Revision Total Knee Arthroplasty With an Imageless, Second-Generation Robotic System.

BACKGROUND: Robotic-assisted total knee arthroplasty is increasingly used in revision total knee arthroplasty (rTKA), with imageless systems recently receiving Food and Drug Administration (FDA) approval. However, there remains a paucity of literature on the use of robotic assistance in revision total knee arthroplasty (TKA). This paper describes the imageless surgical technique for robotic revision TKA using a second-generation robotic system and details both intraoperative and 90-day outcomes. METHODS: This was a retrospective review of 115 robotic revision TKAs from March 2021 to May 2023 at 3 tertiary academic centers. Patient demographics, perioperative surgical data, and 90-day outcomes were collected. Pain and Patient-Reported Outcomes Measurement Information System scores preoperatively and postoperatively were recorded. All-cause reoperations at the final follow-up were detailed. The mean patient age was 65 years (range, 43 to 88), and 58% were women. The mean follow-up time was 13 months (range, 3 to 51). RESULTS: The most common indications for rTKA were instability (n = 37, 32%) and aseptic loosening (n = 42, 37%). There were 83 rTKAs to a posterior-stabilized liner, 22 to a varus-valgus constrained liner, and 5 to a hinged construct. The median polyethylene size was 11 (interquartile range, 10 to 13), and 93% of patients had their joint line restored within 5 millimeters of the native contralateral knee. Within the 90-day postoperative window, there were 8 emergency department visits and 2 readmissions. At the final follow-up, there were 5 reoperations and 2 manipulations under anesthesia. There were 4 patients who required irrigation and debridement after superficial wound dehiscence, and one had an arthrotomy disruption after a fall. CONCLUSIONS: This review demonstrates favorable intraoperative and 90-day outcomes and suggests that imageless robotic surgery is a promising modality in rTKA. Further studies comparing the longitudinal outcomes after robotic and conventional rTKA are warranted.

Current Concepts on the Clinical and Economic Impact of Periprosthetic Joint Infections.

Total joint arthroplasty (TJA) is a common procedure performed throughout the entire world in hopes of alleviating debilitating hip or knee pain. The projected number of TJAs performed in the United States alone is projected to exceed 1.9 million by 2030 and 5 million by 2040. With the significant increase in TJA performed, more periprosthetic joint infections (PJIs) are likely to be encountered. PJIs are a devastating complication of TJA. The economic and clinical burden must be understood and respected to minimize occurrence and allow optimal patient outcomes.

Quantitative Real-Time Polymerase Chain Reaction May Serve as a Useful Adjunct to Conventional Culture in The Detection of Cutibacterium acnes in the Glenohumeral Joint: A Study of 100 Consecutive Patients.

Objectives: Synovial fluid or tissue culture is the current gold standard for diagnosis of infection, but Cutibacterium acnes (C. acnes) is a frequent cause of shoulder PJI and is a notoriously fastidious organism. The purpose of this study was to compare quantitative real-time polymerase chain reaction (qRT-PCR) to standard culture as a more rapid, sensitive means of identifying C. acnes from the glenohumeral joint. We hypothesized that qRT-PCR would be more effective than standard culture at identifying C. acnes and would have greater sensitivity and specificity for detecting infection. Methods: This was a prospective observational study with 100 consecutive patients undergoing arthroscopic or open shoulder surgery with known positive and negative controls. Intraoperatively, synovial fluid and tissue was obtained for C. acnes qRT-PCR and results were blinded to the gold standard microbiology cultures. Results: Clinical review demonstrated 3 patients (3%) with positive cultures, none of which were positive for C. acnes. Of the samples tested by the C. acnes qRT-PCR standard curve, 12.2% of tissue samples and 4.5% of fluid samples were positive. Culture sensitivity was 60.0%, specificity was 100.0%, PPV was 100.0%, and NPV was 97.9%. C. acnes qRT-PCR standard curve sensitivity, specificity, PPV, and NPV was 60.0%, 90.3%, 25.0%, and 97.7% respectively for tissue specimens and 0%, 95.2%, 0%, and 95.2% respectively, for fluid specimens. For combination of culture and tissue qRT-PCR, the sensitivity, specificity, PPV and NPV was 100%, 90.3%, 35.7%, and 100%, respectively. Conclusion: We report that qRT-PCR for C. acnes identified the organism more frequently than conventional culture. While these findings demonstrate the potential utility of qRT-PCR, the likelihood of false positive results of qRT-PCR should be considered. Thus, qRT-PCR may be useful as an adjuvant to current gold standard workup of synovial fluid or tissue culture for the diagnosis of infection.

Refractory Pseudomonas aeruginosa infections treated with phage PASA16: A compassionate use case series.

BACKGROUND: A growing number of compassionate phage therapy cases were reported in the last decade, with a limited number of clinical trials conducted and few unsuccessful clinical trials reported. There is only a little evidence on the role of phages in refractory infections. Our objective here was to present the largest compassionate-use single-organism/phage case series in 16 patients with non-resolving Pseudomonas aeruginosa infections. METHODS: We summarized clinical phage microbiology susceptibility data, administration protocol, clinical data, and outcomes of all cases treated with PASA16 phage. In all intravenous phage administrations, PASA16 phage was manufactured and provided pro bono by Adaptive Phage Therapeutics. PASA16 was administered intravenously, locally to infection site, or by topical use to 16 patients, with data available for 15 patients, mainly with osteoarticular and foreign-device-associated infections. FINDINGS: A few minor side effects were noted, including elevated liver function enzymes and a transient reduction in white blood cell count. Good clinical outcome was documented in 13 out of 15 patients (86.6%). Two clinical failures were reported. The minimum therapy duration was 8 days with a once- to twice-daily regimen. CONCLUSIONS: PASA16 with antibiotics was found to be relatively successful in patients for whom traditional treatment approaches have failed previously. Such pre-phase-1 cohorts can outline potential clinical protocols and facilitate the design of future trials. FUNDING: The study was funded in part by The Israeli Science Foundation IPMP (ISF_1349/20), Rosetrees Trust (A2232), United States-Israel Binational Science Foundation (2017123), and the Milgrom Family Support Program.

Hydrogen Peroxide, Povidone-Iodine and Chlorhexidine Fail to Eradicate Staphylococcus aureus Biofilm from Infected Implant Materials.

Hydrogen peroxide, povidone-iodine, and chlorhexidine are antiseptics that are commonly added to irrigants to either prevent or treat infection. There are little clinical data available that demonstrate efficacy of adding antiseptics to irrigants in the treatment of periprosthetic joint infection after biofilm establishment. The objective of the study was to assess the bactericidal activity of the antiseptics on S. aureus planktonic and biofilm. For planktonic irrigation, S. aureus was exposed to different concentrations of antiseptics. S. aureus biofilm was developed by submerging a Kirschner wire into normalized bacteria and allowing it to grow for forty-eight hours. The Kirschner wire was then treated with irrigation solutions and plated for CFU analysis. Hydrogen peroxide, povidone-iodine, and chlorhexidine were bactericidal against planktonic bacteria with over a 3 log reduction (p < 0.0001). Unlike cefazolin, the antiseptics were not bactericidal (less than 3 log reduction) against biofilm bacteria but did have a statistical reduction in biofilm as compared to the initial time point (p < 0.0001). As compared to cefazolin treatment alone, the addition of hydrogen peroxide or povidone-iodine to cefazolin treatment only additionally reduced the biofilm burden by less than 1 log. The antiseptics demonstrated bactericidal properties with planktonic S. aureus; however, when used to irrigate S. aureus biofilms, these antiseptics were unable to decrease biofilm mass below a 3 log reduction, suggesting that S. aureus biofilm has a tolerance to antiseptics. This information should be considered when considering antibiotic tolerance in established S. aureus biofilm treatment.

An Analysis of Radiographic Leg Length Discrepancy and Hip Offset in Patients at Risk of Developing Osteoarthritis.

Background: Leg length and hip offset are important principles in total hip arthroplasty (THA). Patients may endorse leg length differences (LLD) postoperatively that may be anatomical or functional. The objective of this study was to determine the normal radiographic variation in leg length and hip offset in a preosteoarthritic population without a THA. Methods: A retrospective study was completed using data from the Osteoarthritis Initiative, a prospective longitudinal study. Patients at risk of developing or with early osteoarthritis without inflammatory arthritis or prior THA were included. Measurements were made from full limb length anterior-posterior (AP) radiographs. Multiple linear regression models were employed to predict side-to-side differences in LLD, Δ femoral offset (FO), Δ abductor muscle length (AML), Δ abductor lever arm, and Δ AP pelvic offset. Results: The mean radiographic LLD was 4.6 mm, with 12 mm within 1 standard deviation. No significant differences were detected between LLD and sex, age, body mass index, or height. The median radiographic differences in FO, AML, abductor lever arm, and AP pelvic offset were 3.2 mm, 4.8 mm, 3.6 mm, and 3.3 mm, respectively. Height was predictive of Δ FO, while both height and age were predictive of Δ AML. Conclusions: Radiographic leg length variations in a population without symptomatic or radiographic osteoarthritis exist. FO and AML are dependent on patient characteristics. Preoperative radiographic LLD is not predicted by age, gender, body mass index, or height. It should be stressed that anatomic reconstruction is one of the many goals of arthroplasty and can stand in conflict with the priority and primary goals of stability and fixation, which should be prioritized.

Multiorganizational consensus to define guiding principles for perioperative pain management in patients with chronic pain, preoperative opioid tolerance, or substance use disorder.

Significant knowledge gaps exist in the perioperative pain management of patients with a history of chronic pain, substance use disorder, and/or opioid tolerance as highlighted in the US Health and Human Services Pain Management Best Practices Inter-Agency Task Force 2019 report. The report emphasized the challenges of caring for these populations and the need for multidisciplinary care and a comprehensive approach. Such care requires stakeholder alignment across multiple specialties and care settings. With the intention of codifying this alignment into a reliable and efficient processes, a consortium of 15 professional healthcare societies was convened in a year-long modified Delphi consensus process and summit. This process produced seven guiding principles for the perioperative care of patients with chronic pain, substance use disorder, and/or preoperative opioid tolerance. These principles provide a framework and direction for future improvement in the optimization and care of 'complex' patients as they undergo surgical procedures.

Dose optimization in surgical prophylaxis: sub-inhibitory dosing of vancomycin increases rates of biofilm formation and the rates of surgical site infection.

Antibiotic stewardship is viewed as having great public health benefit with limited direct benefit to the patient at the time of administration. The objective of our study was to determine if inappropriate administration of antibiotics could create conditions that would increase the rates of surgical infection. We hypothesized that sub-MIC levels of vancomycin would increase Staphylococcus aureus growth, biofilm formation, and rates of infection. S. aureus MRSA and MSSA strains were used for all experiments. Bacteria were grown planktonically and monitored using spectrophotometry. Quantitative agar culture was used to measure planktonic and biofilm bacterial burden. A mouse abscess model was used to confirm phenotypes in vivo. In the planktonic growth assay, increases in bacterial burden at » MIC vancomycin were observed in USA300 JE2 by 72 h. Similar findings were observed with ½ MIC in Newman and SH1000. For biofilm formation, USA300 JE2 at » and ½ MIC vancomycin increased biofilm formation by approximately 1.3- and 2.3-fold respectively at 72 h as compared to untreated controls. Similar findings were observed with Newman and SH1000 with a 2.4-fold increase in biofilm formation at ½ MIC vancomycin. In a mouse abscess model, there was a 1.2-fold increase with sub-MIC vancomycin at 3 days post infection. Our study showed that Sub-optimal vancomycin dosing promoted S. aureus planktonic growth and biofilm formation, phenotypic measures of bacterial virulence. This phenotype induced by sub-MIC levels of vancomycin was also observed to increase rates of infection and pathogenesis in our mouse model. Risks of exposure to sub-MIC concentrations with vancomycin in surgical procedures are greater as there is decreased bioavailability in tissue in comparison to other antibiotics. This highlights the importance of proper antibiotic selection, stewardship, and dosing for both surgical prophylaxis and treatment of infection.

How well does intra-operative contact path predict post-operative contact path during activities of daily living after medial unicompartmental knee arthroplasty?

Robot-assisted surgical systems can predict post-operative kinematics based upon intra-operative passive kinematics, planned implant position and alignment, and soft-tissue tension. It is currently unknown how well the intra-operative passive kinematics replicate the post-surgical weight bearing active kinematics. This study compared intra-operative and post-operative tibiofemoral implant contact paths after medial unicompartmental knee arthroplasty (mUKA). Passive intraoperative and active postoperative tibiofemoral contact path data was collected from eight patients who underwent mUKA. Intraoperative contact path data was measured using a navigation system. Postoperative contact path data was measured during walking, chair rise, stair ascent, and stair descent using a biplane radiography system and a validated tracking process. A total of 86 movement trials were included in the analysis. The contact point on the femur implant was up to 9.8 mm more medial and up to 8.3 mm less anterior at low flexion angles during activities of daily living than during passive extension intra-operatively, and the contact point on the tibia implant was up to 13.8 mm less lateral and up to 5.8 mm less posterior at low flexion angles during activities of daily living than during intra-operative passive extension. Femoral contact paths primarily differed between 3° and 42° of flexion; and tibial contact paths differed between 3° and 50° of flexion. This pilot study is the first to compare intra-operative and post-operative weight bearing contact paths. The primary conclusions from this study are that contact points on the femur implant are more medial and less anterior at low flexion angles during activities of daily living than during passive extension intra-operatively, and that the contact points on the tibia implant are less lateral at low flexion angles during activities of daily living than during intra-operative passive extension.

Medial Unicompartmental Knee Arthroplasty Restores Native Knee Kinematics During Activities of Daily Living: A Pilot Study.

The ability of unicompartmental knee arthroplasty (UKA) to restore native knee kinematics during activities of daily living remains unclear. The objectives of this prospective study were to identify changes in knee kinematics after medial UKA (mUKA) and to determine if mUKA restores native knee kinematics during activities of daily living. We hypothesized that kinematics are different between the mUKA knee and contralateral knee before surgery, that mUKA restores native knee kinematics, and that mUKA does not affect lateral compartment dynamic joint space. Nine participants performed walking, chair rise, stair ascent and stair descent within a biplane radiography system before and after mUKA. Bilateral knee kinematics were determined for each activity using a validated tracking process that matched subject-specific bones and implants to the biplane radiographs. Compared to contralateral knee, the pre-UKA knee was more adducted (p ≤ 0.019), and more laterally translated (p ≤ 0.008) during all four activities. Additionally, compared to contralateral knee, pre-UKA knee was less internally rotated (p ≤ 0.044) during chair rise and stair ascent. Lateral compartment dynamic joint space did not change during any activity from pre to post mUKA. Our results indicate that mUKA generally restores native kinematics during activities of daily living without altering lateral compartment dynamic joint space.

Engineered peptide PLG0206 overcomes limitations of a challenging antimicrobial drug class.

The absence of novel antibiotics for drug-resistant and biofilm-associated infections is a global public health crisis. Antimicrobial peptides explored to address this need have encountered significant development challenges associated with size, toxicity, safety profile, and pharmacokinetics. We designed PLG0206, an engineered antimicrobial peptide, to address these limitations. PLG0206 has broad-spectrum activity against >1,200 multidrug-resistant (MDR) ESKAPEE clinical isolates, is rapidly bactericidal, and displays potent anti-biofilm activity against diverse MDR pathogens. PLG0206 displays activity in diverse animal infection models following both systemic (urinary tract infection) and local (prosthetic joint infection) administration. These findings support continuing clinical development of PLG0206 and validate use of rational design for peptide therapeutics to overcome limitations associated with difficult-to-drug pharmaceutical targets.

Risk Stratification and Pain Outcomes Following Revision Total Hip Arthroplasty for Adverse Local Tissue Reactions.

BACKGROUND: Revision total hip arthroplasty (THA) for adverse local tissue reactions (ALTRs) secondary to head-neck taper corrosion is associated with a high complication rate. Diagnosis of ALTR is based on risk stratification using the patient's history and examination, implant risk, serum metal ion levels, and imaging. The purpose of this study was to determine if stratification using similar risk factors is predictive of outcomes following revision THA for metal-on-polyethylene (MoP) ALTR. METHODS: We performed a retrospective review on 141 patients revised for ALTR due to head-neck taper corrosion. Pain outcomes following surgery were analyzed using a generalized linear mixed model. Complications were defined as instability/dislocation, infection, fracture, nerve palsy, leg-length discrepancy, or reoperation. RESULTS: The overall complication rate was 17.7%. The odds of having pain decreased by 44% after revision surgery (Odds Ratio = 0.56, 95% Confidence Interval: 0.324 to 0.952). There was no significant difference in instability/dislocation based on either increased or decreased head-neck offset (P = .67) or magnetic resonance imaging findings of abductor loss, effusion size, and degree of ALTR (P = .73). Increased serum cobalt (P = .31) and chromium (P = .08) levels did not predict complications; however, a decreased cobalt-chromium ratio was associated with postoperative complications (2.8 versus 3.5; P = .002). CONCLUSION: These findings are the first to suggest that patients who have ALTR after MoP THA undergoing revision surgery demonstrated major pain relief. Increasing femoral head offset did not change rates of instability/dislocation. In clinical scenarios where preoperative cobalt-chromium femoral head offsets were greater than available ceramic head offsets, a mandatory decrease in femoral head offset did not increase rates of instability/dislocation.

High Mortality After Total Knee Arthroplasty Periprosthetic Joint Infection is Related to Preoperative Morbidity and the Disease Process but Not Treatment.

BACKGROUND: Periprosthetic joint infection (PJI) mortality rate is approximately 20%. The etiology for high mortality remains unknown. The objective of this study was to determine whether mortality was associated with preoperative morbidity (frailty), sequalae of treatment, or the PJI disease process itself. METHODS: A multicenter observational study was completed comparing 184 patients treated with septic revision total knee arthroplasty (TKA) to a control group of 38 patients treated with aseptic revision TKA. Primary outcomes included time and the cause of death. Secondary outcomes included preoperative comorbidities and Charlson Comorbidity Index (CCMI) measured preoperatively and at various postoperative timepoints. RESULTS: The septic revision TKA cohort experienced earlier mortality compared to the aseptic cohort, with a higher mortality rate at 90 days, 1, 2, and 3 years after index revision surgery (P = .01). There was no significant difference for any single cause of death (P > .05 for each). The mean preoperative CCMI was higher (P = .005) in the septic revision TKA cohort. Both septic and aseptic cohorts experienced a significant increase in CCMI from the preoperative to 3 years postoperative (P < .0001 and P = .002) and time of death (P < .0001 both) timepoints. The septic revision TKA cohort had a higher CCMI 3 years postoperatively (P = .001) and at time of death (P = .046), but not one year postoperatively (P = .119). CONCLUSION: Compared to mortality from aseptic revision surgery, septic revision TKA is associated with earlier mortality, but there is no single specific etiology. As quantified by changes in CCMI, PJI mortality was associated with both frailty and the PJI disease process, but not treatment.

Predictors of Adverse Local Tissue Reaction in a High-Risk Population.

BACKGROUND: Adverse local tissue reaction (ALTR) is a recognized complication of total hip arthroplasty (THA) with metal-on-polyethylene (MoP) bearing surface implants. Specific models of THA implants have been identified as having a higher incidence of ALTR. The purpose of this study is to determine if serum metal levels, patient symptoms, implant factors, and imaging findings can be predictive of ALTR within this high-risk population. METHODS: We retrospectively reviewed an observational cohort of 474 patients who underwent MoP THA and were at increased risk of having ALTR. Patients were stratified based on the presence or absence of ALTR. Patient symptoms, serum metal ions, implant head offset, and imaging findings were compared. RESULTS: Patients with ALTR were more likely to be symptomatic (52.9% vs 9.9%, P < .0001). The presence of ALTR was associated with significantly higher serum cobalt and chromium levels (6.2 ppb vs 3.6 ppb, P < .0001; 2.3 ppb vs 1.2 ppb, P < .0001). Head offsets greater than 4 mm were associated with a higher prevalence of ALTR (53% vs 38%, P = .05). On metal artifact reduction sequence magnetic resonance imaging, patients with ALTR had larger effusions (4.7 cm vs 2.1 cm, P < .001) and a higher incidence of trochanteric bursitis (47% vs 16%, P < .001). CONCLUSIONS: In high-risk MoP implants, serum cobalt and chromium levels are elevated, even in patients without ALTR. A larger femoral head offset is a risk factor for the development of ALTR. Our study suggests that patients presenting with painful THA and elevated metal ions require risk stratification based on patient symptoms, metal artifact reduction sequence magnetic resonance imaging findings, and implant factors.

Staphylococcus aureus genotype variation among and within periprosthetic joint infections.

Staphylococcus aureus is a common organism in orthopedic infections, but little is known about the genetic diversity of strains during an infectious process. Using periprosthetic joint infection (PJI) as a model, a prospective study was designed to quantify genetic variation among S. aureus strains both among and within patients. Whole genome sequencing and multilocus sequence typing was performed to genotype these two populations at high resolution. In nasal cultures, 78% of strains were of clonal complexes CC5, CC8, and CC30. In PJI cultures, only 63% could be classified in these common clonal complexes. The PJI cultures had a larger proportion of atypical strains, and these atypical strains were associated with poor host status and compromised immune conditions. Mutations in genes involved in fibronectin binding (ebh, fnbA, clfA, and clfB) systematically distinguished later PJI isolates from the first PJI isolate from each patient. Repeated mutations in S. aureus genes associated with extracellular matrix binding were identified, suggesting adaptive, parallel evolution of S. aureus during the development of PJI.

Prospective Activity of PLG0206, an Engineered Antimicrobial Peptide, on Chronic Periprosthetic Joint Infection Total Knee Arthroplasty Components Ex Vivo: The Knee Explant Analysis (KnEA) Study.

PLG0206 is an engineered antimicrobial peptide that has completed phase 1 clinical studies. A prospective study was completed on explanted implants from chronic periprosthetic joint infections (n = 17). At a concentration of 1 mg/mL for 15 min, there was a mean 4-log10 reduction (range, 1 to 7) in the bacterial CFU identified from the implants. IMPORTANCE Chronically infected prosthetics of the knee were exposed to PLG0206, an engineered antimicrobial peptide, at a concentration of 1 mg/mL for 15 min. A mean 4-log10 reduction (range, 1 to 7) in the number of bacteria occurred, which may translate to improved clinical outcomes for persons with prosthetic joint infection of the knee.

Synovial Fluid-Induced Aggregation Occurs across Staphylococcus aureus Clinical Isolates and is Mechanistically Independent of Attached Biofilm Formation.

Rapid synovial fluid-induced aggregation of Staphylococcus aureus is currently being investigated as an important factor in the establishment of periprosthetic joint infections (PJIs). Pathogenic advantages of aggregate formation have been well documented in vitro, including recalcitrance to antibiotics and protection from host immune defenses. The objective of the present work was to determine the strain dependency of synovial fluid-induced aggregation by measuring the degree of aggregation of 21 clinical S. aureus isolates cultured from either PJI or bloodstream infections using imaging and flow cytometry. Furthermore, by measuring attached bacterial biomass using a conventional crystal violet assay, we assessed whether there is a correlation between the aggregative phenotype and surface-associated biofilm formation. While all of the isolates were stimulated to aggregate upon exposure to bovine synovial fluid (BSF) and human serum (HS), the extent of aggregation was highly variable between individual strains. Interestingly, the PJI isolates aggregated significantly more upon BSF exposure than those isolated from bloodstream infections. While we were able to stimulate biofilm formation with all of the isolates in growth medium, supplementation with either synovial fluid or human serum inhibited bacterial surface attachment over a 24 h incubation. Surprisingly, there was no correlation between the degree of synovial fluid-induced aggregation and quantity of surface-associated biofilm as measured by a conventional biofilm assay without host fluid supplementation. Taken together, our findings suggest that synovial fluid-induced aggregation appears to be widespread among S. aureus strains and mechanistically independent of biofilm formation. IMPORTANCE Bacterial infections of hip and knee implants are rare but devastating complications of orthopedic surgery. Despite a widespread appreciation of the considerable financial, physical, and emotional burden associated with the development of a prosthetic joint infection, the establishment of bacteria in the synovial joint remains poorly understood. It has been shown that immediately upon exposure to synovial fluid, the viscous fluid in the joint, Staphylococcus aureus rapidly forms aggregates which are resistant to antibiotics and host immune cell clearance. The bacterial virulence associated with aggregate formation is likely a step in the establishment of prosthetic joint infection, and as such, it has the potential to be a potent target of prevention. We hope that this work contributes to the future development of therapeutics targeting synovial fluid-induced aggregation to better prevent and treat these infections.

The Staphylococcus aureus toxin-antitoxin system YefM-YoeB is associated with antibiotic tolerance and extracellular dependent biofilm formation.

The high antibiotic tolerance of Staphylococcus aureus biofilms is associated with challenges for treating periprosthetic joint infection. The toxin-antitoxin system, YefM-YoeB, is thought to be a regulator for antibiotic tolerance, but its physiological role is unknown. The objective of this study was to determine the biofilm and antibiotic susceptibility phenotypes associated with S. aureus yoeB homologs. We hypothesized the toxin-antitoxin yoeB homologs contribute to biofilm formation and antibiotic susceptibility. Disruption of yoeB1 and yoeB2 resulted in decreased biofilm formation in comparison to Newman and JE2 wild-type (WT) S. aureus strains. In comparison to yoeB mutants, both Newman and JE2 WT strains had higher polysaccharide intercellular adhesin (PIA) production. Treatment with sodium metaperiodate increased biofilm formation in Newman WT, indicating biofilm formation may be increased under conditions of oxidative stress. DNase I treatment decreased biofilm formation in Newman WT but not in the absence of yoeB1 or yoeB2. Additionally, WT strains had a higher extracellular DNA (eDNA) content in comparison to yoeB mutants but no differences in biofilm protein content. Moreover, loss of yoeB1 and yoeB2 decreased biofilm survival in both Newman and JE2 strains. Finally, in a neutropenic mouse abscess model, deletion of yoeB1 and yoeB2 resulted in reduced bacterial burden. In conclusion, our data suggest that yoeB1 and yoeB2 are associated with S. aureus planktonic growth, extracellular dependent biofilm formation, antibiotic tolerance, and virulence.