ABSTRACT.Purpose: To compare limbal and pars plana silicone oil removal (SOR) in aphakic eyes and to evaluate the acute effect of silicone oil flow to the corneal endothelium. Methods: Sixteen aphakic patients with silicone oil endotamponade requiring SOR were recruited for this prospective study and randomly scheduled for limbal or pars plana SOR. The central corneal thickness (CCT), visual acuity (VA) and intraocular pressure were measured preoperatively, on the first postoperative day and 4 months after surgery. Endothelial cell density (ECD) was measured preoperatively and at the end of follow-up. The in vitro study was performed on ten enucleated porcine eyes. Corneoscleral discs were prepared and fixed on artificial anterior chamber followed by 2.5-mm limbal incision and 5-ml silicone oil injection in six cases and 5 ml balanced salt solution (BSS) in four cases. Results: The ECD decreased by 239.2 ± 86.7 (13.9%) and 86.7 ± 22.4 cells ⁄ mm 2 (5%) after limbal (n = 8) and pars plana SOR (n = 8), respectively (p < 0.001 for both). The difference between the groups was significant (p < 0.001). A significant increase in CCT and corresponding decrease in VA was noted on the first postoperative day using both procedures. At the end of follow-up, the CCT and VA were comparable to initial values. Postoperative hypotony (£6 mmHg) was observed more frequently after limbal SOR. In the experiment, lamellar abrasions of corneal endothelium were observed after silicone oil injection, whereas no changes were observed after BSS injection. Conclusion: Limbal SOR causes more considerable damage to the corneal endothelium than the pars plana approach because of mechanical abrasion.