The Effect of Platelet-Rich Fibrin (PRF), Plasma Rich in Growth Factors (PRGF), and Enamel Matrix Proteins (Emdogain) on Migration of Human Gingival Fibroblasts

Talebi Ardakani, Mohammad Reza; Meimandi, Mansour; Shaker, Reza; Golmohammadi, Shima

doi:10.30476/dentjods.2019.44917

The Effect of Platelet-Rich Fibrin (PRF), Plasma Rich in Growth Factors (PRGF), and Enamel Matrix Proteins (Emdogain) on Migration of Human Gingival Fibroblasts

Document Type : Original Article

Authors

Mohammad Reza Talebi Ardakani ¹

Mansour Meimandi ²

Reza Shaker ³

Shima Golmohammadi ⁴

¹ Dept. of Periodontics, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

² Dept. of Periodontology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

³ Periodontist, Private Practice, Tehran, Iran.

⁴ Dept. of Periodontics, Lorestan University of Medical Sciences, Khorramabad, Iran.

https://doi.org/10.30476/dentjods.2019.44917

Abstract

Statement of the Problem:Platelet preparations such as plasma rich in growth factors (PRGF), platelet-rich fibrin (PRF), or enamel matrix proteins(Emdogain) are commonly used for soft and hard tissue regeneration. However, their comparative effectiveness has not been extensively studied, and a consensus has yet to be reached on their efficacy.
Purpose:The aim of this in vitro study was to compare the effect of PRF, PRGF, and Emdogain on the proliferation of human gingival fibroblasts (HGF).
Materials and Method: Artificial wounds were made in HGF cell culture after the fibroblasts reached nearly 100% of confluence. The study groups included leukocyte and platelet-rich fibrin (LPRF), pure platelet-rich fibrin (PPRF), PRGF and Emdogain. Stained cells were photographed at 48 h and one week and the percent of wound filling was measured. Statistical analysis was performed by one-way ANOVA and p values < 0.05 were considered statistically significant.
Results: The highest wound filling percentages at both time intervals were observed in the PPRF group, followed by the PRGF. The lowest percentage of wound healing among test groups was observed in the LPRF while Emdogain yielded modest results. However, statistical analysis showed similar wound healing values in PRGF, PPRF, and Emdogain groups.
Conclusion:Within the limitations of this study, PRGF, PPRF, and Emdogain were similarly effective in enhancing the fibroblast proliferation and artificial wound closure.

Keywords

● Platelet

rich plasma ● PRF ● Enamel matrix proteins ● Fibroblast ● Wound Healing

1. Wang HL, Greenwell H, Fiorellini J, Giannobile W, Offenbacher S, Salkin L, et al. Position paper on periodontal regeneration. J Periodontol. 2005; 76: 1601–1622. [PubMed] [Google Scholar]

2. Plachokova AS, van den Dolder J, Stoelinga PJ, Jansen JA. The bone regenerative effect of platelet-rich plasma in combination with an osteoconductive material in rat cranial defects. Clin Oral Implants Res. 2006; 17: 305–311. [PubMed] [Google Scholar]

3. Arnoczky SP, Tarvin GB, Marshall JL. Anterior cruciate ligament replacement using patellar tendon. An evaluation of graft revascularization in the dog. J Bone Joint Surg Am. 1982; 64: 217–224. [PubMed] [Google Scholar]

4. Kawase T, Okuda K, Yoshie H, Burns DM. Cytostaticaction of enamel matrix derivative (EMDOGAIN) on human oral squamous cellcarcinoma-derived SCC25 epithelial cells. J Periodontal Res. 2000; 35: 291–300. [PubMed] [Google Scholar]

5. Ivanovski S. Periodontal regeneration. Aust Dent J. 2009; 54 Suppl 1: S118–S128. [PubMed] [Google Scholar]

6. Cate AR, Deporter DA. The degradative role of the fibroblast in the remodelling and turnover of collagen in soft connective tissue. Anat Rec. 1975; 182: 1–13. [PubMed] [Google Scholar]

7. Götz W, Gerber T, Michel B, Lossdörfer S, Henkel KO, Heinemann F. Immunohistochemical characterization of nanocrystalline hydroxyapatite silica gel (NanoBone(r)) osteogenesis: a study on biopsies from human jaws. Clin Oral Implants Res. 2008; 19: 1016–1026. [PubMed] [Google Scholar]

8. Moharamzadeh K, Brook I, Van Noort R. Biocompat-ibility of Resin-based Dental Materials. Materials. 2009; 2: 514. [Google Scholar]

9. Slavkin HC, Boyde A. Cementun: an epithelial secretory product? . J Dent Res. 1975;53:157. [Google Scholar]

10. Simmer JP, Fincham AG. Molecular mechanisms of dental enamel formation. Crit Rev Oral Biol Med. 1995; 6: 84–108. [PubMed] [Google Scholar]

11. Hammarström L. Enamel matrix, cementum development and regeneration. J Clin Periodontol. 1997; 24(9 Pt 2): 658–668. [PubMed] [Google Scholar]

12. Sculean A, Chiantella GC, Windisch P, Donos N. Clinical and histologic evaluation of human intrabony defects treated with an enamelmatrix protein derivative (Emdogain) Int J Periodontics Restorative Dent. 2000; 20: 374–381. [PubMed] [Google Scholar]

13. Giannobile WV, Somerman MJ. Growth and amelogenin-like factors in periodontal wound healing. A systematic review. Ann Periodontol 2003; 8: 193–204. [PubMed] [Google Scholar]

14. Hammarström L, Heijl L, Gestrelius S. Periodontal regeneration in a buccal dehiscence model in monkeys after application of enamel matrix proteins. J Clin Periodontol. 1997; 24(9 Pt 2): 669–677. [PubMed] [Google Scholar]

15. Gestrelius S, Andersson C, Lidström D, Hammarström L, Somerman M. In vitro studies on periodontal ligament cells and enamel matrix derivative. J Clin Periodontol. 1997; 24(9 Pt 2): 685–692. [PubMed] [Google Scholar]

16. Zeldich E, Koren R, Nemcovsky C, Weinreb M. Enamel matrix derivative stimulates human gingival fibroblast proliferation via ERK. J Dent Res. 2007; 86: 41–46. [PubMed] [Google Scholar]

17. Marx RE, Carlson ER, Eichstaedt RM, Schimmele SR, Strauss JE, Georgeff KR. Platelet-rich plasma: Growth factor enhancement for bone grafts. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1998; 85: 638–646. [PubMed] [Google Scholar]

18. Anitua E. Plasma rich in growth factors: preliminary results of use in the preparation of futuresites for implants. Int J Oral Maxillofac Implants. 1999; 14: 529–535. [PubMed] [Google Scholar]

19. Aghaloo TL, Moy PK, Freymiller EG. Evaluation of platelet-rich plasma in combination with anorganic bovine bone in the rabbit cranium: a pilot study. Int J Oral Maxillofac Implants. 2004; 19: 59–65. [PubMed] [Google Scholar]

20. Kevy S, Jacobson M. Preparation of growth factor enriched autologous platelet gel. Proceeding of the 27th annual meeting of services Biomaterials 2001. Available at: [https://www.nlm.nih.gov/privacy.html. ]

21. Mosesson MW. Fibrinogen and fibrin structure and functions. J Thromb Haemost. 2005; 3: 1894–1904. [PubMed] [Google Scholar]

22. Choukroun J, Diss A, Simonpieri A, Girard MO, Schoeffler C, Dohan SL, et al. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part IV: clinical effects on tissue healing. Oral Surg Oral Med Oral Pathol Oral Radiol Endod . 2006; 101: e56–e60. [PubMed] [Google Scholar]

23. Dohan Ehrenfest, DM Choukroun, J Diss, A Dohan, SL Dohan, AJ Mouhyi, J Gogly. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part I: technologicalconcepts and evolution. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006; 101: e37–e44. [PubMed] [Google Scholar]

24. Schilephake H. Bone growth factors in maxillofacial skeletal reconstruction. Int J Oral Maxillofac Surg. 2002; 31: 469–484. [PubMed] [Google Scholar]

25. Tuan TL, Song A, Chang S, Younai S, Nimni ME. In vitro fibroplasia: matrix contraction, cell growth, and collagen production of fibroblasts cultured in fibrin gels. Exp Cell Res. 1996; 223: 127–134. [PubMed] [Google Scholar]

26. He L, Lin Y, Hu X, Zhang Y, Wu H. A comparative study of platelet-rich fibrin (PRF) and platelet-rich plasma (PRP) on the effect of proliferation and differentiation of rat osteoblasts in vitro. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009; 108: 707–713. [PubMed] [Google Scholar]

27. Liang CC1, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cellmigration in vitro. Nat Protoc. 2007; 2: 329–333. [PubMed] [Google Scholar]

28. Anitua E, Sánchez M, Nurden AT, Nurden P, Orive G, Andía I. New insights into and novel applications for platelet-rich fibrin therapies. Trends Biotechnol. 2006; 24: 227–234. [PubMed] [Google Scholar]

29. Anitua E, Sánchez M, Orive G, Andía I. The potential impact of the preparation rich in growth factors (PRGF) in different medical fields. Biomaterials. 2007; 28: 4551–4560. [PubMed] [Google Scholar]

30. El-Sharkawy H1, Kantarci A, Deady J, Hasturk H, Liu H, Alshahat M, Van Dyke. Platelet-rich plasma: growth factors and pro- and anti-inflammatory properties. J Periodontol. 2007; 78: 661–669. [PubMed] [Google Scholar]

31. Woodall J Jr, Tucci M, Mishra A, Asfour A, Benghuzzi H. Cellular effects of platelet rich plasmainterleukin1 release from prp treatedmacrophages. Biomed Sci Instrum. 2008; 44: 489–494. [PubMed] [Google Scholar]

32. Creeper F, Lichanska AM, Marshall RI, Seymour GJ, Ivanovski S. The effect of platelet-rich plasma on osteoblast and periodontal ligament cell migration, proliferation and differentiation. J Periodontal Res. 2009; 44: 258–265. [PubMed] [Google Scholar]

33. Vahabi S, Vaziri S, Torshabi M, Rezaei Esfahrood Z. Effects of Plasma Rich in Growth Factors and Platelet-Rich Fibrin on Proliferation and Viability of Human Gingival Fibroblasts. J Dent (Tehran) 2015; 12: 504–512. [PMC free article] [PubMed] [Google Scholar]

34. Dohan Ehrenfest DM, Diss A, Odin G, Doglioli P, Hippolyte MP, Charrier JB. In vitro effects of Choukroun's PRF (platelet-rich fibrin) on human gingival fibroblasts, dermal prekeratinocytes, preadipocytes, and maxillofacial osteoblasts in primary cultures. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009; 108: 341–352. [PubMed] [Google Scholar]

35. Liu Y, Kalén A, Risto O, Wahlström O. Fibroblast proliferation due to exposure to a platelet concentrate in vitro is pH dependent. Wound Repair Regen. 2002; 10: 336–340. [PubMed] [Google Scholar]

36. Graziani F, Ivanovski S, Cei S, Ducci F, Tonetti M, Gabriele M. The in vitro effect of different PRP concentrations on osteoblasts and fibroblasts. Clin Oral Implants Res. 2006; 17: 212–219. [PubMed] [Google Scholar]

37. Lopez-Vidriero E, Goulding KA, Simon DA, Sanchez M, Johnson DH. The use of platelet-rich plasma in arthroscopy and sports medicine: optimizing the healing environment. Arthroscopy. 2010; 26: 269–278. [PubMed] [Google Scholar]

38. Gassling VL, Açil Y, Springer IN, Hubert N, Wiltfang J. Platelet-rich plasma and platelet-rich fibrin in human cell culture. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009; 108: 48–55. [PubMed] [Google Scholar]

39. Kawase T, Okuda K, Wolff LF, Yoshie H. Platelet-rich plasma-derived fibrin clot formation stimulates collagen synthesis in periodontal ligament and osteoblastic cells in vitro. J Periodontol. 2003; 74: 858–864. [PubMed] [Google Scholar]

40. Kawase T, Okuda K, Saito Y, Yoshie H. In vitro evidence that the biological effects of platelet-rich plasma on periodontal ligament cells is not mediated solely by constituent transforming-growth factor-beta or platelet-derived growth factor. J Periodontol. 2005; 76: 760–767. [PubMed] [Google Scholar]

41. Dohan Ehrenfest DM, Doglioli P, de Peppo GM, Del Corso M, Charrier JB. Choukroun's platelet-rich fibrin (PRF) stimulates in vitro proliferation and differentiation of human oral bone mesenchymal stem cell in a dose-dependent way. Arch Oral Biol. 2010; 55: 185–194. [PubMed] [Google Scholar]

42. Tidball JG. Inflammatory cell response to acute muscle injury. Med Sci Sports Exerc. 1995; 27: 1022–1032. [PubMed] [Google Scholar]

43. Froum SJ, Wallace SS, Tarnow DP, Cho SC. Effect of platelet-rich plasma on bone growth and osseointegration in human maxillary sinus grafts: three bilateral case reports. Int J Periodontics Restorative Dent. 2002; 22: 45–53. [PubMed] [Google Scholar]

44. Gruber R, Varga F, Fischer MB, Watzek G. Platelets stimulate proliferation of bone cells: involvement of platelet-derived growth factor, microparticles and membranes. Clin Oral Implants Res. 2002; 13: 529–535. [PubMed] [Google Scholar]

45. Kawase T, Okuda K, Momose M, Kato Y, Yoshie H, Burns DM. Enamel matrix derivative (EMDOGAIN) rapidly stimulates phosphorylation of the MAP kinase family and nuclear accumulation of smad2 in both oral epithelial and fibroblastic human cells. J Periodontal Res. 2001; 36: 367–376. [PubMed] [Google Scholar]

46. Keila S, Nemcovsky CE, Moses O, Artzi Z, Weinreb M. In vitro effects of enamel matrix proteins on rat bone marrow cells and gingival fibroblasts. J Dent Res. 2004; 83: 134–138. [PubMed] [Google Scholar]

47. Sanders JE, Chuang A, Swiec GD, Bisch FC, Herold RW, Buxton TB, et al. The effects of enamel matrix derivative and cyclic mechanical strain on human gingival fibroblasts in an in vitro defect healing model. Int J Periodontics Restorative Dent. 2011; 31: 671–678. [PubMed] [Google Scholar]