Intraocularly, fatty acid-binding protein 4 (FABP4) and 5 (FABP5) mainly originate from human ocular choroidal fibroblasts (HOCF), nonpigmented ciliary epithelium (HNPCE) cells have been suggested to be pivotally involved in intraocular pathophysiology. To elucidate the unidentified regulatory mechanisms of gene expression secretion FABPs, effects glucose levels, acids (FAs), peroxisome proliferator-activated receptor (PPAR) modulators were studied. additional biological role laser...

Thyroid-associated orbitopathy (TAO) is a disfiguring periocular connective tissue disease associated with autoimmune thyroid disorders. It potentially blinding condition, for which no effective pharmacological treatment has been established. Despite suggested role played by thyrotropin receptor activation in the pathogenesis of TAO, cellular and molecular events contributing to fibrotic inflammatory process TAO are not fully defined. By developing three-dimensional organoid culture human...

Respiratory dysfunction is a common complication of obesity, conferring cardiovascular morbidity and increased mortality often necessitating mechanical ventilatory support. While impaired lung expansion in the setting adipose mass reduced central response to hypercapnia have been implicated as pathophysiological drivers, impact obesity on respiratory muscles—in particular, diaphragm—has not investigated detail. Here, we demonstrate that chronic high-fat diet (HFD) feeding impairs diaphragm...

Abstract To establish a deepening of the upper eyelid sulcus (DUES) model that can be induced by prostaglandin (PG) analogues, three-dimension (3D) tissue culture was employed. Upon adipogenesis 3T3-L1 organoid, effects either Bimatoprost acid (BIM-A), or PGF2α were examined. During adipogenesis, organoid size, lipid staining BODIPY and expression extracellular matrix (ECM) immunocytochemistry and/or quantitative PCR The size increased remarkably during while such increases significantly...

Purpose To elucidate the molecular etiology of deepening upper eyelid sulcus (DUES) induced by prostaglandin (PG) analogs, a three-dimensional (3D) tissue culture system was employed using human orbital fibroblasts (HOFs). Methods During adipogenesis, changes in HOF 3D organoid sizes, as well their lipids stained BODIPY and expression extracellular matrix (ECM) immunolabeling and/or quantitative PCR, were studied presence or absence either 100-nM bimatoprost acid F2α. Results The size...

To elucidate the currently unknown mechanisms responsible for diverse biological aspects between two-dimensional (2D) and three-dimensional (3D) cultured 3T3-L1 preadipocytes, RNA-sequencing analyses were performed. During a 7-day culture period, 2D- 3D-cultured cells subjected to lipid staining by BODIPY, qPCR adipogenesis related genes, including peroxisome proliferator-activated receptor γ (Pparγ), CCAAT/enhancer-binding protein alpha (Cebpa), Ap2 (fatty acid-binding 4; Fabp4), leptin,...

To study the molecular mechanisms responsible for inducing spatial proliferation of malignant melanomas (MM), three-dimension (3D) spheroids were produced from several MM cell lines including SK-mel-24, MM418, A375, WM266-4, and SM2-1, their 3D architectures cellular metabolisms evaluated by phase-contrast microscopy Seahorse bio-analyzer, respectively. Several transformed horizontal configurations observed within most these spheroids, degree deformity was increased in order: SK-mel-24. An...

Abstract To elucidate molecular pharmacology of Rho-associated coiled-coil containing protein kinase inhibitors (ROCK-i, Ripasudil and Y27632) on their efficiency for aqueous outflow, 2D or 3D cultures a human trabecular meshwork (HTM) were prepared in the presence TGFβ2. Those examined by transendothelial electrical resistance (TEER, 2D), electronic microscopy (EM, 3D), expression extracellular matrix (ECM) including collagen1 (COL1), COL4 COL6, fibronectin (FN) immunolabeling and/or...

To establish appropriate ex vivo models for a glaucomatous trabecular meshwork (TM), two-dimensional (2D) and three-dimensional (3D) cultures of human cells (HTM) were prepared in the presence 250 nM dexamethasone (DEX) or 5 ng/mL TGFβ2, characterized by following analyses; transendothelial electrical resistance (TEER) measurements, FITC dextran permeability, scanning electron microscopy expression extracellular matrix (ECM) including collagen (COL)1, 4 6, fibronectin (FN), α-smooth muscle...

Purpose: The effects of Rho-associated coiled-coil containing protein kinase (ROCK) 1 and 2 inhibitor, ripasudil hydrochloride hydrate (Rip), ROCK2 KD025 or rosiglitazone (Rosi) on two-dimension (2D) three-dimension (3D) cultured human conjunctival fibroblasts (HconF) treated by transforming growth factor (TGFβ2) were studied. Methods: Two-dimension HconF examined transendothelial electrical resistance (TEER, 2D), size stiffness (3D), the expression extracellular matrix (ECM) including...

To elucidate the currently unknown molecular mechanisms responsible for aberrant expression of recoverin (Rec) within cancerous cells, we examined two-dimensional (2D) and three-dimensional (3D) cultures Rec-negative lung adenocarcinoma A549 cells which had been transfected with a plasmid containing human cDNA (A549 Rec) or an empty as mock control MOCK). Using these measured cytotoxicity by several anti-tumor agents (2D), cellular metabolism including mitochondrial glycolytic functions...

Abstract 2D and 3D cultures of 3T3-L1 cells were employed in a study the effects Omidenepag (OMD), interacting with non-prostanoid EP2 receptor, on adipogenesis. Upon adipogenesis, lipid staining, mRNA expression adipogenesis-related genes ( Pparγ, CEBPa , Ap2, Glut4 ) extracellular matrix (ECM) including collagen type 1, 4 6, fibronectin, size physical property organoids compared between groups that had been treated agonists (butaprost OMD) PGF2α. these significantly suppressed staining...

To elucidate the currently unknown molecular mechanisms responsible for similarity and difference during acquirement of resistance against gemcitabine (GEM) paclitaxel (PTX) in patients with pancreatic carcinoma, we examined two-dimensional (2D) three-dimensional (3D) cultures parent MIA PaCa-2 cells (MIA PaCa-2-PA) their GEM cell line PaCa-2-GR) PTX PaCa-2-PR). Using these cells, 3D spheroid configurations cellular metabolism, including mitochondrial glycolytic functions, a Seahorse...

Since the recent discovery of prostaglandin-associated peri-orbitopathy, a great deal interest has developed concerning side effects anti-glaucoma medications toward periocular fatty tissue, especially their adipogenesis. Two- or three-dimension (2D 3D) cultures 3T3-L1 cells were employed to elucidate Rho-associated coiled-coil containing protein kinase inhibitor (ROCK-i) drug, Ripasudil, and other ROCK-i, such as Y27632 on Ultrastructure by electron microscopy physical stiffness...

To characterize our recently established in vitro glaucomatous human trabecular meshwork (HTM) models using dexamethasone (DEX)- or TGF-β2-treated HTM cells, (1) two-dimensional (2D) cultured cells were characterized by means of the real-time cellular metabolism analysis a Seahorse analyzer, and (2) effects mechanical compression stresses toward three-dimensional (3D) spheroids evaluated analyzing gene expression several ECM proteins, inflammatory cytokines, ER stress-related factors those...

The objective of the current study was to elucidate clinicopathological significance and appearance in vitro three-dimension (3D) spheroid models oral malignant tumors that were prepared from four pathologically different squamous cell carcinoma (OSCC; low-grade; SSYP MO-1000, intermediate-grade; LEM2) adenosquamous (OASC; high-grade; Mesimo) obtained patients with stages. To characterize biological these lines themselves, two-dimensional (2D) cultured cells subjected cellular metabolic...

Adipose tissues are closely related to physiological functions and pathological conditions in most organs. Although differentiated 3T3-L1 preadipocytes have been used for vitro adipose studies, the difference cellular characteristics of adipogenic differentiation two-dimensional (2D) culture three-dimensional (3D) remain unclear. In this study, we evaluated gene expression patterns using RNA sequencing metabolic an extracellular flux analyzer with without induction 2D 3D culture. culture,...

Abstract A pan-ROCK-inhibitor, ripasudil (Rip), and a ROCK2 inhibitor, KD025, were used To study the effects of Rho-associated coiled-coil containing protein kinase (ROCK)1 2 on two-dimensional (2D) three-dimensional (3D) cultures TGFβ2-treated human trabecular meshwork (HTM) cells. In presence 5 ng/mL TGFβ2, these inhibitors characterized by transendothelial electrical resistance (TEER), FITC-dextran permeability, size stiffness 3D sphenoids, expression extracellular matrix (ECM) including...

The objective of the current study was to perform a screening drug-induced effects prostaglandin F2α (PGF2α) and EP2 agonist, omidenepag (OMD), using two- three-dimensional (2D 3D) cultures dexamethasone (DEX)-treated human trabecular meshwork (HTM) cells. on 2D monolayers were characterized by measuring transendothelial electrical resistance (TEER) fluorescein isothiocyanate (FITC)–dextran permeability, physical properties 3D spheroids, gene expression extracellular matrix (ECM) molecules,...

The objective of the current study was to examine roles ROCK1 and 2 on spatial architecture human corneal stroma. We examined effects a pan-ROCK inhibitor (pan-ROCK-i), ripasudil, ROCK2 (ROCK2-i), KD025 expression genes that encode for ECM proteins including collagen (COL) 1, 4, 6, fibronectin (FN), their regulators, tissue metalloproteinase (TIMP) 1-4, matrix (MMP) 2, 9 14, ER stress-related factors two- three-dimensional (2D 3D) cultures stroma fibroblasts (HCSFs), physical properties 3D...