Orbital Tissue Research Articles

FRI552 Orbitopathy As The Sole Manifestation Of Hashimoto’s Thyroiditis In A Euthyroid Patient: A Case Report

Abstract Disclosure: C. Labib: None. Z. Saeed: None. M. Trabulsi: None. F. Ramharrack: None. INTRODUCTION: Thyroid associated ophthalmopathy (TAO) is caused by an autoimmune process involving the orbital tissue, however, its etiopathogenesis is still largely unclear. It is common in hyperthyroid patients with Graves’ disease and has also been reported in euthyroid and hypothyroid Graves’ patients with positive thyroid receptor antibodies (TRAb). However, only 5% of patient with Hashimoto’s thyroiditis (HT) develop orbitopathy. Here we report an unusual case of a male patient, clinically and biochemically euthyroid, who had severe bilateral exophthalmos due to underlying Hashimoto’s thyroiditis. CASE PRESENTATION: A 68-year-old male patient was evaluated for worsening bilateral eye protrusion associated with redness and grittiness for many years. The patient was otherwise asymptomatic; denied fatigue, any change in weight, constipation, joint/muscle pain or any other symptoms. On physical examination he was noted to have bilateral moderate periorbital edema, conjunctival injection along with mildly enlarged thyroid. Initial labs showed normal TSH 4.10 mIU/L (0.4-5.0mIU/L) and free T4 0.8 ng/dL (0.8-1.8 ng/dL). Further labs were notable for normal antithyrotropin antibody (TRAb) 1.09 IU/l and normal TSI less than 89%. Anti-thyroid peroxidase antibody (anti-TPO) level was as high as 12 IU/mL (less than 9 IU/mL). Thyroid US showed multinodular goiter with bilateral nodules. A Subsequent FNA was benign. The degree of exophthalmos on the Hertel ophthalmometer was 30 mm bilaterally. Orbital magnetic resonance imaging (MRI) showed proptosis of the left and right globes as well as prominence of intraorbital fat. Extraocular musculature was symmetric bilaterally and only minimally prominent but diffuse. Treatment with teprotumumab infusion was started with improvement in the patient’s symptoms. CASE DISCUSSION: Usually mild eye symptoms are seen in Hashimoto’s thyroiditis such as dryness, blurred vision, upper eyelid retraction and loss of lateral third of eyebrows. In rare cases it can cause orbitopathy similar to that seen in Graves’ disease. Like Graves’ orbitopathy, in addition to reversing the underlying thyroid abnormalities, moderate-to-severe orbitopathy is treated with corticosteroids or teprotumumab. This case sheds the light on exophthalmos being an atypical presentation of Hashimoto’s thyroiditis in a clinically and biochemically euthyroid patient. Physicians should be cognizant of this rare manifestation for prompt recognition and early treatment which can prevent corneal involvement and blindness due to optic nerve compression. Presentation: Friday, June 16, 2023

Evaluation of orbital soft tissue biomechanical parameters in patients with thyroid eye disease using the non-contact Corvis ST.

In thyroid eye disease (TED), all ocular components and adnexa such as extraocular muscles, orbital adipose tissues, eyelids, and tear glands could be affected. This study aimed to study the orbital biomechanical parameters in patients with TED, in terms of differences with healthy individuals and correlation with clinical findings, using Corvis ST (CST, Oculus Wetzlar). In this study, 26 consecutive patients with TED were recruited. Demographic data were collected, and patients with TED were assessed for exophthalmos, intraocular pressure, and clinical activity score. Biomechanical response parameters of one randomly-chosen eye of each patient, including whole eye movement length (WEMl) and time (WEMt), were evaluated by the CST, and data were compared between patients and age- and gender-matched healthy controls. The mean age was 39.88 ± 11.61years old for patients with TED and 34.38 ± 8.57years old for the healthy subjects. Nine out of 26 patients with TED and nine of 26 healthy individuals were male. The median duration of thyroid disease was 36 (IQR 54) months and the median duration of thyroid ophthalmopathy was 27 (IQR 27) months. Four out of 26 patients (7.7%) had active disease. The mean WEMl was 206.15 ± 61.58µm in the TED group and 254.23 ± 64.01μm in the healthy group, the difference of which was statistically significant (p = 0.008). The median of WEMt was 20.90 (1.15) msec in the TED group and 21.45 (0.93) msec in the healthy group (p < 0.001). Also, the mean of WEMl and WEMt were lower in patients with active disease compared to patients with quiescent disease. The CST-derived WEMl was significantly smaller in patients with thyroid eye disease compared to normal subjects. The WEMl and WEMt were relatively shorter in the patients with active TED compared to the patients with quiescent TED, although small numbers of patients with active TED limits took a statistically significant conclusion. WEMl and WEMt might be useful in evaluating the compliance of the orbit in patients with TED.

Cre-loxP System-Based Mouse Model for Investigating Graves' Disease and Associated Orbitopathy.

Background: Graves' disease (GD), one of the most common forms of autoimmune thyroid disorders, is characterized by hyperthyroidism caused by antibodies (Abs) against the extracellular A-subunit of the thyrotropin receptor (TSHR). Various approaches have been used to create mouse models of GD, including transfected fibroblasts and immunization with plasmids or adenoviruses expressing human TSHR A-subunit (hTSHR A-subunit). These models, however, require repeated immunization and produce inconsistent results. In this study, we established a novel Cre-loxP system-based mouse model that is able to generate the hTSHR A-subunit, mimicking human GD, and characterized the histological changes in Graves' orbitopathy (GO) progression after a single injection. Materials and Methods: A Cre-loxP system-based mouse model was constructed by inserting the CAG-loxP-STOP-loxP-hTSHR A-subunit cassette into the Rosa26 locus of the mouse genome. Conditional expression of the hTSHR A-subunit was successfully achieved by intramuscular injection of the transactivator of transcription-Cre recombinase (GD mice). Blood tests for anti-TSHR Abs and the total thyroxine (T4) level were performed. Magnetic resonance imaging (MRI) was used to monitor morphological changes in the eyes. A histological examination of the thyroid gland and retrobulbar tissues was performed to observe pathological changes. Results: Twenty-four (8 control and 16 GD) mice were investigated. All GD mice exhibited higher levels of TSHR Abs compared with the control group. Moreover, more than 80% of the mouse models showed elevated T4 levels accompanied by thyroid goiter. MRI analysis revealed an increased volume of retrobulbar tissue, while immunohistochemical staining of orbital tissues exhibited macrophage infiltration and muscle fibrosis in the GD mice, contrasting with the control group. Conclusions: Our novel mouse model for GD, which showed the histological features of GO, was successfully established using the Cre-loxP system. This animal model offers improved insights and contributes to advancing methodological developments for GD and GO.

Orbital Liposarcoma: A Surveillance, Epidemiology and End Results Database Study.

Orbital liposarcoma is a challenging tumor to treat due to its rarity and high rate of local recurrence, and the role of radiotherapy and chemotherapy remain unclear. Analysis of big data may improve our overall understanding of orbital disease and role of adjuvant therapies. Data were extracted from the Surveillance, Epidemiology and End Results (SEER) Research Plus database from 1975 to 2017. All patients with a diagnosis of liposarcoma (ICD-O3 codes 8850-8858, 8869-8862, 8870, 8880, 8881) were included. Cases were divided into 4 groups by primary site: orbit, retroperitoneum, soft tissue, and other. A total of 16,958 patients were included. Patients with orbital involvement were younger and more likely to be female ( p < 0.05). Among orbital lesions, myxoid liposarcoma was the most common histologic subtype (6/19; 31.6%) followed by well differentiated (5/19; 26.3%). This differed from the distribution of histologic subtypes encountered elsewhere, for which well-differentiated liposarcoma was the most common (retroperitoneum 979/3,136; 31%, soft tissue 3,493/11,671; 30%, and other sites 497/2,132; 23%, p < 0.05). Dedifferentiated histologic subtype was the second most common subtype found in the retroperitoneum (946/3,136; 30%), whereas it was less common in the orbit (2/19; 11%) and soft tissue (1,396/11,671; 12%) ( p < 0.001). Patients with orbital liposarcoma had similar disease-specific mortality compared with soft-tissue location ( p = 0.825) and lower disease-specific mortality compared with retroperitoneal location ( p < 0.001). When all locations were combined, patients with well-differentiated liposarcoma had the lowest disease-specific mortality. Patients with orbital liposarcoma tend to be younger, female, and have a better prognosis than those with retroperitoneal disease, likely due to the lower incidence of dedifferentiated histologic subtype.

Construction of predictive ceRNA network and identification of the patterns of immune cells infiltrated in Graves' ophthalmopathy.

Graves' ophthalmopathy (GO) is a multifactorial disease, and the mechanism of non coding RNA interactions and inflammatory cell infiltration patterns are not fully understood. This study aims to construct a competing endogenous RNA (ceRNA) network for this disease and clarify the infiltration patterns of inflammatory cells in orbital tissue to further explore the pathogenesis of GO. The differentially expressed genes were identified using the GEO2R analysis tool. The Kyoto encyclopedia of genes and genomes (KEGG) and gene ontology analysis were used to analyze differential genes. RNA interaction relationships were extracted from the RNA interactome database. Protein-protein interactions were identified using the STRING database and were visualized using Cytoscape. StarBase, miRcode, and DIANA-LncBase Experimental v.2 were used to construct ceRNA networks together with their interacted non-coding RNA. The CIBERSORT algorithm was used to detect the patterns of infiltrating immune cells in GO using R software. A total of 114 differentially expressed genes for GO and 121 pathways were detected using both the KEGG and gene ontology enrichment analysis. Four hub genes (SRSF6, DDX5, HNRNPC,and HNRNPM) were extracted from protein-protein interaction using cytoHubba in Cytoscape, 104 nodes and 142 edges were extracted, and a ceRNA network was identified (MALAT1-MIR21-DDX5). The results of immune cell analysis showed that in GO, the proportions of CD8+ T cells and CD4+ memory resting T cells were upregulated and downregulated, respectively. The proportion of CD4 memory resting T cells was positively correlated with the expression of MALAT1, MIR21, and DDX5. This study has constructed a ceRNA regulatory network (MALAT1-MIR21-DDX5) in GO orbital tissue, clarifying the downregulation of the proportion of CD4+ stationary memory T cells and their positive regulatory relationship with ceRNA components, further revealing the pathogenesis of GO.

Reconstruction of Medial Orbital Wall Using Autologous Perpendicular Plate of Ethmoid.

The aim of this study is to investigate the feasibility of perpendicular plate of ethmoid as material for the reconstruction of medial orbital wall. The main outcome measurement was preoperative and postoperative orbital volume. The authors performed a study including 17 patients who have isolated medial orbital wall fracture (blow-out fracture). All the patients were fixed the defect using autologous perpendicular plate of ethmoid under endonasal approach. The authors compared the preoperative and postoperative orbital volume difference (unaffected orbit, affected orbit) of all the patients, and observed the improvement of diplopia or ocular motility disorders after operation. All 17 medial orbital wall reconstruction surgeries were successful with no severe postoperative ophthalmic complications. Statistically significant differences were found between the preoperative and postoperative orbital tissue volumes for the affected orbit. There was no statistically significant difference found between the tissue volume of the contralateral unaffected orbit and the affected orbit after reconstruction. And postoperative computed tomography showed the implant is in place and there is no medial rectus incarceration. Autologous perpendicular plate of ethmoid proved to be safe and effective in the reconstruction of medial orbital wall under endonasal approach with cost-effectivence, low complication rate, high biocompatibility, and minimally invasion.

Machine Learning Model with Texture Analysis for Automatic Classification of Histopathological Images of Ocular Adnexal Mucosa-associated Lymphoid Tissue Lymphoma of Two Different Origins

Purpose The purpose of this study was to develop artificial intelligence algorithms that can distinguish between orbital and conjunctival mucosa-associated lymphoid tissue (MALT) lymphomas in pathological images. Methods Tissue blocks with residual MALT lymphoma and data from histological and flow cytometric studies and molecular genetic analyses such as gene rearrangement were procured for 129 patients treated between April 2008 and April 2020. We collected pathological hematoxylin and eosin-stained (HE) images of lymphoma from these patients and cropped 10 different image patches at a resolution of 2048 × 2048 from pathological images from each patient. A total of 990 images from 99 patients were used to create and evaluate machine-learning models. Each image patch of three different magnification rates at ×4, ×20, and ×40 underwent texture analysis to extract features, and then seven different machine-learning algorithms were applied to the results to create models. Cross-validation on a patient-by-patient basis was used to create and evaluate models, and then 300 images from the remaining 30 cases were used to evaluate the average accuracy rate. Results Ten-fold cross-validation using the support vector machine with linear kernel algorithm was identified as the best algorithm for discriminating between conjunctival mucosa-associated lymphoid tissue and orbital MALT lymphomas, with an average accuracy rate under cross-validation of 85%. There were ×20 magnification HE images that were more accurate in distinguishing orbital and conjunctival MALT lymphomas among ×4, ×20, and ×40. Conclusion Artificial intelligence algorithms can successfully distinguish HE images between orbital and conjunctival MALT lymphomas.

Metabolic features of orbital adipose tissue in patients with thyroid eye disease.

Thyroid eye disease (TED) is the most frequent orbital disease in adults and is characterized by the accumulation of orbital adipose tissue (OAT). It can lead to eyelid retraction or even vision loss. Orbital decompression surgery serves as the primary treatment for inactive TED by removing the excess OAT. However, there is a lack of alternative treatments to surgery due to the unclear understanding of the pathogenesis, particularly the metabolic features. Accordingly, our study was implemented to explore the content and features of metabolites of OATs from TED patients. The OATs used in the current study were obtained from the orbital decompression surgery of seven patients with inactive TED. We also collected control OATs from eye surgical samples of five individuals with no history of autoimmune thyroid diseases, TED, or under non-inflammatory conditions. The liquid chromatography mass spectrometer was used for the measurements of the targeted metabolites. Afterwards, we performed differential metabolite assay analysis and related pathway enrichment analysis. In our study, a total of 149 metabolite profiles were detected in all participants. There were significant differences in several metabolite profiles between the TED group and the control group, mainly including uric acid, oxidized glutathione, taurine, dGMP, oxidized glutathione 2, uracil, hexose-phosphate, 1-methylnicotinamide, D-sedoheptulose 1,7-bisphosphate, and uridine 5'-monophosphate (all p-value < 0.05). The TED-related pathways identified included purine metabolism, beta-alanine metabolism, glutathione metabolism (p-values < 0.05). Our study found overlaps and differences including uric acid and uracil, which are in accordance with metabolites found in blood of patients with TED from previous study and several newly discovered metabolite by our study such as hexose-phosphate, 1-methylnicotinamide, D-sedoheptulose 1,7-bisphosphate, compared to those tested from blood, OAT, or urine samples reported in previous studies. The findings of our study shed light on the metabolic features of OAT in individuals with TED. These results may help identify new treatment targets for TED, providing potential avenues for developing alternative treatments beyond ophthalmic surgery.

Comparison of surgical effect in active and inactive Dysthyroid Optic Neuropathy after Endoscopic Transnasal Medial Orbital Decompression.

To evaluate and compare the changes in orbital soft tissue volume and visual function after endoscopic transnasal medial orbital decompression in patients with active and inactive dysthyroid optic neuropathy (DON). This prospective, cohort study recruited 112 patients (112 eyes) with DON who were divided into an active and inactive DON group (56 eyes each) by clinical activity scores. All patients underwent endoscopic transnasal medial orbital decompression. The pre- and post-operative orbital soft tissue volumes were measured with high-resolution computed tomography (CT) using Mimics software. Visual function, including best-corrected visual acuity (BCVA), visual field (VF), and visual evoked potential (VEP), was recorded before and after surgery. Preoperatively, compared with the inactive DON group, the active DON group had greater extraocular muscle volume (EMV) and EMV/orbital volume (OV) ratio, but worse BCVA, VF, and exophthalmos. Postoperatively, although the EMV slightly increased, with the enlarged medial rectus muscle contributing dramatically, the EMV/OV ratio decreased in patients with DON. Besides, visual function including BCVA, VF, VEP and exophthalmos was also improved in both groups after surgery. There were no significant differences in postoperative OV; EMV; EMV/OV ratio; and the BCVA, VF, and VEP parameters between both groups (all P > 0.05). Patients with DON who did not respond well to steroids, regardless of disease activity, may benefit from orbital decompression via the decrease in the proportion of EMV in OV, especially patients with active DON, who showed more improved visual function than patients with inactive DON.

Macular microvasculature in patients with thyroid-associated orbitopathy: a cross-sectional study

PurposeThe aim of this study was to evaluate macular blood flow in patients with thyroid-associated orbitopathy (TAO) as compared to healthy subjects. The inflammatory nature of the disease, as well as the vascular congestion caused by the increase in the volume of orbital soft tissue and extraocular muscles, rationalize the assessment of retinal blood flow changes in these patients.MethodsThis is a cross-sectional study with the convenience sampling method. Macular flow density was assessed using optical coherence tomography angiography (OCTA) and compared between patients with TAO and healthy individuals. We also compared macular flow density in two subgroups of patients based on clinical activity score (CAS).ResultsEighty–five cases, including 30 healthy individuals and 55 patients with TAO, participated. The foveal avascular zone (FAZ) area was significantly larger in the patient group than in the control. Patients with active TAO with CAS 3 or more had significantly larger FAZ areas than those with CAS less than 3 (p = 0.04).ConclusionWe showed that the FAZ area is larger in active TAO patients and can be considered a possible candidate feature for monitoring disease activity and thyroid-associated vasculopathy.

Subcutaneous and Orbital Adipose Tissues: Intrinsic Differences in Carotenoid Contents and Lipidomic Compositions.

The color difference in human subcutaneous fat (SF) and orbital fat (OF) is apparent, but the reasons have been rarely elaborated. We speculate that differences in carotenoid and lipid contents may account for the discrepancy in color. In this study, the intrinsic differences in SF and OF were analyzed using ultrahigh-performance liquid chromatography coupled with Q-Exactive liquid chromatography mass spectrometry/mass spectrometry (UPLC-QE Plus LC-MS/MS). Lipid profiling was performed in an independent batch. The morphology between orbital septum and SF differed statistically in the size of adipocytes and the distribution area of adipocytes. We compared carotenoid contents between two groups (seven samples) and found that lutein was more abundant in SF than that in OF with a p-value of 0.0409, suggesting that lutein could be mainly responsible for the yellow color of adipose tissue. Lipidomic results proved that SF and OF were well differentiated. Totally, 402 lipid features were detected, with 349 features in the positive ion mode and 53 features in the negative ion mode. Features (99.9%) in the positive ion mode and features (98.7%) in the negative ion mode well described various separation patterns in principal component analysis. Thirty-two features selected by variable importance in projection might account for the diversity of compounds in SF and OF. In conclusion, SF and OF differed from each other in carotenoids and lipidome. It is helpful to study the metabolism process of lipid droplets in adipocytes.

Assessment of age-related change of the ocular support system.

To estimate the material stiffness of the orbital soft tissue in human orbits using an inverse numerical analysis approach, which could be used in future studies to understand the behaviour under dynamic, non-contact tonometry or simulate various ophthalmological conditions. Clinical data were obtained for the left eye of 185 Chinese participants subjected to a complete ophthalmic examination, including tests by the Corvis ST and Pentacam. 185 numerical models of the eye globes were built with idealised geometry of the sclera while considering the corneal tomography measured by the Pentacam. The models were extended to include representations of the orbital soft tissue (OST), which were given idealised geometry. The movement of the whole eye in response to an air-puff directed at the central cornea was examined and used in an inverse analysis process to estimate the biomechanical stiffness parameters of the OST. The results indicated a weak correlation of E t with the progression of age, regardless of the stress at which E t was calculated. However, there was evidence of significant differences in E t between some of the age groups. There was statistical evidence of significant differences between E t in the age range 20< years < 43 relative to E t in OST with age ranges 43< years < 63 (p = 0.022) and 63< years < 91 (p = 0.011). In contrast, E t in OST with age ranges 43< years < 63 and 63< years < 91 were not significantly different (p = 0.863). The optimised mechanical properties of the OST were found to be almost four times stiffer than properties of fatty tissue of previous experimental work. This study consolidated previous findings of the role of extraocular muscles on the ocular suppor system. In addition, the rotation of the globe during corvis loading is suggested to be of posterior components of the globe and shall be further investigated.

Bullet in the orbital apex: Explosive situation!

A healthy thirty-seven-year-old male presented with a gunshot injury to the eye. The bullet was lodged in the orbital apex and had caused rupture of the left globe and extensive damage to the orbital soft tissues including the optic nerve, as well as the bones at the apex. This large metallic foreign body needed to be removed as it was causing continuous bleeding and cellulitis; also, its location was precarious. The surgery for tracking and removal of the bullet presented several interesting pointers which are described in this case report.

Treatment strategies for orbital gas-producing necrotizing fasciitis secondary to odontogenic maxillary sinusitis: Technical notes

This paper presents treatment strategies for orbital necrotizing fasciitis (NF) in a case of a 33-year-old male diagnosed with orbital NF, which developed after dental root canal treatment. Although orbital NF is rare, it is rapidly progressive and can easily lead to the loss of tissue and visual function, sometimes to a life-threatening extent. Prompt and adequate treatment has been a challenge yet remains quite essential. In addition to the conventional approach to NF, such as immediate antibiotic administration and drainage, orbital NF patients like this case were often treated by incorporating additional steps, which include: 1) performing minimally invasive but adequate removal of necrotic tissue through intraoperative use of ultrasound equipment and postoperative use of proteolytic enzyme-containing ointment for chemical debridement; 2) managing intraorbital pressure by lateral cantholysis and orbital floor removal (decompression); and 3) maintaining the aerobic conditions of the wound after surgical drainage via orbital wall removal. Thus far, satisfactory results in patients with extensive NF of the orbit, including the presented case, were achieved with regards to preserving periorbital tissues, vision, and ocular motility through a multidisciplinary approach. These should be considered as optional means of preserving the orbital tissue and visual function.

Radiomics analysis of T1WI and T2WI magnetic resonance images to differentiate between IgG4-related ophthalmic disease and orbital MALT lymphoma

BackgroundPreoperative differentiation between IgG4-related orbital disease (IgG4-ROD) and orbital mucosa-associated lymphoid tissue (MALT) lymphoma has a significant impact on clinical decision-making. Our research aims to construct and evaluate a magnetic resonance imaging (MRI)-based radiomics model to assist clinicians to better identify IgG4-ROD and orbital MALT lymphoma and make better preoperative medical decisions.MethodsMR images and clinical data from 20 IgG4-ROD patients and 30 orbital MALT lymphoma patients were classified into a training (21 MALT; 14 IgG4-ROD) or validation set (nine MALT; six IgG4-ROD). Radiomics features were collected from T1-weighted (T1WI) and T2-weighted images (T2WI). Student’s t-test, the least absolute shrinkage and selection operator (LASSO) and principal component analysis (PCA) were conducted to screen and select the radiomics features. Support vector machine (SVM) classifiers developed from the selected radiomic features for T1WI, T2WI and combined T1WI and T2WI were trained and tested on the training and validation set via five-fold cross-validation, respectively. Diagnostic performance of the classifiers were evaluated with area under the curve (AUC) readings of the receiver operating characteristic (ROC) curve, and readouts for precision, accuracy, recall and F1 score.ResultsAmong 12 statistically significant features from T1WI, four were selected for SVM modelling after LASSO analysis. For T2WI, eight of 51 statistically significant features were analyzed by LASSO followed by PCA, with five features finally used for SVM. Combined analysis of T1WI and T2WI features selected two and four, respectively, for SVM. The AUC values for T1WI and T2WI classifiers separately were 0.722 ± 0.037 and 0.744 ± 0.027, respectively, while combined analysis of T1WI and T2WI classifiers further enhanced the classification performances with AUC values ranging from 0.727 to 0.821.ConclusionThe radiomics model based on features from both T1WI and T2WI images is effective and promising for the differential diagnosis of IgG4-ROD and MALT lymphoma. More detailed radiomics features and advanced techniques should be considered to further explore the differences between these diseases.

Simultaneous Effects of a Selective EP2 Agonist, Omidenepag, and a Rho-Associated Coiled-Coil Containing Protein Kinase Inhibitor, Ripasudil, on Human Orbital Fibroblasts.

Purpose: To assess the combined effects of omidenepag (OMD), a selective EP2 agonist, and ripasudil (Rip), an inhibitor of rho-associated coiled-coil containing protein kinases, on the human orbital adipose tissue, two-dimensional (2D) or three-dimensional (3D) cultures of human orbital fibroblasts (HOFs) were employed. Methods: Cellular metabolic functions (2D), physical (3D), lipid staining (3D), and quantitative polymerase chain reaction for adipogenesis-related genes, PPARγ and AP2, and extracellular matrix (ECM) molecules, including collagen (COL)1, 4, and 6, and fibronectin (FN) (3D) were evaluated in the presence of OMD (100 nM) and/or Rip (10 μM). Results: Real-time metabolic analyses revealed that the adipogenic differentiation (DIF+) with OMD significantly shifted an energetic state toward energetic, whereas DIF+ with Rip significantly shifted that toward quiescent. In the case of both drugs upon DIF+, the metabolic effect of OMD was predominant. DIF+ induced enlargement and stiffed 3D spheroid with increased lipid staining and mRNA expression of adipogenesis-related genes, COL4 and COL6, and decreased the expression of COL1. In the presence of OMD and/or Rip to DIF+, (1) the sizes were further increased by Rip and the stiffness was significantly decreased by OMD or Rip and (2) COL4 or AP2 expression was substantially increased by OMD or Rip, respectively. Conclusion: The results presented herein indicate that the metabolic effects of OMD and Rip exerted opposing effects and the effects of OMD toward Ap2 and ECM expressions were distinct from those of Rip, but the effects of OMD toward the physical aspects and adipogenesis of the 3D cultured HOFs were similar to the effects of Rip.

RNA aptamers with specific binding affinity to CD40 (CD40Apt) represents a promising antagonist of the CD40-CD40L signaling for thyroid-associated ophthalmopathy (TAO) treatment in mouse

Thyroid-associated ophthalmopathy (TAO) is the most common autoimmune inflammatory diseases of the orbit. The CD40-CD40L pathway has been regarded as a potential molecular mechanism contributing to the development and progression of TAO, and RNA aptamers with specific binding affinity to CD40 (CD40Apt) represents a promising inhibitor of the CD40-CD40L signaling in TAO treatment. In this study, CD40Apt was confirmed to specifically recognize mouse CD40-positive ortibtal fibroblast. Mouse orbital fibroblasts were isolated from TAO mice model orbital tissues and validated. In TGF-β-induced orbital fibroblast activation model in vitro, CD40Apt administration inhibited TGF-β-induced cell viability, decreased TGF-β-induced α-SMA, Collagen I, Timp-1, and vimentin levels, and suppressed TGF-β-induced phosphorylation of Erk, p38, JNK, and NF-κB. In TAO mice model in vivo, CD40Apt caused no significant differences to the body weight of mice; furthermore, CD40Apt improved the eyelid broadening, ameliorated inflammatory infiltration and the hyperplasia in orbital muscle and adipose tissues in model mice. Concerning orbital fibroblast activation, CD40Apt reduced the levels of CD40, collagen I, TGF-β, and α-SMA in orbital muscle and adipose tissues of model mice. Finally, CD40Apt administration significantly suppressed Erk, p38, JNK, and NF-κB phosphorylation. In conclusion, CD40Apt, specifically binds to CD40 proteins in their natural state on the cell surface with high affinity, could suppress mouse orbital fibroblast activation, therefore improving TAO in mice model through the CD40 and downstream signaling pathways. CD40Apt represents a promising antagonist of the CD40-CD40L signaling for TAO treatment.

Inflammation and Fibrosis in Orbital Inflammatory Disease: A Histopathologic Analysis.

The purpose of this study was to compare the histopathologic inflammation and fibrosis of orbital adipose tissue in orbital inflammatory disease (OID) specimens. In this retrospective cohort study, inflammation, and fibrosis in orbital adipose tissue from patients with thyroid-associated orbitopathy (TAO), granulomatosis with polyangiitis (GPA), sarcoidosis, nonspecific orbital inflammation (NSOI), and healthy controls were scored by 2 masked ocular pathologists. Both categories were scored on a scale of 0 to 3 with scoring criteria based on the percentage of specimens containing inflammation or fibrosis, respectively. Tissue specimens were collected from oculoplastic surgeons at 8 international centers representing 4 countries. Seventy-four specimens were included: 25 with TAO, 6 with orbital GPA, 7 with orbital sarcoidosis, 24 with NSOI, and 12 healthy controls. The mean inflammation and fibrosis scores for healthy controls were 0.0 and 1.1, respectively. Orbital inflammatory disease groups' inflammation (I) and fibrosis (F) scores, formatted [I, F] with respective p -values when compared to controls, were: TAO [0.2, 1.4] ( p = 1, 1), GPA [1.9, 2.6] ( p = 0.003, 0.009), sarcoidosis [2.4, 1.9] ( p = 0.001, 0.023), and NSOI [1.3, 1.8] ( p ≤ 0.001, 0.018). Sarcoidosis had the highest mean inflammation score. The pairwise analysis demonstrated that sarcoidosis had a significantly higher mean inflammation score than NSOI ( p = 0.036) and TAO ( p < 0.0001), but no difference when compared to GPA. GPA had the highest mean fibrosis score, with pairwise analysis demonstrating a significantly higher mean fibrosis score than TAO ( p = 0.048). Mean inflammation and fibrosis scores in TAO orbital adipose tissue samples did not differ from healthy controls. In contrast, the more "intense" inflammatory diseases such as GPA, sarcoidosis, and NSOI did demonstrate higher histopathologic inflammation and fibrosis. This has implications in prognosis, therapeutic selection, and response monitoring in orbital inflammatory disease.

Selective BD2 Inhibitor Exerts Anti-Fibrotic Effects via BRD4/FoxM1/Plk1 Axis in Orbital Fibroblasts From Patients With Thyroid Eye Disease.

We investigated the therapeutic potential of ABBV744, a bromodomain and extra-terminal (BET) inhibitor with selectivity for the second bromodomain (BD2) in thyroid eye disease (TED). The anti-fibrotic effects of ABBV744 and its underlying mechanism were explored in cultured orbital fibroblasts (OFs) from patients with TED. Immunohistochemistry (IHC) and real-time quantitative polymerase chain reaction (RT-qPCR) assays were conducted on orbital connective tissues from TED and controls. RT-qPCR, Western blot, Cell-counting Kit-8 (CCK-8), and 5-ethynyl-2'-deoxyuridine (EdU) cell proliferation assays were conducted on OFs isolated from patients with TED. The expression of BRD4 was upregulated in the orbital tissues of patients with TED relative to controls and in TED OFs stimulated with TGF-β1. Further, we showed that BRD4 modulated the profibrotic process through the interaction with Forkhead Box M1 (FoxM1) and its downstream molecule Polo-like kinase 1 (Plk1) in cultured TED OFs. Inhibition of BRD4 both by BD2 selective inhibitor ABBV744 and pan-BET inhibitor JQ1 exerted anti-fibrotic effects, whereas ABBV744 displayed superior anti-fibrotic effects and acceptable safety compared to JQ1. We conclude that BDR4 may modulate the profibrotic process in OFs of patients with TED via the FoxM1/Plk1 axis, and that selectively targeting BD2 domain of BRD4 may therefore be a potential therapeutic option for treating patients with TED.

Management of blowout fracture with iliac bone graft

Introduction: Blowout fracture is a traumatic deformity of the orbital wall and the repair is indicated when enophthalmos is troublesome to the patient, or when diplopia interferes patient’s activities. Iliac bone has been reported in the immediate and secondary repair of orbital wall deformities following blowout fracture. This case is to report blowout fracture management using iliac bone graft. Case Illustration: A 35 years old man complains pain on his left eye when moving his eye and diplopia in upgazed since 4 months after fell down from stairs. Eye position is orthophoria, visual acuity 6/6 on both eye, anterior and posterior segments are normal, restricted movement to superior on left eye, Hertel exophthalmometry examination on right eye 15 mm and left eye 10 mm (base 90). On CT-scan examination, it was found that an orbital floor fracture was characterized by herniation of the orbital tissue. The patient was diagnosed with left eye enophthalmos, suspect inferior rectus muscle and adnexal entrapment and neglected blow out fracture. LE Repair Orbital Floor Fracture, Release Muscle Rectus Inferior, Exploration (join op with Plastic Reconstructive and Aesthetic Surgery Department) is done to this patient. Conclusion: Iliac bone graft can be used as an alternative in orbital floor reconstruction, since it easy to harvest, low risk to rejected or extruded and can be used in large defect. Optimal outcome of blowout fracture repair can be achieved if the surgical management is done before two weeks after the incidence.