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Biomedical subjects

G Ciapetti

Publications and source records attributed to G Ciapetti.

At least 91 records · Page 5Linked to original sources

Cell culture methods for testing biocompatibility.

Cell culture systems may be of value in testing the biocompatibility of prosthetic materials before they are introduced into clinical use. In recent years, in vitro methods for assaying biomaterials have gained in importance owing to the growing concern over the use of animals for biomaterials testing. Significant effort is therefore being focused toward developing predictive and quantitative, but also simple and reliable, methods of testing using cultured cells. At present, a number of methods for measuring both the cytotoxicity and the specific cytocompatibility of different materials are available. The usefulness of these systems is no longer confined to screening new materials; they can be used to study the mechanisms of action of various materials during tissue/material interaction. This paper reviews the published literature on the use of cell culture models in evaluating biocompatibility and reports on the personal experience of the authors, who have been using cell culture systems for many years and for different purposes.

Biocompatible Materials↗

Production of prostacyclin and fibrinolysis modulators by endothelial cells cultured in the presence of polyethylene terephthalate.

The aim of this study was the evaluation of the in vitro production of prostacyclin, and of tissue plasminogen activator (tPA) and its inhibitor, PAI-1, by human endothelial cells cultured in the presence of polyethylene terephthalate (PET). After a 48 h contact between the cells and the polymer, the concentration of 6-keto-PGF1 alpha, a stable metabolite of prostacyclin, tPA, and PAI-1, was assayed on the supernatants. Contact of the endothelial cells with PET produced a highly significant reduction of 6-keto-PGF1 alpha with respect to control cultures. Tissue plasminogen activator concentration in the supernatants of the cultures in contact with the material was similar to that observed in the controls, while PAI-1 production was significantly reduced. It can be concluded that the contact between endothelial cells and PET determines a reduction in the platelet aggregability and an increase of the fibrinolytic activity due to a decrease in PAI-1, while tPA concentration remains unchanged.

6-Ketoprostaglandin F1 alpha↗

In vitro evaluation of cell/biomaterial interaction by MTT assay.

The tetrazolium-based colorimetric assay (MTT test) measures only in vitro living cells and the results are directly related to the number of viable cultured cells. It has been adopted in immunological investigations, cancer research and, recently, biocompatibility evaluation. We used the MTT method with minor modifications to fit it to an in vitro study of biomaterial-cell interactions. The MTT assay was confirmed to be feasible, rapid and reproducible. Moreover, it showed a good correlation with other in vitro proliferation assays, such as the 3H-thymidine uptake assay. By using the MTT method and the ASTM procedure for extracting biomaterials, we quantified the in vitro cell compatibility of different metals and polymers.

Biocompatible Materials↗

[Assessment of the plasma phase of coagulation in platelet-rich plasma after passage through a special polyester filter].

The authors have examined the alterations in the coagulation pathway of plasma filtered through a polyester filter coated by a hydrophilic polymer. The activation of the coagulation pathway was evaluated by measuring prothrombin time, activated partial thromboplastin time, coagulation factors and fibrinopeptide A. The results obtained demonstrate that the examined filter does not induce significative activation of the coagulation pathway.

Blood Coagulation↗

Metabolic changes in bone in old age.

The authors conducted a study on the effect of aging upon the factors which regulate calcium metabolism. Hormonal and biochemical tests were performed on samples from patients selected on the basis of sex, age, and other characteristics in order to determine the trend of these regulatory factors in middle and old age, with particular attention to osteoporosis in women. The authors propose several theories regarding the pathogenetic mechanisms of osteoporosis on the basis of this study as well as the literature.

Adult↗

[In vitro and in vivo evaluation of the blood-biomaterial interaction].

The Authors, after a short description of the phenomena following the interaction between blood and biomaterials, take into consideration the different methods used in studying the in vitro, in vivo and ex vivo hemocompatibility, as well as the problems involved by each method and its contribution to the knowledge of hemocompatibility. The Authors report their experience on the in vitro study of the effects induced by some biomaterials on platelets and coagulation factors.

Animals↗

In vitro methods to evaluate metal-cell interactions.

The aim of this study was to test different metals, widely employed in constructing prosthetic devices, by in vitro methods. Biological effects of such materials were analyzed through four different assays on human lymphocytes and granulocytes. The lymphocyte proliferation assay gave quantitative results, while the viability test showed the morphological appearance of the cells correlated well with previous results. NK cytotoxicity and granulocyte chemokinesis tests provided interesting data on leucocyte performance when challenged with metals. Therefore the present study adds new basic information on cell behaviour when metal products are present in the body, e.g. around devices implanted in human tissues.

Biocompatible Materials↗

Cellular events in the mechanisms of prosthesis loosening.

The functional restoration of a joint damaged by trauma or disease is obtained by prosthetic surgery. In particular the implantation of hip prostheses is regarded as routine in orthopedic surgery and thorough research has been developed in this field. The prosthetic replacement of the knee and even more so the ankle and elbow occurs less frequently in clinical practice and has been studied less intensively. The results of artifical hip replacement are generally good, both in terms of pain relief and the restoration of satisfactory joint function. Nevertheless, as time passes, a high rate of failures have been recorded due to prosthesis infections, fracture and wearing of the prosthetic components and prosthesis loosening by various causes. The use of ultra-filtered air and laminar flow in operating theatres and antibiotic prophylaxis have dramatically reduced the incidence of infections in total hip arthroplasty. Thanks to the setting up of new stem configurations and the use of superalloys that are highly resistant to fatigue failure, the fracture of the femoral component has been virtually eliminated as a complication of total hip arthroplasty replacements. Loosening is thus the most frequent complication in total hip replacement.

Bone Cements↗

Raman spectroscopy for the study of porous composite alumina coating.

A new acetabular cup prosthesis made of alumina and bioactive glass was studied and evaluated using nondestructive and noninvasive Raman laser spectroscopy. The prosthesis consists of an alumina socket coated with a double layer of alumina beads bonded to the prosthesis with a layer of glass. Raman spectroscopy showed that both the socket and the beads are composed of alpha-Al2O3 (corundum), while the bioactive glass (alkaline and alkaline earth alumosilicate with a small quantity of zirconium oxide) showed this to have an intermediate structure between that of vitreous silica and that of the alkaline and alkaline earth disilicate glasses. The Raman spectra of the contact surfaces between the bioactive glass and the alumina of the assembled prosthesis and of an alumina disk coated with the same glass showed the presence of new bands which may be correlated with a chemical interaction between the components of the prosthesis.

Aluminum↗

Comparative study of the thromboresistance of Dacron combined with various polyurethanes.

In vitro tests were carried out on Dacron samples differently knitted and on Dacron vascular prostheses coated with various urethanes. All the materials were put in contact with human platelet-rich plasma; the subsequent assay of three platelet released substances, i.e. beta-thromboglobulin, platelet factor 4 and thromboxane B2, as well as the quantification of platelet retention, were used to establish the degree of thrombogenicity of the material itself. In some cases Dacron-urethanes composites showed better thromboresistance than any other materials conventionally used in vascular surgery.

Biocompatible Materials↗

Multilayered bead ceramic composite coating for hip prostheses: experimental studies and preliminary clinical results.

The clinical use of a new ceramic composite material made up of multilayered alumina beads and adhered to a ceramic surface with a high-temperature-melting bioglass is proposed. The result is a structure characterised by a 27% porosity and an average pore diameter of 400 microns. The actual structure of the parts as well as their interface interaction were determined by Raman laser. The mechanical resistance of the adherence of this coating for the ceramic substratum was good enough to resist the stress to which it was submitted when used for a new model of prosthetic acetabulum. Experiments with rabbits revealed the good biocompatibility of the composite. The osteoproductive activity of the tissue surrounding the implant led to the gradual filling of the porosity with trabecular structures. The preliminary results of the clinical experience which began in November 1985, confirm the good compatibility of the ceramic composite.

Aluminum Oxide↗

Results of histological grading on 100 cases of hip prosthesis failure.

One hundred cases of hip prosthesis failure were classified on the basis of the different types of tissue reaction occurring around alloplastic material. The results revealed infectious phlogosis in 32% of the cases, phlogosis due to wear in 42%, phlogosis due to allergy in 1% and mixed phlogosis in 25%. The distribution of the type and degree of intensity of the phlogosis, in relation to the duration of the implant, is also highlighted. This new grading technique yields reproducible results.

Biocompatible Materials↗

The effect of injection of powdered biomaterials on mouse peritoneal cell populations.

The authors suggest an experimental model for the assaying, before implantation, of the tissue reaction that wear particles from artificial joints can cause in the human body. Mice were intraperitoneally injected with biomaterial powders in suspension in saline. The materials used were chromium, chromium oxide, nickel, nickel oxide, aluminum, alumina, alumina-titania, cobalt, molybdenum, titanium, silver, zirconium oxide, iron oxide, and stainless steel. Peritoneal lavage was performed a week after injection. A thorough morphological and quantitative analysis of the cell suspension thus obtained was made both immediately after collection and after a 24 h culture.

Animals↗

Biocompatibility testing of prosthetic implant materials by cell cultures.

The main aspects of biocompatibility are discussed first then the methodology used and the results obtained. The cells used included epithelial cells, lymphocytes, fibroblasts, and macrophages. The most commonly used methods were morphological observation, radioactive tracer uptake, and chemotactic migration analysis. It is concluded that cell cultures are a reliable and sensitive method for initial screening in testing the biocompatibility of the materials used in the construction of prosthetic implants.

Animals↗

Effects of chromium extract on cytokine release by mononuclear cells.

We have evaluated the effects of chromium extract on the release by peripheral blood mononuclear cells (PBMCs) of cytokines favouring bone resorption. Furthermore, we have evaluated whether the chromium effects could be correlated with the activation and proliferation of PBMCs. Cell cultures were maintained in serum-free medium (AIM-V), in order to avoid the interference of exogenous growth factors. Increasing concentrations of chromium extract, ranging between 3 and 100%, were added to culture medium. Cytokine release (IL-1beta, TNFalpha, IL-6, GM-CSF and IFNgamma) was assessed on both PBMCs cultured with AIM-V only (unstimulated PBMC) and PBMCs cultured with AIM-V plus phytohaemagglutinina (PHA-stimulated PBMC). The activation and proliferation of PBMCs were evaluated by assessing DNA synthesis and soluble IL-2 receptor release, in order to determine whether an IL-2-dependent immune response can be induced by chromium. Our results show that in unstimulated PBMCs chromium ions slightly increased the release of pro-inflammatory cytokines, such as TNFalpha and IL-6, even though the increase is not significant. On the contrary, the different concentrations of chromium extract significantly inhibited the response to PHA stimulation, as shown by the decrease in IL-6 and sIL-2r release, and by the influence on cell viability and DNA synthesis. Both these effects are undesirable and support hypotheses on the biological effects of chromium. The continuous release of chromium from the implant could induce in PBMCs the release of bone-resorbing cytokines, which in the long term could be responsible for irreversible tissue damage. Moreover, chromium seems to inhibit the IL-2-dependent response of PBMCs, so that they are not able to trigger an efficient cell-mediated immune response.

Bone Resorption↗

Adhesive protein expression on human endothelial cells after in vitro contact with woven Dacron.

In this research adhesive proteins are studied in order to evaluate the interference of woven Dacron in the endothelialization process and in the ability of endothelial cells to bind circulating leucocytes. Endothelial cells from human umbilical vein (HUVEC) were put in contact with woven Dacron for 24 h. PECAM-1, ELAM-1, ICAM-1 and VCAM-1 expression was then evaluated by flow cytometry, using indirect immunofluorescence reaction with monoclonal antibodies. The study of adhesive proteins was completed with the quantitative determination of surface antigens expressed as the antibody binding capacity (ABC). Antigenic density was calculated by the DAKO QFIT calibration system for indirect immunofluorescence. After contact with woven Dacron no significant change was observed in the percentage of positive cells or in the fluorescence intensity of the adhesins. No significant variation was also noted by calculating the surface antigen density by means of calibration fluorospheres. It can be concluded that the material examined does not significantly affect leucocyte adhesion to the endothelium.

Biocompatible Materials↗