Background: Because of advances in technology, the number of orthopedic surgeries, mainly hip and knee replacement surgeries, has increased, with a total of 150,000 prosthetic surgeries estimated per year in the United States and 400,000 worldwide., Methods: We used an exploratory cross-sectional study, with a quantitative approach to determine the microbial load in instruments used in orthopedic surgeries, quantifying and identifying the microbial growth genus and species, according to the surgical potential of contamination that characterizes the challenge faced by the Material and Sterilization Center at the Institute of Orthopedics and Traumatology of Hospital das Clinicas of the School of Medicine of the University of Sao Paulo, Brazil.The orthopedic surgical instruments were immersed, after their use, in sterilized distilled water, sonicated in an ultrasonic washer, and posteriorly agitated. Subsequently, the wash was filtrated through a 0.45-mum membrane and incubated in aerobic and anaerobic mediums and in medium for fungi and yeasts., Results: In clean surgeries, 47% of the instruments were contaminated; in contaminated surgeries, 70%; and, in infected surgeries, 80%. Regardless of the contamination potential of the surgeries, the highest quantitative incidence of microorganism recovery was located in the 1 to 100 colony-forming unit range, and 13 samples presented a microbial growth potential >300 colony-forming units. Regardless of the contamination potential of the surgeries, there was a convergence in the incidence of negative-coagulase Staphylococcus growth (28%, clean surgeries; 32%, contaminated surgeries; and 29%, infected surgeries) and Staphylococcus aureus (28%, contaminated surgeries; and 43%, infected surgeries)., Conclusion: Most of the microorganisms recovered from the analyzed instruments (78%) were vegetative bacteria that presented their death curve at around 80 degrees C, characterizing a low challenge considering the processes of cleaning and sterilization currently employed by the Material and Sterilization Center. Fewer microorganisms were recovered from instruments used in clean surgeries in comparison with those used in contaminated and infected surgeries., (2010 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.)