The GM-CSF/CCL17 pathway in obesity-associated osteoarthritic pain and disease in mice.

OBJECTIVES: We have previously identified a granulocyte macrophage-colony stimulating factor (GM-CSF)/C-C motif ligand 17 (CCL17) pathway in monocytes/macrophages, in which GM-CSF regulates the formation of CCL17, and it is important for an experimental osteoarthritis (OA) model. We explore here additional OA models, including in the presence of obesity, such as a requirement for this pathway. DESIGN: The roles of GM-CSF, CCL17, CCR4, and CCL22 in various experimental OA models, including those incorporating obesity (eight-week high-fat diet), were investigated using gene-deficient male mice. Pain-like behavior and arthritis were assessed by relative static weight distribution and histology, respectively. Cell populations (flow cytometry) and cytokine messenger RNA (mRNA) expression (qPCR) in knee infrapatellar fat pad were analyzed. Human OA sera were collected for circulating CCL17 levels (ELISA) and OA knee synovial tissue for gene expression (qPCR). RESULTS: We present evidence that: i) GM-CSF, CCL17, and CCR4, but not CCL22, are required for the development of pain-like behavior and optimal disease in three experimental OA models, as well as for exacerbated OA development due to obesity, ii) obesity alone leads to spontaneous knee joint damage in a GM-CSF- and CCL17-dependent manner, and iii) in knee OA patients, early indications are that BMI correlates with a lower Oxford Knee Score (r = -0.458 and p = 0.0096), with elevated circulating CCL17 levels (r = 0.2108 and p = 0.0153) and with elevated GM-CSF and CCL17 gene expression in OA synovial tissue. CONCLUSIONS: The above findings indicate that GM-CSF, CCL17, and CCR4 are involved in obesity-associated OA development, broadening their potential as targets for possible treatments for OA.

Retrospective Analysis of Oral and Maxillofacial Biopsies: An Institutional Study.

AIM: To determine the preponderance and the distribution of pathological lesions in oral and maxillofacial region reported in and around the Madurai city. STUDY DESIGN: Retrospective study on the oral and maxillofacial biopsies taken during 11 years period from 2007 to 2018 in a CSI Dental College, Madurai, Tamilnadu. A total of 1000 cases were analyzed and 904 were selected. The parameters recorded were age, sex, area, and the histopathological report obtained. Descriptive statistics was used for analysis. RESULT: The most common soft tissue lesion was traumatic fibroma (16.1%) and the prevalent hard tissue lesion was periapical cyst (12.7%). The more common oral malignant tumour observed was squamous cell carcinoma (7.3%) and benign tumor was ameloblastoma (5.3%). The frequently affected sites were mandible (36.1%) and buccal mucosa (19.8%). There was more predilection among male than females. The frequency of lesion is common in age group of 31-60 years. A considerable similarity was observed between age, type of lesion and its location. CONCLUSIONS: This study evaluated chronic periapical lesions were the most common hard tissue lesions and fibromas in soft tissue category. The idea of this histopathological study concludes on the requirement for biopsy to rule out type of the lesion and its severity to start up the earlier management.

Modeling early stage atherosclerosis in a primary human vascular microphysiological system.

Novel atherosclerosis models are needed to guide clinical therapy. Here, we report an in vitro model of early atherosclerosis by fabricating and perfusing multi-layer arteriole-scale human tissue-engineered blood vessels (TEBVs) by plastic compression. TEBVs maintain mechanical strength, vasoactivity, and nitric oxide (NO) production for at least 4 weeks. Perfusion of TEBVs at a physiological shear stress with enzyme-modified low-density-lipoprotein (eLDL) with or without TNFα promotes monocyte accumulation, reduces vasoactivity, alters NO production, which leads to endothelial cell activation, monocyte accumulation, foam cell formation and expression of pro-inflammatory cytokines. Removing eLDL leads to recovery of vasoactivity, but not loss of foam cells or recovery of permeability, while pretreatment with lovastatin or the P2Y11 inhibitor NF157 reduces monocyte accumulation and blocks foam cell formation. Perfusion with blood leads to increased monocyte adhesion. This atherosclerosis model can identify the role of drugs on specific vascular functions that cannot be assessed in vivo.

Oxygen consumption in human, tissue-engineered myobundles during basal and electrical stimulation conditions.

During three-dimensional culture of skeletal muscle in vitro, electrical stimulation provides an important cue to enhance skeletal muscle mimicry of the in vivo structure and function. However, increased respiration can cause oxygen transport limitations in these avascular three-dimensional constructs, leading to a hypoxic, necrotic core, or nonuniform cell distributions in larger constructs. To enhance oxygen transport with convection, oxygen concentrations were measured using an optical sensor at the inlet and outlet of an 80 µl fluid volume microphysiological system (MPS) flow chamber containing three-dimensional human skeletal muscle myobundles. Finite element model simulations of convection around myobundles and oxygen metabolism by the myobundles in the 80 µl MPS flow chamber agreed well with the oxygen consumption rate (OCR) at different flow rates, suggesting that under basal conditions, mass transfer limitations were negligible for flow rates above 1.5 µl s-1. To accommodate electrodes for electrical stimulation, a modified 450 µl chamber was constructed. Electrical stimulation for 30 min increased the measured rate of oxygen consumption by the myobundles to slightly over 2 times the basal OCR. Model simulations indicate that mass transfer limitations were significant during electrical stimulation and, in the absence of mass transfer limitations, electrical stimulation induced about a 20-fold increase in the maximum rate of oxygen consumption. The results indicate that simulated exercise conditions increase respiration of skeletal muscle and mass transfer limitations reduce the measured levels of oxygen uptake, which may affect previous studies that model exercise with engineered muscle.

Genetically Encoding Albumin Binding into Chemotherapeutic-loaded Polypeptide Nanoparticles Enhances Their Antitumor Efficacy.

We report the development of drug-encapsulating nanoparticles that bind endogenous albumin upon intravenous injection and evaluate their in vivo performance in a murine as well as canine animal model. The gene encoding a protein-G derived albumin binding domain (ABD) was fused to that of a chimeric polypeptide (CP), and the ABD-CP fusion was recombinantly synthesized by bacterial expression of the gene. Doxorubicin (DOX) was conjugated to the C-terminus of the ABD-CP fusion, and conjugation of multiple copies of the drug to one end of the ABD-CP triggered its self-assembly into ∼100 nm diameter spherical micelles. ABD-decorated micelles exhibited submicromolar binding affinity for albumin and also preserved their spherical morphology in the presence of albumin. In a murine model, albumin-binding micelles exhibited dose-independent pharmacokinetics, whereas naked micelles exhibited dose-dependent pharmacokinetics. In addition, in a canine model, albumin binding micelles resulted in a 3-fold increase in plasma half-life and 6-fold increase in plasma exposure as defined by the area under the curve (AUC) of the drug, compared with naked micelles. Furthermore, in a murine colon carcinoma model, albumin-binding nanoparticles demonstrated lower uptake by the reticuloendothelial system (RES) system organs, the liver and spleen, that are the main target organs of toxicity for nanoparticulate delivery systems and higher uptake by the tumor than naked micelles. The increased uptake by s.c. C26 colon carcinoma tumors in mice translated to a wider therapeutic window of doses ranging from 20 to 60 mg equivalent of DOX per kg body weight (mg DOX equiv·kg-1 BW) for albumin-binding ABD-CP-DOX micelles, as compared to naked micelles that were only effective at their maximum tolerated dose of 40 mg DOX equiv·kg-1 BW.