Lead perforation is a rare but serious complication of pacemaker implantations, and in the present study the associated tissue failure was investigated by means of in-vitro penetration of porcine and bovine ventricular tissue. Rectangular patches from the right ventricular free wall and the interventricular septum were separated, bi-axially stretched and immersed in physiological salt solution at 37( composite function)C before load displacement curves of in total 891 penetrations were recorded. To this end flat-bottomed cylindrical punches of different diameters were used, and following mechanical testing the penetration sites were histological analyzed using light and electron microscopes. Penetration pressure, i.e. penetration force divided by punch cross-sectional area decreased slightly from 2.27(SD 0.66) to 1.76(SD0.46)N/mm(2) for punches of 1.32 to 2.30 mm in diameter, respectively. Deep penetration formed cleavages aligned with the local fiber orientation of the tissue, and hence, a mode-I crack developed, where the crack faces were wedged open by the advancing punch. The performed study derived novel failure data from ventricular tissue due to deep penetration and uncovered associated failure mechanisms. This provides information to derive mechanical failure models, which are essential to enrich our current understanding of failure of soft biological tissues and to guide medical device development.