Cartilage Signal Research Articles

Triboelectric nanogenerator based on well-dispersed and oxide-free liquid metal-doped conductive hydrogel as self-powered wearable sensor for respiratory and thyroid cartilage signal monitoring

The poor dispersibility and easy surface oxidation of liquid metal (LM) significantly reduce the conductivity, mechanical property and stability of the composite hydrogels, thereby affecting their sensing performance. Herein, a well-dispersed and oxide-free LM-doped conductive double network (DN) hydrogel (CMC/P(AA-co-DAC)/NaCl-LM) is successfully prepared by employing bio-macromolecules (carboxymethyl cellulose (CMC) and double-bond functionalized CMC (CMC-AGE crosslinker) encapsulated oxide-free LM droplet at acidic condition as the first network and poly(acrylic acid-co-acryloyloxyethyltrimethyl ammonium chloride) (P(AA-co-DAC)) as the second network under the presence of NaCl. In this hydrogel, the multiple interactions and the synergistic effect between two interpenetrating networks insures multifunctional features of the hydrogels. As a result, the composite hydrogel exhibits superior mechanical properties (stress of 389.81 kPa and strain of 2866.44%) and high conductivity (5.27S/m). Subsequently, a triboelectric nanogenerator (CPL-TENG) is constructed using the composite hydrogel as electrode material, which displayed an excellent electrical out performance (VOC of 223.32V, ISC of 9.78 μA, QSC of 82.33 nC, and power density of 4.30W/m2) and highly-sensitive (pressure sensitivity of 0.268 kPa-1). Eventually, the as-fabricated CPL-TENG as a self-powered sensor for respiratory signal monitoring and machine learning-assisted language recognition is described and its great potential in wearable electronics and energy supply devices is foreseen.

7434 Modulation of FGFR-FGF2-FGF23 Signaling Partially Rescues Osteoarthritis in Hyp Mice Homolog of Human X- Linked Hypophosphatemia

Abstract Disclosure: M.M. Hurley: None. L. Xiao: None. Human with X-linked hypophosphatemia (XLH) and its Hyp mice homolog develop several phenotypic abnormalities due to phosphate wasting mediated by abnormal Fibroblast Growth Factor Receptor-1 (FGFR1) and Fibroblast Growth Factor 23 (FGF23) signaling. XLH and Hyp mice also develop severe osteoarthropathy (OA) whose etiology is not fully defined. We previously reported that Fibroblast growth factor 2 (FGF2) regulates FGF23, specifically, mice overexpressing the high molecular weight FGF2 isoforms (HMWFGF2) in osteoprogenitors phenocopy Hyp mice including development of OA that worsens with age. Since we also reported that Hyp mice overexpress HMWFGF2 isoforms in osteoblasts and osteocytes, we examined whether the knee OA phenotype in Hyp mice was associated with aberrant FGFR1, FGF2, FGF23 signaling in knee joint articular cartilage and subchondral bone and whether in vivo FGFR tyrosine kinase inhibitor BGJ398 would modulate the OA phenotype in knee joints of Hyp mice. Ten weeks-old Control and Hyp mice were treated with Vehicle or BGJ398 for 12 weeks. Safranin-O staining of knee articular cartilage for proteoglycan revealed decreased staining in Hyp vehicle-treated mice that was partially restored to the cartilage thickness of Control mice in the Hyp-BGJ398-treated group. Significantly increased degradative enzyme matrix metalloproteinase-13 (MMP-13) in Hyp vehicle-treated knee articular cartilage was significantly reduced by BGJ398. Immunostaining revealed significantly increased phosphorylated FGFR1, FGF2 and FGF23 in both articular cartilage and subchondral bone. While BGJ398 significantly reduced immunostaining for phosphoFGFR1 and FGF2 in both articular cartilage and subchondral bone, it significantly reduced FGF23 in subchondral bone but not in articular cartilage. Significantly increased phospho-ERK1/2 labeling in Hyp-vehicle treated articular cartilage was partially rescued by BGJ398. This study demonstrates that increased FGFR1/FGF2 signaling contributes to OA in Hyp mice that can be partially rescued by FGF receptor inhibitor via reduction in MAPK signaling. Presentation: 6/1/2024

Cbfβ regulates Wnt/β-catenin, Hippo/Yap, and Tgfβ signaling pathways in articular cartilage homeostasis and protects from ACLT surgery-induced osteoarthritis.

As the most common degenerative joint disease, osteoarthritis (OA) contributes significantly to pain and disability during aging. Several genes of interest involved in articular cartilage damage in OA have been identified. However, the direct causes of OA are poorly understood. Evaluating the public human RNA-seq dataset showed that CBFB (subunit of a heterodimeric Cbfβ/Runx1, Runx2, or Runx3 complex) expression is decreased in the cartilage of patients with OA. Here, we found that the chondrocyte-specific deletion of Cbfb in tamoxifen-induced Cbfbf/f;Col2a1-CreERT mice caused a spontaneous OA phenotype, worn articular cartilage, increased inflammation, and osteophytes. RNA-sequencing analysis showed that Cbfβ deficiency in articular cartilage resulted in reduced cartilage regeneration, increased canonical Wnt signaling and inflammatory response, and decreased Hippo/Yap signaling and Tgfβ signaling. Immunostaining and western blot validated these RNA-seq analysis results. ACLT surgery-induced OA decreased Cbfβ and Yap expression and increased active β-catenin expression in articular cartilage, while local AAV-mediated Cbfb overexpression promoted Yap expression and diminished active β-catenin expression in OA lesions. Remarkably, AAV-mediated Cbfb overexpression in knee joints of mice with OA showed the significant protective effect of Cbfβ on articular cartilage in the ACLT OA mouse model. Overall, this study, using loss-of-function and gain-of-function approaches, uncovered that low expression of Cbfβ may be the cause of OA. Moreover, Local admission of Cbfb may rescue and protect OA through decreasing Wnt/β-catenin signaling, and increasing Hippo/Yap signaling and Tgfβ/Smad2/3 signaling in OA articular cartilage, indicating that local Cbfb overexpression could be an effective strategy for treatment of OA.

A loss of primary cilia by a reduction in mTOR signaling correlates with age-related deteriorations in condylar cartilage.

Age-related deterioration of condylar cartilage is an etiological factor in temporomandibular joint-osteoarthritis (TMJ-OA). However, its underlying mechanism remains unknown. Therefore, we examined age-related changes and the relationship between mTOR signaling and primary cilia in condylar cartilage to determine the intrinsic mechanisms of age-related TMJ-OA. Age-related morphological changes were analyzed using micro-computed tomography and safranin O-stained histological samples of the mandibular condyle of C57BL/6J mice (up to 78weeks old). Immunohistochemistry was used to assess the activity of mTOR signaling, primary cilia frequency, and Golgi size of condylar chondrocytes. Four-week-old mice receiving an 11-week series of intraperitoneal injections of rapamycin, a potent mTOR signaling inhibitor, were used for the histological evaluation of the condylar cartilage. The condylar cartilage demonstrated an age-related reduction in cartilage area, including chondrocyte size, cell density, and cell size distribution. The Golgi size, primary cilia frequency, and mTOR signaling also decreased with age. Rapamycin injections resulted in both diminished cartilage area and cell size, resembling the phenotypes observed in aged mice. Rapamycin-injected mice also exhibited a smaller Golgi size and lower primary cilia frequency in condylar cartilage. We demonstrated that a loss of primary cilia due to a decline in mTOR signaling was correlated with age-related deteriorations in condylar cartilage. Our findings provide new insights into the tissue homeostasis of condylar cartilage, contributing to understanding the etiology of age-related TMJ-OA.

ABNORMAL CHONDROCYTE MORPHOLOGY, CLUSTERING, AND CELL SWELLING IN A SCRAPED HUMAN CARTILAGE MODEL OF OSTEOARTHRITIS

AbstractObjectivesOsteoarthritis (OA) is a complex joint disorder characterised by the loss of extracellular matrix (ECM) leading to cartilage degeneration. Changes to cartilage cell (chondrocyte) behaviour occur including cell swelling, the development of fine cytoplasmic processes and cell clustering leading to changes in cell phenotype and development of focal areas of mechanically-weak fibrocartilaginous matrix[1]. To study the sequence of events in more detail, we have investigated the changes to in situ chondrocytes within human cartilage which has been lightly scraped and then cultured with serum.MethodsHuman femoral heads were obtained with Ethical permission and consent from four female patients (mean age 74 yrs) undergoing hip arthroplasty following femoral neck fracture. Osteochondral explants of macroscopically-normal cartilage were cultured as a non-scraped control, or scraped gently six times with a scalpel blade and both maintained in culture for up to 2wks in Dulbecco's Modified Eagle's Medium (DMEM) with 25% human serum (HS). Explants were then labelled with CMFDA (5-chloromethylfluorescein-diacetate) and PI (propidium iodide) (10μM each) to identify the morphology of living or dead chondrocytes respectively. Explants were imaged using confocal microscopy and in situ chondrocyte morphology, volume and clustering assessed quantitatively within standardised regions of interest (ROI) using Imaris® imaging software.ResultsWithin 2wks of culture with HS, chondrocyte volume increased significantly from 412±9.3µm3 (unscraped) at day 0 to 724±16.6 µm3 (scraped) [N(n) = 4(380)] (P=0.0002). Chondrocyte clustering was a prominent feature of HS culture as the percentage of clusters in the cell population increased with scraping from 4.8±1.4% to 14.9±3.9% [N(n) = 4(999)] at week 2 (P=0.0116). In addition, the % of the chondrocyte population within clusters increased from approximately 38% to 60%, and the number of cells per cluster increased significantly from 3.2±0.08 to 4±0.22 (P=0.031). The development of abnormal ‘fibroblastic-like’ chondrocyte morphology demonstrating long (>5µm) cytoplasmic processes also occurred, however the time course of this was more variable. For some samples, clustering occurred before abnormal morphology, but for others the opposite occurred. Typically, by the second week, 17±2.64% of the cell population had processes and this increased to 22±4.02% [N(n) = 4(759)] with scraping.ConclusionsScraping the cartilage will remove surface constituents including lubricants (e.g. lubricin, hyaluronic acid, phospholipids), extracellular matrix constituents (collagen, proteoglycans – potentially the ‘lamina splendens’) and cells (chondrocytes and mesenchymal stromal cells (MSCs)). Although we do not know which of these component(s) is important, the effect is to dramatically increase the permeation of serum factors into the cartilage matrix and signal the development of cytoplasmic processes, cell clustering and swelling. It is notable that these cellular changes are similar to those occurring in early OA[1]. This raises the interesting possibility that scraped cartilage cultured with human serum recapitulates some of the changes to in situ chondrocytes during early stages of cartilage degeneration and as such, could be a useful model for following the deleterious changes to matrix metabolism.Declaration of Interest(b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project.

IL-6 Reduces Spheroid Sizes of Osteophytic Cells Derived from Osteoarthritis Knee Joint via Induction of Apoptosis

Osteophytes in osteoarthritis (OA) joints contribute to restriction of joint movement, joint pain, and OA progression, but little is known about osteophyte regulators. Examination of gene expression related to cartilage extracellular matrix, endochondral ossification, and growth factor signaling in articular cartilage and osteophytes obtained from OA knee joints showed that several genes such as COL1A1, VCAN, BGLAP, BMP8B, RUNX2, and SOST were overexpressed in osteophytes compared with articular cartilage. Ratios of mesenchymal stem/progenitor cells, which were characterized by co-expression of CD105 and CD166, were significantly higher in osteophytic cells than articular cells. A three-dimensional culture method for cartilage and osteophyte cells was developed by modification of cultures of self-assembled spheroid cell organoids (spheroids). These spheroids cultured in the media for mesenchymal stem cells containing transforming growth factor-β3 showed characteristic morphologies and gene expression profiles of articular cartilage and osteophytes, respectively. The effects of IL-1β, tumor necrosis factor-α, and IL-6 on the spheroids of articular and osteophytic cells were studied. To the best of our knowledge, they provide the first evidence that IL-6 suppresses the spheroid size of osteophytic cells by inducing apoptosis and reducing extracellular matrix molecules. These data show that IL-6 is the suppressor of osteophyte growth and suggest that IL-6 expression and/or activity are implicated in the regulation of osteophyte formation in pathologic joints.

Chondral Injury Associated With ACL Injury: Assessing Progressive Chondral Degeneration With Morphologic and Quantitative MRI Techniques.

Anterior cruciate ligament (ACL) injuries are associated with a risk of post-traumatic osteoarthritis due to chondral damage. Magnetic resonance imaging (MRI) techniques provide excellent visualization and assessment of cartilage and can detect subtle and early chondral damage. This is often preceding clinical and radiographic post-traumatic osteoarthritis. Morphologic and quantitative MRI techniques can assess early and progressive degenerative chondral changes after acute ACL injury. Prospective longitudinal cohort. Level 3. Sixty-five participants with acute unilateral ACL injuries underwent bilateral knee MRI scans within 1 month of injury. Fifty-seven participants presented at 6 months, while 54 were evaluated at 12 months. MRI morphologic evaluation using a modified Noyes score assessed cartilage signal alteration, chondral damage, and subchondral bone status. Quantitative T1ρ and T2 mapping at standardized anatomic locations in both knees was assessed. Participant-reported outcomes at follow-up time points were recorded. Baseline Noyes scores of MRI detectable cartilage damage were highest in the injured knee lateral tibial plateau (mean 2.5, standard error (SE) 0.20, P < 0.01), followed by lateral femoral condyle (mean 2.1, SE 0.18, P < 0.01), which progressed after 1 year. Longitudinal prolongation at 12 months in the injured knees was significant for T1ρ affecting the medial and lateral femoral condyles (P < 0.01) and trochlea (P < 0.01), whereas T2 values were prolonged for medial and lateral femoral condyles (P < 0.01) and trochlea (P < 0.01). The contralateral noninjured knees also demonstrated T1ρ and T2 prolongation in the medial and lateral compartment chondral subdivisions. Progressive chondral damage occurred despite improved patient-reported outcomes. After ACL injury, initial and sustained chondral damage predominantly affects the lateral tibiofemoral compartment, but longitudinal chondral degeneration also occurred in other compartments of the injured and contralateral knee. Early identification of chondral degeneration post-ACL injury using morphological and quantitative MRI techniques could enable interventions to be implemented early to prevent or delay PTOA.

Clinical and radiological efficacy of single-dose intra-articular high-molecular-weight hyaluronic acid in knee osteoarthritis.

While visco-supplementation is being used for the treatment of knee osteoarthritis (OA), the published reports vary widely in benefits afforded by this treatment. It was therefore proposed to assess the objective parameters along with subjective outcomes. Our study assessed the radiological and clinical efficacy of single-dose high-molecular-weight intra-articular hyaluronic acid (HMW-IAHA) injection in knee OA. This interventional cohort study was conducted in a calculated sample size of 44 patients with knee OA. Visual analog scale (VAS) and knee OA and outcome score (KOOS) were used for clinical assessment, and whole organ magnetic resonance imaging score (WORMS) for radiological assessment. The subjects were given a single dose of HMW-IAHA injection, 90 mg/3 ml, and were assessed on day 0 and day 90. Statistical Package for Social Sciences (SPSS) software was used. At the day 90 follow-up, there was an improvement in mean ± standard deviation values of VAS score (day 0: 8.53 ± 0.81, day 90: 5.97 ± 0.87), KOOS score (day 0: 27.33 ± 15.18, day 90: 57.26 ± 14.26), and the cartilage signal and morphology in the medial femorotibial joint (day 0: 11.02 ± 6.26 and day 90: 10.91 ± 6.22) and patellofemoral joint (day 0: 10.35 ± 4.36 and day 90: 10.28 ± 4.39) compartments. There was a decrease in synovitis score from 2.3 ± 1.61 to 1.3 ± 1.3 in the medial femorotibial joint compartment and total WORMS score (day 0: 66.57 ± 36.06, day 90: 65.14 ± 35.62). A single dose of intra-articular injection with high-molecular-weight hyaluronic acid produces improvement in the clinical symptoms and quality of life as well as is effective in maintaining the articular cartilage integrity and reducing synovial inflammation.

Inhibition of histone lysine demethylase 6A promotes chondrocytic activity and attenuates osteoarthritis development through repressing H3K27me3 enhancement of Wnt10a.

Histone hypermethylation represses gene transcription, which affects cartilage homeostasis or joint remodeling. Trimethylation of lysine 27 of histone 3 (H3K27me3) changes epigenome signatures, regulating tissue metabolism. This study aimed to investigate whether loss of H3K27me3 demethylase Kdm6a function affected osteoarthritis development. We revealed that chondrocyte-specific Kdm6a knockout mice developed relatively long femurs and tibiae as compared to wild-type mice. Kdm6a deletion mitigated osteoarthritis symptoms, including articular cartilage loss, osteophyte formation, subchondral trabecular bone loss, and irregular walking patterns of destabilized medial meniscus-injured knees. In vitro, loss of Kdm6a function compromised the loss in expression of key chondrocyte markers Sox9, collagen II, and aggrecan and improved glycosaminoglycan production in inflamed chondrocytes. RNA sequencing showed that Kdm6a loss changed transcriptomic profiles, which contributed to histone signaling, NADPH oxidase, Wnt signaling, extracellular matrix, and cartilage development in articular cartilage. Chromatin immunoprecipitation sequencing uncovered that Kdm6a knockout affected H3K27me3 binding epigenome, repressing Wnt10a and Fzd10 transcription. Wnt10a was, among others, functional molecules regulated by Kdm6a. Forced Wnt10a expression attenuated Kdm6a deletion-induced glycosaminoglycan overproduction. Intra-articular administration with Kdm6a inhibitor GSK-J4 attenuated articular cartilage erosion, synovitis, and osteophyte formation, improving gait profiles of injured joints. In conclusion, Kdm6a loss promoted transcriptomic landscapes contributing to extracellular matrix synthesis and compromised epigenetic H3K27me3-mediated promotion of Wnt10a signaling, preserving chondrocytic activity to attenuate osteoarthritic degeneration. We highlighted the chondroprotective effects of Kdm6a inhibitor for mitigating the development of osteoarthritic disorders.

Zinc finger E-box binding homebox 2 alleviated experimental osteoarthritis in rats

ABSTRACT Aims Osteoarthritis (OA) is a chronic inflammatory disease, characterized by degradation of extracellular matrix, apoptosis of chondrocytes and inflammation in articular cartilage. Zinc finger E-box binding homebox 2 (ZEB2), a transcription repressor, has been demonstrated with anti-inflammatory role in some cells. The analysis from GEO data reveals that ZEB2 expression is upregulated in articular cartilage of OA patients and experimental OA rodents. This study aims to confirm the function of ZEB2 in OA process. Material and methods The experimental OA was induced by anterior cruciate ligament transaction (ACLT) in rats, and the adenovirus loaded with ZEB2 coding sequence was intra-articularly injected into rats (1 × 10 PFU). The primary articular chondrocytes were stimulated by interleukin-1β (IL-1β) (10 ng/ml) to mimic the osteoarthritic injury, and transfected with the adenovirus carrying ZEB2 coding or silencing sequence. The apoptosis, content of extracellular matrix, inflammation, and the activity of NFκB signaling in chondrocytes and cartilage were determined. Results ZEB2 was highly expressed in osteoarthritic cartilage tissues and IL-1β-treated chondrocytes. The overexpression of ZEB2 restrained ACLT- or IL-1β administration-induced apoptosis, matrix degradation and inflammation in vivo or in vitro, evidenced by changed levels of cleaved caspase-3/PARP, collagen-II, aggrecan, matrix metalloproteinase 3/13, tumor necrosis factor-α and interleukin-6. Additionally, the phosphorylation of NFκB p65, IκBα and IKKα/β, and the nuclear translocation of p65 was blocked by ZEB2, implying inactivation of this signaling. Conclusions ZEB2 mitigated osteoarthritic symptoms in rats and chondrocytes, and NFκB signaling might be involved. These findings may provide novel insights for clinical treatment of OA.

Cartilage-specific deficiency of clock gene Bmal1 accelerated articular cartilage degeneration in osteoarthritis by up-regulation of mTORC1 signaling.

Although a growing body of studies recently demonstrated that circadian clock gene Bmal1 plays an important role in cartilage development and homeostasis, evidence regarding the contribution of Bmal1 in articular cartilage of OA progression is still unclear. In the present study, we investigated the direct role of Bmal1 in articular cartilage homeostasis during OA progression using tamoxifen-induced cartilage-specific knockout mice. We found that the expression of BMAL1 was decreased in OA-damaged and aging cartilage tissues. Cartilage-specific deletion of Bmal1 promoted cartilage degradation and chondrocyte apoptosis, and inhibited chondrocyte anabolism in OA mice, leading to acceleration of articular cartilage degeneration and osteophyte formation during OA progression. Mechanistic study indicated that loss of Bmal1 resulted in hyperactivation of mammalian target of rapamycin complex 1(mTORC1) signaling in OA cartilage, and pharmacological inhibition of mTORC1 signaling pathway by rapamycin alleviated partially Bmal1 ablation-induced cartilage degradation and chondrocyte apoptosis in ex vivo OA model. Therefore, our results provide the evidence of a vital role for Bmal1 in cartilage degradation in post-traumatic OA by partially regulating the mTORC1 signaling.

Integrin αVβ3 Signaling in the Progression of Osteoarthritis Induced by Excessive Mechanical Stress

Osteoarthritis (OA) is believed to be linked with cartilage degeneration, subchondral bone sclerosis, and synovial inflammation that lead to joint failure, and yet treatment that can effectively reverse the pathological process of the disease still not exists. Recent evidence suggests excessive mechanical stress (eMS) as an essential role in the pathogenesis of OA. Increased levels of integrin αVβ3 have been detected in osteoarthritic cartilage and were previously implicated in OA pathogenesis. However, the role of integrin αVβ3 in the process of eMS-induced OA remains unclear. Here, histologic and proteomic analyses of osteoarthritic cartilage in a rat destabilization of the medial meniscus model demonstrated elevated expression of integrin αVβ3 as well as more serious cartilage degeneration in the medial weight-bearing area. Furthermore, results of in vitro study demonstrated that eMS led to a significant increase of integrin αVβ3 expression and phosphorylation of downstream signaling molecules such as FAK and ERK, as well as upregulated expressions of inflammatory and degradative mediators. In addition, we found that inhibition of integrin αVβ3 could alleviate chondrocyte inflammation triggered by eMS both in vivo and in vitro. Our findings suggest a central role for upregulation of integrin αVβ3 signaling in OA pathogenesis and demonstrate that activation of integrin αVβ3 signaling in cartilage contributes to inflammation and joint destruction in eMS-induced OA. Taken together, our data presented here provide a possibility for targeting integrin αVβ3 signaling pathway as a disease-modifying therapy.

Tailoring Brachytherapy by Identification of Fibrosis vs. Tumor: Evaluating Acute and Chronic Fibrosis via UTE MRI in Gynecological Cancer

<h3>Purpose/Objective(s)</h3> Radiation injury triggers inflammation, fibroblast migration, and deposition of collagen (acute fibrosis) that later remodels into denser matrices forming scar (chronic fibrosis) at the previous location of tumor. Non-contrast enhanced Short-Inversion-Time (STIR) ultra-short echo time (UTE) MRI has been used to quantify collagen fractions in collagen/aqueous-solution mixtures and to evaluate acute fibrosis. Late-gadolinium enhanced (LGE) IR UTE MRI is a technique that has been used to evaluate chronic scar in the heart. The role of these UTE MR techniques to monitor evolving fibrosis over the course of radiation in cancer has not been studied. We hypothesize that the percentage of fibrosis relative to initial tumor volume increases over the course of external beam radiation prior to brachytherapy in gynecological cancer. <h3>Materials/Methods</h3> <i>Subjects:</i> Cervical and vaginal cancer patients undergoing concurrent chemoradiation were included. MR images were acquired pre- and 1 day post 45 Gy EBRT. <i>Image Acquisition</i>: Imaging was performed on 1.5T MRI using a prototype stack of spirals dual echo (TE_1,2=50, 2690µs) STIR UTE sequence with TI=60, or 80ms. To evaluate acute (diffuse) fibrosis, which has a short-TE, TE₂ images were subtracted from TE₁ images. Chronic fibrosis (necrotic scar) was evaluated using TE₁ of LGE IR UTE, TI=200ms, acquired ≥10 min after contrast injection. <i>Image Analysis:</i> All images were rigidly registered to the treatment planning CT. EBRT CTV contours were propagated onto the MR images. STIR-UTE images were segmented using region-growing. Acute fibrosis (F_A) and chronic fibrosis (F_C) contours were segmented on the non-contrast STIR UTE and on the LGE UTE, respectively. Fibrosis signals were normalized between successive scans based on the femoral heads' cartilage signal. Percentages of the F_A and F_C within the CTV were determined and compared pre- and post-EBRT using a paired t-test. <h3>Results</h3> In five cervical cancer and two vaginal cancer patients, F_A was observed in the irradiated tumor and lymph nodes. The extent of baseline F_A vs. F_C was not significantly different (p=.2) (Table 1). Baseline F_A was significantly lower than post-treatment F_A within the CTV (pre-treatment/post-treatment: 11±11%/34±15%, p=.01). F_C was significantly higher (pre-treatment/post-treatment: 5±2%/18±10%, p=.01). Post-EBRT, there was a significant difference between F_A and F_C within the CTV (p=.01) suggesting a faster increase in acute fibrosis as compared to scar, which requires more time to mature. <h3>Conclusion</h3> We observed an increase in F_A and F_C post-EBRT within the CTV. F_A can serve as an early indicator of F_C. Identifying regions of fibrosis has the potential to guide brachytherapy treatment planning. Future work will focus on precisely prescribing high-doses of brachytherapy to remnant tumor.

Comparison of Arthroscopic, X-ray and MRI T2 Mapping Findings for Varus Ankle Osteoarthritis

Category: Ankle Arthritis; Arthroscopy Introduction/Purpose: We have previously reported the indication criteria for supramalleolar osteotomy for varus ankle osteoarthritis using our arthroscopic classification. However arthroscopy is invasive, MRI T2 mapping (T2M) is also useful for noninvasive preoperative assessment of articular cartilage condition. The purpose of this study is to investigate the relationship among these. Methods: The subjects were 19 patients with 19 joints, 3 males and 16 females, mean age 65.2 years (45-78), 3 joints in stage 2, 8 joints in stage 3a, and 8 joints in stage 3b according to radiographic classification by Takakura et al. Measurements were taken on the coronal section of the talar dome. To evaluate the arthroscopic findings, we used our five grades classification, which is based on the percentage of eburnation areas on the articular surface, we measured the percentage of the area of the missing joint space to the width of the talar dome in X-rays and the ratio of the area of cartilage signal loss to the width of the talar dome in T2M. We determined how well the radiographic and T2M findings match the arthroscopic findings. Results: The results showed that there were 2 joints in grade 2, 15 joints in grade 3, and 2 joints in grade 4 in arthroscopic classification. The concordance rate between arthroscopy and X-rays measurements was 100% (2 joints/2 joints) for grade 2, 46.7% (7/15) for grade 3, and 0% (0/2) for grade 4. In all 10 unmatched joints, X-rays underestimated the condition of the articular cartilage compared to arthroscopic findings. The concordance rate between arthroscopy and T2M measurements was 50% (1/2) for grade 2, 80% (12/15) for grade 3, and 100% (2/2) for grade 4. Two joints were underestimated and two were overestimated by T2M compared to the arthroscopic findings. Conclusion: Compared to arthroscopic findings, X-rays tended to underestimate the condition of articular cartilage, while T2M tended to overestimate it, but in general, T2M showed a higher concordance rate with the arthroscopic findings than X-rays. T2M is more useful than X-rays as a tool for cartilage evaluation to determine the operative method for varus ankle osteoarthritis.

Artemisinin relieves osteoarthritis by activating mitochondrial autophagy through reducing TNFSF11 expression and inhibiting PI3K/AKT/mTOR signaling in cartilage

Osteoarthritis (OA) is a widespread chronic degenerative joint disease characterized by the degeneration of articular cartilage or inflamed joints. Our findings indicated that treatment with artemisinin (AT) downregulates the protein levels of MMP3, MMP13, and ADAMTS5, which are cartilage degradation-related proteins in OA, and inhibits the expression of inflammatory factors in interleukin-1β (IL-1β)-stimulated chondrocytes. However, the mechanism of the role of AT in OA remains unclear. Here, we performed gene sequencing and bioinformatics analysis in control, OA, and OA + AT groups to demonstrate that several mRNA candidates were enriched in the PI3K/AKT/mTOR signaling pathway, and TNFSF11 was significantly downregulated after AT treatment. TNFSF11 was downregulated in the OA + AT group, whereas it was upregulated in rat OA tissues and OA chondrocytes. Therefore, we confirmed that TNFSF11 was the target gene of AT. In addition, our study revealed that AT relieved cartilage degradation and defection by activating mitochondrial autophagy via inhibiting the PI3K/AKT/mTOR signaling pathway in IL-1β-induced chondrocytes. Furthermore, an OA model was established in rats with medial meniscus destabilization. Injecting AT into the knee joints of OA rat alleviated surgical resection-induced cartilage destruction. Thus, these findings revealed that AT relieves OA by activating mitochondrial autophagy by reducing TNFSF11 expression and inhibiting PI3K/AKT/mTOR signaling.

Proteolysis of fibrillin-2 microfibrils is essential for normal skeletal development.

The embryonic extracellular matrix (ECM) undergoes transition to mature ECM as development progresses, yet few mechanisms ensuring ECM proteostasis during this period are known. Fibrillin microfibrils are macromolecular ECM complexes serving structural and regulatory roles. In mice, Fbn1 and Fbn2, encoding the major microfibrillar components, are strongly expressed during embryogenesis, but fibrillin-1 is the major component observed in adult tissue microfibrils. Here, analysis of Adamts6 and Adamts10 mutant mouse embryos, lacking these homologous secreted metalloproteases individually and in combination, along with in vitro analysis of microfibrils, measurement of ADAMTS6-fibrillin affinities and N-terminomics discovery of ADAMTS6-cleaved sites, identifies a proteostatic mechanism contributing to postnatal fibrillin-2 reduction and fibrillin-1 dominance. The lack of ADAMTS6, alone and in combination with ADAMTS10 led to excess fibrillin-2 in perichondrium, with impaired skeletal development defined by a drastic reduction of aggrecan and cartilage link protein, impaired BMP signaling in cartilage, and increased GDF5 sequestration in fibrillin-2-rich tissue. Although ADAMTS6 cleaves fibrillin-1 and fibrillin-2 as well as fibronectin, which provides the initial scaffold for microfibril assembly, primacy of the protease-substrate relationship between ADAMTS6 and fibrillin-2 was unequivocally established by reversal of the defects in Adamts6-/- embryos by genetic reduction of Fbn2, but not Fbn1.

Imaging in retropubic cartilaginous cyst

The retropubic cartilaginous cyst is a benign slow-growing degenerative lesion arising from the fibrocartilage of the pubic symphysis. Degeneration is secondary to multiparity or pelvic trauma. The symptoms are commonly urinary tract-related and vary depending on the location. A confident diagnosis can be made on MRI demonstrating typical continuity with the pubic symphyseal joint, pubic diastasis, and cartilage signal. Management is typically conservative, with large lesions and failed conservative treatment requiring excision.

Association of Mucoid Degeneration of the ACL with Medial Tibiofemoral Osteoarthritis Progression at MR Imaging

Abstract Background Using MRI, ACL mucoid degeneration is defined as a thickened ACL with increased signal intensity on all MR pulse sequences, with discrete fibers easily distinguished on fatsaturated T2-weighted or fat-saturated proton-density (PD)-weighted images but poorly differentiated on T1-weighted or non-fat-saturated PD-weighted images. Objective To assess the prevalence of ACL mucoid degeneration in a population of patients referred for routine knee MRI, and its association with age and structural joint damage. Patients and Methods Our study is a retrospective study conducted at the radiology department of Ain Shams University hospitals and Ain Shams University Specialized Hospital including 81 cases of knees with ACL mucoid degeneration by MRI and no sex predilection. Cases and controls were scored with respect to independent articular features: cartilage signal and morphology, subarticular bone marrow abnormality, subarticular cysts, subarticular bone attrition, marginal osteophytes and medial meniscal integrity. Results Patients with ACL mucoid degeneration were older than patients with a normal ACL, without statistically significant sex difference. Knees with ACL mucoid degeneration had statistically significant medial meniscal injuries and cartilage damage involving the central and posterior MTFC compared to control knees with a normal ACL frequency matched for age, sex and MR field strength. Conclusion Our study proved that there is a strong association between ACL mucoid degeneration and cartilage damage in MTFC.

Magnetic resonance imaging of focal fibrocartilaginous dysplasia — findings derived from a three-dimensional gradient echo sequence

Focal fibrocartilaginous dysplasia is a rare benign bone lesion of young children that causes deformities in the extremities. However, the pathogenesis and treatments have not been defined and the MR manifestations have been less well described. To describe the MR manifestations of focal fibrocartilaginous dysplasia, especially on the T1-W three-dimensional (3-D) volumetric interpolated breath-hold examination (VIBE) sequence. In this retrospective study, the authors reviewed the MR and radiographic images, pathology and medical records of 21 cases of focal fibrocartilaginous dysplasia. All cases were evaluated by spin-echo MRI sequence. Among them, 17 cases were evaluated by T1-W 3-D VIBE sequence. The cohort consisted of 13 boys and 8 girls ages 4-75months. In 14 cases, focal fibrocartilaginous dysplasia was located in the tibia, 3 in the femur and 4 in the ulna. MRI 3-D VIBE sequence findings showed all cases had hypointense fiber band structures in the bone defect areas. The fibrous bands in the lower extremities ended in the epiphysis or epiphyseal plate, and in the upper extremities the epiphysis or carpal bone. Ten cases had hyperintensities that might represent cartilage composition. Four cases had cartilage signals that were continuous with the epiphyseal cartilage. MR spin-echo sequence findings showed that bone marrow edema of the adjacent joint was observed in eight cases, enlargement of the epiphyseal plate in three cases and medial meniscus injury in five cases. The 3-D VIBE sequence reveals useful details in focal fibrocartilaginous dysplasia.

Gli1+ progenitors mediate bone anabolic function of teriparatide via Hh and Igf signaling.

SUMMARYTeriparatide is the most widely prescribed bone anabolic drug in the world, but its cellular targets remain incompletely defined. The Gli1+ metaphyseal mesenchymal progenitors (MMPs) are a main source for osteoblasts in postnatal growing mice, but their potential response to teriparatide is unknown. Here, by lineage tracing, we show that teriparatide stimulates both proliferation and osteoblast differentiation of MMPs. Single-cell RNA sequencing reveals heterogeneity among MMPs, including an unexpected chondrocyte-like osteoprogenitor (COP). COP expresses the highest level of Hedgehog (Hh) target genes and the insulin-like growth factor 1 receptor (Igf1r) among all cell clusters. COP also expresses Pth1r and further upregulates Igf1r upon teriparatide treatment. Inhibition of Hh signaling or deletion of Igf1r from MMPs diminishes the proliferative and osteogenic effects of teriparatide. The study therefore identifies COP as a teriparatide target wherein Hh and insulin-like growth factor (Igf) signaling are critical for the osteoanabolic response in growing mice.