ABSTRACT
Nanofibers are thought to enhance cell adhesion, growth, and function. We demonstrate that the choice of building blocks in self-assembling nanofiber systems can be used to control cell behavior. The use of 2 D-coated, self-assembled nanofibers in controlling lens epithelial cells, fibroblasts, and mesenchymal stem cells was investigated, focusing on gene and protein expression related to the fibrotic response. To this end, three nanofibers with different characteristics (morphology, topography, and wettability) were compared with two standard materials frequently used in culturing cells, TCPS, and a collagen type I coating. Cell metabolic activity, cell morphology, and gene and protein expression were analyzed. The most hydrophilic nanofiber with more compact network consisting of small fibers proved to provide a beneficial 2 D environment for cell proliferation and matrix formation while decreasing the fibrotic/stress behavior in all cell lines when compared with TCPS and the collagen type I coating. This nanofiber demonstrates the potential to be used as a biomimetic coating to study the development of fibrosis through epithelial-to-mesenchymal transition. This study also shows that nanofiber structures do not enhance cell function by definition, because the physico-chemical characteristics of the nanofibers influence cell behavior as well and actually can be used to regulate cell behavior toward suboptimal performance. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2252-2265, 2017.
Subject(s)
Coated Materials, Biocompatible/chemistry , Epithelial Cells/cytology , Fibroblasts/cytology , Mesenchymal Stem Cells/cytology , Nanofibers/chemistry , Tissue Scaffolds/chemistry , Cell Adhesion , Cell Line , Cell Proliferation , Cells, Cultured , Coated Materials, Biocompatible/adverse effects , Epithelial Cells/metabolism , Epithelial Cells/pathology , Epithelial-Mesenchymal Transition , Fibroblasts/metabolism , Fibroblasts/pathology , Fibrosis/etiology , Fibrosis/metabolism , Fibrosis/pathology , Gene Expression Regulation , Humans , Hydrophobic and Hydrophilic Interactions , Mesenchymal Stem Cells/metabolism , Mesenchymal Stem Cells/pathology , Nanofibers/adverse effects , Nanofibers/ultrastructure , Tissue Scaffolds/adverse effectsABSTRACT
PURPOSE: To moderate the capsular opacification (CO) response after lens surgery, an experimental study was performed in which nanofibre-based hydrogels (nanogels) with different ratios of attached peptides were applied to provide extracellular matrix-related cues for lens epithelial cells (LECs) in a porcine eye model. METHODS: The lens content was removed, and the capsules were refilled with nanogel. Lenses were divided into two groups, the first group (n = 34) was refilled with nanogels containing different ratios of two laminin-derived peptides (IKVAV + YIGSR), and the latter group (n = 26) was refilled with nanogel combinations of a fibronectin-derived and a type IV collagen-derived peptide (RGDS + DGEA). Two lenses were refilled with culture medium to investigate the effect of the medium on LECs. After refilling, lenses were extracted and cultured for 3 weeks. Lens epithelial cells (LECs) were assessed for morphology and alpha-smooth muscle actin (αSMA) expression using confocal laser scanning microscopy. RESULTS: Differences were seen in cell morphology between lenses refilled with nanogels with IKVAV + YIGSR and RGDS + DGEA peptides. In nanogels with IKVAV + YIGSR peptides, differences in LEC morphology were largest when ratios between the peptides were unequal, whereas LEC responses from the RGDS + DGEA refilled groups showed variation in LEC morphology dependent on the total quantity of mixed-in peptides. The culture medium did not induce proliferation or transformation of LECs. CONCLUSIONS: Ratios and concentrations of cell adhesion-mediating peptides both can direct the LEC response, depending on the adhesion molecules of origin, by influencing LEC proliferation and transformation. Nanogels with incorporated peptides may be tuned towards CO prevention.
Subject(s)
Capsule Opacification/prevention & control , Lens Capsule, Crystalline/drug effects , Peptides/pharmacology , Polyethylene Glycols/pharmacology , Polyethyleneimine/pharmacology , Actins/metabolism , Animals , Cataract Extraction , Collagen Type IV/chemistry , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Fibronectins/chemistry , Hydrogels/chemistry , Laminin/chemistry , Lens Capsule, Crystalline/metabolism , Nanogels , Peptides/chemistry , Polyethylene Glycols/chemistry , Polyethyleneimine/chemistry , Sus scrofaABSTRACT
Nanofiber-based hydrogels (nanogels) with different, covalently bound peptides were used as an extracellular environment for lens epithelial cells (LECs) in order to modulate the capsular opacification (CO) response after lens surgery in a porcine eye model. Lenses were divided into 15 groups (n = 4 per group), the lens content was removed and the empty capsules were refilled with nanogel without peptides and nanogels with 13 combinations of 5 different peptides: two laminin-derived, two fibronectin-derived, and one collagen IV-derived peptide representing cell adhesion motifs. A control group of 4 lenses was refilled with hyaluronan. After refilling, lenses were extracted from the porcine eye and cultured for three weeks. LECs were assessed for morphology and alpha smooth muscle actin (αSMA) expression using confocal laser scanning microscopy. Compared to hyaluronan controls, lenses filled with nanogel had less CO formation, indicated by a lower αSMA expression (P = 0.004). Microscopy showed differences in morphological cell response within the nanogel refilled groups. αSMA expression in these groups was highest in lenses refilled with nanogel without peptides (9.54 ± 11.29%). Overall, LEC transformation is reduced by the presence of nanogels and the response is improved even further by incorporation of extracellular matrix peptides representing adhesion motifs. Thus, nanomaterials targeting biological pathways, in our case interactions with integrin signaling, are a promising avenue toward reduction of CO. Further research is needed to optimize nanogel-peptide combinations that fully prevent CO.
Subject(s)
Capsule Opacification/prevention & control , Epithelial Cells/cytology , Epithelial-Mesenchymal Transition/drug effects , Extracellular Matrix Proteins/administration & dosage , Hydrogels , Lens Capsule, Crystalline/cytology , Oligopeptides/administration & dosage , Actins/metabolism , Animals , Biomarkers/metabolism , Capsule Opacification/pathology , Collagen Type IV/administration & dosage , Collagen Type IV/chemical synthesis , Drug Delivery Systems , Extracellular Matrix Proteins/chemical synthesis , Fibronectins/administration & dosage , Fibronectins/chemical synthesis , Fluorescent Antibody Technique, Indirect , Laminin/administration & dosage , Laminin/chemical synthesis , Lens, Crystalline/cytology , Nanofibers , Oligopeptides/chemical synthesis , Organ Culture Techniques , Sus scrofaABSTRACT
A convergent synthesis of racemic 5-amino[6]hexahelicene is described. Cross-coupling reactions are used to assemble a pentacyclic framework, and a metal-catalyzed ring-closure comprises the final step. The enantiomers were separated by means of chromatography and the absolute configurations were assigned by comparison of the CD spectra with hexahelicene. The t1/2 â value for racemization at 210 °C was approximately 1â hour. Scanning tunneling microscopy (STM) measurements were carried out on enantiopure and racemic samples of aminohelicene on Au(111) under ultrahigh vacuum (UHV) conditions.
ABSTRACT
Posterior capsular opacification (PCO) is a common complication of cataract surgery. The development of PCO is due to a combination of the processes of proliferation, migration, and transdifferentiation of residual lens epithelial cells (LECs) on the lens capsule. In the past decades, various forms of PCO prevention have been examined, including adjustments of techniques and intraocular lens materials, pharmacological treatments, and prevention by interfering with biological processes in LECs. The only method so far that seems effective is the implantation of an intraocular lens with sharp edged optics to mechanically prevent PCO formation. In this review, current knowledge of the prevention of PCO will be described. We illustrate the biological pathways underlying PCO formation and the various approaches to interfere with the biological processes to prevent PCO. In this type of prevention, the use of nanotechnological advances can play a role.
Subject(s)
Capsule Opacification/prevention & control , Posterior Capsule of the Lens/pathology , Capsule Opacification/etiology , Cataract Extraction/adverse effects , Cell Movement , Cell Proliferation , Epithelial Cells/pathology , Humans , Lens, Crystalline/pathologyABSTRACT
The design, synthesis and biological evaluation of novel triazolyl p38α MAPK inhibitors with improved water solubility for formulation in cationic liposomes (SAINT-O-Somes) targeted at diseased endothelial cells is described. Water-solubilizing groups were introduced via a 'click' reaction of functional azides with 2-alkynyl imidazoles and isosteric oxazoles to generate two small libraries of 1,4-disubstituted 1,2,3-triazolyl p38α MAPK inhibitors. Triazoles with low IC50 values and desired physicochemical properties were screened for in vitro downregulation of proinflammatory gene expression and were formulated in SAINT-O-Somes. Triazolyl p38α MAPK inhibitor 88 (IC50=0.096 µM) displayed the most promising in vitro activity.
Subject(s)
Imidazoles/chemical synthesis , Protein Kinase Inhibitors/chemical synthesis , Triazoles/chemistry , Triazoles/chemical synthesis , p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors , Human Umbilical Vein Endothelial Cells , Humans , Imidazoles/chemistry , Imidazoles/metabolism , Liposomes/chemistry , Protein Binding , Protein Kinase Inhibitors/chemistry , Protein Kinase Inhibitors/metabolism , Solubility , Structure-Activity Relationship , Triazoles/metabolism , Water/chemistry , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
H(3)R inverse agonists based on an aminopropoxy-phenyloxazoline framework constitute highly valuable druglike lead compounds that display efficacy in a mouse model of recognition memory.
Subject(s)
Histamine H3 Antagonists/pharmacology , Oxazoles/pharmacology , Receptors, Histamine H3/chemistry , Animals , CHO Cells , Caco-2 Cells , Cell Line , Cricetinae , Cricetulus , Drug Design , Drug Inverse Agonism , Histamine H3 Antagonists/chemical synthesis , Histamine H3 Antagonists/chemistry , Humans , Imidazoles/chemistry , Mice , Oxazoles/chemical synthesis , Oxazoles/chemistry , Prefrontal Cortex/drug effects , Rats , Rats, Wistar , Receptors, Histamine H3/metabolism , Structure-Activity RelationshipABSTRACT
Research on the therapeutic applications of the histamine H3 receptor (H3R) has traditionally focused on antagonists/inverse agonists. In contrast, H3R agonists have received less attention despite their potential use in several disease areas. The lower availability of H3R agonists not only hampers their full therapeutic exploration, it also prevents an unequivocal understanding of the structural requirements for H3R activation. In the light of these important issues, we present our findings on 4-benzyl-1H-imidazole-based H3R agonists. Starting from two high throughput screen hits (10 and 11), the benzyl side chain was altered with lipophilic groups using combinatorial and classical chemical approaches (compounds 12-31). Alkyne- or oxazolino-substituents gave excellent affinities and agonist activities up to the single digit nM range. Our findings further substantiate the growing notion that basic ligand sidechains are not necessary for H 3R activation and reveal the oxazolino group as a hitherto unexplored functional group in H3R research.
Subject(s)
Histamine Agonists/chemical synthesis , Imidazoles/chemical synthesis , Oxazoles/chemical synthesis , Receptors, Histamine H3/metabolism , Animals , CHO Cells , Combinatorial Chemistry Techniques , Cricetinae , Cricetulus , Cytochrome P-450 Enzyme System/metabolism , Drug Design , Guinea Pigs , Histamine Agonists/chemistry , Histamine Agonists/pharmacology , Humans , Imidazoles/chemistry , Imidazoles/pharmacology , In Vitro Techniques , Intestines/drug effects , Intestines/physiology , Models, Molecular , Muscle Contraction/drug effects , Muscle, Smooth/drug effects , Muscle, Smooth/physiology , Oxazoles/chemistry , Oxazoles/pharmacology , Protein Binding , Radioligand Assay , Structure-Activity RelationshipABSTRACT
In this study, the piperidine ring of immepip and its analogues was replaced by a rigid heterocyclic pyridine ring. Many compounds in the series exhibit high affinity and agonist activity at the human histamine H(3) receptor. Particularly, the 4-pyridinyl analogue of immepip (1c, immethridine) is identified as a novel potent and highly selective histamine H(3) receptor agonist (pK(i) = 9.07, pEC(50) = 9.74) with a 300-fold selectivity over the closely related H(4) receptor.