ABSTRACT
BACKGROUND: Injection laryngoplasty is an option for treatment of dysphonia following vocal fold paralysis. Modified fibrin glue with suspended chondrocytes might be a favorable cell-based material for permanent vocal fold medialization. METHODS: We compared fibrin glue with suspended chondrocytes to collagen and hyaluronic acid gels concerning alteration of vocal fold vibration and correct intralaryngeal placement after intralaryngeal injection into porcine larynges. Viscoelastic properties of the materials were analyzed by means of a parallel plate rheometer. RESULTS: Fibrin glue with cells was comparable to collagen and hyaluronic acid with respect to amplitudes, symmetry, and periodicity of vocal fold vibration. Application and positioning of fibrin glue with suspended chondrocytes were technically undemanding and comparable with controls. Complex stress modulus of fibrin glue with suspended cells was comparable to that of collagen gel. CONCLUSIONS: Fibrin glue with suspended chondrocytes seems suitable for the indication of injection laryngoplasty and holds promise for permanent vocal fold medialization.
Subject(s)
Chondrocytes/transplantation , Fibrin Tissue Adhesive/administration & dosage , Tissue Engineering/methods , Viscoelastic Substances/administration & dosage , Vocal Cord Paralysis/therapy , Animals , Cells, Cultured , Collagen/administration & dosage , Humans , Hyaluronic Acid/administration & dosage , Injections , Models, Animal , Swine , Viscosupplements/administration & dosageABSTRACT
Glioblastomas are characterized by an intense local invasiveness that limits surgical resection. One mechanism by which glioma cells enforce their migration into brain tissue is reorganization of tumour associated extracellular matrix (ECM). Collagen XVI is a minor component of connective tissues. However, in glioblastoma tissue it is dramatically upregulated compared to the ECM of normal cortex. The aim of this study is to delineate tumour cell invasion and underlying mechanisms involving collagen XVI by using a siRNA mediated collagen XVI knockdown model in U87MG human glioblastoma cells. Knockdown of collagen XVI resulted in decreased invasiveness in Boyden chamber assays, and in a reduction of focal adhesion contact numbers per cell. Gene expression was upregulated for protocadherin 18 and downregulated for kindlin-1 and -2. Proliferation was not affected while flow cytometric analysis demonstrated reduced ß1-integrin activation in collagen XVI knockdown cells. We suggest that in glioblastoma tissue collagen XVI may impair the cell-cell interaction in favour of enhancement of invasion. The modification of the ß1-integrin activation pattern through collagen XVI might be a molecular mechanism to further augment the invasive phenotype of glioma cells. Elucidating the underlying mechanisms of glioma cell invasion promoted by collagen XVI may provide novel cancer therapeutic approaches in neurooncology.
Subject(s)
Central Nervous System Neoplasms/pathology , Collagen/antagonists & inhibitors , Glioma/pathology , Cadherins/metabolism , Cell Adhesion/physiology , Cell Line, Tumor , Cell Movement , Central Nervous System Neoplasms/metabolism , Collagen/genetics , Collagen/metabolism , Down-Regulation , Extracellular Matrix/metabolism , Glioma/metabolism , Humans , Integrin beta1/metabolism , Membrane Proteins/metabolism , Neoplasm Invasiveness , Neoplasm Proteins/metabolism , Protocadherins , RNA Interference , RNA, Small Interfering/metabolism , Up-RegulationABSTRACT
Type XVI collagen belongs to the family of fibril-associated collagens with interrupted triple helices (FACIT). Recently, high affinity to integrin alpha1beta1 has been shown allowing cells expressing those integrins to attach and spread on recombinant type XVI collagen. Here, we show that type XVI collagen is overexpressed in dysplastic areas of mucosal epithelium from oral squamous cell carcinoma (OSCC) patients. Induction of its expression in OSCC cell lines (COLXVI cells) leads to an increased expression of Kindlin-1. Moreover, we demonstrate a significantly increased Kindlin-1/beta1-integrin interaction. Additionally, we detected a higher number of activated beta1-integrins in COLXVI cells and found a neo-expression of alpha1 integrin subunit on these cells. FACS analysis revealed a significantly higher amount of COLXVI cells in S-phase and G2/M-phase 6h after synchronisation leading to a markedly higher proliferation activity. Blocking beta1-integrins with a specific antibody resulted in reduced proliferation of COLXVI cells. In summary, we demonstrate that overexpression of type XVI collagen in aberrant oral keratinocytes leads to Kindlin-1 induction, increased Kindlin-1/beta1-integrin interaction, integrin activation and subsequently to a proliferative cellular phenotype.
Subject(s)
Carcinoma, Squamous Cell/metabolism , Cell Proliferation , Collagen/metabolism , Mouth Neoplasms/metabolism , Cell Line, Tumor , Gene Expression/genetics , Humans , Integrin alpha1/metabolism , Integrin beta1/metabolism , Male , Membrane Proteins/genetics , Membrane Proteins/metabolism , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Protein Binding/physiology , S Phase/physiology , TransfectionABSTRACT
In Crohn's disease (CD) the stress-shield of intestinal subepithelial myofibroblasts (ISEMF) provided by intact tissue is disturbed due to inflammation and thus, cells start with remodelling activities. This is characterized by increased numbers of collagen-producing ISEMF causing an uncontrolled, irreversible wound-healing response to the chronic inflammation of the gastrointestinal tract. Reconstitution of the original ECM leads ISEMF to exit this cycle. In contrast, during fibrosis, ISEMF persist. It is known that ISEMF produce and deposit collagen types I, III, IV and V; however synthesis and the role of fibrillar peripheral molecules like collagen type XVI have not been addressed yet. Here, we have analyzed the distribution of collagen XVI in the normal and inflamed bowel wall, its gene and protein expression by ISEMF of different inflammation stages, the cell-matrix interactions in different phases of the inflammatory process and their effect on cell spreading, proliferation and migration. Collagen XVI is deposited in the submucosa of the intestinal wall where it co-localizes with fibrillin-1 and integrin alpha1. ISEMF reveal increasing gene and protein expression of collagen XVI concurrent to increasing inflammation. ISEMF reveal more mature focal adhesion contacts when seeded on collagen XVI resulting in an extensive cell spreading. This involves recruitment of alpha1beta1 integrin, which shows increased cell surface expression on ISEMF in late stages of inflammation. We assume that collagen XVI promotes persistence of ISEMF in the normal and, even stronger in the inflamed bowel wall by stabilizing focal adhesion contacts via cell-matrix interaction preferentially through recruitment of alpha1ss1 integrin into the tips of the focal adhesion contacts.
Subject(s)
Collagen/metabolism , Crohn Disease/metabolism , Extracellular Matrix/metabolism , Focal Adhesions/metabolism , Biopsy , Blotting, Western , Cell Adhesion/physiology , Cell Proliferation , Collagen/genetics , Collagen/ultrastructure , Crohn Disease/pathology , Extracellular Matrix/pathology , Extracellular Matrix/ultrastructure , Fibroblasts , Flow Cytometry , Focal Adhesions/pathology , Focal Adhesions/ultrastructure , Humans , Microscopy, Confocal , Microscopy, Electron, Scanning , Microscopy, Fluorescence , RNA/chemistry , RNA/genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The poor prognosis of glioblastoma patients is related to diffuse brain invasion and interaction of tumor cells with extracellular matrices (ECM). We describe expression and function of the FACIT-collagen XVI in glioblastomas. We found upregulation of collagen XVI mRNA as well as protein in glioblastomas as compared to normal cortex. SiRNA knockdown resulted in decreased cell adhesion whereas increased adhesion was observed on surfaces coated with collagen XVI. The migration of glioblastoma cells on this substrate remained unchanged. Our results demonstrate de-novo expression of collagen XVI in glioblastomas as part of the tumor specific remodeling of the ECM.
Subject(s)
Cell Adhesion , Cell Movement , Collagen/biosynthesis , Fibril-Associated Collagens/biosynthesis , Glioblastoma/pathology , Cell Line, Tumor , Collagen/genetics , Extracellular Matrix/metabolism , Fibril-Associated Collagens/genetics , Glioblastoma/metabolism , Humans , Up-RegulationSubject(s)
Collagen/metabolism , Extracellular Matrix/chemistry , Ganglia, Spinal , Nerve Regeneration/physiology , Neurons/metabolism , Protein Isoforms/metabolism , Animals , Collagen/genetics , Female , Ganglia, Spinal/chemistry , Ganglia, Spinal/cytology , Ganglia, Spinal/pathology , In Situ Hybridization , Mice , Neurons/cytology , Protein Isoforms/genetics , Sciatic Nerve/cytology , Sciatic Nerve/metabolism , Sciatic Nerve/pathologyABSTRACT
Fracture repair recapitulates in adult organisms the sequence of cell biological events of endochondral ossification during skeletal development and growth. After initial inflammation and deposition of granulation tissue, a cartilaginous callus is formed which, subsequently, is remodeled into bone. In part, bone formation is influenced also by the properties of the extracellular matrix of the cartilaginous callus. Deletion of individual macromolecular components can alter extracellular matrix suprastructures, and hence stability and organization of mesenchymal tissues. Here, we took advantage of the collagen IX knockout mouse model to better understand the role of this collagen for organization, differentiation and maturation of a cartilaginous template during formation of new bone. Although a seemingly crucial component of cartilage fibrils is missing, collagen IX-deficient mice develop normally, but are predisposed to premature joint cartilage degeneration. However, we show here that lack of collagen IX alters the time course of callus differentiation during bone fracture healing. The maturation of cartilage matrix was delayed in collagen IX-deficient mice calli as judged by collagen X expression during the repair phase and the total amount of cartilage matrix was reduced. Entering the remodeling phase of fracture healing, Col9a1(-/-) calli retained a larger percentage of cartilage matrix than in wild type indicating also a delayed formation of new bone. We concluded that endochondral bone formation can occur in collagen IX knockout mice but is impaired under conditions of stress, such as the repair of an unfixed fractured long bone.
