Presoaking Grafts in Vancomycin Does Not Impair Graft-Bone Healing in a Rat Anterior Cruciate Ligament Reconstruction Model.

BACKGROUND: The vancomycin presoaking technique (wherein grafts are treated with a vancomycin solution [VS] for anterior cruciate ligament reconstruction [ACLR]) reduces the infection rate after ACLR. However, the effects of this technique on graft-bone healing have not been fully elucidated. PURPOSE: To investigate the effects of vancomycin presoaking on graft-bone healing in a rat ACLR model. STUDY DESIGN: Controlled laboratory study. METHODS: Long flexor digitorum longus tendons were obtained from 9 Wistar rats, and each was randomly allocated to the normal saline (NS) or VS groups. The grafts were immersed in sterile saline for 30 minutes in the NS group and in a 5-mg/mL VS in the VS group. The presence of time-zero graft bacterial contamination was confirmed, and the grafts were incubated in Fluidised Thioglycollate Broth for 2 weeks. ACLR was performed on the right knees of 65 male Wistar rats using the flexor digitorum longus tendons. Each graft was similarly treated. Biomechanical testing, micro-computed tomography, and histological evaluations were performed 4 and 12 weeks postoperatively. RESULTS: The VS group showed significantly reduced graft contamination at time zero (P = .02). The mean maximum loads to failure were 13.7 ± 8.2 N and 11.6 ± 4.8 N in the NS and VS groups, respectively, at 4 weeks (P = .95); and 23.2 ± 13.2 N and 30.4 ± 18.0 N in the NS and VS groups, respectively, at 12 weeks (P = .35). Regarding micro-computed tomography, the mean bone tunnel volumes were 3.76 ± 0.48 mm3 and 4.40 ± 0.58 mm3 in the NS and VS groups, respectively, at 4 weeks (P = .41); and 3.51 ± 0.38 mm3 and 3.67 ± 0.35 mm3 in the NS and VS groups, respectively, at 12 weeks (P = .54). Histological semiquantitative examination revealed no clear between-group differences at any time point. CONCLUSION: Presoaking grafts in vancomycin in a rat ACLR model demonstrated no discernible adverse effects on short- and midterm biomechanical, radiological, and histological investigations. CLINICAL RELEVANCE: The findings provide guidance for surgeons when considering this technique.

Augmentation of a Nonanatomical Repair of a Medial Meniscus Posterior Root Tear With Centralization Using Three Knotless Anchors May Be Associated With Less Meniscal Extrusion and Better Compressive Load Distribution in Mid-Flexion Compared With Non-Anatomical Root Repair Alone in a Porcine Knee Model.

PURPOSE: To investigate the biomechanics of the centralization augmentation using knotless soft anchors to a nonanatomical transtibial pull-out root repair in a porcine medial meniscus posterior root tear (MMPRT) model. METHODS: Porcine knee joints (N = 10) were used to perform one of the following procedures: (1) intact; (2) MMPRT; (3) nonanatomical root repair; (4) nonanatomical root repair with centralization using 2 anchors: anchors were inserted at the posterior medial collateral ligament (MCL) border and 10 mm anterior to the posterior MCL border; and (5) nonanatomical root repair with centralization using 3 anchors: another anchor was placed 10 mm posterior to the posterior MCL border. Contact area on the medial meniscus (MM), contact pressure in the MM and tibial cartilage, and MM extrusion were evaluated at 30°, 45°, 60°, and 90° knee flexions under 200 N compressive force. RESULTS: MM extrusion at the posterior MCL border was significantly reduced after root repair with centralization using 3 anchors than after root repair alone at 30° (-0.063 mm vs 1.5 mm, P = .017), 45° (0.21 mm vs 1.7 mm, P = .018), and 60° (0.78 mm vs 2.3 mm, P = .019). There were no significant differences in MM extrusion between the root repair alone and root repair with centralization using 2 anchors at all flexion angles. The contact area in the middle and posterior MM was significantly greater after centralization with 3 anchors than after root repair alone at all flexion angles (except the posterior MM at 90°). The mean contact pressure in the tibial cartilage was significantly lower after centralization with 3 anchors than after root repair at all angles. CONCLUSIONS: Augmentation of a nonanatomical repair of a medial meniscus posterior root tear with centralization using three knotless anchors may be associated with less meniscal extrusion and better compressive load distribution between 30° and 60° flexion compared with nonanatomical root repair alone in a porcine model. CLINICAL RELEVANCE: This biomechanical study at time zero suggests that the addition of centralization using 3 knotless anchors may reduce MM extrusion and restore the load-distributing function of the MM.

Effect of disinfectants and manual wiping for processing the cell product changeover in a biosafety cabinet.

Introduction: The process of cell product changeover poses a high risk of cross-contamination. Hence, it is essential to minimize cross-contamination while processing cell products. Following its use, the surface of a biosafety cabinet is commonly disinfected by ethanol spray and manual wiping methods. However, the effectiveness of this protocol and the optimal disinfectant have not yet been evaluated. Here, we assessed the effect of various disinfectants and manual wiping methods on bacterial removal during cell processing. Methods: The hard surface carrier test was performed to evaluate the disinfectant efficacy of benzalkonium chloride with a corrosion inhibitor (BKC + I), ethanol (ETH), peracetic acid (PAA), and wiping against Bacillus subtilis endospores. Distilled water (DW) was used as the control. A pressure sensor was employed to investigate the differences in loading under dry and wet conditions. The pre-spray for wiping was monitored by eight operators using a paper that turns black when wet. Chemical properties, including residual floating proteins, and mechanical properties, such as viscosity and coefficient of friction, were examined. Results: In total, 2.02 ± 0.21-Log and 3.00 ± 0.46-Log reductions from 6-Log CFU of B. subtilis endospores were observed for BKC + I and PAA, respectively, following treatment for 5 min. Meanwhile, wiping resulted in a 0.70 ± 0.12-Log reduction under dry conditions. Under wet conditions, DW and BKC + I showed 3.20 ± 0.17-Log and 3.92 ± 0.46-Log reductions, whereas ETH caused a 1.59 ± 0.26-Log reduction. Analysis of the pressure sensor suggested that the force was not transmitted under dry conditions. Evaluation of the amount of spray by eight operators showed differences and bias in the spraying area. While ETH had the lowest ratio in the protein floating and collection assays, it exhibited the highest viscosity. BKC + I had the highest friction coefficient under 4.0-6.3 mm/s; however, that of BKC + I decreased and became similar to the friction coefficient of ETH under 39.8-63.1 mm/s. Conclusions: DW and BKC + I are effective for inducing a 3-Log reduction in bacterial abundance. Moreover, the combination of optimal wet conditions and disinfectants is essential for effective wiping in specific environments containing high-protein human sera and tissues. Given that some raw materials processed in cell products contain high protein levels, our findings suggest that a complete changeover of biosafety cabinets is necessary in terms of both cleaning and disinfection.

Biomechanical analysis of a centralization procedure for extruded lateral meniscus after meniscectomy in porcine knee joints.

The recently developed arthroscopic centralization for lateral meniscal extrusion has obtained satisfactory short-term clinical and radiological results and improves the meniscus biomechanical properties. However, the effectiveness of treatment for meniscus extrusion after partial meniscectomy still requires elucidation. This study investigated the effect of centralization with modifications from a mechanical viewpoint. Porcine knee joints (N = 6) were set in a universal tester under the following conditions: (1) Intact; (2) Meniscectomy: Inner half of the posterior half meniscus was removed; (3) Extrusion: Posterior meniscus was dislocated laterally by transecting the posterior root and the meniscotibial ligament; (4) Centralization-1: Centralization procedure using one anchor; (5) Centralization-2: Centralization procedure using two anchors; and (6) Centralization-ad: Centralization with capsular advancement using two anchors. Load distributions and contact pressure in the meniscus and tibial cartilage were evaluated with an axial compressive force of 200 N. After meniscectomy, the tibial cartilage load increased and that of the medial margin of the posterior part of the meniscus decreased. When the meniscus was extruded, the load was concentrated only on the tibial cartilage. Centralization-1 increased the load on the meniscus, while Centralization-2 further increased the meniscus load but decreased the tibial cartilage load. Centralization-ad further decreased the load on the tibial plateau. The average contact pressure of the tibial cartilage was significantly higher in the Extrusion group than in the Intact group or the Centralization-ad group. From a biomechanical viewpoint, centralization with capsular advancement was the most effective of the tested procedures for treatment for an extruded meniscus after partial meniscectomy.

Relationship between medial meniscus extrusion and cartilage measurements in the knee by fully automatic three-dimensional MRI analysis.

BACKGROUND: We developed a fully automatic three-dimensional knee MRI analysis software that can quantify meniscus extrusion and cartilage measurements, including the projected cartilage area ratio (PCAR), which represents the ratio of the subject's actual cartilage area to their ideal cartilage area. We also collected 3D MRI knee data from 561 volunteers (aged 30-79 years) from the "Kanagawa Knee Study." Our purposes were to verify the accuracy of the software for automatic cartilage and meniscus segmentation using knee MRI and to examine the relationship between medial meniscus extrusion measurements and cartilage measurements from Kanagawa Knee Study data. METHODS: We constructed a neural network for the software by randomly choosing 10 healthy volunteers and 103 patients with knee pain. We validated the algorithm by randomly selecting 108 of these 113 subjects for training, and determined Dice similarity coefficients from five other subjects. We constructed a neural network using all data (113 subjects) for training. Cartilage thickness, cartilage volume, and PCAR in the medial femoral, lateral femoral, medial tibial, and lateral tibial regions were quantified by using the trained software on Kanagawa Knee Study data and their relationship with subject height was investigated. We also quantified the medial meniscus coverage ratio (MMCR), defined as the ratio of the overlapping area between the medial meniscus area and the medial tibial cartilage area to the medial tibial cartilage area. Finally, we examined the relationship between MMCR and PCAR at middle central medial tibial (mcMT) subregion located in the center of nine subregions in the medial tibial cartilage. RESULTS: Dice similarity coefficients for cartilage and meniscus were both approximately 0.9. The femoral and tibial cartilage thickness and volume at each region correlated with height, but PCAR did not correlate with height in most settings. PCAR at the mcMT was significantly correlated with MMCR. CONCLUSIONS: Our software showed high segmentation accuracy for the knee cartilage and meniscus. PCAR was more useful than cartilage thickness or volume since it was less affected by height. Relations ips were observed between the medial tibial cartilage measurements and the medial meniscus extrusion measurements in our cross-sectional study. TRIAL REGISTRATION: UMIN, UMIN000032826 ; 1 September 2018.

2-Methacryloyloxyethyl Phosphorylcholine Polymer Coating Inhibits Bacterial Adhesion and Biofilm Formation on a Suture: An In Vitro and In Vivo Study.

Initial bacterial adhesion to medical devices and subsequent biofilm formation are known as the leading causes of surgical site infection (SSI). Therefore, inhibition of bacterial adhesion and biofilm formation on the surface of medical devices can reduce the risk of SSIs. In this study, a highly hydrophilic, antibiofouling surface was prepared by coating the bioabsorbable suture surface with poly(2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate) (PMB). The PMB-coated and noncoated sutures exhibited similar mechanical strength and surface morphology. The effectiveness of the PMB coating on the suture to suppress adhesion and biofilm formation of methicillin-resistant Staphylococcus aureus and methicillin-susceptible Staphylococcus aureus was investigated both in vitro and in vivo. The bacterial adhesion test revealed that PMB coating significantly reduced the number of adherent bacteria, with no difference in the number of planktonic bacteria. Moreover, fluorescence microscopy and scanning electron microscopy observations of adherent bacteria on the suture surface after contact with bacterial suspension confirmed PMB coating-mediated inhibition of biofilm formation. Additionally, we found that the PMB-coated sutures exhibited significant antibiofouling effects in vivo. In conclusion, PMB-coated sutures demonstrated bacteriostatic effects associated with a highly hydrophilic, antibiofouling surface and inhibited bacterial adhesion and biofilm formation. Therefore, PMB-coated sutures could be a new alternative to reduce the risk of SSIs.

The effect of a centralization procedure for extruded lateral meniscus on load distribution in porcine knee joints at different flexion angles.

BACKGROUND: Meniscal extrusion results in loss of the ability to resist hoop strain and biomechanical overload on the joint articular surface. A centralization technique has been developed to overcome these problems. In this study, we analyzed the biomechanics of the extruded and centralized lateral meniscus (LM) in porcine knee joints at different flexion angles. METHODS: Porcine knee joints (n = 8) were set in the universal tester and each knee was tested under the following states: 1) intact; 2) extrusion-meniscal extrusion was created by resecting the posterior root of the LM and posterior synovial capsule; and 3) centralization-centralization was performed by two anchors inserted in the lateral tibial plateau. Deviation distance of the meniscus, contact pressure, and contact area in the anterior LM, middle LM, posterior LM, and the contact pressure of the tibial cartilage were evaluated with an axial compressive force of 200 N at knee flexion angles of 30°, 45°, 60°, and 90°. RESULTS: The deviation distance of LM significantly increased in extrusion but was restored to the intact status after centralization at all angles. Both the contact pressure and area significantly decreased in extrusion and were restored after centralization close to the intact status in the anterior and middle LM; in the posterior LM, however, decreased contact pressure and area were not restored after centralization. The contact pressure of the tibial cartilage increased significantly in extrusion but decreased close to the intact status after centralization. CONCLUSIONS: This centralization procedure could reduce extrusion of the LM and restore the load-distributing function of the anterior-middle LM. However, the procedure itself could not restore hoop function in cases where the defect lies in the posterior LM.

Biomechanical analysis of the centralization procedure for extruded lateral menisci with posterior root deficiency in a porcine model.

PURPOSE: The purpose of this study was to investigate the biomechanical properties of load distribution following a centralization procedure for extruded lateral menisci with posterior root deficiency in a porcine model. METHODS: Six porcine knee joints were analyzed in a universal tester, as follows: 1) Intact; 2) Extrusion (meniscus extrusion was created by resecting the posterior root of the lateral meniscus, as well as the posterior synovial capsule); and 3) Centralization (two anchors were inserted at the lateral tibial plateau, and the meniscus was sutured to secure it close to the original position). Meniscus extrusion was evaluated using two markers put on the posterior cruciate ligament and the lateral meniscus, and the load distribution were assessed using a pressure mapping sensor system after applying a loading force of 200 N to the knee joint. RESULTS: Distance between two markers (mm, Average; 95% CI) was larger in the extrusion group (21.9; 17.8, 25.6) than in the intact (18.1; 15.1, 22.7) or the centralization (15.3; 12.9, 18.0) groups. The contact area (mm2) in the middle of the meniscus was significantly smaller in the extrusion group (45.8; 18.5, 73.2) than in the intact (85.7; 72.1, 99.2) or the centralization (98.3; 88.8, 107.8) groups. The maximum contact pressure (MPa) in the tibial plateau was significantly higher in the extrusion group (0.37; 0.35, 0.40) than in the intact (0.29; 0.21, 0.37) or the centralization (0.29; 0.22, 0.36) groups. CONCLUSIONS: The centralization procedure enabled a reduction of the meniscus extrusion in the lateral meniscus with posterior root deficiency and restored the maximum load and contact pressure to values close to those of the normal knee joint.

Comparison of High-Hydrostatic-Pressure Decellularized Versus Freeze-Thawed Porcine Menisci.

The meniscus functions as a load distributor and secondary stabilizer in the knee, and the loss of the meniscus increases the risk of osteoarthritis. Freeze-thawed menisci are used in clinical practice to replace defective menisci; however, the disadvantages of freeze-thawed tissues include disease transmission and immune rejection. In this study, we decellularized menisci using high hydrostatic pressure (HHP) and compared the decellularized menisci with freeze-thawed menisci. Porcine menisci were either pressurized at 1,000 MPa for 10 min and then washed with DNase solution or frozen at -80°C for 2 days and thawed. These menisci then underwent in vitro histological, biochemical, and biomechanical comparisons with native menisci. The HHP-treated and freeze-thawed menisci were also subcutaneously implanted in a pig, and later harvested for histological analysis. The numbers of histologically detected cells were significantly lower and the amount of biochemically detected DNA was approximately 100-fold lower in HHP-treated than in native and freeze-thawed menisci. The compression strength of the HHP-decellularized menisci was decreased after 1 and 50 cycles at 20% strain but was unchanged in the freeze-thawed menisci. After implantation, the numbers of multinucleated giant cells were significantly lower around the HHP-treated menisci than around the freeze-thawed menisci. Recellularization of the HHP-decellularized menisci was confirmed. Thus, although the HHP-decellularized menisci were mechanically inferior to the freeze-thawed meniscus in vitro, they were immunologically superior. Our study is the first to demonstrate the use of HHP for decellularization of the meniscus. © 2019 The Authors. Journal of Orthopaedic Research® published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 37:2466-2475, 2019.