Quantitative assessment of the response of osteoblast- and macrophage-like cells to particles of Ni-free Fe-base alloys.

In the present study, the effect of mechanically alloyed particles of new FeAlCr alloys developed for potential applications as surgical implants has been tested on osteoblast- and macrophage-like cells and compared to particles of the Ti6Al4V alloy, for which there is a good clinical experience. After microstructural characterisation of the particles, cells were cultured with particles for 24-48 h using three different concentrations of particles, and the response of cells was quantified by assessment of viability, proliferation, and morphology. Mineralisation by osteoblasts was verified after 21 days. The amount of aluminium and chromium ions in the culture medium of macrophages was measured by graphite furnace atomic absorption and phagocytosis of particles assessed by light microscopy. Viability and proliferation of osteoblast- and macrophage-like cells were substantially unaffected by the presence of particles of the new alloys, which were phagocytosed according to their size. Aluminium and chromium ions were released in the culture medium, but no direct correlation with the cell behaviour was found. In vitro mineralisation was achieved by osteoblasts in due time. The new alloys are well tolerated in in vitro systems, and, due to their chemical and mechanical characteristics, they are under development for surgical implants.

In vitro assessment of phagocytosis of bovine collagen by human monocytes/macrophages using a spectrophotometric method.

The use of a wound dressing with covering and haemostatic properties significantly improves wound healing. In this study, a lyophilized bovine collagen sponge used for the treatment of wounds and ulcerae has been tested in a cell culture system. Phagocytosis of collagen fragments by human blood monocytes/macrophages has been investigated. For the assessment of collagen ingestion by mononuclear phagocytes, a picrosirius dye specific for collagen molecules has been used. By adapting this histochemical technique to microplate cell culture system, replicate monocyte cultures are assayed. Collagen content is determined by evaluating spectrophotometrically at 540 nm the absorbance of a sirius red/picric acid solution. Using this simple and sensitive method, the phagocytosis of bovine collagen by LPS-stimulated monocytes/macrophages has been ascertained.

Assessment of metal extract toxicity on human lymphocytes cultured in vitro.

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.

[Assessment of various immunologic parameters in silicone implant carriers]. / Valutazione di alcuni parametri immunologici in pazienti portatrici di protesi di silicone.

The Authors evaluated some immunological parameters in women carrying silicone gel-filled breast implants for over one year. Peripheral blood samples from 22 patients were examined in order to assess both the antigenic pattern of lymphocyte subpopulations by cytofluorimetric analysis, and the cell proliferation of PHA-stimulated lymphocytes by the uptake of tritiated thymidine. These tests were performed at the time of the sample withdrawal and after in vitro reexposure to silicone extract for 48-72h. Changes in lymphocyte subpopulations and functional response were observed when patients were divided into groups according to the type of surgery, i.e. breast augmentation or reconstruction, or to the degree of periprosthetic capsular contracture. These results suggest the possibility of an interaction between silicone and the immune system, which cannot be disregarded for the explanation of the silicone related complications.

Assessment of viability and proliferation of in vivo silicone-primed lymphocytes after in vitro re-exposure to silicone.

The functional response of peripheral blood lymphocytes isolated from 22 patients with silicone gel-filled breast implants was assessed after in vitro re-exposure to silicone. Using cell culture test methods to quantify proliferation and viability and/or activation of lymphocyte microcultures, i.e., the uptake of tritiated thymidine (3H-TdR uptake test) and the reduction of formazan salts (MTT assay), interesting data were obtained. Peripheral blood lymphocytes purified from patients wearing silicone gel-filled breast implants react in vitro to silicone showing a statistically significant increase of both proliferation and viability, while healthy subjects do not respond on in vitro exposure to silicone. Differences resulted even more statistically significant when patients were divided into two groups depending on the type of surgery they underwent: patients with breast augmentation for aesthetic reasons seem to have an increased responsiveness in vitro to silicone compared to patients who experienced a reconstructive surgery of the breast. Although they are still preliminary, being referred to a limited population, these results suggest that the lymphocytes of patients with silicone gel-filled breast implants could be sensitized in vivo toward silicone; the re-exposure of these cells to silicone leads to a higher functional response which could be looked for by using quantitative in vitro test methods.

[Assessment of the plasma phase of coagulation in platelet-rich plasma after passage through a special polyester filter]. / Valutazione della fase plasmatica della coagulazione in plasma ricco di piastrine dopo passaggio attraverso un particolare filtro in poliestere.

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.