Biomedical subjects

L Roncali

Publications and source records attributed to L Roncali.

At least 55 records · Page 3Linked to original sources

PubMed1999

Localization of factor VIII-related antigen in the endothelium of the chick embryo chorioallantoic membrane.

In this study, by using a polyclonal antibody against factor VIII-related antigen (FVIII-RA), we have examined the expression of FVIII-RA in the blood and lymphatic vessels of the chick embryo chorioallantoic membrane (CAM). The antibody marked the endothelium of blood and lymphatic vessels starting from day 8 of incubation and the cytoplasm of the allantoic epithelial cells. The application of this antibody may be useful for quantifying neovascularization in response to various angiogenic stimuli applied to the CAM.

Allantois↗

PubMed1999

Endogenous and exogenous fibroblast growth factor-2 modulate wound healing in the chick embryo chorioallantoic membrane.

Re-epithelization and the formation of a granulation tissue consisting of inflammatory cells, newly formed blood vessels, and fibroblasts embedded in a loose collagenous extracellular matrix, are critical events occurring during wound healing. In this study, utilizing the chick embryo chorioallantoic membrane (CAM) as an in vivo model of wound healing, we investigated the role of endogenous and exogenous fibroblast growth factor-2 (FGF-2) in the wound healing reparative processes. The results showed that: (1) neutralizing anti-FGF-2 antibodies (400 ng/embryo) decreased significantly the rate of wound healing (occurring only in 25% of specimens) when applied close to the edge of the wound, causing a significant decrease of microvessel and fibroblast density, and of an inflammatory macrophage infiltrate in the wounded area; (2) conversely, the application of exogenous recombinant FGF-2 (1.0 microg/embryo) greatly accelerated the wound repair occurring approximately 24h earlier than in untreated CAMs, stimulating angiogenesis, fibroblast proliferation, and macrophage infiltration. These findings demonstrate the role of FGF-2 in wound healing of the CAM and suggest that CAM, usually employed as an in vivo assay to study angiogenesis, can also be utilized as an in vivo model for the easy, rapid, and economic screening of molecules potentially able to affect the wound healing process.

Journal Article↗

PubMed1999

Angiogenesis and endothelium phenotype expression in embryonic adrenal gland and cerebellum grafted onto chorioallantoic membrane.

Vascularization and endothelial phenotype were investigated in embryonic tissues grafted onto chorioallantoic membrane (CAM) by means of immunocytochemistry and electron microscopy. Single grafts of adrenal gland or cerebellum and double grafts of adrenal gland plus cerebellum were performed, using tissues from chick or quail embryos as donors and CAMs of chick embryos as recipients. Vessels of quail origin were discriminated from those of chick origin by the anti-MB1 monoclonal antibody, specific for antigenic determinants of the quail endothelial cells. The cerebellum endothelia were distinguished from the adrenal and CAM endothelia by a polyclonal antibody against the isoform 1 of the glucose transporter (GLUT1), which is a marker of barrier-provided brain vessels. The observations, carried out, 6 days after implantation, revealed the new-growth of microvessels from the CAM into the grafted tissues, and vice versa, in both single and double transplants. In addition, in the double grafts, adrenal-derived vessels were seen to grow into the cerebellum and cerebellum-native vessels into the adrenal tissue. The combined immunocytochemical and electronmicroscopical study demonstrated that the adrenal, fenestrated sinusoids and the cerebellar, barrier-provided capillaries maintain their original phenotype when they grow within the non-native tissues. The conventional theory on the endothelial responsiveness to environmental signals has been discussed and some concluding remarks have been made.

Journal Article↗

PubMed1999

Bone marrow angiogenesis and mast cell density increase simultaneously with progression of human multiple myeloma.

Immunohistochemical, cytochemical and ultrastructural data showing vivid angiogenesis and numerous mast cells (MCs) in the bone marrow of 24 patients with active multiple myeloma (MM) compared with 34 patients with non-active MM and 22 patients with monoclonal gammopathy of undetermined significance (MGUS) led us to hypothesize that angiogenesis parallels progression of MM, and that MCs participate in its induction via angiogenic factors in their secretory granules.

Adult↗

PubMed1999

Temporal expression of the matrix metalloproteinase MMP-2 correlates with fibronectin immunoreactivity during the development of the vascular system in the chick embryo chorioallantoic membrane.

In this study we have examined in the chick embryo chorioallantoic membrane (ChAM) the expression of the matrix metalloproteinase 2 (MMP-2) and have correlated this parameter with the expansion of the ChAM vasculature and with the expression of 3 extracellular matrix components (fibronectin, laminin and type IV collagen), which differentially modulate angiogenesis. In the early phases of ChAM development, between d 6 and d 8 of incubation, when the increase of the ChAM vasculature is maximal, higher values of MMP-2 and, respectively, of fibronectin immunoreactive area, are detectable. These results indicate that MMP-2 activity and fibronectin expression are 2 strictly related components of angiogenesis occurring in vivo.

Animals↗

PubMed1999

In vivo angiogenic activity of urokinase: role of endogenous fibroblast growth factor-2.

In vitro experimental evidences suggest that the proteolytic degradation of the extracellular matrix (ECM) by activation of the urokinase-type plasminogen activator (uPA)/plasmin system may affect growth factor activity and bioavailability. However, no direct in vivo observations were available to support this hypothesis. Here we demonstrate that endothelial GM 7373 cells overexpressing human uPA (uPA-R5 cells) cause the release of (125)I-labeled fibroblast growth factor-2 (FGF2) from endothelial ECM in a plasmin-dependent manner. Accordingly, uPA-R5 cells are angiogenic in vivo when applied on the top of the chorioallantoic membrane (CAM) of the chick embryo. In contrast, mock-transfected Neo2 cells are unable to release ECM-bound (125)I-FGF2 and are poorly angiogenic. Neovascularization elicited by uPA-R5 cells is significantly reduced by neutralizing anti-FGF2 antibodies to values similar to those observed in Neo2 cell-treated CAMs. Accordingly, purified human uPA stimulates neovascularization of the CAM in the absence of an inflammatory response. The angiogenic activity of uPA is significantly inhibited by neutralizing anti-FGF2 antibodies or by pretreatment with phenylmethylsulfonyl fluoride. The non-catalytic, receptor-binding amino-terminal fragment of uPA is instead non angiogenic. Taken together, the data indicate that uPA is able to induce angiogenesis in vivo via a plasmin-dependent degradation of ECM that causes the mobilization of stored endogenous FGF2.

Allantois↗

PubMed1998

Glucose transporter GLUT1 localization in human foetus telencephalon.

The endothelial cells of the mature cerebral microvessels, provided with barrier devices (blood-brain barrier, BBB), selectively express the glucose transporter isoform 1 (GLUT1). Presence and localization of the GLUT1 were studied by immunogold silver staining (IGSS) labelling on ultrathin sections of foetal human telencephalon tissue embedded in Lowicryl HM20 according to the progressive lowering of temperature (PLT) method. In the microvascular endothelial cells of the human telencephalon GLUT1 molecules are detected at the 12th gestational week and their expression is increased at the 18th week. In both ages, the transporter is mainly localized on the ablumenal and lateral endothelial cell membranes, and at 18 weeks a greater number of GLUT1 antigenic sites are also seen at the lumenal membrane. Our findings demonstrate both the expression and subcellular localization of GLUT1 be developmentally regulated and suggest an early functioning of the BBB-GLUT1 transporter in the developing human brain.

Blood-Brain Barrier↗

PubMed1998

ICAM 1 expression and fluid phase endocytosis of cultured brain microvascular endothelial cells following exposure to interferon beta-1a and TNFalpha.

We have studied the effect of interferon (IF) beta-1a on the basal and TNFalpha-induced intercellular adhesion molecule 1 (ICAM 1) expression and fluid phase endocytosis (FPE) of horseradish peroxidase in cultured rat brain microvascular endothelial cells. Neither basal ICAM 1 expression nor basal FPE were significantly affected by 24-72 h exposure to 1000 U/ml IFbeta-1a. ICAM 1 induction and FPE enhancement caused by 100 U/ml TNFalpha for 24 h was not influenced by simultaneous administration of 1000 U/ml IFbeta-1a. Treatment of cultures with IFbeta-1a for 48 h followed by 24-h coincubation with TNFalpha (100 U/ml) and IFbeta-1a (1000 U/ml) resulted in significant downregulation of TNFalpha-induced ICAM 1 expression and FPE. Downregulation of TNFalpha-induced ICAM 1 expression was not observed when combined treatment with TNFalpha (100 U/ml) and IFbeta-1a (1000 U/ml) for 24 h was followed by 48 h exposure to IFbeta-1a. We concluded that the blood-brain barrier endothelium may be a target of IFbeta-1a. Further, these in vitro findings may correlate with the results of recent clinical trials indicating that chronic treatment of relapsing remitting multiple sclerosis with IFbeta-1a prevents both clinical exacerbations and the appearance on Magnetic Resonance Imaging of new lesions enhanced by gadolinium which is taken up by increased transendothelial fluid phase vesicular transport.

Animals↗

PubMed1998

Systemic sclerosis stimulates angiogenesis in the chick embryo chorioallantoic membrane.

Skin biopsies from patients with systemic sclerosis (SSc) were investigated for their angiogenic activity by using the chick embryo chorioallantoic membrane (CAM) assay. Ten samples of SSc and 10 of normal skin from age- and sex-matched subjects were grafted onto the CAM, and the angiogenic response in pathological and control implants was assessed on histological sections by a planimetric point-count method 4 days after grafting. The vascular counts in the area underlying the SSc were significantly higher than those of normal skin and a dense mononuclear cell infiltrate was detectable around the blood vessels in pathological specimens. These results suggest that SSc may promote angiogenesis, perhaps leading to the release of several angiogenic factors. Moreover, the role played in the angiogenic response by the inflammatory cells forming the cellular infiltrate is suggested by this study.

Adult↗

PubMed1998

Ultrastructural localization of lectin binding sites in the developing brain microvasculature.

The temporo-spatial patterning of lectin-binding sites was examined by lectin histochemistry and quantitative methods in the microvasculature of the optic tectum of 9-, 14-, 20-day-old embryos and 30-day-old chickens. Horseradish peroxidase and colloidal-gold-labelled lectins were used for detection of beta-D-galactose (RCA-I, Ricinus communis agglutinin-I) and of N-acetylglucosamine and sialic residues (WGA, Wheat germ agglutinin) at light and electron microscopical levels. At the light microscopical level, RCA-I and WGA binding sites were detectable in the early embryonic capillaries in a diffuse staining pattern; in later embryonic stages and in adult animals, RCA-I labelling became located on the abluminal surface of the vessels, while WGA staining was detected on the luminal surface. Ultrastructurally, gold labelling for RCA-I was seen intracytoplasmically in endothelial cells in 9-day-old embryos. In 14-to 20-day-old embryos and in chickens, binding sites for RCA-I were detected in endothelial tight junctions and basement membranes. In contrast, labelling of the gold-coupled WGA lectin was distributed almost exclusively on the luminal endothelial surface already in early embryos. The results indicate that the endothelial cells of the optic tectum acquire functional polarity early in their development and that glycoconjugates containing beta-D-galactose residues are involved in the biochemical composition of the tight junctions and basement membrane, which are considered to be key structures in blood-brain barrier (BBB) differentiation.

Animals↗

PubMed1998

Vascularization of embryonic adrenal gland grafted onto chorioallantoic membrane.

Vascularization and endothelial phenotype expression were analysed in embryonic adrenal tissue grafted onto chorioallantoic membrane (CAM), by means of routine light microscopy and immunocytochemical staining, and of electron microscopy. Adrenal gland tissue from chick or quail embryos (donors) was grafted onto CAMs of chick or quail embryos (host). Vessels of chick origin were discriminated from those of quail origin by monoclonal antibodies, anti-MB1, specific for quail endothelial and haemopoietic cells, and QCPN, which labels quail cell nuclei. Vessels of adrenal type were distinguished from those of CAM-type by their ultrastructural endothelial phenotype - porous in the former and continuous in the latter. The observations carried out 6 days after implantation indicate that the adrenal gland develops and differentiates according to a virtually normal histological pattern. As regards the adrenal and CAM vascularization, the grafting procedure elicits angiogenic events consisting in the formation of peripheral anastomoses between the graft and the CAM original microvasculature and in new-growth of vessels from the CAM into the grafted tissue and vice versa. As to the endothelial phenotype, the ultrastructural results demonstrate that besides its own native vasculature, the adrenal tissue contains vessels with continuous endothelium and the CAM mesenchyme is supplied by adrenal-type, fenestrated vessels.

Adrenal Glands↗

PubMed1998

Do mast cells help to induce angiogenesis in B-cell non-Hodgkin's lymphomas?

Morphological and morphometric data showing a higher number of mast cells (MCs) in the stroma of B-cell non-Hodgkin's lymphomas (B-NHL) than in benign lymphadenopathies are presented in support of the suggestion that angiogenesis during the progression of B-NHL may be partly mediated by angiogenic factors in their secretory granules.

Humans↗

PubMed1998

Expression of tenascin is related to angiogenesis in pre-eclampsia.

METHODS: Ten samples of decidua basalis from pre-eclamptic women and 10 from healthy primigravid women subjected to caesarean section (control tissues) were investigated immunohistochemically for changes in angiogenesis and expression of tenascin, an extracellular matrix protein thought to modulate angiogenesis. In addition, pre-eclamptic and control samples were grafted onto the chick embryo chorioallantoic membrane (CAM) to study their possible angiogenic activity in vivo. RESULTS: Although the microvessel area was low in control samples, it increased significantly in the pre-eclamptic decidua. In parallel, the area covered by tenascin staining underwent a significant expansion that was highly correlated with the microvessel area. Angiogenic activity was assessed both macroscopically and microscopically on histological sections 4 days after grafting. In contrast to control samples, pre-eclamptic decidua induced an intense capillary growth, and numerous capillaries converging towards the implant and forming a spoke-wheel pattern were evident. Histological examination using a planimetric point-count method showed that microvessel counts in the CAM area under and around the pre-eclamptic implants were significantly higher than those of control samples. Again, the extracellular matrix of nearby microvessels was strongly immunoreactive with tenascin. CONCLUSION: It is suggested that, unlike normal decidua, pre-eclamptic decidua displays an intense, autonomous angiogenic activity, probably mediated by tenascin in addition to its own angiogenic factors.

Allantois↗

PubMed1998

Lipoprotein (a) induces angiogenesis on the chick embryo chorioallantoic membrane.

BACKGROUND: Both lipoprotein (a) [Lp(a)] and angiogenesis have been shown to be associated with initiation and progression of atherosclerotic plaque. Lp(a) and two neutralizing anti-Lp(a) antibodies were investigated for their capacity to affect the vasoproliferative processes of the chick embryo chorioallantoic membrane (CAM), a useful model for such an investigation. METHODS: Gelatin sponges loaded with Lp(a) alone or together with anti-Lp(a) antibodies, or with vehicle alone, phosphate-buffered saline (PBS), were implanted in vivo onto the CAM at incubation day 8. Four days later, sponges and the adjacent CAM tissues were assessed for the extent of angiogenesis in terms of microvessel counts. RESULTS: Lp(a)-loaded sponges gave significantly higher counts than those loaded with the LP(a)-anti-Lp(a) antibodies complex, which overlapped those treated with PBS. The angiogenic response was similar to that obtained with basic fibroblast growth factor, a well known angiogenic molecule. CONCLUSION: These data suggest that Lp(a) is capable of inducing angiogenesis in vivo, which might account for its ability to enhance and support atherosclerosis.

Allantois↗

PubMed1998

Angiogenesis in hepatocellular carcinoma: an experimental study in the chick embryo chorioallantoic membrane.

Ten samples of human hepatocellular carcinoma and three of a laceration injure of the liver (controls) were grafted onto the chick embryo chorioallantoic membrane (CAM) to investigate their possible angiogenic activity. The angiogenic response in pathological and control implants was assessed on histologic sections by a morphometric method, 4 days after grafting. The vascular count in the CAMs treated with the pathological implants was significantly higher compared to control ones and the angiogenic response induced by pathological implants was comparable to that of a well known angiogenic molecule, namely basic fibroblast growth factor. The role played in vasoproliferative response by angio-genic cytokines released by tumor cells, by CAM extracellular matrix and by the perivascular mononuclear cells was supported by this study.

Allantois↗

PubMed1998

Role of basic fibroblast growth factor in the formation of the capillary plexus in the chick embryo chorioallantoic membrane. An in situ hybridization, immunohistochemical and ultrastructural study.

The chick embryo chorioallantoic membrane (CAM) is supplied by an extensive capillary network. We have previously demonstrated that a Mr 16,000 basic fibroblast growth factor (FGF2)-like molecule is present in the CAM. At present, no data are available on the cellular source(s) of FGF2 in the CAM. In this work, CAM has been investigated by in situ hybridization with the aim to identify the source(s) of endogenous FGF2 during development. The immunohistochemical expression of fibronectin, laminin and type IV collagen in the CAM extracellular matrix (ECM) and the ultrastructural relationships between chorionic epithelium and the underlying capillary plexus were also studied. Our findings strongly suggest that FGF2 regulates the development of the capillary plexus by two sequential steps. In an early paracrine phase, chorionic epithelial cells secrete FGF2, thus eliciting an angiogenic response in the undifferentiated mesodermal blood vessels. In response to this paracrine signalling, the newly formed endothelial cells move through a permissive ECM and migrate beneath the chorion. Here, they synthesize an autocrine supply of FGF2 necessary to further proliferate and differentiate, thus originating the capillary plexus.

Allantois↗

PubMed1998

Free laminin in the extracellular matrix of B-cell non Hodgkin's lymphomas.

The authors show that in the perivascular stroma of B-cell non-Hodgkin's lymphomas (B-NHL) is present a granular, speckled pattern of expression of laminin, similar to the 'free-laminin' firstly described by Lugassy et al. (1997) in human melanoma. The role of this form of laminin may be to promote the migration of tumor cells, which express a variety of binding-laminin surface proteins, whose expression is significantly increased in several human cancer models.

Extracellular Matrix↗

PubMed1998

Astroglia-microvessel relationship in the developing human telencephalon.

The telencephalon of 12 and 18 week-old human foetuses was examined for evidence of astroglia-microvessel relationship. Immature astroglia cells (radial glia and astroblasts) and astrocytes were immunostained using antibodies to the cytoskeletal proteins vimentin (VIM) and glial fibrillary acidic protein (GFAP). The microvessels were detected using an antibody to the blood-brain barrier (BBB)-specific glucose transporter GLUT1. Two extracellular matrix (ECM) glycoproteins, laminin (LM), an endothelial-derived molecule, and tenascin-C (TN-C), a glia-derived molecule, were also analyzed. In the two stages examined, VIM- and GFAP-positive fibers of the radial glia establish close relationships with the radial and periventricular microvessels, which are GLUT1-positive and lined by an LM-positive basal lamina-like matrix. At the 18th week, also radial glia transitional forms and immature astrocytes exhibit extensive contacts with the microvasculature. A TN-C-rich ECM is revealed around the vascular plexus of ventricular zones at the 12th week, and around the newly growing radial microvessels and the microvessel branching sites at the 18th week. The observations taken as a whole, suggest that during the telencephalon morphogenesis the immature astroglia cells play a role in the early establishment of the distribution pattern of the neural microvessels and in their growth and maturation.

Abortion, Spontaneous↗