Risk factors for delayed bone union in opening wedge high tibial osteotomy.

OBJECTIVES: The purpose of this study was to investigate the relationship between patient demographics and potential intraoperative factors and delayed bone union in opening wedge high tibial osteotomy (OWHTO). PATIENTS AND METHODS: A retrospective review of 65 patients (37 females, 28 males; mean age: 60.1±10.1 years; range, 44 to 77 years) who underwent OWHTO using an angle-stable implant with beta-tricalcium phosphate gap filling between September 2016 and October 2019 was conducted. The osteotomy site was divided into five zones from the lateral hinge on anteroposterior radiographs, and we defined the zone in which bone healing was observed. The bone union area was assessed according to this definition at three, six, nine, and 12 months after surgery, and bone union was defined as union at the fourth zone or greater. A generalized estimating equations approach was employed to investigate longitudinal data pertaining to bone union area as a dependent variable. In addition, the association of bone union at six months postoperatively and predictors were evaluated using cross-sectional statistical methods. The categorical predictors included in the models were smoking, diabetes, hinge fracture, and autologous osteophyte grafting. The continuous variables included in the models were age, body mass index, opening gap width, and plate position. RESULTS: Smoking (odds ratio [OR]=0.478, p<0.01), large opening gap width (OR=0.941, p=0.014), and anterior plate placement (OR=0.971, p<0.01) were significantly associated with decreased bone union area. Union rate at six months in smokers was significantly lower compared to nonsmokers (16.6% and 67.8%, respectively; OR=0.10, p=0.023). Area under the curve in the receiver operating characteristic analysis for bone union at six months was 0.60 for gap width and 0.63 for plate placement. CONCLUSION: Smoking, large opening gap width, and anterior plate placement are risk factors for delayed bone union after OWHTO. Surgeons should avoid anterior placement of the plate and carefully consider other options for smokers and those who require a large correction.

The association of the medial meniscus covering ratio with knee cartilage thickness at 15 medial compartment subregions as found by three-dimensional analysis using MRI.

Purpose: The correlation of cartilage thickness measured by three-dimensional (3D) magnetic resonance imaging (MRI) and the medial meniscal coverage ratio (MMCR), which presented pathology of the medial meniscus extrusion (MME) in 3D MRI, has not yet been elucidated. The study's purpose was to retrospectively verify whether the average cartilage thickness calculated by the automatic MRI 3D analysis system for each subregion was correlated with MMCR. Methods: A total of 60 patients underwent medial meniscus repair or high tibial osteotomy to treat their medial knee osteoarthritis. Cartilage thickness and MMCR were automatically calculated using 3D MRI software. The MMCR was defined as the ratio of the area covered by the meniscus within the medial tibial cartilage area to the total medial tibial cartilage area. The association between MMCR and the average cartilage thickness at 15 subregions in the medial femoral condyle (MFC) and medial tibial plateau (MTP) was evaluated using Spearman's rank correlation coefficient. Results: Kellgren-Lawrence grade exhibited a negative correlation with MMCR and a positive correlation with MME width. Cartilage thickness in the MTP had a moderately positive correlation with MMCR at four subregions and a weakly positive correlation at another subregion. Cartilage thickness in the MFC showed a moderately positive correlation with MMCR at five subregions and a weakly positive correlation at one subregion. Conclusions: Cartilage thickness calculated by automatic MRI 3D analysis system had a positive correlation with MMCR for all subregions of the anterior and middle subregions in the MFC and for five regions of nine subregions of the anterior and middle subregions in the MTP. Level of evidence: Level II, cross-sectional study (diagnosis).

Evaluation of cleaning methods for change-over after the processing of cell products to avoid cross-contamination risk.

Introduction: Cell-processing facilities face the risk of environmental bacteria contaminating biosafety cabinets during processing, and manual handling of autologous cell products can result in contamination. We propose a risk- and evidence-based cleaning method for cross-contamination, emphasizing proteins and DNA. Methods: The transition and residual risks of the culture medium were assessed by measuring both wet and dried media using fluorescence intensity. Residual proteins and DNA in dried culture medium containing HT-1080 cells were analyzed following ultraviolet (UV) irradiation, wiping, and disinfectant treatment. Results: Wet conditions showed a higher transition to distilled water (DW), whereas dry conditions led to higher residual amounts on SUS304 plates. Various cleaning methods for residual culture medium were examined, including benzalkonium chloride with a corrosion inhibitor (BKC + I) and DW wiping, which demonstrated significantly lower residual protein and DNA compared to other methods. Furthermore, these cleaning methods were tested for residual medium containing cells, with BKC + I and DW wiping resulting in an undetectable number of cells. However, in some instances, proteins and DNA remained. Conclusions: The study compared cleaning methods for proteins and DNA in cell products, revealing their advantages and disadvantages. Peracetic acid (PAA) proved effective for nucleic acids but not proteins, while UV irradiation was ineffective against both proteins and DNA. Wiping emerged as the most effective method, even though traceability remained challenging. However, wiping with ETH was not effective as it caused protein immobilization. Understanding the characteristics of these cleaning methods is crucial for developing effective contamination control strategies.

Serial Assessment of Factors Associated With Patient-Reported Outcomes up to 2 Years After ACL Reconstruction Using Autologous Hamstring Tendon: Results From the TMDU MAKS Study.

Background: Understanding the factors related to patient-reported outcomes (PROs) after anterior cruciate ligament reconstruction (ACLR) can lead to more effective interventions. Purpose/Hypothesis: The purpose of this study was to identify factors associated with PRO scores after ACLR. It was hypothesized that concomitant meniscal treatment and postoperative range of motion (ROM) would be associated with early postoperative PRO scores and that postoperative physical findings would be associated with 2-year postoperative PRO scores. Study Design: Cohort study; Level of evidence, 2. Methods: We examined the data from the Tokyo Medical and Dental University Multicenter Arthroscopic Knee Surgery (TMDU MAKS) Study for patients who underwent primary ACLR with autologous hamstring tendon grafts; 1252 patients in the TMDU MAKS Study were eligible for inclusion. The International Knee Documentation Committee (IKDC) subjective score, Knee injury and Osteoarthritis Outcome Score (KOOS), and Lysholm score of the patients at 3 months (n = 675), 1 year (n = 660), and 2 years (n = 375) postoperatively were assessed using multiple regression to evaluate the strength of the relationship between PRO scores and the following predictor variables: patient-specific factors, treatment-specific factors, and physical findings. Results: Improvement in all PROs significantly exceeded the minimal important change at 1 and 2 years postoperatively. Older age and female sex were predictive of lower PRO scores up to 2 years postoperatively. Concurrent meniscal treatment and limited range of knee motion were predictive of lower PRO scores at 3 months and 1 year postoperatively. A tighter knee (ie, less anterior translation) on the injured side compared with the contralateral knee was predictive of lower KOOS-Quality of Life and IKDC scores at 2 years postoperatively. At all 3 postoperative time points, greater side-to-side difference in knee ROM was predictive of lower PRO scores. Conclusion: Inferior PRO scores were associated with concomitant meniscal treatment and limited postoperative ROM until 1 year postoperatively and with older age and female sex up to 2 years postoperatively. PRO scores were associated with knee stability at all assessed time points.

Steep medial posterior tibial slope angle and ramp lesion are independent risk factors for an increase in quantitatively measured pivot shift in patients with ACL-deficient knees.

Background: Delayed anterior cruciate ligament (ACL) reconstruction often causes residual anterolateral rotatory instability (ALRI) and consequent knee osteoarthritis, warranting the recommendation of early ACL reconstruction within 6 months after injury. Nonetheless, some cases show notable instability, even shortly after injury. The purpose of this study was to identify risk factors for an increase in quantitatively measured pivot shift in patients with ACL-deficient knees within 6 months after injury. Methods: Patients with primary ACL reconstruction within 6 months after injury and quantitative triaxial accelerometer measurements of preoperative pivot shift were included. Descriptive statistics were calculated for 11 independent variables (age, gender, time from injury to surgery, KT-1000, knee extension angle, lateral and medial posterior tibial slope angle, medial and lateral meniscus tears, ramp lesion and Tegner active scale). A single regression analysis was performed on the 11 items and acceleration during the pivot shift, and a multiple regression analysis was performed for items with p value less than 0.1. Results: Overall, 111 patients met the inclusion criteria. Single regression analysis showed that medial posterior tibial slope angle, medial meniscus tear and ramp lesion were significantly correlated with acceleration during the pivot shift (p < 0.001, p = 0.04 and p = 0.001). Multiple regression analysis identified medial posterior tibial slope angles and ramp lesions as independent factors for increased preoperative pivot shift (p = 0.005 and 0.01). Conclusion: A steep medial posterior tibial slope angle and ramp lesion were independent risk factors for increased quantitatively measured pivot shift in patients with ACL-deficient knees. Level of Evidence: Level IV, case series study.

A novel methodology utilizing microchip implants to monitor individual activity and body temperature for assessing knee pain in group-housed rats.

The pain assessment in animals is challenging as they cannot verbally express the site and severity of pain. In this study, we tried a small implantable actimeter, "Nanotag", to monitor spontaneous locomotor activity and body temperature in animals suffering from a chemical-induced rat knee arthritis as compared to naïve and steroid-treated rats. Nanotag could detect the decrease in locomotor activity quickly after the arthritis induction and anti-inflammation analgesic treatment by intra-articular injection of steroid significantly improved locomotor activity. These changes were in the same line with those of a conventional knee pain evaluation method (incapacitance test). Nanotag can be utilized as the non-interventional, continuous, and completely objective monitoring the amount of pain in rat knee arthritis model. This traditional yet innovative method may be universally applicable to various pain models and species, making it a worthwhile device for research across diverse fields.

Synovial mesenchymal stem cells secrete more lubricin than adipose mesenchymal stem cells after injection into rat osteoarthritis knees.

Intra-articular injection of mesenchymal stem cells (MSCs) is envisioned as a solution for knee osteoarthritis (OA). Although synovial MSCs (SyMSCs) are promising for cartilage regeneration, the clinical choice is usually adipose MSCs (AdMSCs). However, the similarities/differences in the mode of action between SyMSCs and AdMSCs remain unclear. Here, we compared factors secreted by human SyMSCs and AdMSCs after injection into OA knees. Human SyMSCs or AdMSCs were injected into the knees of rat partial meniscectomy models. The next day, the knee joints were collected to analyze the distribution of injected MSCs and transcriptome changes in the human MSCs and rat synovium. Non-injected MSCs were mixed with rat synovium as a control. After injection, no difference was apparent in intra-articular distribution of the SyMSCs or AdMSCs. RNA sequencing demonstrated an enrichment of cytokine-cytokine receptor interaction-related genes in both human SyMSCs and AdMSCs after injection. Differentially expressed genes (DEGs) specific to SyMSCs were associated with cartilage matrix synthesis and homeostasis. PCR analysis of the matrisome-related DEGs showed significantly higher expression of PRG4 in SyMSCs than in AdMSCs after injection. Immunostaining also confirmed a significantly greater expression of lubricin by SyMSCs than by AdMSCs. These findings indicate that SyMSCs will be a more promising treatment for OA.

Cartilage thickness in the medial compartment of the knee joint evaluated by MRI three-dimensional analysis correlates with weight-bearing line ratio and joint line convergence angle.

PURPOSE: The correlation between cartilage thickness evaluated by 3D MRI and knee alignment has not been elucidated. The study's purpose was to retrospectively verify whether average cartilage thickness calculated by the automatic MRI 3D analysis system for each subregion was correlated with knee alignment. METHODS: A total of 53 patients underwent medial meniscus repair or high tibial osteotomy for medial knee osteoarthritis. Cartilage thickness was automatically calculated using 3D MRI software. Knee alignment, weight-bearing line ratio (WBLR), joint line convergence angle (JLCA), medial proximal tibial angle (MPTA), and lateral distal femoral angle (LDFA) were calculated on a weight-bearing long-leg radiograph using digital planning software. The association between knee alignment and the average cartilage thickness at 18 subregions in the medial femoral condyle (MFC) and medial tibial plateau (MTP) was evaluated using Spearman's rank correlation coefficient. RESULTS: Cartilage thickness of the MFC had moderately positive correlations with WBLR at four subregions and weak correlations at two subregions. Cartilage thickness of the MTP had moderately positive correlations with WBLR at four subregions. Cartilage thickness of the MFC had moderately negative correlations with JLCA at six subregions. Cartilage thickness of the MTP had moderately negative correlations with JLCA at four subregions and a weak correlation at one subregion. Conversely, cartilage thickness had weak correlations with MPTA or LDFA. CONCLUSIONS: In knees with pathological conditions in the medial compartment, cartilage thicknesses by 3D MRI had positive correlations with WBLR and JLCA at almost all subregions of the MFC and at the anterior-middle and central-external of the MTP. Treatment strategies should be considered in light of these regions. LEVEL OF EVIDENCE: Cross-sectional study (diagnosis); Level of evidence, 2.

Biomechanical analysis of load distribution in porcine hip joints at different acetabular coverages.

BACKGROUND: Developmental dysplasia of the hip causes secondary osteoarthritis. Finite element analysis suggests high hip joint contact pressure in patients with hip dysplasia and a reduction in contact pressure after periacetabular osteotomy. However, few biomechanical studies have examined the load distribution in the hip joint. This study aimed to investigate the biomechanical properties of load distribution in porcine hip joints at different acetabular coverages. METHODS: Six porcine hip joints were analyzed using three models: 1) neutral coverage, 2) 15° under-coverage (defined as dysplasia model), and 3) 15° over-coverage created by varying the acetabular coverage. The load distribution was assessed using a pressure-mapping sensor system after applying a loading force of 100 N to the hip joint. RESULTS: In the dysplasia model, the load was concentrated at the acetabular rim; in the neutral and over-coverage models, it was dispersed. The average contact pressure was significantly higher in the dysplasia model than in the neutral coverage model ([0.42 vs. 0.3 MPa]; p = 0.004). The contact area was significantly smaller in the dysplasia model than in the neutral coverage model ([250.7 vs. 345.0 mm2]; p = 0.004). No significant differences were observed in contact pressure or area between the neutral and over-coverage models. CONCLUSIONS: Insufficient acetabular coverage in the dysplasia model demonstrated higher contact pressure and smaller contact area than the neutral model. Conversely, the contact pressure and area in the over-coverage model did not differ significantly from those in the normal model. Therefore, surgeons should note that acetabular coverage overcorrection has limited effect; normalization is crucial during periacetabular osteotomy.

Equivalent Clinical Outcomes Between Anatomical Alignment Versus Mechanical Alignment of Simultaneous Bilateral Total Knee Arthroplasty Using a Posterior-Stabilized Prosthesis During an Average Follow-Up of Five Years: A Prospective Randomized Clinical Trial.

BACKGROUND: This study aimed to investigate the differences in clinical outcomes between anatomical alignment and mechanical alignment in each knee, respectively, in patients who underwent bilateral total knee arthroplasty. METHODS: Bilateral simultaneous total knee arthroplasty using a posterior-stabilized prosthesis was performed between June 2016 and May 2018, with one knee undergoing anatomical alignment and the contralateral knee undergoing mechanical alignment, which was randomized. There were 80 knees in 40 patients (mean age 75 years, range 60 to 87, with 4 men and 36 women) followed up for at least 2 years (mean follow-up 4.9 years, range 2 to 7 years). On the anatomical side, the distal femoral cut was set 2° more valgus to the mechanical axis, and the tibial cut was set to 2 or 3° varus using a specific guide. Implant position and lower extremity alignment were measured using postoperative radiographs and computed tomography images. The range of motion before and after surgery, Knee Society score, patient satisfaction, postoperative 2011 Knee Society questionnaire, and Knee Injury and Osteoarthritis Outcome Scores were evaluated. RESULTS: A significant difference was observed in joint line orientation (0.6 versus 2.9°, P < .001), but not in the postoperative hip-knee-ankle angle between 2 groups. There was one knee in the mechanical alignment group that required revision surgery at 5 years due to implant loosening. Before surgery, the anatomical alignment group had a significantly lower knee extension angle (-7.2 versus -6.0°, P = .035) and Knee Society Score (46.1 versus 49.1, P = .046). No significant difference was observed between the groups except for the 2011 Knee Society Questionnaire satisfaction scores (26.7 versus 25.7, P = .035). More participants preferred the anatomical alignment side postoperatively. CONCLUSIONS: Anatomical and mechanical alignments had similar clinical outcomes in patients undergoing bilateral total knee arthroplasty at a mean of 4.9 years follow-up. LEVELS OF EVIDENCE: Level Ⅰ (Randomized control study).

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.

Variations in knee cartilage thickness: Fully automatic three-dimensional analysis of MRIs from five manufacturers.

BACKGROUND: Measurements of knee cartilage thickness derived from MR images are attractive biomarkers for osteoarthritis research. Although some cross-sectional multivendor studies exist, none have employed fully automatic three-dimensional MRI analysis. Our objective was to evaluate the variations in knee cartilage thickness measurements obtained using automated methods and MRI instruments from five different vendors. METHODS: The subjects were 10 healthy volunteers aged 22-60 years. MRI models with 3 Tesla strength from five different companies were used. Cartilage thickness was quantified fully automatically for seven regions. We hypothesized that "the MRI model influences cartilage thickness measurements." Inter-measurement error, defined as the absolute difference between the targeted and median thicknesses determined by the five MRI models, was analyzed using histograms. The factors generating the largest inter-measurement error were also examined. RESULTS: No exceptional trends attributable to a specific instrument model were observed, and the p-value from the Kruskal-Wallis test exceeded 0.05 in all seven regions. Therefore, the study hypothesis was rejected. Of the 350 measurements, the inter-measurement error was ≤0.05 mm in 53 %, ≤0.10 mm in 75 %, and ≤0.20 mm in 95 %. Analysis of the medial tibial cartilage, which had the largest inter-measurement error, revealed mis-extraction of synovial fluid as cartilage. CONCLUSIONS: The choice of MRI model did not influence cartilage thickness measurements. Overall, 95 % of the inter-measurement errors were within 0.20 mm. The greatest error resulted from mis-extracting synovial fluid as cartilage.

ARID5B is a negative modulator of IL-6 production in rheumatoid arthritis synovial fibroblasts.

Recent single-cell RNA-sequencing analysis of rheumatoid arthritis (RA) synovial tissues revealed the heterogeneity of RA synovial fibroblasts (SFs) with distinct functions such as high IL-6 production. The molecular mechanisms responsible for high IL-6 production will become a promising drug target of RASFs to treat RA. In this study, we performed siRNA screening of 65 transcription factors (TFs) differentially expressed among RASF subsets to identify TFs involved in IL-6 production. The siRNA screening identified 7 TFs including ARID5B, a RA risk gene, that affected IL-6 production. Both long and short isoforms of ARID5B were expressed and negatively regulated by TNF-α in RASFs. The siRNA knockdown and lentiviral overexpression of long and short isoforms of ARID5B revealed that the long isoform suppressed IL-6 production stimulated with TNF-α. eQTL analysis using 58 SFs demonstrated that RA risk allele, rs10821944, in intron 4 of the ARID5B gene had a trend of eQTL effects to the expression of long isoform of ARID5B in SFs treated with TNF-α. ARID5B was found to be a negative modulator of IL-6 production in RASFs. The RA risk allele of ARID5B intron may cause high IL-6 production, suggesting that ARID5B will become a promising drug target to treat RA.

Scanning electron microscopy analysis of synovial and adipose mesenchymal stem cells adhering to cartilage.

Introduction: Mesenchymal stem cells (MSCs) are increasingly used for intra-articular injections in the treatment of knee osteoarthritis. The aim of this study was to use scanning electron microscopy (SEM) to compare the morphological characteristics of synovial and adipose MSCs. Methods: Synovium and adipose tissues were concurrently harvested from eight patients with knee osteoarthritis. Suspensions of both synovial and adipose MSCs were examined to identify the presence of microspikes. In addition to this study, the MSC suspensions in four patients were applied to abraded porcine cartilage discs and observed 10 s, 10 min, and 1 h later. Results: The median percentage of cells exhibiting microspikes was 14% for synovial MSC suspensions and 13% for adipose MSC suspensions; this difference was not statistically significant (n = 8). No notable differences were detected in the number of adherent cells or in the proportion of cells displaying microspikes or pseudopodia. Strong correlations were found between the proportion of cells with pseudopodia and the number of attached cells for both synovial (r = 0.92, n = 12) and adipose (r = 0.86, n = 12) MSCs, with no significant difference in the correlation coefficients between the two groups. Conclusion: SEM analysis revealed no obvious differences in morphological characteristics during MSC adhesion to cartilage for either synovial or adipose MSCs.

Real-Time Virtual Sonography-Guided Ossicle Removal in Unresolved Osgood-Schlatter Disease.

Osgood-Schlatter disease (OSD) reduces participation in sports activities for adolescents, and, in unresolved cases of the disease in which daily life or sports activity after skeletal maturity are affected, surgical intervention to remove the ossicle is performed to alleviate symptoms. We present a real-time virtual sonography (RVS)-guided ossicle removal in unresolved OSD. The knee joint angle is set at 20°, which is the same position used in magnetic resonance imaging. A 1.5-cm longitudinal skin incision is made at the medial side of the tuberosity. An ultrasound probe covered with a sterile sleeve is then placed longitudinally at the level of tuberosity. The forceps is inserted to peel the patellar tendon off the posterior side of the ossicle. Then the anterior side of the ossicle is peeled off in both the transverse and longitudinal views. When the ossicle is unstable enough, forceps are used to grasp the ossicle and carefully remove it. RVS can see magnetic resonance imaging information in the surrounding area beyond what can be detected by the ultrasound probe, and RVS enables easy determination of the anatomical position of the ossicle, and removal of the ossicle is achieved with low invasiveness.

Histological and biochemical changes in a rat rotator cuff tear model with or without the subacromial bursa.

The subacromial bursa (SAB) plays an important role in the tendon healing process. Based on previous reports, co-culture of the rotator cuff (RC) and SAB have been shown to increase the tendon-related gene expressions, inflammatory cytokines, and tensile strength. However, the nature of the specific biochemical alterations during the inflammatory and repair phases of tendon healing with or without the SAB remain unknown. Using a full-thickness RC tear rat model, we determined how the presence or absence of the SAB alters the histological characteristics and gene expressions. After 3 and 6 weeks, tissues were collected for histological and real-time quantitative polymerase chain reaction (RT-qPCR) evaluations. Results showed greater cell density at 3 weeks, neovascularization and tendon thickening at 6 weeks with SAB preservation. Immunostaining revealed significant increases in type 3 collagen (COL3) expression at 6 weeks with SAB preservation. The RT-qPCR results showed that SAB preservation induced significant increases in the expression of scleraxis, matrix metalloproteinase-13 (MMP-13), interleukin-1ß (IL-1ß), and inducible nitric oxide synthase (iNOS) at 3 weeks and significant increases in COL3, IL-10, and arginase-1 (Arg-1) at 6 weeks. An RC tear undergoes more appropriate inflammatory and repair phases during the tendon healing process when the SAB is retained.

Operator-derived particles and falling bacteria in biosafety cabinets.

Introduction: To ensure the sterility of cell products that cannot undergo conventional sterilization processes, it is imperative to establish and maintain a clean room environment, regulated through environmental monitoring, including particle counts. Nevertheless, the impact of particles generated by operators as potential contaminants remains uncertain. Thus, in this study, we conducted an accelerated test to assess the correlation between particles generated by operators and airborne bacteria, utilizing biosafety cabinets within a typical laboratory setting. These biosafety cabinets create a controlled environment with air conditioning and high-efficiency particulate air (HEPA) filters, offering fundamental data relevant to cell production. Materials and methods: We conducted a simulation followed by real-time experiments involving human operations to explore the quantity of particles, particle sizes, and the percentage of bacteria within these particles. This investigation focused on conditions with heightened particle generation from operators within a biosafety cabinet. The experiment was conducted on operators wearing textile and non-woven dustless clothing within biosafety cabinets. It entailed tapping the upper arms for a duration of 2 min. Results: Observations under biosafety cabinet-off conditions revealed the presence of various particles and falling bacteria in textile clothing. In contrast, no particles or falling bacteria were detected in operators wearing dustless clothing within biosafety cabinets. Notably, a correlation between 5 µm particles and colony-forming units in textile clothing was identified through this analysis. The ratio of falling bacteria to the total number of particles within the biosafety cabinet was 0.8 ± 0.5 % for textile clothing, while it was significantly lower at 0.04 ± 0.2 % for dustless clothing. Conclusion: This study demonstrated that the number of particles and falling bacteria varied depending on the type of clothing and that quantitative data could be used to identify risks and provide basic data for operator education and evidence-based control methods in aseptic manufacturing areas. Although, this study aims to serve as an accelerated test operating under worst-case conditions, the results need to make sure the study range in general research.

Noninvasive total counting of cultured cells using a home-use scanner with a pattern sheet.

The inherent variability in cell culture techniques hinders their reproducibility. To address this issue, we introduce a comprehensive cell observation device. This new approach enhances the features of existing home-use scanners by implementing a pattern sheet. Compared with fluorescent staining, our method over- or underestimated the cell count by a mere 5%. The proposed technique showcased a strong correlation with conventional methodologies, displaying R2 values of 0.91 and 0.99 compared with the standard chamber and fluorescence methods, respectively. Simulations of microscopic observations indicated the potential to estimate accurately the total cell count using just 20 fields of view. Our proposed cell-counting device offers a straightforward, noninvasive means of measuring the number of cultured cells. By harnessing the power of deep learning, this device ensures data integrity, thereby making it an attractive option for future cell culture research.

Characteristics of the synovial microenvironment and synovial mesenchymal stem cells with hip osteoarthritis of different bone morphologies.

BACKGROUND: Variations in bone morphology in patients with hip osteoarthritis (HOA) can be broadly categorized into three types: atrophic, normotrophic, and hypertrophic. Despite the investigations examining clinical elements, such as bone morphology, pain, and range of motion, our understanding of the pathogenesis of HOA remains limited. Previous studies have suggested that osteophytes typically originate at the interface of the joint cartilage, periosteum, and synovium, potentially implicating synovial mesenchymal stem cells (SMSCs) in the process. This study aimed to investigate the potential factors that drive the development of bone morphological features in HOA by investigating the characteristics of the synovium, differentiation potential of SMSCs, and composition of synovial fluid in different types of HOA. METHODS: Synovial tissue and fluid were collected from 30 patients who underwent total hip arthroplasty (THA) with the variable bone morphology of HOA patients. RNA sequencing analysis and quantitative reverse transcription-polymerase chain reaction (RT-qPCR) were performed to analyse the genes in the normotrophic and hypertrophic synovial tissue. SMSCs were isolated and cultured from the normotrophic and hypertrophic synovial tissues of each hip joint in accordance with the variable bone morphology of HOA patients. Cell differentiation potential was compared using differentiation and colony-forming unit assays. Cytokine array was performed to analyse the protein expression in the synovial fluid. RESULTS: In the RNA sequencing analysis, 103 differentially expressed genes (DEGs) were identified, predominantly related to the interleukin 17 (IL-17) signalling pathway. Using a protein-protein interaction (PPI) network, 20 hub genes were identified, including MYC, CXCL8, ATF3, NR4A1, ZC3H12A, NR4A2, FOSB, and FOSL1. Among these hub genes, four belonged to the AP-1 family. There were no significant differences in the tri-lineage differentiation potential and colony-forming capacity of SMSCs. However, RT-qPCR revealed elevated SOX9 expression levels in synovial tissues from the hypertrophic group. The cytokine array demonstrated significantly higher levels of CXCL8, MMP9, and VEGF in the synovial fluid of the hypertrophic group than in the normotrophic group, with CXCL8 and MMP9 being significantly expressed in the hypertrophic synovium. CONCLUSION: Upregulation of AP-1 family genes in the synovium and increased concentrations of CXCL8, MMP9, and VEGF were detected in the synovial fluid of the hypertrophic group of HOA patients, potentially stimulating the differentiation of SMSCs towards the cartilage and thereby contributing to severe osteophyte formation.

Association of AI-determined Kellgren-Lawrence grade with medial meniscus extrusion and cartilage thickness by AI-based 3D MRI analysis in early knee osteoarthritis.

The associations among Kellgren-Lawrence (KL) grade, medial meniscus extrusion (MME), and cartilage thickness in knee osteoarthritis (OA) remain insufficiently understood. Our aim was to determine these associations in early to moderate medial tibiofemoral knee OA. We included 469 subjects with no lateral OA from the Kanagawa Knee Study. KL grade was assessed using artificial intelligence (AI) software. The MME was measured by MRI, and the cartilage thickness was evaluated in 18 subregions of the medial femorotibial joint by another AI system. The median MME width was 1.4 mm in KL0, 1.5 mm in KL1, 2.4 mm in KL2, and 6.0 mm in KL3. Cartilage thinning in the medial femur occurred in the anterior central subregion in KL1, expanded inwardly in KL2, and further expanded in KL3. Cartilage thinning in the medial tibia occurred in the anterior and middle external subregions in KL1, expanded into the anterior and middle central subregions in KL2, and further expanded in KL3. The absolute correlation coefficient between MME width and cartilage thickness increased as the KL grade increased in some subregions. This study provides novel insights into the early stages of knee OA and potentially has implications for the development of early intervention strategies.