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Biomedical subjects

A Facchini

Publications and source records attributed to A Facchini.

At least 181 records · Page 10Linked to original sources
PubMedDate not supplied

[Tissue engineering applications: cartilage lesions repair by the use of autologous chondrocytes].

Promising new therapies based on tissue engineering have been recently developed for cartilage repair. The association of biomaterials with autologous chondrocytes expanded in vitro can represent a useful tool to regenerate this tissue. The scaffolds utilised in such therapeutical applications should provide a pre-formed three-dimensional shape, prevent cells from floating out of the defect, have sufficient mechanical strength, facilitate uniform spread of cells and stimulate the phenotype of transplanted cells. Hyaff-11 is a hyaluronic-acid based biodegradable polymer, that has been shown to provide successful cell carrier for tissue-engineered repair. From our findings we can state that human chondrocytes seeded on Hyaff-11 are able to maintain in vitro the characteristic of differentiated cells, expressing and producing collagen type II and aggrecan which are the main markers of cartilage phenotype, down-regulating collagen type I. Moreover, it seems to be a useful scaffold for cartilage repair both in animal models and clinical trials in humans, favouring the formation of a hyaline-like tissue. In the light of these data, we can hypothesise, for the future, the use of autologous chondrocyte transplantation together with gene therapy as a treatment for rheumatic diseases such as osteoarthritis.

Animals↗
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Relationship between serum RANTES levels and radiological progression in rheumatoid arthritis patients treated with methotrexate.

OBJECTIVE: The aim of this study was to evaluate the relationship between serum chemokines and the clinical and radiological response to a one-year course of methotrexate (MTX) in patients suffering from rheumatoid arthritis (RA). METHODS: Twenty out-patients suffering from active RA entered a one-year open prospective study on the effects of low dose MTX therapy. Plain radiographs of the hands and feet were taken at study entry and at the end of the follow-up, and were compared for the number of eroded joints. Serum levels of both C-X-C and C-C chemokines were obtained before the initation of MTX and after 6 and 12 months of treatment. RESULTS: The levels of serum RANTES before treatment were significantly higher in RA patients than in the controls and returned to normal levels after one year of treatment. Serum levels of the other chemokines were either in the normal range or undetectable. Twelve patients (60%) did not show any new eroded joints at the end of the follow-up period and were considered as radiological responders (RR). Serum levels of GRO-alpha and RANTES after 6 months of treatment were significantly higher among the patients with radiological progression than in RR patients. CONCLUSIONS: We observed high levels of serum RANTES in a series of RA patients during the active stage of the disease. MTX treatment significantly lowered the serum levels of RANTES, GRO-alpha and MCP-1. High levels of serum RANTES or GRO-alpha after 6 months of MTX treatment seem to be predictive of radiological erosions after one year.

Acute Disease↗
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Expression of different chemokines by human osteosarcoma cells in response to tumor necrosis factor-alpha.

We investigated the expression of different chemokines in the surnatants and inside the cells of four human osteosarcoma cell lines. HOS, U-2 OS, MG63 and Saos-2 cells were cultured for 24, 48, 72 hours both in basal conditions and after stimulus with TNF alpha. Human stromal cells were used as control. IL-8 and MCP-1 are present in higher concentration in the surnatants in contrast to RANTES which is present primarily inside the cells. IL-8 and MCP-1 are not totally expressed by all the human osteosarcoma cell lines in unstimulated conditions, but became detectable after TNF alpha treatment. In general, this cytokine stimulated the production and release of the three chemokines.

Cell Division↗
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Ly-6A/E gene is widely expressed among transformed nonhematopoietic cells. Autocrine modulation by interferon.

Expression of the Ly-6A/E gene by transformed cells was investigated in 14 cell cultures of C57BL/6 and BALB/c mouse origin derived from spontaneous or chemically-induced non-hematopoietic tumours and from cells transformed by SV40 virus. Histologic types included carcinomas, sarcomas and a melanoma. Thirteen out of 14 cell cultures expressed membrane Ly-6A/E antigens; only the B16-A melanoma was negative, but it expressed Ly-6A/E after incubation with IFN-gamma. The effect of in vitro permanence was studied on early (< 10) and late (> 20) passages of B16-A melanoma and MN/MCA1 fibrosarcoma. Late passage B16-A cells were Ly-6A/E-negative and refractory to induction of Ly-6A/E (but not of H-2 antigens) by IFN-alpha, IFN-beta, or IFN-gamma; MN/MCA1 maintained a high expression of Ly-6A/E, but no increase was induced by IFNs. It was found that both early and late in vitro passages of MN/MCA1 actively produced IFN-alpha/beta. The analysis of cells of fibroblastic origin revealed a significant correlation between IFN release in the culture medium and Ly-6A/E levels. Culture of fibrosarcoma cells in the presence of an anti-IFN-alpha/beta serum reduced Ly-6A/E expression, thus indicating the existence of an autocrine loop.

Animals↗
PubMedDate not supplied

Tissue engineering for cartilage repair: in vitro properties of a hyaluronan-derivative.

Association of biomaterials with autologous cells can provide a new generation of implantable devices for cartilage and bone repair. Such scaffolds should provide a performed three-dimensional shape, prevent cells from floating out of the defect, have sufficient mechanical strength, facilitate uniform spread of cells, and stimulate the phenotype of transplanted cells. Hyaff-11 is a recently developed hyaluronic-acid based biodegradable polymer, that has been shown to provide successful cell scaffolds for tissue-engineered repair. The aim of this study was to evaluate in vitro the potential of Hyaff-11 to support the growth of human chondrocytes and to maintain their original phenotype. Our data indicate that human chondrocytes seeded on Hyaff-11 express and produce collagen type II and aggrecan and downregulate the production of collagen type I. These results provide an in vitro demonstration of therapeutic potential of Hyaff-11 as a delivery vehicle in tissue-engineered repair of articular cartilage defects.

Adolescent↗
PubMedDate not supplied

Establishment of a new human immortalized chondrocyte cell line.

Chondrocytes from human adult articular healthy cartilage were transfected in primary culture with a plasmid containing two human papilloma virus type 16 early function genes: E6 and E7, using the highly efficient cationic liposome-mediated (lipofection) procedure. The transfection was verified by reverse transcriptase-polymerase chain reaction analysis of E7 mRNA and by immunofluorescence localization of the E7 protein in the cell cytoplasm. The established chondrocyte cell line was examined in monolayer and in two culture conditions that were described to re-induce differentiated characteristics: culturing in a serum-free defined medium and seeding on a hyaluronan-based three-dimensional biomaterial. Immortalized cells were able to re-express the main markers of chondrocyte phenotype, both at mRNA and protein levels, under the two defined cultured conditions used. The cell line that we obtained may be a useful tool for increasing our knowledge of the genetic and biochemical events involved in the processes of cartilage growth and differentiation, and of the etiopathogenesis of many rheumatic diseases.

Cell Line↗
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Hyaluronan-based biomaterial (Hyaff-11) as scaffold to support mineralization of bone marrow stromal cells.

Various techniques are widely used to repair bone defects, association of hyaluronan-based biodegradable polymers (Hyaff-11) with bone marrow stromal cells (BMSC) promises to provide successful cell scaffolds for tissue-engineered repair of bone tissue. We evaluate in vitro and in vivo the potential of Hyaff-11 to facilitate mineralization of BMSC. Rat BMSC were seeded on Hyaff-11 and their differentiation were assessed at different time points. Osteogenic differentiation was investigated in vitro analysing the expression of alkaline phosphatase and osteocalcin. Mineralization of bone defects was evaluated also in vivo implanting Hyaff-11 scaffold combined with BMSC in large segmental radius defects. In vitro, we found a decrease expression of alkaline phosphatase and an increase of osteocalcin. In vivo, our data showed that mineralization was induced and basic fibroblast growth factor contributed to this process. These results provide a demonstration to therapeutic potential of Hyaff-11 as appropriate carrier vehicle for differentiation and mineralization of BMSC and for the repair of bone defects.

Animals↗
PubMedDate not supplied

Vascular endothelial growth factor activities on osteoarthritic chondrocytes.

OBJECTIVE: Evaluation of the role of VEGF in cartilage pathophysiology. METHODS: VEGF release from chondrocytes in the presence of IL-1beta, TGFbeta and IL-10 was detected by immunoassay. VEGF receptor -1 and -2 expression and VEGF ability to modulate caspase -3 and cathepsin B expression were detected by immunohistochemistry on cartilage biopsies and cartilage explants. VEGF effects on chondrocyte proliferation was analysed by a fluorescent dye that binds nucleic acids. RESULTS: VEGF production by osteoartritis (OA) chondrocytes was significantly reduced by IL-1beta while it was increased in the presence of TGFbeta. Cartilage VEGFR-1 immunostaining was significantly downregulated in 'early' OA patients compared to normal controls (NC). VEGFR-2 expression was negligible both in OA and in NC. VEGF decreased the expression of caspase-3 and cathepsin B, whereas it did not affect proliferation. CONCLUSION: VEGF is able to down-modulate chondrocyte activities related to catabolic events involved in OA cartilage degradation.

Adult↗
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Antibodies to hepatocyte derived antigens and liver damage in patients after bone marrow transplantation.

The follow-up sera from 33 patients who received allogenic bone marrow transplantation (BMT) were tested by ELISA for antibodies to liver derived antigens present in a macromolecular, lipid associated, protein complex (LSP). Anti-LSP were found in 20% of patients with graft-vs-host disease (1 with acute GVHD, 3 with chronic GVHD), but in none of the 13 patients without GVHD. Lymphocytotoxic antibodies (LCTA) were frequently detected with similar prevalence in patients with and without GVHD. In the 4 anti-LSP +ve cases, LCTA serum levels did not follow the trend of anti-LSP antibody levels. Liver biopsy was performed in 10 patients with clinical and biochemical sign of GVHD and showed features of hepatic GVHD in six cases; although piecemeal necrosis was present in 3 cases, anti-LSP were detected only in one patient. An anti-LSP response can be elicited in patients after BMT only during GVHD, with no relationship to LCTA and without a close association with signs of periportal hepatocyte damage.

Adolescent↗