Main Article Content

Abstract

Highlights:



  1. Sodium hydroxide causing platelet rich fibrin stimulates limbal stem cell proliferation in chemical trauma.

  2. Limbal stem cell niche and influences limbal stemness was repaired by Platelet Rich Fibrin.


 


Abstract:


Chemical injuries of the eye produce extensive damage to the ocular surface and limbal stem cells, resulting in permanent unilateral or bilateral visual impairment. Alkali injuries occur more frequently than acid injuries. Platelets are a rich source of potential wound healing, promoting polypeptide growth factors. This study aimed to investigate the effect of platelet-rich fibrin (PRF) lysates on limbal stem cell proliferation, which was exposed to sodium hydroxide that resembled limbal stem cell deficiency due to chemical trauma. Confluent rabbit (Oryctolagus cuniculus) limbal stem cells wounded using 20µL of 0.00625 M sodium hydroxide (pH 13) were treated with platelet-rich fibrin lysates (PRF) (0, 5, and 10%). PRF lysates were prepared from peripheral rabbit blood according to Choukroun's method without using anticoagulant and foreign factors for platelet activation. The proliferation of limbal stem cells was measured by a 3-(4,5-dimethylthiazol-2-yl)-2.50 diphenyl tetrazolium bromide (MTT) colorimetric assay at 24, 48, and 72 hours after exposure to sodium hydroxide. Proliferation significantly increased limbal stem cells with PRF lysates 5% (p<0.01) and 10% (p<0.01) group compared with the control (PRF 0%). There was no significant difference between PRF lysates of 5% and 10% (p>0.01). The highest proliferation of limbal stem cells was found in the PRF lysates 5% group after 48 hours (100.24%). PRF stimulated limbal stem cell proliferation in chemical trauma caused by the sodium hydroxide model. PRF repaired the limbal stem cell niche and influenced the limbal stemness. The present findings warrant further research on PRF as a novel alternative treatment for limbal stem cell deficiency.

Keywords

Platelet Rich Fibrin Lysates Limbal Stem Cells Limbal Stem Cells Deficiency Proliferation Sodium Hydroxide

Article Details

How to Cite
Prabawati, W. E. ., Suhendro, G. ., & Retnowati, E. . (2022). A First Step to Novel Approach for Treating Alkali Injury of the Cornea: Effect of Platelet Rich Fibrin Lysates on Cultured Rabbit (Oryctolagus cuniculus) Limbal Stem Cell Proliferation Exposed by Sodium Hydroxide. Folia Medica Indonesiana, 58(2), 150–155. https://doi.org/10.20473/fmi.v58i2.36321

References

  1. Anderson D, Ellies P, Pires R, et al (2001). Amniotic membrane transplantation for partial limbal stem cell deficiency. Br. J. Opththalmology 85, 567–575.
  2. Cakmak H, Can G, Can M, et al (2017). A novel graft option after pterygium excision: platelet-rich fibrin for conjunctivoplasty. Eye 31, 1606–1612.
  3. Castro-Muñozledo F (2015). The mammalian limbal stem cell niche: A complex interaction between cells, growth factors and extracellular matrix. In: Biology in Stem Cell Niche. Springer, Switzerland, pp. 23–56.
  4. Duan X, Lin Z, Lin X, et al (2017). Study of platelet-rich fibrin combined with rat periodontal ligament stem cells in periodontal tissue regeneration. J. Cell. Mol. Med. 22, 1047–1055.
  5. Fan T, Xu B, Zhao J, et al (2011). Establishment of an untransfected human corneal epithelial cell line and its biocompability with denuded amniotic membrane. Int. J. Ophthalmol. 4, 228–234.
  6. Freire V, Andollo N, Extebarria J, et al (2014). Corneal wound healing promoted by 3 blood derivatives: An in vitro and in vivo comparative study. Cornea J. 33, 614–620.
  7. Hu N, Zhang Y, Gu H, et al (2012). Effect of bone marrow mesenchymal stem cells on cell proliferation and growth factor expression of limbal epithelial cells in vitro. Ophthalmic Res. 48, 82–88.
  8. Kadar T, Dachier S, Horwitz V, et al (2013). Development of limbal stem cell deficiency following chemical injury-pathogenesis and therapeutic strategies. US Opththalmic Rev. 6, 101–104.
  9. Kocaba V, Damour O, Auxenfans C, et al (2016). Limbal stem cell deficiency management. A review. J. Fr. Ophtalmol. 39, 791–803.
  10. Mark P, Kleinsorge M, Gaebel R, et al (2013). Human mesenchymal stem cells display reduced expression of CD 105 after culture in serum-free medium. Stem Cells Int. 2013, 1–8.
  11. Naik B, Karunakar P, Jayadev M, et al (2013). Role of platelet-rich fibrin in wound healing: A critical review. J. Conserv. Dent. 16, 284–293.
  12. Nakatsu N, González S, Mei H, et al (2014). Human limbal mesenchymal cells support the growth of human corneal epithelial stem/ progenitor cells. Investig. Ophthalmol. Vis. Sci. 55, 6953–6959.
  13. Nguyen H, Nguyen M, Trinh V, et al (2016). Platelet-rich fibrin influences on proliferation and migration of human gingival fibroblast. Int. J. Exp. Dent. Sci. 5, 83–88.
  14. Pattamatta U, Willcox M, Stapleton F, et al (2009). Bovine lactoferrin stimulates human corneal epithelial alkali wound healing in vitro. Investig. Ophthalmol. Vis. Sci. 50, 1636–1643.
  15. Sacchetti M, Rama P, Bruscolini A, et al (2018). Limbal stem cell transplantation: Clinical results, limits, and perspectives. Stem Cells Int. 2018, 1–13.
  16. Saeed M, El-Rahman M, Helal M, et al (2017). Efficacy of human platelerich fibrin exudate vs fetal bovine serum on proliferation and differentiation of dental pulp stem cells. Int. J. Stem Cell 11, 38–47.
  17. Soffer E, Ouhayoun J, Anagnostou F (2003). Fibrin sealants and platelet preparations in bone and periodontal healing. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. 95, 521–528.
  18. Wu C, Lee S, Tsai C, et al (2012). Platelet-rich fibrin increases cell attachment, proliferation and collagen-related protein expression of human osteoblasts. Aust. Dent. J. 57, 207–212.