Abstract
Purpose To compare two platelet-rich plasma (PRP) preparation methods (double spin (D-PRP) and TriCell PRP (T-PRP)) for eye drops, concentration yields of platelets and other cells, release of growth factors, and efficacy of the de-epithelization rabbit model. Methods PRP was extracted by D-PRP and T-PRP from 30 ml blood from healthy adults. After extraction, platelets and leukocytes were counted. D-PRP and T-PRP were preserved during A: 1 h storage at room temperature, B: 1 h storage at −20°C, C: 24 h storage at 4°C, and D: 24 h storage at −20°C. Platelet-derived growth factor (PDGF) was measured. Freezing/thawing PRP eye drops and control were instilled in the de-epithelization rabbit model four times per day for 5 days. Histology was compared between eyes treated with control, D-PRP, and T-PRP. Results 14 ml of D-PRP and 4 ml of T-PRP were extracted from 30 ml whole blood samples. D-PRP and T-PRP had 41.36 ± 8.43 × 104 and 67.02 ± 13.55 × 104 platelets and 0.41 ± 0.24 × 103/ml and 10.09 ± 4.29 × 103/ml leucocytes, respectively. In the four storage methods, PDGF concentrations in T-PRP were higher than those in D-PRP eye drops. Freezing/thawing D-PRP and T-PRP increased PDGF concentrations. Histology showed corneal epithelium thickness in T-PRP-treated eyes after healing (38.41 ± 9.10 μm) was significantly higher than that in control-treated (27.77 ± 4.76 μm) and D-PRP-treated eyes (18.32 ± 3.14 μm) (P < 0.05). There was no corneal damage with inflammation in corneal stroma and epithelium of all tested groups. Electron microscopy revealed strong adhesion between cell junctions in T-PRP-treated eyes. Conclusions Freezing/thawing of PRP extracted with the T-PRP kit may result in high platelet and leukocyte concentration and produce high PDGF concentration. PRP eye drops including leucocytes could increase thickness of corneal epithelium without corneal inflammation.
Highlights
Ocular surface disorders caused by infection, inflammation, and injury may cause a delay in wound healing of the corneal tissue such as the epithelium and stroma, resulting in visual disturbance and persistent pain
Eye drop of platelet-rich plasma (PRP) derived from human blood of one healthy volunteer (D-PRP or T-PRP) or phosphate buffer solution (PBS) as the control was instilled in rabbit cornea four times per day after epithelium removal
We found that the concentration of all growth factors in the T-PRP stock conditions was higher than that in D-PRP
Summary
Ocular surface disorders caused by infection, inflammation, and injury may cause a delay in wound healing of the corneal tissue such as the epithelium and stroma, resulting in visual disturbance and persistent pain. Eye drops derived from blood and body derivatives are used due to the stimulation of cellular proliferation and migration via the supply of an active mixture of growth factors and cytokines at the ocular surface. As these derivatives can be obtained from the patient’s own peripheral blood, there are few side effects and immune reaction. We hypothesized PRP extracted using kits would be more effective compared to PRP prepared by spin methods, due to the higher concentration of growth factors. We measured and compared the growth factors and efficacy in vitro in PRP eye drops produced using the PRP extraction kit and conventional methods
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