Purpose: To evaluate the role of corneal structural resistance as a surgical failure factor in deep lamellar keratoplasty (DLK). Method: A total of 10 eyes of 10 patients underwent DLK at the Ophthalmic Hospital in Rome. The big bubble technique was performed for deep stromal dissection by air injection. Seven patients were affected by advanced keratoconus and corneal thinning ranging from 441 to 235 µm. Two patients were affected by central corneal opacity from herpetic keratitis, and one patient suffered from corneal leucoma caused by bacterial keratitis. Clinical follow-up comprising final astigmatism and visual acuity findings were evaluated with a minimum follow-up of 12 weeks. Results: DLK was successfully performed in eight eyes, five of which were affected by moderate to advanced keratoconus and three by post-infective corneal opacity. In these patients preoperative ultrasonic pachymetry ranged between 441 and 287 µm. In the remaining two patients a perforation of the Descemet’s membrane (DM) occurred while attempting to separate it from the overlying stroma by the big bubble technique, requiring a penetrating keratoplasty (PK) to be performed. In both cases preoperative ultrasonic pachymetry was below 250 µm. Both perforations occurred at a different site than the needle site and at the operative time of the big-bubble injection. Discussion: An ultrastructurally weakened DM may suffer a loss of resistance to a stressing force, becoming unable to tolerate the big bubble technique, and thus being perforated. Since the weakening of the DM is related to end-stage keratoconus corneal thinning, the preoperative corneal thickness rather than the surgeon’s ability can play a major role in surgical failure of DLK. Our study reveals a very high risk of perforation of the DM when pre-operative total pachymetry is below limit of 250 µm.