Single cell transcriptomics in human osteoarthritis synovium and in silico deconvoluted bulk RNA sequencing.

OBJECTIVES: To reveal the heterogeneity of different cell types of osteoarthritis (OA) synovial tissues at a single-cell resolution, and determine by novel methodology whether bulk-RNA-seq data could be deconvoluted to create in silico scRNA-seq data for synovial tissue analyses. METHODS: OA scRNA-seq data (102,077 synoviocytes) were provided by 17 patients undergoing total knee arthroplasty; 9 tissues with matched scRNA-seq and bulk RNA-seq data were used to evaluate six in silico gene deconvolution tools. Predicted and observed cell types and proportions were compared to identify the best deconvolution tool for synovium. RESULTS: We identified seven distinct cell types in OA synovial tissues. Gene deconvolution identified three (of six) platforms as suitable for extrapolating cellular gene expression from bulk RNA-seq data. Using paired scRNA-seq and bulk RNA-seq data, an "arthritis" specific signature matrix was created and validated to have a significantly better predictive performance for synoviocytes than a default signature matrix. Use of the machine learning tool, Cell-type Identification By Estimating Relative Subsets of RNA Transcripts x (CIBERSORTx), to analyze rheumatoid arthritis (RA) and OA bulk RNA-seq data yielded proportions of T cells and fibroblasts that were similar to the gold standard observations from RA and OA scRNA-seq data, respectively. CONCLUSION: This novel study revealed heterogeneity of synovial cell types in OA and the feasibility of gene deconvolution for synovial tissue.

Alpha defensin-1 attenuates surgically induced osteoarthritis in association with promoting M1 to M2 macrophage polarization.

OBJECTIVE: Macrophages play an important part in the pathogenesis of osteoarthritis (OA). Our objective was to determine the effects of α-defensin-1 on macrophage polarization and consequently OA. METHODS: OA synovial tissue and synovial fluid were assessed for the presence of M1 (CD68+CD16+CD206-) and M2 (CD68+CD206+CD16-) macrophages by flow cytometry. M0, M1, and M2 macrophages were co-cultured with OA chondrocytes to determine their influence on chondrogenic phenotype. Polarization of THP-1 activated monocytes from M1 to M2 in response to α-defensin-1 was evaluated by flow cytometry, RT-PCR and RNA sequencing. Effects of intra-articular α-defensin-1 in vivo were evaluated in a rat meniscal/ligamentous injury (MLI) model. RESULTS: The quantity of M1 exceeded M2 polarized macrophages in human OA synovial tissue (mean difference 26.1% [13.6-38.6%], P < 0.001) and fluid (mean difference 10.5% [5.0-16.1%], P = 0.003). M1 to M2 polarization in vitro was most effectively promoted with 10 ng/mL α-defensin-1. Compared with untreated macrophages, the α-defensin-1 polarized macrophages modified co-cultured OA chondrocytes from a pro-catabolic state to a pro-anabolic (regenerative-like) state based on expression of COL2A1, ACN, MMP3, MMP13 and ADAMTS5. Intra-articular α-defensin-1 decreased severity of cartilage damage and synovitis in the MLI rat model. RNAseq analyses suggested insulin and Toll-like receptor signaling pathways in the chondroprotective α-defensin-1 mechanism of action. CONCLUSION: α-defensin-1 promotes M1 to M2 macrophage polarization in vitro, has beneficial effects on chondrocytes indirectly via M2 macrophage polarization, and attenuates the severity of OA in vivo, suggesting it might be a candidate treatment for OA.

[Risk factors of allogeneic red blood transfusion following enhanced recovery hip and knee arthroplasty].

Objective: To Explore the current status and risk factors of perioperative allogenic red blood cell transfusion following enhanced recovery hip and knee joint arthroplasty. Methods: Patients who have taken their primary unilateral total hip and knee arthroplasty (THA, TKA) or simultaneous primary THA were retrospectively included from January, 2019 to December, 2019 in West China Hospital. The baseline characteristics were compared between patients with allogeneic transfusion and those without. And logistic regression were used to analyze the risk factors of perioperative allogeneic red blood cell transfusion. Results: A total of 2 034 patients (2072 arthroplasties) were included, 705 males and 1 329 females, aged (60±24) years. Of all, 1 137 patients received primary THA (38 simultaneous THA), 897 patients received primary unilateral TKA. Eleven (0.54%) patients received allogeneic red blood cell transfusion, and the mean volume was (2.6±1.2) U. Deep venous thrombosis occurred in 2 patients (0.09%) undergoing primary TKA. The transfusion rate in primary THA patients was 0.79% (9/1 137), and 0.22% (2/897) in TKA. Lower preoperative hemoglobin level (P=0.041) and more hematological comorbidities (P=0.005) were detected in transfused patients. And logistic analysis further revealed that preoperative substandard hemoglobin level was the most important risk factor for transfusion (OR=5.663, P=0.018). Conclusions: Under the intervention of enhanced recovery after surgery concept and modern blood management strategies, the transfusion requirement has been significantly reduced following primary joint arthroplasty. Pre-operative hemoglobin level should be an important threshold for perioperative blood management.

The antifibrinolytic and anti-inflammatory effects of multiple doses of oral tranexamic acid in total knee arthroplasty patients: a randomized controlled trial.

Essentials Perioperative blood loss and inflammatory response can significantly affect recovery after surgery. We studied the effects of multiple-dose oral tranexamic acid on blood loss and inflammatory response. A postoperative four-dose regimen brought about maximum reduction in postoperative blood loss. A postoperative four-dose regimen reduced inflammatory response and promoted early rehabilitation. SUMMARY: Background Tranexamic acid (TXA) can reduce blood loss and the inflammatory response at multiple doses in total knee arthroplasty patients. However, the optimal regimen has not been determined. Objectives To identify the most effective regimen for achieving maximum reductions in blood loss and the inflammatory response. Patients/Methods Two hundred and seventy-five patients were randomized to receive a placebo (group A), a single 2-g oral dose of TXA 2 h preoperatively followed by 1 g of oral TXA 3 h postoperatively (group B), a single dose followed by 1 g of oral TXA 3 h and 7 h postoperatively (group C), a single dose followed by 1 g of oral TXA 3 h, 7 h and 11 h postoperatively (group D), or a single dose followed by 1 g of oral TXA 3 h, 7 h, 11 h and 15 h postoperatively (group E). The primary outcome was total blood loss on postoperative day (POD) 3. Secondary outcomes included a decrease in the hemoglobin level, coagulation parameters, inflammatory marker levels, and thromboembolic complications. Results Groups D and E had significantly lower blood loss and smaller decreases in hemoglobin level than groups A, B, and C, with no significant difference on POD 3 between groups D and E. Significantly enhanced coagulation was identified for the four multiple-dose regimens; however, all thromboelastographic parameters remained within normal ranges. Group E had the lowest inflammatory marker levels and pain, and the greatest range of motion. No thromboembolic complications were identified. Conclusion The four-dose regimen yielded the maximum reductions in blood loss and inflammatory response, improved analgesia, and promoted early rehabilitation. Further studies are required to ensure that these findings are reproducible.

Blood-conserving efficacy of multiple doses of oral tranexamic acid associated with an enhanced-recovery programme in primary total knee arthroplasty: a randomized controlled trial.

Aims: The aim of this study was to identify the most effective regimen of multiple doses of oral tranexamic acid (TXA) in achieving maximum reduction of blood loss in total knee arthroplasty (TKA). Patients and Methods: In this randomized controlled trial, 200 patients were randomized to receive a single dose of 2.0 g of TXA orally two hours preoperatively (group A), a single dose of TXA followed by 1.0 g orally three hours postoperatively (group B), a single dose of TXA followed by 1.0 g three and nine hours postoperatively (group C), or a single dose of TXA followed by 1.0 g orally three, nine, and 15 hours postoperatively (group D). All patients followed a routine enhanced-recovery protocol. The primary outcome measure was the total blood loss. Secondary outcome measures were hidden blood loss (HBL), reduction in the level of haemoglobin, the rate of transfusion and adverse events. Results: Groups C (661.1 ml, sd 262.4) and D (597.7 ml, sd 219.6) had significantly lower mean total blood loss compared with groups A and B. The mean HBL was significantly lower in groups B (699.2 ml), C (533.1 ml) and D (469.9 ml) than in group A (p = 0.006, p < 0.001, and p < 0.001, respectively). Groups C (2.22 ml, sd 0.91) and D (2.04 ml, sd 0.95) had a lower reduction in the level of haemoglobin than groups A and B. However, there were no differences between groups C and D in relation to the three parameters. Conclusion: The addition of two or three postoperative doses of TXA to one preoperative dose produced a significant reduction in blood loss. The two-dose postoperative regimen is the least necessary regimen for clinical efficacy in primary unilateral TKA. The three-dose regimen produced maximum reduction of blood loss. Cite this article: Bone Joint J 2018;100-B:1025-32.

[Gait analysis at the early stage after direct anterior approach in total hip arthroplasty].

OBJECTIVE: To evaluate the result of operation and gait analysis at the early stage after direct anterior approach (DAA) in total hip arthroplasty (THA). METHODS: In this study, 20 patients who suffered from necrosis of femoral head or developmental dysplasia of the hip were scheduled to undergo THA. The basic information and visual analogue scale (VAS) score, Harris score before and after surgery were recorded. All of the patients finished the gait analysis before the surgery and 6 weeks and 12 weeks after the surgery, the data were compared with those of normal adult people. RESULTS: Their hospital stay after the operation was 3.3 d, the VAS score after the operation was no more than 4 points, the positions of prosthesis were satisfactory, and there was no dislocation. The gait analysis results contained step speed, stride, the range of motion (ROM) of hip and knee. The step speed before the surgery (preoperation, Pre) was 0.64 m/s, 6 weeks after the surgery (6W) was 0.77 m/s, 12 weeks after the surgery (12W) was 1.07 m/s, and the control group was 1.19 m/s. The stride at Pre, 6W, 12W, and control group were 43.15 steps/min, 51.42 steps/min, 55.52 steps/min, and 57.15 steps/min, respectively. The ROM of hip joint at Pre, 6W, 12W, and control group were 31.00°, 39.62°, 40.40°, and 45.67°, respectively. The ROM of knee joint at Pre, 6W, 12W, and control group were 50.52°, 59.28°, 67.29°, and 70.42°, respectively. The results of the gait analysis showed that the gait recovery after the direct anterior total hip arthroplasty was very fast and at the 12th week after surgery the gait of the patients was close to the normal adult people. CONCLUSION: The direct anterior approach is one of the choosable approach of the THA, and this kind of surgery has a better recovery of gait after the operation, and at the end of 12 weeks after the surgery the gait is very close to the normal adult people. But we also need more studies to prove this conclusion.

Both systemic and local lipopolysaccharide (LPS) burden are associated with knee OA severity and inflammation.

OBJECTIVE: The microbiome is recognized as a new frontier in medicine with connections to a variety of diseases. We aimed to evaluate the association of lipopolysaccharide (LPS), a key pro-inflammatory product of the microbiome, with severity of inflammation, symptoms and radiographic abnormalities of knee osteoarthritis (OA). DESIGN: LPS was measured using a recombinant Factor C (rFC) assay, carefully optimized for systemic and synovial fluid (SF) analyses. LPS binding protein (LBP) was tested in both serum and SF of 25 patients (31 knees) from the Etarfolatide cohort for association with OA phenotypic outcomes. Models were adjusted for age, gender and body mass index. RESULTS: Based on LPS spike-and-recovery, both serum and SF dilutions of 0.1% were required to achieve recovery rates of at least 75% in all test specimens. Low coefficients of variation (CVs) (<10%) were achieved with both serum and SF dilutions <0.2%. Serum LPS and LBP were associated with the abundance of activated macrophages in the knee joint capsule and synovium. SF LPS and LBP were associated with the abundance of activated macrophages in the synovium. Serum LPS, LBP and SF LPS were associated with knee osteophyte severity. SF LPS was positively associated with knee joint space narrowing (JSN) severity and total WOMAC score. SF LBP was positively associated with self-reported knee pain score. CONCLUSION: These data strongly support a role for LPS in the pathogenesis and severity of structural abnormalities and symptoms of knee OA.

The arthropathic and functional impairment features of adult Kashin-Beck disease patients in Aba Tibetan area in China.

OBJECT: The purpose of the study was to analyze the features of arthropathic changes and functional impairments as well as the correlations between them for adult patients suffered with Kashin-Beck disease (KBD) in Aba Tibetan area of Sichuan Province, China. METHOD: Nine hundred and eighty-nine adult KBD patients in Aba KBD prevalence area were investigated. The arthropathic changes including arthritic pain (evaluated by visual analog pain score (VAS)), deformity, limited range of joint motion (ROM), as well as daily living and working function were examined, evaluated and analyzed. RESULTS: Ninety-two percent of patients suffered with multiple affected joints in both upper and lower extremities. The most frequently affected joints were knee (86.1%) and hand (77.2%). The most painful joints were knee (VAS 7.1 ± 1.9) and elbow (VAS 6.8 ± 2.1). Joint deformities most frequently represented as enlargement of interphalangeal joints (93.2%). Limitation of ROM occurred most frequently in hand (76.7%) and elbow (38.4%). Multiple linear regression analysis revealed that only joint pain (regression coefficient: -0.504, 95% confidence interval (CI): -0.820-0.188, P ＜ 0.001) and ROM (regression coefficient: 0.017, 95% CI: 0.011-0.024, P ＜ 0.001) were independent risk factors affecting daily living and working function. CONCLUSION: Most adult patients suffered with multiple affected joints in both upper and lower limbs. The elbow, hand and knee were the most frequently and severely affected joints. The pain and limited ROM were the independent risk factors of daily living and working function.

Relationship between the angle of vertebral screws and spinal lateral angulation after fixation of thoracolumbar fractures via an anterior approach.

This study investigated possible contributors to lateral spinal angulation after surgical fixation of thoracolumbar fractures via an anterior approach. We retrospectively examined lateral angulation in 172 cases of thoracolumbar fractures treated in this manner. The coronal Cobb angle and angles of the screws relative to the endplates were determined from radiographs. The patients completed the Short Form 36, Oswestry Disability Index, Japanese Orthopaedic Association Back Pain Evaluation Questionnaire, and Visual Analogue Scale at the final follow-up visit. The mean coronal Cobb angle was 0.75° ± 3.91° (-14.25° to 14.55°) preoperatively, 3.17° ± 4.07° (-8.18° to 14.01°) immediately postoperatively, and 3.46° ± 4.21° (-1.05° to 17.27°) at the final follow-up visit. The superior posterior and inferior anterior screws were more parallel to their respective endplates when the approach was made ≥2 vs ≤1 vertebral levels above the fracture (P < 0.001). Lateral angulation was more likely when the approach was made ≤1 vs ≥2 levels above the fracture (P < 0.001). The coronal Cobb angle differed significantly (P < 0.01) between patients with lumbar and thoracic fractures. The immediate postoperative coronal Cobb angle correlated tightly with the sum of the screw angles (superior plus inferior posterior and/or inferior plus superior anterior). Lateral angulation may occur after surgical fixation of thoracic and lumbar fractures via an anterior approach. Non-parallelism between the vertebral screws and their corresponding endplates may predict postoperative lateral spinal angulation. Postoperative lateral angulation does not correlate with low back pain, quality of life, or preoperative lateral angulation.

Targeted antiosteosarcoma methotrexate-bisphosphonate conjugate induces apoptosis of osteosarcoma cells in vitro.

OBJECTIVES: To investigate the effect of targeted antiosteosarcoma methotrexate-bisphosphonate conjugate on growth inhibition and apoptosis in human osteosarcoma MG-63 cells. MATERIALS AND METHODS: MG-63 cells were treated with various concentrations of methotrexate-bisphosphonate conjugate and apoptosis was monitored via an MTT assay, cell morphology, TUNEL assay and flow cytometry analysis. RESULTS: The survival rate of MG-63 cells treated for 24 to 96 hours with 2000 mg/ml or more of methotrexate-bisphosphonate conjugate decreased significantly. Cells treated with conjugate showed typical apoptotic features using inverted phase contrast microscopy and fluorescence staining, and the majority of cells demonstrated a positive result in the TUNEL assay. Karyopyknosis and crescent aggregation of chromatin were observed in conjugate-treated cells by electron microscopy. Flow cytometry of MG-63 cells treated with methotrexate-bisphosphonate conjugate showed a time and dose-dependent increase in apoptosis (p < 0.05). CONCLUSIONS: A targeted antiosteosarcoma methotrexate-bisphosphonate conjugate induces apoptosis in human osteosarcoma MG-63 cells. This new conjugate is a valuable experimental tool for the therapy of osteosarcoma.

Is there reduced polyethylene wear and longer survival when using a mobile-bearing design in total knee replacement? A meta-analysis of randomised and non-randomised controlled trials.

The purpose of this study was to undertake a meta-analysis to determine whether there is lower polyethylene wear and longer survival when using mobile-bearing implants in total knee replacement when compared with fixed-bearing implants. Of 975 papers identified, 34 trials were eligible for data extraction and meta-analysis comprising 4754 patients (6861 knees). We found no statistically significant differences between the two designs in terms of the incidence of radiolucent lines, osteolysis, aseptic loosening or survival. There is thus currently no evidence to suggest that the use of mobile-bearing designs reduce polyethylene wear and prolong survival after total knee replacement.

Preventive effects of supplemental selenium and selenium plus iodine on bone and cartilage development in rats fed with diet from Kashin-Beck disease endemic area.

The purpose of this study was to investigate the effects of supplemental selenium and selenium plus iodine on bone and growth plate cartilage histology and serum biochemistic parameters in rats. Ninety-six Wistar rats were randomly divided into the following four groups: group A, the rats fed with normal diet; group B, fed with diet from Kashin-Beck disease (KBD) endemic area; group C, fed with diet from KBD endemic area supplemented with selenium; and group D, fed with diet from KBD endemic area supplemented with selenium and iodine. After 4, 8, and 12 weeks, bone and cartilage samples were collected from the rats and were examined for morphological changes in the tibial growth zone and for changes in the plate cartilage and metaphysic. Compared to the rats fed with diet from the KBD endemic area, the rats fed with the supplemental selenium or selenium plus iodine exhibited diminished necrosis of the chondrocytes in the growth plate. In the groups of rats receiving supplemental selenium and selenium plus iodine, the bone volume/tissue volume ratio (BV/TV), the trabecular thickness (Tb.Th), and the trabecular number were increased, while the trabecular separation was decreased. In the 12th week of the experiment, BV/TV and Tb.Th were significantly increased in the selenium plus iodine group compared to the selenium group. It is concluded that feeding the diet from the KBD endemic area caused necrosis of chondrocytes and dysfunctions of bone development similar to the pathological changes that are seen in KBD. Selenium and iodine protected chondrocytes in growth plate and promoted the formation of trabecular bone. The effects of selenium plus iodine on bone formation were more obvious than those of selenium alone.

Circumferential osteotomy of the medial acetabular wall in total hip replacement for the late sequelae of childhood septic arthritis of the hip.

We performed 52 total hip replacements in 52 patients using a cementless acetabular component combined with a circumferential osteotomy of the medial acetabular wall for the late sequelae of childhood septic arthritis of the hip. The mean age of the patients at operation was 44.5 years (22 to 66) and the mean follow-up was 7.8 years (5 to 11.8). The mean improvement in the Harris Hip Score was 29.6 points (19 to 51) at final follow-up. The mean cover of the acetabular component was 98.5% (87.8% to 100%). The medial acetabular wall was preserved with a mean thickness of 8.3 mm (1.7 to 17.4) and the mean length of abductor lever arm increased from 43.4 mm (19.1 to 62) to 54.2 mm (36.5 to 68.6). One acetabular component was revised for loosening and osteolysis 4.5 years postoperatively, and one had radiolucent lines in all acetabular zones at final review. Kaplan-Meier survival was 94.2% (95% confidence interval 85.8% to 100%) at 7.3 years, with revision or radiological loosening as an end-point when two hips were at risk. A cementless acetabular component combined with circumferential medial acetabular wall osteotomy provides favourable results for acetabular reconstruction in patients who present with late sequelae of childhood septic hip arthritis.

Beta-CTX and ICTP act as indicators of skeletal metastasis status in male patients with non-small cell lung cancer.

Bone metastasis is common in lung cancer patients and associated with reduced quality of life and reduced overall and median survival, so the early detection of bone metastasis and monitoring of its status is very important for clinicians. Serum bone-specific alkaline phosphatase (BAP), osteocalcin (OC), beta isomer of C-terminal telopeptide of type I collagen (beta-CTX) and cross-linked C-terminal telopeptide of type I collagen (ICTP) were compared with regard to their usefulness as indicators of bone metastasis in lung cancer. The serum concentrations of the 4 markers were measured by commercially available tests in 96 male patients with non-small cell lung cancer and 30 male patients with other pulmonary diseases. The levels of both â-CTX and ICTP were significantly higher in 61 lung cancer patients with bone metastases than in 35 lung cancer patients without bone metastases (both p<0.001), and significantly correlated with the extent of bone disease. Although ICTP had a better sensitivity and accuracy than beta-CTX (75.4% vs 65.6% and 72.9% vs 68.8%, respectively), they had a similar area under the receiver operating characteristic curve (0.85 vs 0.83). These results support the use of beta-CTX and ICTP as an adjunct tool for the diagnosis and screening of bone metastasis in lung cancer.