Objective We determine the main physical effects of pneumoperitoneum on tumour dissemination and port-site metastases occurrence.Design A prospective randomised study in rats.Methods A human ovarian cancer cell line (IGR-OV1) was xenografted in nude rats. Seven days after cancer inoculation, surgery was performed. Rats were randomised in two main groups and underwent either: gasless laparoscopy (n 20) CO2 laparoscopy with pneumoperitoneum pressure at 4 mmHg (n 20), or 8 mmHg (n 20) with in each case, increasing operative duration: 30,60,90 or 120 minutes (®ve rats for each time). Animals were killed seven days after the intervention. Main outcome measures Tumour dissemination and frequency of port-site metastases.Results Tumour dissemination was not in¯uenced by gas pressures or duration of procedure. The rate of rats with at least one port-site metastasis (one or two) was similar in all groups: gasless: n 10/20; 4 mmHg CO2: 5/20; 8 mmHg CO2: 7/20,(P 0.26). The number of port-site metastases were signi®cantly higher in the gasless group compared with the 4 mmHg CO2 group (15/40 (37.5%) vs 5/40 (12.5%), P 0.01). Difference was not signi®cant between the 8mmHg group and the gasless group (9/40(22.5%) vs 15/40(37.5%), P 0.14) or the 4mmHg group (9/40(37.5%) vs 5/40 (12.5%), P 0.24). Duration of procedures had no signi®cant in¯uence on port-site metastases rate (P . 0.05). Conclusions Unlike previous animal studies, port-site metastases were more frequent with gasless laparoscopy than with CO2 pneumoperitoneum. Local peritoneal factors could play an important role in port-site metastases mechanism.