Year 2020, Volume 6 , Issue 6, Pages 587 - 599 2020-11-04

Micro- and nanoscale characterization of different natural biomaterials for ocular surface regeneration

Gamze DERELİ CAN [1] , Atakan TEVLEK [2] , Mehmet Erol CAN [3] , Elif ÖNCÜ [4] , Halil Murat AYDIN [5] , Nurullah CAGİL [6]

Objectives: This study aims to characterize the widely used biological derived membranes in clinics in terms of micro-nano scale mechanical and morphological properties. Within this scope, advanced platelet-rich fibrin (A-PRF), leucocyte-and platelet-rich fibrin (L-PRF) and human amniotic membrane were studied in this research study.

Methods: Nano-indentation, optical coherence tomography (OCT), scanning electron microscopy (SEM), and in vitro degradation test were performed for material characterization.

Results: The nano-indentation test revealed significantly higher modulus of elasticity and hardness values in A-PRF group, while OCT presented significantly higher thickness measurements when compared L-PRF. A loose 3D architecture formation due to the large pores formed by means of large fiber diameter were observed in A-PRF group. Besides, platelets were observed among the large fibers in A-PRF membranes on the contrary of L-PRF membranes. Low fiber diameter and high cellular separation were recorded in L-PRF group due to the high centrifugal force application. Therefore, it was observed that the platelets were located mostly on the surface of the membranes in L-PRF. The loose 3D architecture of A-PRF membranes is thought to release growth factors for a longer period of time, ensuring cellular integrity. On the other hand, degradation test results indicated that amniotic membranes degrade to about 85% in one week, while L-PRF and A-PRF were lost their initial weights approximately 31% and 40%, respectively.

Conclusions: This comparative characterization study of three different natural biomaterials used in a wide range of clinical applications, from dentistry to ophthalmology, was thought to guide surgeons on the selection of site-specific material.

Leucocyte-and platelet-rich fibrin, advanced platelet-rich fibrin, amniotic membrane, nano-indentation, optical coherence tomography
  • 1. Lin X, Huang J, Shi Y, Liu W. Tissue engineering and regenerative medicine in applied research: a year in review of 2014. Tissue Eng Part B Rev 2015;21:177-86.
  • 2. Burnouf T, Goubran HA, Chen T-M, Ou K-L, El-Ekiaby M, Radosevic M. Blood-derived biomaterials and platelet growth factors in regenerative medicine. Blood Rev 2013;27:77-89.
  • 3. 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;28:4551-60.
  • 4. Dohan Ehrenfest DM, Rasmusson L, Albrektsson T. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). Trends Biotechnol 2009;27:158-67.
  • 5. Kang Y-H, Jeon SH, Park J-Y, Chung J-H, Choung Y-H, Choung H-W, et al. Platelet-rich fibrin is a Bioscaffold and reservoir of growth factors for tissue regeneration. Tissue Eng Part A 2011;17:349-59.
  • 6. Dohan DM, Choukroun J, Diss A, Dohan SL, Dohan AJJJ, Mouhyi J, et al. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part I: technological concepts and evolution. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2006;101:37-44.
  • 7. Dohan DM, Choukroun J, Diss A, Dohan SL, Dohan AJJJ, Mouhyi J, et al. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part II: Platelet-related biologic features. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2006;101:45-50.
  • 8. Sadeghi-Ataabadi M, Mostafavi-Pour Z, Vojdani Z, Sani M, Latifi M, Talaei-Khozani T. Fabrication and characterization of platelet-rich plasma scaffolds for tissue engineering applications. Mater Sci Eng C Mater Biol Appl 2017;71:372-80.
  • 9. Ghanaati S, Booms P, Orlowska A, Kubesch A, Lorenz J, Rutkowski J, et al. Advanced platelet-rich fibrin: a new concept for cell-based tissue engineering by means of inflammatory cells. J Oral Implantol 2014;40:679-89.
  • 10. Kobayashi E, Flückiger L, Fujioka-Kobayashi M, Sawada K, Sculean A, Schaller B, et al. Comparative release of growth factors from PRP, PRF, and advanced-PRF. Clin Oral Investig 2016;20:2353-60.
  • 11. Dohan Ehrenfest DM, Pintyo NR, Pereda A, Jimenez P, Del Corso M, Kang BS, et al. The impact of the centrifuge characteristics and centrifugation protocols on the cells, growth factors and fibrin architecture of a leukocyte- and platelet-rich fibrin (L-PRF) clot and membrane. Platelets 2018;29:171-84.
  • 12. Can ME, Çakmak HB, Dereli Can G, Ünverdi H, Toklu Y, Hücemenoğlu S. A novel technique for conjunctivoplasty in a rabbit model: platelet-rich fibrin membrane grafting. J Ophthalmol 2016;2016:1965720.
  • 13. Can ME, Dereli Can G, Cagil N, Cakmak HB, Sungu N. Urgent therapeutic grafting of platelet-rich fibrin membrane in descemetocele. Cornea 2016;35:1245-9.
  • 14. Dohan Ehrenfest DM, Del Corso M, Diss A, Mouhyi J, Charrier J-B. Three-dimensional architecture and cell composition of a Choukroun’s platelet-rich fibrin clot and membrane. J Periodontol 2010;81:546-55.
  • 15. Khorshidi H, Raoofi S, Bagheri R, Banihashemi H. Comparison of the Mechanical Properties of Early Leukocyte- and Platelet-Rich Fibrin versus PRGF/Endoret Membranes. Int J Dent 2016;2016:1849207.
  • 16. Sam G, Vadakkekuttical RJ, Amol NV. In vitro evaluation of mechanical properties of platelet-rich fibrin membrane and scanning electron microscopic examination of its surface characteristics. J Indian Soc Periodontol 2015;19:32-6.
  • 17. Dohan Ehrenfest DM, Del Corso M, Inchingolo F, Sammartino G, Charrier J-B. Platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) in human cell cultures: growth factor release and contradictory results. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010;110:418-21.
  • 18. Fontana S, Olmedo DG, Linares JA, Guglielmotti MB, Crosa ME. Effect of platelet-rich plasma on the peri-implant bone response: an experimental study. Implant Dent 2004;13:73-8.
  • 19. Sharma A, Pradeep AR. Autologous platelet-rich fibrin in the treatment of mandibular degree II furcation defects: a randomized clinical trial. J Periodontol 2011;82:1396-403.
  • 20. Simonpieri A, Choukroun J, Del Corso M, Sammartino G, Dohan Ehrenfest DM. Simultaneous sinus-lift and implantation using microthreaded implants and leukocyte- and platelet-rich fibrin as sole grafting material: a six-year experience. Implant Dent 2011;20:2-12.
  • 21. Ji B, Sheng L, Chen G, Guo S, Xie L, Yang B, et al. The combination use of platelet-rich fibrin and treated dentin matrix for tooth root regeneration by cell homing. Tissue Eng Part A 2015;21:26-34.
  • 22. Dohan Ehrenfest DM, Bielecki T, Jimbo R, Barbé G, Del Corso M, Inchingolo F, et al. Do the fibrin architecture and leukocyte content influence the growth factor release of platelet concentrates? An evidence-based answer comparing a pure platelet-rich plasma (P-PRP) gel and a leukocyte- and platelet-rich fibrin (L-PRF). Curr Pharm Biotechnol 2012;13:1145-52.
  • 23. Dohan Ehrenfest DM, Diss A, Odin G, Doglioli P, Hippolyte M-P, Charrier J-B. 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;108:341-52.
  • 24. Bielecki T, Dohan Ehrenfest DM. Platelet-rich plasma (PRP) and Platelet-Rich Fibrin (PRF): surgical adjuvants, preparations for in situ regenerative medicine and tools for tissue engineering. Curr Pharm Biotechnol 2012;13:1121-30.
  • 25. Dohan Ehrenfest DM, Andia I, Zumstein MA, Zhang C-Q, Pinto NR, Bielecki T. Classification of platelet concentrates (Platelet-Rich Plasma-PRP, Platelet-Rich Fibrin-PRF) for topical and infiltrative use in orthopedic and sports medicine: current consensus, clinical implications and perspectives. Muscles Ligaments Tendons J 2014;4:3-9.
  • 26. Cieslik-Bielecka A, Choukroun J, Odin G, Dohan Ehrenfest DM. L-PRP/L-PRF in esthetic plastic surgery, regenerative medicine of the skin and chronic wounds. Curr Pharm Biotechnol 2012;13:1266-77.
  • 27. Braccini F, Tardivet L, Dohan Ehrenfest DM. [The relevance of Choukroun’s Platelet-Rich Fibrin (PRF) during middle ear surgery: preliminary results]. Rev Laryngol Otol Rhinol (Bord) 2009;130:175-80. [Article in French]
  • 28. Litwiniuk M, Grzela T. Amniotic membrane: new concepts for an old dressing. Wound Repair Regen 2014;22:451-6.
  • 29. Zhao B, Liu J-Q, Zheng Z, Zhang J, Wang S-Y, Han S-C, et al. Human amniotic epithelial stem cells promote wound healing by facilitating migration and proliferation of keratinocytes via ERK, JNK and AKT signaling pathways. Cell Tissue Res 2016;365:85-99.
  • 30. Russo V, Tammaro L, Di Marcantonio L, Sorrentino A, Ancora M, Valbonetti L, et al. Amniotic epithelial stem cell biocompatibility for electrospun poly(lactide-co-glycolide), poly(ε-caprolactone), poly(lactic acid) scaffolds. Mater Sci Eng C Mater Biol Appl 2016;69:321-9.
  • 31. Mohammadi AA, Johari HG, Eskandari S. Effect of amniotic membrane on graft take in extremity burns. Burns 2013;39:1137-41.
  • 32. Arai N, Tsuno H, Okabe M, Yoshida T, Koike C, Noguchi M, et al. Clinical application of a hyperdry amniotic membrane on surgical defects of the oral mucosa. J Oral Maxillofac Surg 2012;70:2221-8.
  • 33. Kesting MR, Wolff K-D, Nobis CP, Rohleder NH. Amniotic membrane in oral and maxillofacial surgery. Oral Maxillofac Surg 2014;18:153-64.
  • 34. Jerman UD, Veranič P, Kreft ME. Amniotic membrane scaffolds enable the development of tissue-engineered urothelium with molecular and ultrastructural properties comparable to that of native urothelium. Tissue Eng Part C Methods 2014;20:317-27.
  • 35. Malhotra C, Jain AK. Human amniotic membrane transplantation: different modalities of its use in ophthalmology. World J Transplant 2014;4:111-21.
  • 36. Mamede AC, Carvalho MJ, Abrantes AM, Laranjo M, Maia CJ, Botelho MF. Amniotic membrane: from structure and functions to clinical applications. Cell Tissue Res 2012;349:447-58.
  • 37. Lai J-Y. Photo-cross-linking of amniotic membranes for limbal epithelial cell cultivation. Mater Sci Eng C Mater Biol Appl 2014;45:313-9.
  • 38. Lai J-Y. Carbodiimide cross-linking of amniotic membranes in the presence of amino acid bridges. Mater Sci Eng C Mater Biol Appl 2015;51:28-36.
  • 39. Hatakeyama I, Marukawa E, Takahashi Y, Omura K. Effects of platelet-poor plasma, platelet-rich plasma, and platelet-rich fibrin on healing of extraction sockets with buccal dehiscence in dogs. Tissue Eng Part A 2014;20:874-82.
  • 40. Miron RJ, Fujioka-Kobayashi M, Bishara M, Zhang Y, Hernandez M, Choukroun J. Platelet-rich fibrin and soft tissue wound healing: a systematic review. Tissue Eng Part B Rev 2017;23:83-99.
Primary Language en
Subjects Ophthalmology
Journal Section Original Articles

Orcid: 0000-0002-7642-0269
Author: Gamze DERELİ CAN (Primary Author)
Institution: Department of Ophthalmology, University of Health Sciences, Bursa Yüksek İhtisas Training and Research Hospital, Bursa, Turkey
Country: Turkey

Orcid: 0000-0003-0601-8642
Author: Atakan TEVLEK
Institution: Department of Bioengineering, Hacettepe University, Institute of Science and Engineering, Ankara, Turkey
Country: Turkey

Orcid: 0000-0002-3026-6681
Author: Mehmet Erol CAN
Institution: Department of Ophthalmology, University of Health Sciences, Bursa Yüksek İhtisas Training and Research Hospital, Bursa, Turkey
Country: Turkey

Orcid: 0000-0002-5774-4604
Author: Elif ÖNCÜ
Institution: Department of Periodontology, Necmettin Erbakan University Faculty of Dentistry, Konya, Turkey
Country: Turkey

Orcid: 0000-0002-0732-1785
Author: Halil Murat AYDIN
Institution: Department of Environmental Engineering, Hacettepe University, Bioengineering Division and Centre for Bioengineering, Ankara, Turkey
Country: Turkey

Orcid: 0000-0002-3845-066X
Author: Nurullah CAGİL
Institution: Department of Ophthalmology, Yıldırım Beyazıt University School of Medicine, Ankara, Turkey
Country: Turkey


Application Date : April 7, 2019
Acceptance Date : October 13, 2019
Publication Date : November 4, 2020

EndNote %0 The European Research Journal Micro- and nanoscale characterization of different natural biomaterials for ocular surface regeneration %A Gamze Dereli̇ Can , Atakan Tevlek , Mehmet Erol Can , Elif Öncü , Halil Murat Aydın , Nurullah Cagi̇l %T Micro- and nanoscale characterization of different natural biomaterials for ocular surface regeneration %D 2020 %J The European Research Journal %P -2149-3189 %V 6 %N 6 %R doi: 10.18621/eurj.550522 %U 10.18621/eurj.550522