Transforming growth factor-β2-mediated mesenchymal transition in lens epithelial cells is repressed in the absence of RAGE

Transforming growth factor-β2 (TGFβ2)-mediated epithelial to mesenchymal transition (EMT) in lens epithelial cells (LECs) has been implicated in fibrosis associated with secondary cataracts. In this study, we investigated whether the receptor for advanced glycation end products (RAGE) plays a role in TGFβ2-mediated EMT in LECs. Unlike in the LECs from wild-type mice, TGFβ2 failed to elicit an EMT response in LECs from RAGE knockout mice. The lack of RAGE also diminished TGFβ2-mediated Smad signaling. In addition, treatment with TGFβ2 increased IL-6 levels in LECs from wild-type mice but not in those from RAGE knockout mice. Treatment of human LECs with the RAGE inhibitor FPS-ZM1 reduced TGFβ2-mediated Smad signaling and the EMT response. Unlike that in wild-type lenses, the removal of fiber cell tissue in RAGE knockout lenses did not result in elevated levels of α-smooth muscle actin (α-SMA), fibronectin (FN), and integrin β1 in capsule-adherent LECs. Taken together, these results suggest that TGFβ2 signaling is intricately linked to RAGE. Targeting RAGE could be explored as a therapeutic strategy against secondary cataracts.

Features of the local cytokine profile in patients with secondary corneal dystrophy of various etiologies

Goal. To study the features of the local cytokine profile of patients with secondary corneal dystrophy of various etiologies. Material and methods. The study was conducted among 4 patients: 1 – with bullous kerathopathy, 1 – with secondary postherpetic corneal dystrophy, 1 – with secondary keratectasia and 1 – with secondary corneal dystrophy of neurogenic origin. The quantitative content of cytokines in the lacrimal fluid was studied: interleukin (IL) 1β, 6, 4, 10, transforming growth factor β2 (TGF-β2). Results. All patients had in 3.0-4.3-fold increase in IL-1β and a 2.0-4.1-fold increase in IL-6. In bullous kerathopathy and secondary corneal dystrophy of neurogenic origin, there was in 1.1-and 1.2-fold increase in IL-4 in both cases, and a slight increase in IL-10 and TGF-β2. At the same time, in secondary keratectasia and secondary postherpetic corneal dystrophy, there was a decrease in the level of these cytokines: IL-4 by 1.7 and 1.3 times, respectively, IL-10 by 1.2 times in both cases, and TGF-β2 by 1.5 and 1.3 times, respectively. Conclusion. The study of the local cytokine profile in patients with secondary corneal dystrophy of different etiologies indicates the presence of similar aspects of pathogenesis, which is of some interest and deserves further study. The obtained results open up prospects for the development of personalized cytokine therapy for secondary corneal dystrophy of various etiologies. Key words: secondary corneal dystrophy, immunology, cytokines.

Role of rhBMP-7, Fibronectin, And Type I Collagen in Dental Implant Osseointegration Process: An Initial Pilot Study on Minipig Animals

Background: The biological factors involved in dental implant osseointegration need to be investigated to improve implant success. Methods: Twenty-four implants were inserted into the tibias of six minipigs. Bone samples were obtained at 7, 14, and 56 days. Biomolecular analyses evaluated mRNA of BMP-4, -7, Transforming Growth Factor-β2, Interleukin-1β, and Osteocalcin in sites treated with rhBMP-7, Type 1 Collagen, or Fibronectin (FN). Inflammation and osteogenesis were evaluated by histological analyses. Results: At 7 and 14 days, BMP-4 and BMP-7 increased in the sites prepared with rhBMP-7 and FN. BMP-7 remained greater at 56 days in rhBMP-7 and FN sites. BPM-4 at 7 and 14 days increased in Type 1 Collagen sites; BMP-7 increased from day 14. FN increased the TGF-β2 at all experimental times, whilst the rhBMP-7 only did so up to 7 days. IL-1β increased only in collagen-treated sites from 14 days. Osteocalcin was high in FN-treated sites. Neutrophilic granulocytes characterized the inflammatory infiltrate at 7 days, and mononuclear cells at 14 and 56 days. Conclusions: This initial pilot study, in a novel way, evidenced that Type 1 Collagen induced inflammation and did not stimulate bone production; conversely FN or rhBMP-7 showed neo-osteogenetic and anti-inflammatory properties when directly added into implant bone site.

Modulation of the Systemic Immune Response in Suckling Rats by Breast Milk TGF-β2, EGF and FGF21 Supplementation

Breast milk is a rich fluid containing bioactive compounds such as specific growth factors (GF) that contribute to maturation of the immune system in early life. The aim of this study was to determine whether transforming growth factor-β2 (TGF-β2), epidermal growth factor (EGF) and fibroblast growth factor 21 (FGF21), compounds present in breast milk, could promote systemic immune maturation. For this purpose, newborn Wistar rats were daily supplemented with these GF by oral gavage during the suckling period (21 days of life). At day 14 and 21 of life, plasma for immunoglobulin (Ig) quantification was obtained and spleen lymphocytes were isolated, immunophenotyped and cultured to evaluate their ability to proliferate and release cytokines. The main result was obtained at day 14, when supplementation with EGF increased B cell proportion to reach levels observed at day 21. At the end of the suckling period, all GF increased the plasma levels of IgG1 and IgG2a isotypes, FGF21 balanced the Th1/Th2 cytokine response and both EGF and FGF21 modified splenic lymphocyte composition. These results suggested that the studied milk bioactive factors, mainly EGF and FGF21, may have modulatory roles in the systemic immune responses in early life, although their physiological roles remain to be established.

Altered expression of exosomal miRNAs from primary human trabecular meshwork cells induced by transforming growth factor-β2

Background Primary open-angle glaucoma (POAG) is tightly related with extracellular matrix (ECM) remodeling of human trabecular meshwork cells (HTMCs). Transforming growth factor-β2 (TGF-β2) can induce ECM remodeling. MicroRNAs (miRNAs) serve as a potential therapeutic target in influencing the development of glaucoma by regulating TGF-β2. Recent studies also have found that exosomes may be involved in the regulation of intraocular pressure in patients of glaucoma. Therefore, the aim of the current study was to investigate the exosomal miRNAs expression changes derived from HTMCs treated with TGF-β2. Methods Exosomes were isolated from HTMCs supernatant cultured for 24 h with TGF-β2. The morphology of exosome pellets was examined by transmission electron microscopy. Nanoparticle tracking analysis used to demonstrate the particle size distribution. Total exosomal RNAs were extracted for subsequent miRNA gene chip analysis to investigate differentially expressed miRNAs between the control cells and treatment cells. Gene Ontology (GO) annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were used to predict potential target and validate possible functions of the exosomal miRNAs. Results There were 23 miRNAs up-regulated and 3 miRNAs down-regulated. Go annotation and KEGG pathway enrichment analysis revealed that 469,102 and 94 GO terms involved in biological processes, cellular components and molecular function for the possible functions of the 26 miRNAs. Conclusions These findings indicate that TGF-β2 may alter exosomal miRNAs expression to participate in the pathogenesis of POAG in which multiple molecular functions and pathways are involved. They may provide significant information for potential biomarkers for POAG diagnosis, clinical treatment and potential prognosis.