e Periprosthetic infection (PI) causes significant morbidity and mortality after fixation and joint arthroplasty and has been extensively linked to the formation of bacterial biofilms. Poly(methyl methacrylate) (PMMA), as a cement or as beads, is commonly used for antibiotic release to the site of infection but displays variable elution kinetics and also represents a potential nidus for infection, therefore requiring surgical removal once antibiotics have eluted. Absorbable cements have shown improved elution of a wider range of antibiotics and, crucially, complete biodegradation, but limited data exist as to their antimicrobial and antibiofilm efficacy. Synthetic calcium sulfate beads loaded with tobramycin, vancomycin, or vancomycin-tobramycin dual treatment (in a 1:0.24 [wt/wt] ratio) were assessed for their abilities to eradicate planktonic methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus epidermidis relative to that of PMMA beads. The ability of the calcium sulfate beads to prevent biofilm formation over multiple days and to eradicate preformed biofilms was studied using a combination of viable cell counts, confocal microscopy, and scanning electron microscopy of the bead surface. Biofilm bacteria displayed a greater tolerance to the antibiotics than their planktonic counterparts. Antibiotic-loaded beads were able to kill planktonic cultures of 10 6 CFU/ml, prevent bacterial colonization, and significantly reduce biofilm formation over multiple days. However, established biofilms were harder to eradicate. These data further demonstrate the difficulty in clearing established biofilms; therefore, early preventive measures are key to reducing the risk of PI. Synthetic calcium sulfate loaded with antibiotics has the potential to reduce or eliminate biofilm formation on adjacent periprosthetic tissue and prosthesis material and, thus, to reduce the rates of periprosthetic infection. P eriprosthetic infection (PI) is a serious complication of total joint arthroplasty with high rates of associated morbidity (1, 2), and a growing body of data suggests that bacterial biofilms are the underlying cause (3-9). Within a biofilm, bacteria display a Ն1,000-fold tolerance to antibiotics than their planktonic counterparts (10) and significant resistance to innate and adaptive host immunity (11). Moreover, biofilms associated with orthopedic hardware are typically difficult to culture using conventional clinical microbiological methods, and the lack of a definitive diagnosis may result in an underestimate of infection rates (12, 13). Consequently, the underlying infection is difficult to diagnose and treat (6, 7), and often the only effective intervention is the twin strategy of thorough debridement and prostheses removal (14).Existing prevention strategies include the use of antibiotic-loaded poly(methyl methacrylate) (PMMA) cement spacers or beads to elevate local antibiotic levels at the surgical site. Studies have demonstrated a significant reduction in infection rates using antibiotic-impregnated ce...