The use of Platelet Rich Fibrin Membrane in Rabbit Conjunctival Tissue Damage

Abstract

 

Purpose

 

To investigate the use of platelet rich fibrin membrane (PRFM) in conjunctival tissue damage in an animal model.

 

Methods

 

The study was made on 24 eyes of 24 rabbits. All rabbits underwent unilateral (right) conjunctival damage. Twelve eyes of 12 rabbits was sutured PRFM between the two ends of damage and the other 12 eyes of 12 rabbits served as the control. To prepare PRFM, 5 mL of fresh blood from the femoral vein of a rabbit was collected into 10 mL glass-coated tubes without anticoagulants. Samples were immediately centrifuged at 2700 rpm (approximately 400 g) for 12 minutes using a Hettich EBA-20 centrifuge. The PRFM clots were concentrated between the red blood cell corpuscles at the bottom of the centrifuge tubes, and acellular plasma called platelet-poor plasma (PPP) at the top of the tubes. PPP was collected by pipetting the supernatant of the centrifuged blood preparation. After PPP removal, using forceps, PRFM clots were mechanically separated from red blood cells and gently compressed using PRFM box to drain the remaining fluid. The rabbits were euthanized, and the eyes were enucleated 1, 3, 7 and 28 days after surgery to perform histopathological examination.

 

Results

 

After enucleation all specimens were taken from the wound area.These specimens were stained with hematoxylin-eosin and immunohistochemical which inclueded vascular endothelial growth factor (VEGF), platelet derived growth factor (PDGF), Transforming growth factor-β (TGF-β) and Smooth muscle antibody (SMA). In PRFM applied group tissue was regenerated primary conjunctival tissue and there was no scar formation observed at the end of day 28.In control group wound healing and fibrosis was observed to continue (Fig.1). After 28 day VEGF, PDGF' TGF- β and SMA were significantly stained in control group than PRFM group in immunohistochemical staining (Fig.2).

 

Conclusions

 

Autologous PRFM is a fibrin matrix in which platelet cytokines, growth factors, and cells are trapped and may be released after a certain time and that can serve as a resorbable membrane. PRFM provides a scaffold material for conjunctival tissue regeneration. It may stimulate conjunctival healing and act as a barrier membrane against the damage area.

 

 

Normal conjunctival tissue in PRFM group (left), in control group inflamation and vascular proliferation are continued(right)

 

 

There is no immunostaining with PDGF in PRFM group(left) and strong staining in control group(right)

 

Keywords: 474 conjunctiva • 637 pathology: experimental • 765 wound healing