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The aim of this research was to evaluate the effect of polyethylene terephthalate (Woven Dacron) on the expression of endothelial integrins. Human umbilical vein endothelial cells were cultured on the material for 24 h. The integrins VLA-2 (α2β1-CD49b/CD29), receptor for laminin and collagen, VLA-5 (α5β1-CD49e/CD29), receptor for fibronectin, VLA-6 (α6β1-CD49f/CD29), receptor for laminin, and αVβ3-CD51/CD61 (receptor for vitronectin) were evaluated by flow cytometry. After contact with polyethylene terephthalate, a slight but significant decrease in the percentage of both CD29 and CD49e positive cells was observed, which suggests a lower number of cells expressing the fibronectin receptor α5β1. Moreover, a significant increase in the mean channel for CD49b and for the vitronectin receptor CD51/CD61 was observed. The reduction in the fibronectin receptor could account for the poor endothelialization observed in vivo on polyethylene terephthalate. The increased expression of the vitronectin receptor, favoring the migration of smooth muscle cells, could give some information about the pathogenesis of intimal hyperplasia, which is a complication of vascular grafts. © 2001 Kluwer Academic Publishers

In order to evaluate if carbon coated polyethylene terephthalate (C-PET) could favor inflammatory reactions, the expression of interleukin-6 (IL-6) by cultured human umbilical vein endothelial cells was tested in vitro. The cultures were put in contact with C-PET for 1, 24, 48 and 72 h. The same cells cultured on tissue culture-treated polystyrene without biomaterials were tested as the negative control; the same cells incubated with LPS were the positive control. The level of IL-6 in the conditioned medium was tested by enzyme immunoassay; the mRNA expression was evaluated by RT-PCR with specific primers. The cultures incubated with C-PET produced non significantly different amount of IL-6 compared to the negative control and did not induce the expression of IL-6 specific mRNA. LPS induced a significantly higher release of IL-6 in the medium and the expression of mRNA after 24, 48 and 72 h. We conclude that C-PET does not stimulate the synthesis of IL-6 and therefore does not favor inflammatory reaction through the release of this cytokine. © 2001 Kluwer Academic Publishers

Deposition and aggregation of lachrymal proteins on the contact lens surface can promote bacterial adherence. Lysozyme is the major tear protein and is also mainly responsible for the formation of protein deposits on contact lenses. Nonsteroidal anti-inflammatory drugs (NSAID) prevent protein aggregation. The effect of a water-soluble NSAID drug on bacterial adherence to high-water-content/ionic disposable contact lenses was examined in a radiolabeling study. Dose-related inhibition of adherence of Staphylococcus aureus, S. epidermidis, and Pseudomonas aeruginosa on both pretreated lenses and after adding the drug to the medium was investigated. When the drug was added to the media, maximal inhibition of S. aureus adherence was observed in trypticase soy broth (59–98% at the lower and higher drug concentrations, respectively); inhibition progressively decreased in calf aqueous humor (48–75%), lysozyme (34–63%), and saline (12–20%) solutions. Inhibition of adherence varied with the three bacterial species; it was maximal with S. aureus, intermediate with S. epidermidis, and minimal with P. aeruginosa. When lenses were pretreated with the drug, consistent, and even higher, inhibitory effects were observed. The results suggest that water-soluble NSAIDs could be used in preventive treatments for conjunctivae and corneal infections in contact lens wearers, and may provide a clue as to which compounds might inhibit protein interaction and bacterial adhesion. © 1998 John Wiley & Sons, Inc. J. Biomed Mater Res, 42, 1–5, 1998.

The mutagenic potential of three commercially available glass–ionomer cements used in dentistry was examined. The cement components were mixed according to the manufacturers indications and set for two defined times: 1 h or, alternatively, 1 wk. Cements B and C set spontaneously; in the case of cement A, the manufacturer suggests the use of a lamp to trigger also a photopolymerization. Photopolymerization, however, was not used. Ames tests were performed on the dimethyl sulphoxide extracts of cements by using Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537, TA 1538 and TA 102. Cement A showed mutagenicity only against TA 1537 strain, either in the presence or absence of metabolic activation with microsomial fraction S9. The other two cements showed no mutagenic potential. We conclude that glass–ionomer cements are, on the whole, safe materials from the viewpoint of genotoxicity, and hypothesize that the mutagenicity observed in cement A could depend on its polymerization performed without light activation. © 1998 Chapman & Hall

The authors examined the modifications of some markers of platelet activation after contact with biomaterials. Glycoprotein GMP-140 (CD62) was evaluated by flow cytometry; β-thromboglobulin (β-TG) and thromboxane B2 (TXB2) were determined by radioimmunoassay. Polyethylene terephthalate (PET) induced a remarkable platelet adhesion and a significant increase in β-TG and TXB2, with no increase in CD62 on the nonadherent platelets. Pyrolytic carbon-coated PET (PC) did not induce platelet adhesion after 15 min of contact, but a significant increase in CD62 was detected. After 30 min a significant increase in platelet adhesion as well as the release of β-TG and TXB2 were noted. The increase was lower than that observed for uncoated PET, and after 30 min of contact with PC the increase no longer was observed. © 1997 John Wiley & Sons, Inc. J Biomed Mater Res, 36, 289–294, 1997.

The ability of some biomaterials to activate plasma coagulation system was examined in vitro. After contact of platelet-rich plasma with biomaterials, some markers of the thrombin formation, i.e., fragment 1 + 2 and fibrinopeptide A, and some inhibitors of the blood coagulation mechanism were tested. Fragment 1 + 2 and fibrinopeptide A were found to be increased by all of the materials, though to a different extent. In particular, fragment 1 + 2 and fibrinopeptide A were significantly increased upon contact with polybutylene terephthalate and with collagen coated polyethylene terephthalate, respectively. Also antithrombin III was shown to decrease following exposure to biomaterials, but statistical significance was found only for polyethylene terephthalate and polyvinylacetate. As a results of this wide range of variability in the parameters, it is advisable to explore the plasma coagulation system with a multiparametric approach in which thrombin formation and coagulation inhibitors are thoroughly investigated.

The effects of orthopaedic cements on the proliferation and cell cycle of in vitro cultured MG63 osteoblast-like cells were examined. Five different cements were mixed and extracted at different time intervals (15 and 60 min, 6, 24 and 48 h). Cell proliferation inhibition (CPI) was evaluated after 72 h culture as the toxicity parameter. As for the toxicity degree, the extracts were considered to have ‘high toxicity’ (CPI ⩾ 50%), ‘medium toxicity’ (50% > CPI > 25%), ‘slow toxicity’ (CPI ⩽ 25%) and ‘no toxicity’ (CPI = 0). Cell cycle phases of MG63 cells were evaluated at 24, 48 and 72 h by flow cytometry; the DNA content was assessed using the propidium iodide uptake and the percentage of cells in the S phase was determined using 5′-bromodeoxyuridine uptake. According to our results, the toxicity is inversely correlated with the time interval between polymerization and extract preparation, and is different according to the cement type. For some cements the effects are still observed 48 h after polymerization. The damaging effect is not linked to a specific phase of the cell cycle, nor does it hamper the restarting of cell proliferation at 72 h.

The effects of endodontic cements on the cell cycle of MG63 osteoblasts cultured in vitro have been examined. Three groups of compounds were tested. Group I encompassed zinc oxide- and eugenol-based cements (Tubliseal, Argoseal, N2), group II consisted of cements with a phenol group other than eugenol (AH26, Forfenan, Methode R/R), and group III included CaOH-based cements (Biocalex, Endocalex). The cell cycle of MG63 cells was analyzed by flow cytometry; the DNA content was evaluated by means of the propidium iodide uptake method, whereas the proportion of cells in the S phase was defined by the incorporation of bromodeoxyuridine later revealed by a specific antibody. The results showed that some root canal sealers could hamper the periapex healing processes by inhibiting the cell proliferation through a selective action on different phases of the cell cycle.

This research aims at evaluating the expression of some adhesive proteins on endothelial cell surface after contact with polyethylene terephthalate coated with pyrolytic carbon (PET + PC). Twenty-two different cultures of human umbilical vein endothelial cells (HUVECs) were put in contact with PET + PC. Both HUVECs grown without the biomaterial and HUVECs incubated with endotoxin were used as control. After 24 h, platelet endothelial cell adhesion molecule-1 (PECAM-1), endothelial leucocyte adhesion molecule-1 (ELAM-1), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were evaluated on the cells by monoclonal antibodies and flow cytometry. In agreement with the literature, after 24 h the culture incubated with endotoxin determined a significant increase in the percentage of positive cells for ELAM-1, a significant increase in fluorescence intensity for ICAM-1, and a significant increase in the percentage of positive cells and fluorescence intensity for VCAM-1. After 24 h of culture with PET + PC, no significant variations in the antigens examined were observed. This demonstrates that such material does not activate in vitro the proteins involved in the adhesion between leucocytes and endothelium or in the adhesion between endothelial cells themselves.

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-PGF1α, 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-PGF1α 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. © 1993 John Wiley & Sons, Inc.

We evaluated the haemocompatibility of a filter of poly(butylene terephthalate) for packed red cell transfusions, which had undergone a physicochemical treatment (corona treatment) in order to reduce the filling time. Leucocyte, erythrocyte and platelet counts were carried out and haematological and coagulation parameters were examined before and after filtration. Moreover, some biochemical plasma parameters were evaluated, which are normally checked in hospitalized patients. The results obtained show that the filter treatment reduces the filling time, does not reduce the efficiency in retaining leucocytes and platelets,and does not induce alterations in filtered red blood cells. However, the treatment prolongs the activated partial thromboplastin time (APTT) and reduces the activity of some factors of the intrinsic coagulation pathway. On the other hand, it does not induce modifications in the biochemical parameters examined.

The authors examined the cytotoxicity, the capability of removing platelets and activating coagulation of two kinds of polybutyleneterephthalate. They are used in the making of filters able to remove leukocytes and platelets from packed red cell units to be transfused. The first filter was made of woven-non woven polybutyleneterephthalate, while the second one was made of the same biomaterial, which had previously undergone a chemico-physical modification in order to increase its wettability. The cytotoxicity of polybutyleneterephthalate, with and without modification, was evaluated by means of a screening test on in-vitro cultures. The test evaluates the cytotoxic effect of the chemicals which can be released in the culture medium by the examined materials. The activation of both platelets and the plasmatic pathway of coagulation and fibrinolysis process was evaluated by determining β-thromboglobulin and platelet factor 4, as well as by assaying some metabolites of arachidonic acid, fibrinopeptide A, protein C, plasminogen, α2-antiplasmin and the degradation products of fibrinogen and fibrin. The results showed that the chemico-physical modification carried out on the polybutyleneterephthalate filter did not change the material cytocompatibility, while determining a higher adhesion of platelets on its surface, without inducing the activation of the plasmatic pathway of coagulation.

In order to evaluate in vitro the suitability of various prosthetic materials for endothelial seeding, human endothelial cells derived from the umbilical vein were placed in direct contact with a variety of polymers. As a control, endothelial cells were cultured in the absence of material. After 24, 48, 72 and 96 hours, the cells were counted and a viability test with neutral red was performed. Assays of 6-keto prostaglandinFla (6-keto-PGF1a), tissue plasminogen activator (tPA) and plasminogen activator inhibitor 1 (PAI-1) were carried out on the supernatants. The cell counting technique demonstrated growth inhibition in the cell populations in contact with Woven Dacron® and Double Velour Dacron® in comparison with control cell cultures. Vital staining with neutral red, always sharply positive in the controls, was weak in the cells placed in contact with the materials. The 6-keto-PGF1a concentration in the supernatant was similar to the control level in the populations in contact with Woven Dacron® and Double Velour Dacron®. The tPA synthesis was higher in the cells exposed to the assayed materials compared to control values, while the PAI-1 concentration in the supernatant was lower in the cultures in contact with all materials.

Cell culture techniques are usually used in the field of biomaterials research and development in order to detect toxic components. Morphological assays are the most widely used methods and give the very first information about the biological compatibility of materials. Cell function assays give more quantitative data, but the comparison of data between different laboratories is difficult. Some of the cell culture methods that are used for biocompatibility studies are described briefly here, and results from our laboratory are reported. Despite some inherent limitations of the cell culture techniques, they are an accurate and reliable method of predicting the biological compatibility of materials to be implanted in vivo.

In order to evaluate if carbon coated polyethylene terephthalate (C-PET) could favor inflammatory reactions, the expression of interleukin-6 (IL-6) by cultured human umbilical vein endothelial cells was tested in vitro. The cultures were put in contact with C-PET for 1, 24, 48 and 72 h. The same cells cultured on tissue culture-treated polystyrene without biomaterials were tested as the negative control; the same cells incubated with LPS were the positive control. The level of IL-6 in the conditioned medium was tested by enzyme immunoassay; the mRNA expression was evaluated by RT-PCR with specific primers. The cultures incubated with C-PET produced non significantly different amount of IL-6 compared to the negative control and did not induce the expression of IL-6 specific mRNA. LPS induced a significantly higher release of IL-6 in the medium and the expression of mRNA after 24, 48 and 72 h. We conclude that C-PET does not stimulate the synthesis of IL-6 and therefore does not favor inflammatory reaction through the release of this cytokine.

The mutagenic potential of three commercially available glass-ionomer cements used in dentistry was examined. The cement components were mixed according to the manufacturers indications and set for two defined times: 1 h or, alternatively, 1 wk. Cements B and C set spontaneously; in the case of cement A, the manufacturer suggests the use of a lamp to trigger also a photopolymerization. Photopolymerization, however, was not used. Ames tests were performed on the dimethyl sulphoxide extracts of cements by using Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537, TA 1538 and TA 102. Cement A showed mutagenicity only against TA 1537 strain, either in the presence or absence of metabolic activation with microsomial fraction S9. The other two cements showed no mutagenic potential. We conclude that glass-ionomer cements are, on the whole, safe materials from the viewpoint of genotoxicity, and hypothesize that the mutagenicity observed in cement A could depend on its polymerization performed without light activation.

The aim of this research was to evaluate the effect of polyethylene terephthalate (Woven Dacron) on the expression of endothelial integrins. Human umbilical vein endothelial cells were cultured on the material for 24 h. The integrins VLA-2 (alpha2beta1-CD49b/CD29), receptor for laminin and collagen, VLA-5 (alpha5beta1-CD49e/CD29), receptor for fibronectin, VLA-6 (alpha6beta1-CD49f/CD29), receptor for laminin, and alphaVbeta3-CD51/CD61 (receptor for vitronectin) were evaluated by flow cytometry. After contact with polyethylene terephthalate, a slight but significant decrease in the percentage of both CD29 and CD49e positive cells was observed, which suggests a lower number of cells expressing the fibronectin receptor alpha5beta1. Moreover, a significant increase in the mean channel for CD49b and for the vitronectin receptor CD51/CD61 was observed. The reduction in the fibronectin receptor could account for the poor endothelialization observed in vivo on polyethylene terephthalate. The increased expression of the vitronectin receptor, favoring the migration of smooth muscle cells, could give some information about the pathogenesis of intimal hyperplasia, which is a complication of vascular grafts.

Deposition and aggregation of lachrymal proteins on the contact lens surface can promote bacterial adherence. Lysozyme is the major tear protein and is also mainly responsible for the formation of protein deposits on contact lenses. Nonsteroidal anti-inflammatory drugs (NSAID) prevent protein aggregation. The effect of a water-soluble NSAID drug on bacterial adherence to high-water-content/ionic disposable contact lenses was examined in a radiolabeling study. Dose-related inhibition of adherence of Staphylococcus aureus, S. epidermidis, and Pseudomonas aeruginosa on both pretreated lenses and after adding the drug to the medium was investigated. When the drug was added to the media, maximal inhibition of S. aureus adherence was observed in trypticase soy broth (59-98% at the lower and higher drug concentrations, respectively); inhibition progressively decreased in calf aqueous humor (48-75%), lysozyme (34-63%), and saline (12-20%) solutions. Inhibition of adherence varied with the three bacterial species; it was maximal with S. aureus, intermediate with S. epidermidis, and minimal with P. aeruginosa. When lenses were pretreated with the drug, consistent, and even higher, inhibitory effects were observed. The results suggest that water-soluble NSAIDs could be used in preventive treatments for conjunctivae and corneal infections in contact lens wearers, and may provide a clue as to which compounds might inhibit protein interaction and bacterial adhesion.

The aim of the research was the evaluation of plasmatic protein adsorption on untreated polybutylene terephthalate, corona treated polybutylene terephthalate and polyvinylacetate coated corona treated polybutylene terephthalate.Total proteins, albumin, immunoglobulins, fibrinogen, insulin and ostecalcin were determined on plasma after contact with these polymers.Unsignificant variations were observed for the assayed proteins.The conclusions are drawn that the tested materials do not adsorbe significantly the most important plasmatic proteins.

The authors examined the modifications of some markers of platelet activation after contact with biomaterials. Glycoprotein GMP-140 (CD62) was evaluated by flow cytometry; beta-thromboglobulin (beta-TG) and thromboxane B2 (TXB2) were determined by radioimmunoassay. Polyethylene terephthalate (PET) induced a remarkable platelet adhesion and a significant increase in beta-TG and TXB2, with no increase in CD62 on the nonadherent platelets. Pyrolytic carbon-coated PET (PC) did not induce platelet adhesion after 15 min of contact, but a significant increase in CD62 was detected. After 30 min a significant increase in platelet adhesion as well as the release of beta-TG and TXB2 were noted. The increase was lower than that observed for uncoated PET, and after 30 min of contact with PC the increase no longer was observed.

The aim of this study was to evaluate endothelin-1 and prostacyclin production by human endothelial cells cultured in the presence of polyethylene terephthalate and collagen-coated PET. Cell counting and the assay of endothelin-1 and 6-keto-prostaglandin F1 alpha, stable metabolite of prostacyclin, were carried out after 48 hour contact of the cells with the examined materials. Endothelial cell contact with uncoated PET caused a significant reduction in cell number, a significant increase in the production of endothelin-1 and a not significant increase in 6-keto-prostaglandin F1 alpha. The endothelial cell contact with collagen-coated PET caused a highly significant decrease in cell number and a not significant decrease in endothelin-1 and 6-keto-prostaglandin F1 alpha. It was concluded that PET causes both a decrease in cell number and a remarkable increase in endothelin-1. On the contrary, collagen-coated PET determines a decrease in cell number and a slight reduction of endothelin-1 and 6-keto-prostaglandin F1 alpha.

Five commercially available bone cements were analysed by high-performance liquid chromatography for detecting the residual content of an accelerator, the amine N,N-dimethyl-p-toluidine (DMPT), after curing. It was found that the concentration of DMPT in aqueous extracts decreases with time, being almost absent 7 days after curing. Differences were noticed among the cements; residual DMPT is higher in cements prepared with higher content of the amine. It is verified that DMPT's toxic effect on cell cultures is dose-related; a delay in the cell replication cycle is induced in vitro. Damage is reversible, thus justifying the low bone cement toxicity that is clinically ascertained.

The authors have evaluated adhesive protein expression and cytokine production by human umbilical vein endothelial cells cultured in contact with polyethylene terephthalate (PET). ELAM-1, ICAM-1 and VCAM-1 expression was determined by flow cytometry; the concentration of interleukin-1α (IL-1α), interleukin-6 (IL-6), granulocyte colony stimulating factor (G-CSF) and granulocyte-macrophage colony stimulating factor (GM-CSF) in the supernatant was determined by enzyme immunoassay. The contact with PET determined a significant increase in ELAM-1 expression and insignificant increase in cytokine production, demonstrating that PET had a limited capability to stimulate endothelial cells in a pro-inflammatory sense.

Cell viability and growth for cytotoxicity evaluation of materials for prosthetic devices has been tested using various methods. The aim of this study was to extend the choice of reliable methods to quantify cytotoxicity of materials in vitro. By measuring both viability and growth of cells exposed to biomaterials in vitro, two different parameters are analysed and quantified upon reading of the absorbance of coloured solutions in a spectrophotometer. Neutral red uptake and amido black staining of cells have been used for cell viability and cell number measurement, respectively: they have been found to be well correlated with the number of surviving cells. These methods have been adjusted to a 96-well microplate cell culture system and re-evaluated as simple and reliable methods for the quantitative assessment of biomaterial effect on cells.

In this study the toxic effects of chromium, nickel, and cobalt extracts on in vitro cultured lymphocytes were evaluated. Graphite furnace atomic absorption spectrometry was used to measure the ion concentration. After serial dilution of the extracts, the viability of lymphocytes at 24, 48, and 72 h was estimated by flow cytometry, including propidium iodide staining and light scatter property assessment, and by MTT reduction test. The results of the investigation allowed us to conclude that 1) standardization of the procedure for preparing extracts is fundamental to obtaining repeatability of results; 2) the toxicity of an extract cannot be evaluated with a single viability assay; a combination of functional and structural tests is required; 3) when methods based on enzymatic reactions are performed, e.g. MTT test, it is advisable to replace the extract containing metal ions with fresh medium in order to avoid any interference with viability testing; 4) the amount of Co and Ni in the extract is similar, but the Cr release is very poor; 5) the lower toxicity of Cr extract probably is due to the lower ion concentration; 6) the assessment of 50% cytotoxic concentration (TC50) allows quantification of materials toxicity and comparison of various metals; and 7) the determination of a noncytotoxic concentration, i.e., a concentration lower than TC10, is required for subsequent investigation of cell functions because such studies can be carried out only on viable cell population.

The ability of some biomaterials to activate plasma coagulation system was examined in vitro. After contact of platelet-rich plasma with biomaterials, some markers of the thrombin formation, i.e., fragment 1 + 2 and fibrinopeptide A, and some inhibitors of the blood coagulation mechanism were tested. Fragment 1 + 2 and fibrinopeptide A were found to be increased by all of the materials, though to a different extent. In particular, fragment 1 + 2 and fibrinopeptide A were significantly increased upon contact with polybutylene terephthalate and with collagen coated polyethylene terephthalate, respectively. Also antithrombin III was shown to decrease following exposure to biomaterials, but statistical significance was found only for polyethylene terephthalate and polyvinylacetate. As a results of this wide range of variability in the parameters, it is advisable to explore the plasma coagulation system with a multiparametric approach in which thrombin formation and coagulation inhibitors are thoroughly investigated.

The authors have examined the response of osteoblastic cells in contact in vitro with different acrylic cements used in orthopaedic surgery. The solid and liquid components of the cements were mixed in sterile environments according to the instructions provided by the manufacturers, and placed in contact with the cells 1/2, 1, 4, 6 and 24 hours after triggering of polymerization. The experimental model, that tends to define the kinetics of the release of toxic substances from acrylic cements, allowed to verify 24 hours after polymerization that the three cements had lost their toxic effects, which instead had been present to varying degrees for the first 1-6 hours. Cytomorphological observations did not reveal cellular damage, which instead was very clear when functional tests were used.

The authors evaluated several immunological parameters in patients with loosened hip arthroplasty in order to determine the state of sensitization with regard to the metallic constituents of the prosthesis. The results obtained by in vivo epicutaneous testing did not reveal a correlation between loosening and sensitization, as the patch test was positive in only 1 out of 16 cases examined. An evaluation of the lymphocyte subpopulation on peripheral blood demonstrated that patients with prostheses in Cr, Co, Mo, Ni alloy present significant lymphopenia, with a reduction in subpopulations CD4 and CD8 and a decrease in in vitro cytotoxic activity. The meaning of these modifications could be interpreted to be either a toxic effect products released from the implant, or as the recruitment of lymphocytes in the site of loosening, due to lymphocyte sequestration phenomena consequent to a cell-mediated hypersensitivity reaction.

The haemocompatibility of polyethylene terephthalate (Dacron ® ) coated with pyrolytic carbon was examined in vitro , evaluating its capability of inducing adhesion and platelet activation, and of modifying the intrinsic coagulation pathway. Platelet adhesion was evaluated by counting platelets before and after in vitro contact of human plasma with the material under examination. Platelet activation was evaluated by determining platelet factor 4 (PF4) and thromboxane B 2 . Intrinsic coagulation pathway alterations were studied by determining activated partial thromboplastin time (APTT) and the activity of single factors. The results obtained show that pyrolytic carbon-coated Dacron induces platelet adhesion, reduction in platelet volume and lower increase in thromboxane production than that obtained after contact with uncoated Dacron. Pyrolytic carbon-coated Dacron does not induce PF4 release, contrary to uncoated Dacron induces a significant release. Moreover, pyrolytic carbon-coated Dacron, induces a significant extension of APTT by reducing the activity of intrinsic pathway factors, particularly factor XI.

We evaluated the role of the immune system in the pathogenesis of the periprosthetic capsular contracture, the most frequently occurring complication following the implant of silicone prostheses. Peripheral blood samples from 22 patients with silicone-gel-filled implants were examined. In all cases a capsule was felt by palpation, and it was classified according to the Baker scale. Ten patients (group 1) had a Baker 2 contracture, and 12 (group 2) had severe contracture rated 3 and 4. The cells positive to antigens CD3, CD4, CD8, HLA-DR, CD19, CD25, CD57, CD16, and CD14, and the cytotoxic activity of the lymphocytes on target cells K562 were assessed by cytofluorimetric analysis. At time 0 there were no statistically significant differences between patients and normal subjects, nor between the two groups. At 48 h, the group 2 patients had a number/mm3 of cells CD57 + significantly higher than both group 1 and control group (P < .05). In group 1 patients, the cytotoxic activity was similar to that of normal subjects, whereas in group 2 it was significantly increased, in respect to both the controls (P < .05) and group 1 (P < .001). In all groups, the contact of the lymphocytes with the silicone extract did not modify either the antigen expression or the lymphocyte functional activity. On the basis of these results we hypothesize the involvement of the immune system in the formation of the capsular contracture around the prosthesis. © 1995 John Wiley & Sons, Inc.

The antibiotic sensitivity of 19 Gram-positive bacterial strains (11 Staphylococcus aureus and 8 Staphylococcus epidermidis) and 16 Gram-negative strains (8 Escherichia coli and 8 Proteus species) was evaluated after contact with stainless steel and with some metals compounding the alloys used for prosthetic devices. The hypothesis was that the resistance to antibiotic therapy of infections associated with prosthetic implants is also due to a modification in the sensitivity of microorganisms. The results, compared to those obtained from control tests, showed only slight variations in the antibiotic sensitivity of the strains put in contact with the metals. In Gram-positive strains, after contact with metals, the increase in sensitivity occurred more frequently than the reduction. In Gram-negative strains, the decrease in sensitivity was more frequent than the increase. Proteus strains showed sensitivity variations more frequently than Escherichia coli strains. Titanium and nickel induced the highest number of variations, both in Gram-positive and Gram-negative strains.

We evaluated the alterations in number, functionality and release reaction of the platelets contained in plasma, filtered through a polyester filter and coated with a hydrophilic polymer. The alterations in number were examined by counting before and after filtration. The morphological modifications were studied by determining the mean platelet volume. The functional alterations were analysed by a platelet aggregation test, induced by ADP and collagen. The presence of products from the release reaction in filtered plasma was studied using radioimmunoassays of beta-thromboglobulin, platelet factor 4 and thromboxane B2. The results obtained showed that the filtration of plasma through the material did not determine a significant platelet adhesion, did not alter the volume or the functionality of the platelets and induced no release reaction.

Infection is one of the major complications in the use of prosthetic devices. Various Authors investigated the causes which favour such infection. We examined the changes in the resistance to antibiotics after contact with the metals used in the construction of prostheses, under the hypothesis that the persistence of the infection is due to a modification in the sensitivity of microorganisms. Compared with the results obtained from control tests, more or less evident variations of sensitivity to antibiotics were found in some strains which were put in contact with some metals.

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. β-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.

The amount of fluoride release from dental cements necessary for an anticariogenic effect is not established: moreover, the possible toxic effects due to high fluoride and aluminum release are not well known and the results are still controversial. The aim of our study was to evaluate fluoride (F) and aluminum (Al) release from dental cements using a 'standardized approach' according to the end-use of the materials, i.e. biocompatibility testing. Two polyacid-modified resin composites of recent application, commonly called compomers (Dyract and Dyract Cem), were compared with two conventional acid-based (Fuji I, Ketac-Cem) and two resin-modified (Vitremer, Vitrebond) glass-ionomer cements (GICs). All types of cement are used in dentistry and are commercially available. Extracts of the cements into minimum essential medium, after setting over a 1-h (group A) and 1-week (group B) period, were performed. The extraction conditions were rigorously standardized. Mean values +/- standard deviation of F- and Al-levels in such extracts were measured and were expressed as microg g(-1) (micrograms of ions per gram of cement). A great difference in the amount of ion release, both F and Al, was shown among the tested materials. The GICs, as well as Ketac-Cem, released more F and Al than the compomers. All of the materials released the greatest proportion of ions when the extraction was performed in the first hour after mixing (group A). Al- and F-values showed a highly significant positive correlation, independently from the curing time. We conclude that the biological assessment of dental cements can be performed only if a pre-evaluation of the leachables is obtained by applying a standardized protocol which allows a useful comparison between the different materials.

This study concerns the activation of the plasmatic pathway of coagulation induced by an experimental filter for the transfusion of packed red cells. A physical-chemical treatment was performed on the filter in order to reduce the priming time. For this purpose, the assay of fibrinopeptide A and protein C was carried out after a contact in vitro between normal plasma and the material used in the construction of the filter. As a control, a commercial filter was used, made of the same material without the physical-chemical treatment. The obtained results show clearly that the physical-chemical treatment does not cause the activation of the plasmatic pathway of coagulation.