Small change in the arithmetic hip-knee-ankle angle during unicompartmental knee arthroplasty improves early postoperative functional outcomes.

INTRODUCTION: The optimal lower-limb alignment after unicompartmental knee arthroplasty (UKA) remains controversial. This study aimed to investigate the optimal lower-limb alignment for functional improvement in the early post-UKA period. We hypothesized that a small change (Δ) in the arithmetic hip-knee-ankle (aHKA) angle during surgery would result in better postoperative knee function. MATERIALS AND METHODS: This single-centered, retrospective study analyzed 91 patients (91 knees) who underwent UKA from April 2021 and December 2022. Preoperative and postoperative standing whole-leg radiographs were used to evaluate the mechanical HKA angle and aHKA angle. The aHKA angle was calculated from the medial proximal tibial angle (MPTA) and lateral distal femoral angle (LDFA). We defined restored aHKA angle as a postoperative aHKA angle within ± 3° of the preoperative aHKA angle. Functional improvement was evaluated using the preoperative and one-year postoperative Knee Society Scoring 2011 (KSS 2011). A multivariate regression analysis was performed to investigate the optimal lower-limb alignment for functional improvement. RESULT: Postoperative restored aHKA angle (p = 0.020) was the only significant factor for improved KSS 2011 scores. Postoperative restored aHKA angle was obtained in 64 patients (70%). ΔMPTA (p < 0.001) and ΔLDFA (p = 0.037) were significant factors associated with a postoperative restored aHKA angle. CONCLUSIONS: UKA restores the native knee, including resurfacing constitutional alignment and medial collateral ligament tension. Minimal change in the aHKA angle during UKA improves the functional outcomes of the knee during the early postoperative period, consistent with the minimally invasive surgical concept of UKA.

Life Course Epidemiology of Hip Osteoarthritis in Japan: A Multicenter, Cross-Sectional Study.

BACKGROUND: The incidence of developmental dysplasia of the hip (DDH) in Japanese newborns has reduced drastically following a primary prevention campaign initiated around 1972 to 1973; this perinatal education campaign promoted maintaining the hips of newborns in the naturally flexed-leg position. The purpose of the present study was to describe the life course epidemiology of hip osteoarthritis (OA) in adolescent and adult patients and to assess its association with exposure to the primary prevention campaign for DDH. METHODS: We included new patients with hip OA diagnosed from January 1, 2022, to December 31, 2022, at 12 core hospitals (8 special-function hospitals and 4 regional medical care support hospitals). The trend in the percentage of hips with a history of DDH treatment in childhood was estimated with use of a centered moving average using the birth year of the patient. We compared the prevalence of severe subluxation (Crowe type II, III, or IV) between patients with secondary hip OA due to hip dysplasia who were born in or before 1972 and those who were born in or after 1973. RESULTS: Overall, 1,095 patients (1,381 hips) were included. The mean age at the time of the survey was 63.5 years (range, 15 to 95 years). A total of 795 patients (1,019 hips; 73.8% of hips) were diagnosed with secondary OA due to hip dysplasia. Approximately 13% to 15% of hips among patients born from 1963 to 1972 had a history of DDH treatment in childhood; however, the percentage decreased among patients born in or after 1973. The prevalence of severe subluxation (Crowe type II, III, or IV) among patients born in or after 1973 was 2.4%, which was significantly less than that among patients born in or before 1972 (11.1%; odds ratio, 0.20; p < 0.001). CONCLUSIONS: As of 2022, secondary hip OA due to hip dysplasia is still responsible for most new cases of adolescent and adult hip OA seen in core hospitals in Japan. However, the perinatal education campaign initiated 50 years ago, which utilized a population approach and advocated for maintaining the hips of newborns in the naturally flexed-leg position, may have improved the environmental factors of DDH, as indicated by the apparently reduced need for treatment of DDH in childhood and the associated severe subluxation. This may result in a reduced need for challenging hip surgery later in life. LEVEL OF EVIDENCE: Prognostic Level III . See Instructions for Authors for a complete description of levels of evidence.

The sourcil roundness index is a useful measure for quantifying acetabular concavity asphericity.

This study aimed to clarify the clinical utility of the sourcil roundness index (SRI), a novel index for quantifying the asphericity of the acetabular concavity, by determining (1) the difference in the SRI between dysplastic and normal hips and (2) the correlation between the SRI and radiographic parameters of hip dysplasia. We reviewed standing anteroposterior pelvic radiographs of 109 dysplastic and 40 normal hips. The SRI was determined as the ratio of the distance from the medial edge of the sourcil to the most concave point of the acetabular sourcil (A) to the distance from the medial to the lateral edge of the sourcil (B). The formula for SRI is (A/B) × 100-50 (%), with an SRI of 0% indicating a perfectly spherical acetabulum, and higher SRI values indicating a more aspherical shape. The median SRI was greater in patients with hip dysplasia than in normal hips (5.9% vs. - 1.4%; p < 0.001). Furthermore, the median SRI was greater in the severe dysplasia subgroup (18.9%) than in the moderate (3.5%) and borderline-to-mild (- 1.3%) dysplasia subgroups (p < 0.05). Quantification of acetabular concavity asphericity by the SRI showed that dysplastic hips had a more lateral acetabular concave point than normal hips, and that the severity of hip dysplasia had an effect on the acetabular concavity asphericity.

Evaluation of the anterior acetabular coverage with a false profile radiograph considering appropriate range of positioning.

This study aimed to (1) set a reference value for anterior center edge angle (ACEA) for preoperative planning of periacetabular osteotomy (PAO), (2) investigate the effects of pelvic rotation and inclination from false profile (FP) radiographs on the measured ACEA, and (3) determine the "appropriate range of positioning" for FP radiograph. This single-centered, retrospective study analyzed 61 patients (61 hips) who underwent PAO from April 2018 and May 2021. ACEA was measured in each digitally reconstructed radiography (DRR) image of the FP radiograph reconstructed in different degrees of pelvic rotation. Detailed simulations were performed to determine the "appropriate range of positioning" (0.67 < ratio of the distance between the femoral heads to the diameter of the femoral head < 1.0). The vertical-center-anterior (VCA) angle was measured on the CT sagittal plane considering the patient-specific standing positions, and its correlation with the ACEA was investigated. The reference value of ACEA was determined by receiver operating characteristic (ROC) curve analysis. The ACEA measurement increased by 0.35° for every 1° pelvic rotation approaching the true lateral view. The pelvic rotation with the "appropriate range of positioning" was found at 5.0° (63.3-68.3°). The ACEA on the FP radiographs showed a good correlation with the VCA angle. The ROC curve revealed that an ACEA < 13.6° was associated with inadequate anterior coverage (VCA < 32°). Our findings suggest that during preoperative PAO planning, an ACEA < 13.6° on FP radiographs indicates insufficient anterior acetabular coverage. Images with the "appropriate positioning" can also have a measurement error of 1.7° due to the pelvic rotation.

Protocol for the RETHINK study: a randomised, double-blind, parallel-group, non-inferiority clinical trial comparing acetaminophen and NSAIDs for treatment of chronic pain in elderly patients with osteoarthritis of the hip and knee.

INTRODUCTION: In patients with chronic pain, oral analgesics are essential treatment options to manage pain appropriately, improve activities of daily living abilities and achieve a higher quality of life (QOL). It is desirable to select analgesics for elderly patients based on comparative data on analgesic effect and risk of adverse events; however, there are few comparative studies so far. The purpose of this study is to determine whether the efficacy and safety of acetaminophen are non-inferior to non-steroidal anti-inflammatory drugs (NSAIDs) in the treatment of chronic pain associated with osteoarthritis of the hip and knee in elderly patients. METHODS AND ANALYSIS: This study is a multicentre, randomised controlled, double-blind, parallel-group study to compare the analgesic effect and adverse events between acetaminophen or NSAIDs (loxoprofen or celecoxib). A total of 400 elderly patients with osteoarthritis of the hip and knee will be recruited from five institutions in Japan. Patients of 65 years or older with osteoarthritis-related pain will be registered and randomly assigned to acetaminophen, loxoprofen or celecoxib with 2:1:1 allocation. The primary endpoint is change in the Brief Pain Inventory (BPI) item 3 (worst pain) score from baseline to week 8. The secondary endpoints are BPI item 3 score change from baseline to week 4, health-related QOL measured by Short Form-8 Health Survey, and occurrence of adverse events including gastrointestinal disorders and abnormal liver function. Data will be analysed in accordance with a predefined statistical analysis plan. ETHICS AND DISSEMINATION: This study protocol was approved by the Kyushu University Hospital Certified Institutional Review Board for Clinical Trials on 28 January 2021 (KD2020004) and the chief executive of each participating hospital. The results of the study will be submitted to international peer-reviewed journals, and the main findings will be presented at international scientific conferences. TRIAL REGISTRATION NUMBER: jRCTs071200112.

The influence of bone marrow edema for the assessment of the boundaries of necrotic lesions in patients with osteonecrosis of the femoral head.

This study aimed to investigate the influence of bone marrow edema (BME) for the assessment of the boundaries of necrotic lesions using unenhanced and contrast-enhanced (CE) magnetic resonance (MR) images in patients with osteonecrosis of the femoral head (ONFH). We retrospectively reviewed 72 consecutive hips in 55 patients of ONFH that were Association Research Circulation Osseous (ARCO) stage III or higher and underwent both unenhanced and contrast-enhanced MR imaging between January 2005 and February 2016. The degree of extension of BMEs, and the boundaries of the necrotic lesions were compared using unenhanced and CE MR images on both mid coronal and mid oblique-axial slices. Forty-two percent of the coronal T1 images, 40% of the coronal fat-saturated T2 images, and 48% of the oblique-axial T1 images showed differences in the boundaries of necrotic lesion, by comparison with those of CET1-weighted MR images. The boundaries of necrotic lesions were clearly detected in all hips on CE coronal slices and 97% of all hips on CE oblique-axial slices. The BME grade in the difference group was significantly higher than in the non-difference group on the coronal plane (P = 0.0058). There were significant differences between the BME grade and duration from the onset of hip pain to MR imaging examination. Multivariate analyses revealed that the duration from the onset to MR imaging examination in both coronal (P = 0.0008) and oblique-axial slices (P = 0.0143) were independently associated with differences in the boundary of necrotic lesion between T1 and CET1-weighted MR images. Our findings suggest that unenhanced MR image may be insufficient for a precise assessment of the boundaries of the necrotic lesions for ONFH cases in the early phase of subchondral collapse due to the diffuse BME.

Multiple Imputation to Salvage Partial Respondents: Analysis of the Forgotten Joint Score-12 After Total Hip Arthroplasty.

BACKGROUND: Missing responses are common when Asian patients complete the Forgotten Joint Score-12 (FJS-12), which is widely used to evaluate total hip arthroplasty (THA). We aimed to provide orthopaedic researchers with a solution for handling missing values in such patient-reported outcome measures (PROMs). METHODS: Patients who had undergone primary THA between 1998 and 2016 (n = 1,021) were investigated in 2020. The FJS-12 and 9 other PROMs, including questions related to Asian lifestyle activities, were administered. Risk factors for missing FJS-12 items were investigated. Partial respondents were matched with complete respondents; then, in each pair, the items not completed by the partial respondent were deleted from the responses of the complete respondent. Predictive mean matching (PMM) was performed in an attempt to recover the deleted items, using 65 sets of imputation models. After the missing values had been imputed, we explored patient characteristics that affected the FJS-12, using data from all complete and partial respondents. RESULTS: A total of 652 patients responded to the survey (393 complete and 193 partial respondents). Partial respondents were older, more often female, and less active. Older respondents were more likely to skip items involving the bed, while those who reported a better ability to sit in the seiza style (traditional Japanese floor sitting) were more likely to skip items about chair sitting. The imputed FJS-12 value exhibited excellent reliability (intraclass correlation coefficient for agreement with the true scores, 0.985). FJS-12 values of complete respondents were significantly higher than those of respondents with 4 to 11 missing items (51.6 versus 32.8, p < 0.001). Older age was associated with higher FJS-12 values, which was revealed only via analysis of the multiply imputed data sets (p < 0.001). CONCLUSIONS: Analysis of only complete FJS-12 responses after THA resulted in a nonresponse bias, preferentially excluding older, female, and less active individuals and those with a traditional floor living style. Multiple imputation could provide a solution to scoring and analyzing PROMs with missing responses by permitting the inclusion of partial respondents. LEVEL OF EVIDENCE: Therapeutic Level III . See Instructions for Authors for a complete description of levels of evidence.

Tapered wedge stems decrease early postoperative subsidence following cementless total hip arthroplasty in Dorr type C femurs compared to fit-and-fill stems.

BACKGROUND: To compare the degree of stem subsidence between two different femoral component designs and to determine the risk factors associated with stem subsidence after cementless total hip arthroplasty (THA) in Dorr type C femurs. METHODS: We retrospectively reviewed 104 consecutive hips in 100 patients with Dorr type C proximal femoral morphology who underwent primary cementless THA using a fit-and-fill stem or a tapered wedge femoral stem at our institution between January 2012 and June 2021. A fit-and-fill stem was used in 55 hips and a tapered wedge stem was used in 49 hips. Radiologically, the distance between the apex of the major trochanter and the stem shoulder were measured at three different time points (immediately [0W], one week [1W], and six weeks [6W] after surgery) and the degrees of stem subsidence were assessed by comparing the distance between 0 and 1W, 1W and 6W, and 0W and 6W, respectively. RESULTS: The mean degree of stem subsidence (0W vs. 1W) was 0.24 mm (standard deviation [SD] 0.36) in the fit-and-fill stem group, and 0.23 mm (SD 0.41) in the tapered wedge stem group. There was no significant difference between the two groups (P = 0.4862). However, the mean degrees of subsidence were significantly higher in the fit-and-fill stem group (1W vs. 6W, 0.38 mm [SD 0.68]; 0W vs. 6W, 0.65 mm [SD 0.87]) than in the tapered wedge stem group (1W vs. 6W, 0.16 mm [SD 0.32]; 0W vs. 6W, 0.24 mm [SD 0.38]) (P < 0.05 for both). In addition, the rates of > 3 mm subsidence (in which instability can be observed) were 18.2% (10 of 55 hips) and 2.0% (1 of 49 hips), respectively. There was also a significant difference between the two stems (P = 0.0091). Multivariate analysis demonstrated that fit-and-fill stem was a risk factor for > 3 mm subsidence after THA in Dorr type C femurs (P = 0.0050). CONCLUSION: Our findings suggest that the tapered wedge stem is more suitable for Dorr type C femurs than the fit-and-fill stem to avoid early postoperative subsidence in cementless THA.

Evaluation of optimal implant alignment in total hip arthroplasty based on postoperative range of motion simulation.

BACKGROUND: Dislocation after total hip arthroplasty is a frequent cause of revision surgery. This study was performed to determine the optimal implant alignment in total hip arthroplasty by simulating the postoperative range of motion. METHODS: All operations were performed via posterolateral approach using combined anteversion of the stem and cup technique. Maximum range of motion without implant impingement was simulated in 79 replaced hips using postoperative computed tomography and the achievement of the required range of motion defined by previous studies was assessed. Optimal cup and stem alignment for impingement-free range of motion were statistically determined using the receiver operator coefficient curve. FINDINGS: Cup inclination and anteversion, stem anteversion, and combined anteversion were 37.6°, 20.1°, 26.2°, and 46.3°, respectively. Maximum range of motion in flexion, extension, internal rotation at 90° of flexion, and external rotation were 131.8°, 42.3°, 56.4°, and 64.5°, respectively. Flexion >110°, extension >30°, internal rotation >30° at 90° of flexion, and external rotation >30° were fulfilled by 96%, 86%, 92%, and 96% of all replaced hips, respectively. Optimal implant alignment for impingement-free range of motion was 34°-43° of cup inclination, 18°-26° of cup anteversion, 17°-29° of stem anteversion, and 35°-56° of combined anteversion. Both cup and stem anteversion showed significant relationship with postoperative range of motion. INTERPRETATION: Surgeons could gain valuable insights into optimal cup and stem alignment to perform postoperative range of motion simulations.

Abductor recovery after muscle-sparing periacetabular osteotomy using a lateral approach.

To decrease hip abductor dysfunction after periacetabular osteotomy using a lateral/trochanteric approach, we aimed to modify transposition osteotomy of the acetabulum (TOA) to not cut the greater trochanter and abductor-iliac crest detachment. We subsequently compared abductor muscle strength recovery between TOAs with [conventional TOA (C-TOA)] and without [modified TOA (M-TOA)] trochanteric osteotomy. C-TOA and M-TOA were performed in 27 and 34 hips, respectively. Hip abduction, flexion and knee extension muscle strength were measured preoperatively and at 3, 5, 10, 24 and 52 weeks postoperatively. The muscle strength ratio of the affected and contralateral lower limbs was compared between the C-TOA and M-TOA groups. Neither the mean Merle d'Aubigné-Postel score at the final follow-up nor the postoperative center-edge angle showed significant differences between the M-TOA and C-TOA groups (15.7 versus 16.4 points; P = 0.25 and 38.5° versus P = 0.62 and 39.8°, respectively). The mean muscle strength ratios of hip abduction at 5, 12 and 24 weeks postoperatively were significantly higher in the M-TOA group than in the C-TOA group (0.62 versus 0.39, 0.76 versus 0.59 and 0.94 versus 0.70; P = 0.03, 0.04 and 0.01, respectively). There were no significant differences between groups at Postoperative Week 52 (P = 0.36). Discomfort at the greater trochanter was observed in 18 hips (66.7%) in the C-TOA group but only in 4 hips (11.2%) in the M-TOA group. In conclusion, M-TOA is less invasive than C-TOA and allows an earlier recovery of abductor muscle strength without significant correction loss.

Interleukin-4 repairs wear particle induced osteolysis by modulating macrophage polarization and bone turnover.

Periprosthetic osteolysis remains as a major complication of total joint replacement surgery. Modulation of macrophage polarization with interleukin-4 (IL-4) has emerged as an effective means to limit wear particle-induced osteolysis. The aim of this study was to evaluate the efficacy of local IL-4 delivery in treating preexisting particle-induced osteolysis. To this end, recently established 8 week modification of murine continuous femoral intramedullary particle infusion model was utilized. Subcutaneous infusion pumps were used to deliver polyethylene (PE) particles into mouse distal femur for 4 weeks to induce osteolysis. IL-4 was then added to the particle infusion for another 4 weeks. This delayed IL-4 treatment (IL-4 Del) was compared to IL-4 delivered continuously (IL-4 Cont) with PE particles from the beginning and to the infusion of particles alone for 8 weeks. Both IL-4 treatments were highly effective in preventing and repairing preexisting particle-induced bone loss as assessed by µCT. Immunofluorescence indicated a significant reduction in the number of F4/80 + iNOS + M1 macrophages and increase in the number of F4/80 + CD206 + M2 macrophages with both IL-4 treatments. Reduction in the number of tartrate resistant acid phosphatase + osteoclasts and increase in the amount of alkaline phosphatase (ALP) + osteoblasts was also observed with both IL-4 treatments likely explaining the regeneration of bone in these samples. Interesting, slightly more bone formation and ALP + osteoblasts were seen in the IL-4 Del group than in the IL-4 Cont group although these differences were not statistically significant. The study is a proof of principle that osteolytic lesions can be repaired via modulation of macrophage polarization.

* Murine Model of Progressive Orthopedic Wear Particle-Induced Chronic Inflammation and Osteolysis.

Periprosthetic osteolysis and subsequent aseptic loosening of total joint replacements are driven by byproducts of wear released from the implant. Wear particles cause macrophage-mediated inflammation that culminates with periprosthetic bone loss. Most current animal models of particle-induced osteolysis are based on the acute inflammatory reaction induced by wear debris, which is distinct from the slowly progressive clinical scenario. To address this limitation, we previously developed a murine model of periprosthetic osteolysis that is based on slow continuous delivery of wear particles into the murine distal femur over a period of 4 weeks. The particle delivery was accomplished by using subcutaneously implanted osmotic pumps and tubing, and a hollow titanium rod press-fit into the distal femur. In this study, we report a modification of our prior model in which particle delivery is extended to 8 weeks to better mimic the progressive development of periprosthetic osteolysis and allow the assessment of interventions in a setting where the chronic particle-induced osteolysis is already present at the initiation of the treatment. Compared to 4-week samples, extending the particle delivery to 8 weeks significantly exacerbated the local bone loss observed with µCT and the amount of both peri-implant F4/80+ macrophages and tartrate-resistant acid phosphatase-positive osteoclasts detected with immunohistochemical and histochemical staining. Furthermore, systemic recruitment of reporter macrophages to peri-implant tissues observed with bioluminescence imaging continued even at the later stages of particle-induced inflammation. This modified model system could provide new insights into the mechanisms of chronic inflammatory bone loss and be particularly useful in assessing the efficacy of treatments in a setting that resembles the clinical scenario of developing periprosthetic osteolysis more closely than currently existing model systems.

NF-κB decoy oligodeoxynucleotide mitigates wear particle-associated bone loss in the murine continuous infusion model.

UNLABELLED: Total joint replacement is a cost-effective surgical procedure for patients with end-stage arthritis. Wear particle-induced chronic inflammation is associated with the development of periprosthetic osteolysis. Modulation of NF-κB signaling in macrophages, osteoclasts, and mesenchymal stem cells could potentially mitigate this disease. In the current study, we examined the effects of local delivery of decoy NF-κB oligo-deoxynucleotide (ODN) on wear particle-induced bone loss in a murine continuous femoral particle infusion model. Ultra-high molecular weight polyethylene particles (UHMWPE) with or without lipopolysaccharide (LPS) were infused via osmotic pumps into hollow titanium rods placed in the distal femur of mice for 4weeks. Particle-induced bone loss was evaluated by µCT, and immunohistochemical analysis of sections from the femur. Particle infusion alone resulted in reduced bone mineral density and trabecular bone volume fraction in the distal femur. The decoy ODN reversed the particle-associated bone volume fraction loss around the implant, irrespective of the presence of LPS. Particle-infusion with LPS increased bone mineral density in the distal femur compared with particle-infusion alone. NF-κB decoy ODN reversed or further increased the bone mineral density in the femur (3-6mm from the distal end) exposed to particles alone or particles plus LPS. NF-κB decoy ODN also inhibited macrophage infiltration and osteoclast number, but had no significant effects on osteoblast numbers in femurs exposed to wear particles and LPS. Our study suggests that targeting NF-κB activity via local delivery of decoy ODN has great potential to mitigate wear particle-induced osteolysis. STATEMENT OF SIGNIFICANCE: Total joint replacement is a cost-effective surgical procedure for patients with end-stage arthritis. Chronic inflammation is crucial for the development of wear particle-associated bone loss. Modulation of NF-κB signaling in macrophages (pro-inflammatory cells), osteoclasts (bone-resorbing cells), and osteoblasts (bone-forming cells) could potentially mitigate this disease. Here we demonstrated that local delivery of decoy NF-κB oligo-deoxynucleotide (ODN) mitigated ultra-high molecular weight polyethylene (UHMWPE) wear particle induced bone loss in a clinically relevant murine model. The protective effects of decoy ODN was associated with reduced macrophage infiltration and osteoclast activation, but had no significant effects on osteoblast numbers. Our study suggests that targeting NF-κB activity via local delivery of decoy ODN has great potential to mitigate wear particle-induced bone loss.

The Absence of Hydroxyapatite Coating on Cementless Acetabular Components Does Not Affect Long-Term Survivorship in Total Hip Arthroplasty.

BACKGROUND: Hydroxyapatite (HA) has been applied to joint prostheses as a bioactive coating to prolong their durability. However, HA benefits for cup survival in total hip arthroplasty (THA) remain controversial. In this study, we compared the survival of cups with and without HA coating during a minimum follow-up of 18 years. METHODS: In total, 183 THA cases in 163 patients were analyzed, including 73 cups with HA coating (HA(+) group) and 110 without HA coating (HA(-) group); otherwise, the cups had identical titanium-sprayed rough surfaces and were fixed with screws. In both groups, the same conventional polyethylene liners were applied. Zirconia and alumina ceramic heads were used in the HA(+) and HA(-) groups, respectively. We retrospectively analyzed cup survival based on cup revision for any reason or aseptic loosening as end points. RESULTS: In total, 7 and 8 revisions were performed in the HA(-) and HA(+) groups with survival rates of 86.3% and 90.1%, respectively. Among them, 3 cups in the HA(-) group and 1 cup in the HA(+) group were revised for aseptic loosening in 20 years (survival rates 94.1% and 98.7%, respectively). No significant difference was detected in cup survival rates between the groups based on both end points. There were no stem revisions during the observation period. CONCLUSION: The results suggested that HA coating did not have either beneficial or adverse effects on the long-term cup survival in primary cementless THA.

Establishment of Green Fluorescent Protein and Firefly Luciferase Expressing Mouse Primary Macrophages for In Vivo Bioluminescence Imaging.

Macrophages play a key role in tissue homeostasis as well as in a range of pathological conditions including atherosclerosis, cancer, and autoimmunity. Many aspects of their in vivo behavior are, however, poorly understood. Bioluminescence imaging (BLI) with green fluorescent protein (GFP) and firefly luciferase (FLUC) labelled autologous reporter macrophages could potentially offer a powerful tool to study macrophage biology, but this approach has been hindered by the relative difficulty of efficient gene transfer into primary macrophages. Here we describe a straightforward method for producing large numbers of GFP/FLUC expressing mouse primary macrophages utilizing lentivirus vector, cyclosporine, and a double infection strategy. Using this method we achieved up to 60% of macrophages to express GFP with correspondingly high FLUC signal. When injected into the circulation using a mouse model of local biomaterial induced inflammation and osteolysis, macrophages were initially detectable within the lungs, followed by systemic homing to the local area of chronic inflammation in the distal femur. In addition, transduced macrophages maintained their ability to assume M1 and M2 phenotypes although the GFP/FLUC expression was altered by the polarizing signals. These reporter macrophages could prove to be valuable tools to study the role of macrophages in health and disease.

NF-κB decoy oligodeoxynucleotide inhibits wear particle-induced inflammation in a murine calvarial model.

Wear particles induce periprosthetic inflammation and osteolysis through activation of nuclear factor kappa B (NF-κB), which up-regulates the downstream target gene expression for proinflammatory cytokines in macrophages. It was hypothesized that direct suppression of NF-κB activity in the early phases of this disorder could be a therapeutic strategy for preventing the inflammatory response to wear particles, potentially mitigating osteolysis. NF-κB activity can be suppressed via competitive binding with double stranded NF-κB decoy oligodeoxynucleotides (ODNs) that blocks this transcription factor from binding to the promoter regions of targeted genes. In this murine calvarial study, clinically relevant polyethylene particles (PEs) with/without ODN were subcutaneously injected over the calvarial bone. In the presence of PE particles, macrophages migrated to the inflammatory site and induced tumor necrosis factor alpha (TNF-α) and receptor activator of nuclear factor kappa B ligand (RANKL) expression, resulting in an increase in the number of osteoclasts. Local injections of ODN mitigated the expression of TNF-α, RANKL, and induced the expression of two anti-inflammatory, antiresorptive cytokines: interleukin-1 receptor antagonist and osteoprotegerin. Local intervention with NF-κB decoy ODN in early cases of particle-induced inflammation in which the prosthesis is still salvageable may potentially preserve periprosthetic bone stock.

Modulation of mouse macrophage polarization in vitro using IL-4 delivery by osmotic pumps.

Modulation of macrophage polarization is emerging as promising means to mitigate wear particle-induced inflammation and periprosthetic osteolysis. As a model for continuous local drug delivery, we used miniature osmotic pumps to deliver IL-4 in order to modulate macrophage polarization in vitro from nonactivated M0 and inflammatory M1 phenotypes towards a tissue regenerative M2 phenotype. Pumps delivered IL-4 into vials containing mouse bone marrow macrophage (mBMM) media. This conditioned media (CM) was collected at seven day intervals up to four weeks (week 1 to week 4 samples). IL-4 concentration in the CM was determined by ELISA and its biological activity was assayed by exposing M0 and M1 mBMMs to week 1 or week 4 CM. The IL-4 concentration in the CM approximated the mathematically calculated amount, and its biological activity was well retained, as both M0 and M1 macrophages exposed to either the week 1 or week 4 CM assumed M2-like phenotype as determined by qRT-PCR, ELISA, and immunocytochemistry. The results show that IL-4 can be delivered using osmotic pumps and that IL-4 delivered can modulate macrophage phenotype. Results build a foundation for in vivo studies using our previously validated animal models and provide possible strategies to locally mitigate wear particle-induced macrophage activation and periprosthetic osteolysis.

NF-κB decoy oligodeoxynucleotide enhanced osteogenesis in mesenchymal stem cells exposed to polyethylene particle.

Excessive generation of wear particles after total joint replacement may lead to local inflammation and periprosthetic osteolysis. Modulation of the key transcription factor NF-κB in immune cells could potentially mitigate the osteolytic process. We previously showed that local delivery of ultrahigh-molecular-weight polyethylene (UHMWPE) particles recruited osteoprogenitor cells and reduced osteolysis. However, the biological effects of modulating the NF-κB signaling pathway on osteoprogenitor/mesenchymal stem cells (MSCs) remain unclear. Here we showed that decoy oligodeoxynucleotide (ODN) increased cell viability when primary murine MSCs were exposed to UHMWPE particles, but had no effects on cellular apoptosis. Decoy ODN increased transforming growth factor-beta 1 (TGF-ß1) and osteoprotegerin (OPG) in MSCs exposed to UHMWPE particles. Mechanistic studies showed that decoy ODN upregulated OPG expression through a TGF-ß1-dependent pathway. By measuring the alkaline phosphatase activity, osteocalcin levels, Runx2 and osteopontin expression, and performing a bone mineralization assay, we found that decoy ODN increased MSC osteogenic ability when the cells were exposed to UHMWPE particles. Furthermore, the cellular response to decoy ODN and UHMWPE particles with regard to cell phenotype, cell viability, and osteogenic ability was confirmed using primary human MSCs. Our results suggest that modulation of wear particle-induced inflammation by NF-κB decoy ODN had no adverse effects on MSCs and may potentially further mitigate periprosthetic osteolysis by protecting MSC viability and osteogenic ability.

Exposure of polyethylene particles induces interferon-γ expression in a natural killer T lymphocyte and dendritic cell coculture system in vitro: a preliminary study.

Two major issues in total joint arthroplasty are loosening of implants and osteolysis caused by wear particle-induced inflammation. Wear particles stimulate the release of pro-inflammatory cytokines, chemokines, and other inflammatory mediators from macrophages and other cells. Although the biological response of macrophages to wear debris is well established, the role of other cell types such as natural killer T lymphocytes (NKT) and dendritic cells (DCs) is limited. Here we show that ultra-high molecular weight polyethylene (UHMWPE) particles stimulate NKT cells to secrete Interferon-γ (IFN-γ); coculture with DCs further enhanced IFN-γ secretion. Furthermore, UHMWPE particles did not stimulate NKT cells to secrete IL-4, while the NKT cell natural ligand α-galactosylceramide (α-GalCer) treatment in the coculture system significantly enhanced both IFN-γ and IL-4 expression by NKT cells. Comparatively, NKT cells and/or DCs exposed to polymethylmethacrylate particles did not stimulate IFN-γ or IL-4 expression. Mouse bone marrow derived macrophage polarization by lipopolysaccharide and conditioned medium from NKT cells and/or DCs exposed to UHMWPE particles increased tumor necrosis factor-α (TNF-α), but reduced arginase-1 expression in macrophages. The current findings indicate that UHMWPE particles stimulate NKT cells/DCs to produce pro-inflammatory cytokines; this pathway is a novel therapeutic target to mitigate wear particle induced peri-prosthetic osteolysis.

Suppression of wear-particle-induced pro-inflammatory cytokine and chemokine production in macrophages via NF-κB decoy oligodeoxynucleotide: a preliminary report.

Total joint replacement (TJR) is very cost-effective surgery for end-stage arthritis. One important goal is to decrease the revision rate, mainly because TJR has been extended to younger patients. Continuous production of ultra-high molecular weight polyethylene (UHMWPE) wear particles induces macrophage infiltration and chronic inflammation, which can lead to periprosthetic osteolysis. Targeting individual pro-inflammatory cytokines directly has not reversed the osteolytic process in clinical trials, owing to compensatory up-regulation of other pro-inflammatory factors. It is hypothesized that targeting the important transcription factor NF-κB could mitigate the inflammatory response to wear particles, potentially diminishing osteolysis. In the current study, NF-κB activity in mouse RAW 264.7 and human THP1 macrophage cell lines, as well as primary mouse and human macrophages, was suppressed via competitive binding with double strand decoy oligodeoxynucleotide (ODN) containing an NF-κB binding element. It was found that macrophage exposure to UHMWPE particles induced multiple pro-inflammatory cytokine and chemokine expression, including TNF-α, MCP1, MIP1α and others. Importantly, the decoy ODN significantly suppressed the induced cytokine and chemokine expression in both murine and human macrophages, and resulted in suppression of macrophage recruitment. The strategic use of decoy NF-κB ODN, delivered locally, could potentially diminish particle-induced periprosthetic osteolysis.