Delineation of VEGF-regulated genes and functions in the cervix of pregnant rodents by DNA microarray analysis
cells; Endocrinology & Metabolism; endothelial growth-factor; estrogen; factor expression; nitric-oxide; parturition; rat; Reproductive Biology; tissue; tumor angiogenesis; vascular-permeability
Background: VEGF-regulated genes in the cervices of pregnant and non-pregnant rodents (rats and mice) were delineated by DNA microarray and Real Time PCR, after locally altering levels of or action of VEGF using VEGF agents, namely siRNA, VEGF receptor antagonist and mouse VEGF recombinant protein. Methods: Tissues were analyzed by genome-wide DNA microarray analysis, Real-time and gel-based PCR, and SEM, to decipher VEGF function during cervical remodeling. Data were analyzed by EASE score (microarray) and ANOVA (Real Time PCR) followed by Scheffe's F-test for multiple comparisons. Results: Of the 30,000 genes analyzed, about 4,200 genes were altered in expression by VEGF, i.e., expression of about 2,400 and 1,700 genes were down- and up-regulated, respectively. Based on EASE score, i.e., grouping of genes according to their biological process, cell component and molecular functions, a number of vascular- and non-vascular-related processes were found to be regulated by VEGF in the cervix, including immune response (including inflammatory), cell proliferation, protein kinase activity, and cell adhesion molecule activity. Of interest, mRNA levels of a select group of genes, known to or with potential to influence cervical remodeling were altered. For example, real time PCR analysis showed that levels of VCAM-1, a key molecule in leukocyte recruitment, endothelial adhesion, and subsequent trans-endothelial migration, were elevated about 10 folds by VEGF. Further, VEGF agents also altered mRNA levels of decorin, which is involved in cervical collagen fibrillogenesis, and expression of eNO, PLC and PKC mRNA, critical downstream mediators of VEGF. Of note, we show that VEGF may regulate cervical epithelial proliferation, as revealed by SEM. Conclusion: These data are important in that they shed new insights in VEGF's possible roles and mechanisms in cervical events near-term, including cervical remodeling.
Mowa C N; Li T B; Jesmin S; Folkesson H G; Usip S E; Papka R E; Hou G C
Reproductive Biology and Endocrinology
2008
2008-12
Journal Article
<a href="http://doi.org/10.1186/1477-7827-6-64" target="_blank" rel="noreferrer noopener">10.1186/1477-7827-6-64</a>
Characterization of the cellular origin of a tissue-engineered human phalanx model by in situ hybridization
cells; Cell Biology; expression; endothelial growth-factor; cartilage; joints
Tissue-engineered models of human phalanges have previously been fabricated from a combination of bovine periosteum, cartilage, tendon, and biodegradable polyglycolic acid and poly-L-lactic acid scaffolds. Resulting constructs implanted in athymic mice for more than 40 weeks developed new bone, cartilage, and tendon and became vascularized, but cell types comprising the constructs were unidentified. The origin of cells in middle phalanx models implanted for 20 weeks in nude mice has been studied by in situ hybridization analyzing species-specific gene expression. Oligonucleotide probes homologous to species-specific gene sequences of bovine type 11 and X collagen, aggrecan, bone sialoprotein, biglycan, and osteopontin, and mouse decorin were labeled with S-35 and hybridized to respective serial sections of bovine tissue, mouse tissue, and phalanx constructs. In situ hybridization showed positive message and tissue-specific localization for all bovine-specific probes examined within cartilaginous and midshaft portions of constructs and negative message for the mouse-specific decorin probe. These data show that osteoblasts and chondrocytes comprising constructs are derived exclusively from their original bovine sources over 20 weeks of implantation. Defining the cellular origin of the models lends insight into their biological, chemical, and physical nature and their growth and development. Maintenance of their initial genotype is crucial for future application of the models in augmenting impaired human phalanges and related tissues.
Chubinskaya S; Jacquet R; Isogai N; Asamura S; Landis W J
Tissue Engineering
2004
2004-07
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1089/1076327041887862" target="_blank" rel="noreferrer noopener">10.1089/1076327041887862</a>
A New Bioartificial Pancreas Utilizing Amphiphilic Membranes For The Immunoisolation Of Porcine Islets A Pilot Study In The Canine
biocompatibility; endothelial growth-factor; Engineering; mice; networks; oxygen-tension; pig islets; rat; survival; transplantation; transplantation; vascularization
We have developed a replaceable bioartificial pancreas to treat diabetes utilizing a unique cocontinous amphiphilic conetwork membrane created for macroencapsulation and immunoisolation of porcine islet cells (PICs). The membrane is assembled from hydrophilic poly(N,N-dimethyl acrylamide) and hydrophobic/oxyphilic polydimethylsiloxane chains crosslinked with hydrophobic/oxyphilic polymethylhydrosiloxane chains. Our hypothesis is that this membrane allows the survival of xenotransplanted PICs in the absence of prevascularization or immunosuppression because of its extraordinarily high-oxygen permeability and small hydrophilic channel dimensions (3-4 nm). The key components are a 5-10 mu m thick semipermeable amphiphilic conetwork membrane reinforced with an electrospun nanomat of polydimethylsiloxane-containing polyurethane, and a laser-perforated nitinol scaffold to provide geometric stability. Devices were loaded with PICs and tested for their ability to maintain islet viability without prevascularization, prevent rejection, and reverse hyperglycemia in three pancreatectomized dogs without immunosuppression. Tissue tolerance was good and there was no systemic toxicity. The bioartificial pancreas protected PICs from toxic environments in vitro and in vivo. Islets remained viable for up to 3 weeks without signs of rejection. Neovascularization was observed. Hyperglycemia was not reversed, most likely because of insufficient islet mass. Further studies to determine long-term islet viability and correction of hyperglycemia are warranted. ASAIO Journal 2009; 55:400-405.
Grundfest-Broniatowski S F; Tellioglu G; Rosenthal K S; Kang J; Erdodi G; Yalcin B; Cakmak M; Drazba J; Bennett A; Lu L; Kennedy J P
Asaio Journal
2009
2009-07
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1097/MAT.0b013e3181a8deba" target="_blank" rel="noreferrer noopener">10.1097/MAT.0b013e3181a8deba</a>