation

Citation: Andrea Cristina Baptista Coelho de Faria., et al. “Gene Expression of the RANK/RANKL/OPG System on a Three-Dimensional Culture of Human Periodontal Fibroblasts Under Continuous Compression”. Acta Scientific Dental Sciences 5.10 (2021): 91-99.

Copyright

Copyright: © 2021 Andrea Cristina Baptista Coelho de Faria., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.