Biocompatibility of platinum-metallized silicone rubber: in vivo and in vitro evaluation

Silicone rubber is commonly used for biomedical applications, including implanted cuff electrodes for both recording and stimulation of peripheral nerves. This study was undertaken to evaluate the consequences of a new platinum metallization method on the biocompatibility of silicone rubber cuff electrodes. This method was introduced in order to allow the manufacture of spiral nerve cuff electrodes with a large number of contacts. The metallization process, implying silicone coating with poly(methyl methacrylate) (PMMA), its activation by an excimer laser and subsequent electroless metal deposition, led to a new surface microtexture. The neutral red cytotoxicity assay procedure was first applied in vitro on BALB/c 3T3 fibroblasts in order to analyze the cellular response elicited by the studied material. An in vivo assay was then performed to investigate the tissue reaction after chronic subcutaneous implantation of the metallized material. Results demonstrate that silicone rubber biocompatibility is not altered by the new platinum metallization method.     

Biocompatibility of platinum-metallized silicone rubber: in vivo and in vitro evaluation

Silicone rubber is commonly used for biomedical applications, including implanted cuff electrodes for both recording and stimulation of peripheral nerves. This study was undertaken to evaluate the consequences of a new platinum metallization method on the biocompatibility of silicone rubber cuff electrodes. This method was introduced in order to allow the manufacture of spiral nerve cuff electrodes with a large number of contacts. The metallization process, implying silicone coating with poly(methyl methacrylate) (PMMA), its activation by an excimer laser and subsequent electroless metal deposition, led to a new surface microtexture. The neutral red cytotoxicity assay procedure was first applied in vitro on BALB/c 3T3 fibroblasts in order to analyze the cellular response elicited by the studied material. An in vivo assay was then performed to investigate the tissue reaction after chronic subcutaneous implantation of the metallized material. Results demonstrate that silicone rubber biocompatibility is not altered by the new platinum metallization method.     

In vivo tissue reactivity of radiation-cured silicone rubber implants.

Silicone rubber implants are clinically used in large numbers and elicit a milk tissue reaction. An occasional patient develops an accentuated reaction, an observation which has stimulated clinicians to try to understand this process more fully. The chemistry of medical silicone implants, including quantitative composition, is reviewed. This in vitro laboratory study show less tissue reaction-cured silicone with SiO2 filler. A proposed system for fabrication and curing of a silicone implant, with the qualities of strength and diminished tissue reactivity, is discussed.     

Synthesis and in vitro antimicrobial and antioxidant activities of quaternary ammonium chitosan modified with nisin

Nisin had been grafted onto quaternary ammonium chitosan (QCS) through an enzyme-catalyzed reaction to enhance its limited antimicrobial activity. QCS was synthesized by incorporating N-(3-chloro-2-hydroxypropyl) trimethyl ammonium chloride (CHPTAC) onto chitosan’s primary amine group. The modification had been confirmed by FT-IR and ¹H NMR spectroscopy. Degree of substitution (DS) of QCS–nisin could be controlled by adjusting the reaction conditions. The synthesized compounds were screened in vitro to evaluate their antimicrobial and antioxidant activities. The results suggested that QCS–nisin significantly suppressed the growth of both gram-positive bacteria and gram-negative bacteria; The antioxidant effects on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, hydroxyl radical and hydrogen peroxide (H₂O₂) proved to be enhanced with increasing DS and concentration. In addition, QCS–nisin showed excellent moisture absorption and retention properties; MTT assay exhibited that QCS–nisin revealed low cytotoxicity effects on cultured NIH-3T3 fibroblasts. These results suggest that QCS–nisin would appear to be a promising candidate for wound dressing application.     

In vivo degradation of silicone rubber poppets in prosthetic heart valves.

Dynamic shear modulus G' was measured throughout the volume of three nonvariant silicone rubber poppets which were recovered from aortic prosthetic heart valves that had been implanted for 4 days, 52 days, and 8 years. Similar measurements were obtained for two unused silicone rubber poppets. Although the recovered poppets exhibited no obvious physical evidence of damage, the silicone rubber had undergone in vivo degradation throughout the poppet volume as indicated by decreases in modulus. The measurements also indicate that the poppet surface degrades at a rate faster than the core. Further, comparison with data reported in the literature suggests that the surfaces of variant poppets degrade at a rate faster than the surfaces of nonvariant poppets.     

In vitro and in vivo antioxidant activity of ambroxol

Abstract In addition to a mucolytic action, ambroxol has antioxidant and anti-inflammatory properties. The antioxidant effects of ambroxol were studied both in vitro and in vivo. In vitro methods, such as (1) inhibition of hyaluronic acid degradation induced by hydroxy radicals and (2) standard lipid peroxidation assay in rat liver mitochondria and gastric mucosa, induced by tert-butyl hydroperoxide, were used. The in vivo approach was based on the study of the protective effect of pretreatment with ambroxol in a rat model of gastric corpus and antral lesions, induced by indomethacin. The inhibition of the degradation of hyaluronic acid was measured as a change of its viscosity; ambroxol (1,000 µl/l) reduced the degradation by 93%. Lipid peroxidation with tert-butyl hydroperoxide as a source of radicals was followed by the formation of thiobarbituric acid reactive substances. Ambroxol (10 mmol/l) inhibited lipid peroxidation by 96% in the rat liver mitochondria, and by 74% in the gastric mucosa. In vivo, ambroxol was administered p.o. at a dose of 10, 30, and 50 mg/kg, at 5, 30, and 60 min prior to indomethacin administration. The highest inhibition of the number of corpus gastric lesions and lowering of the lesion index (38% and 62%, respectively) was shown after the administration of 50 mg/kg, 30 min before indomethacin administration. Antral lesions were inhibited to a lesser extent by the same dose of ambroxol, administered 30 min before indomethacin treatment. Inhibition of the number of antral lesions reached 27% and the total area of the gastric damage was even larger (the ulcer index reached -5%).     

Antimicrobial activity of silicone rubber used in hydrocephalus shunts, after impregnation with antimicrobial substances.

Colonisation of cerebrospinal fluid shunts by coagulate-negative staphylococci (Staphylococcus albus) is a serious problem. Because of its possible role in prevention of the condition, the antimicrobial activity of silicone rubber after impregnation with antimicrobial drugs was studied. The method of impregnation used and test methods were found to be important. Formaldehyde-urea condensates gave no activity. Gentamicin sulphate gave activity which was short-lived. Sodium and diethanolamine fusidates and clindamycin hydrochloride gave prolonged activity. A method of impregnation was developed which could be applied to commercially available shunts before use.     

In vitro and in vivo antioxidant activity of ambroxol

In addition to a mucolytic action, ambroxol has antioxidant and anti-inflammatory properties. The antioxidant effects of ambroxol were studied both in vitro and in vivo. In vitro methods, such as (1) inhibition of hyaluronic acid degradation induced by hydroxy radicals and (2) standard lipid peroxidation assay in rat liver mitochondria and gastric mucosa, induced by tert-butyl hydroperoxide, were used. The in vivo approach was based on the study of the protective effect of pretreatment with ambroxol in a rat model of gastric corpus and antral lesions, induced by indomethacin. The inhibition of the degradation of hyaluronic acid was measured as a change of its viscosity; ambroxol (1,000 microl/l) reduced the degradation by 93%. Lipid peroxidation with tert-butyl hydroperoxide as a source of radicals was followed by the formation of thiobarbituric acid reactive substances. Ambroxol (10 mmol/l) inhibited lipid peroxidation by 96% in the rat liver mitochondria, and by 74% in the gastric mucosa. In vivo, ambroxol was administered p.o. at a dose of 10, 30, and 50 mg/kg, at 5, 30, and 60 min prior to indomethacin administration. The highest inhibition of the number of corpus gastric lesions and lowering of the lesion index (38% and 62%, respectively) was shown after the administration of 50 mg/kg, 30 min before indomethacin administration. Antral lesions were inhibited to a lesser extent by the same dose of ambroxol, administered 30 min before indomethacin treatment. Inhibition of the number of antral lesions reached 27% and the total area of the gastric damage was even larger (the ulcer index reached -5%).     

Inorganic coatings for cardiovascular stents: In vitro and in vivo studies

The in-vitro and in-vivo biocompatibility of two oxides (TiO and ZrO) and diamond-like carbon (D) coated stents has been assessed and compared with uncoated stainless steel (St) stents. In vitro studies demonstrated that both fibrinogen adsorption and platelet adhesion were significantly higher on D coating compared to those on oxide coatings and uncoated stainless steel. In addition TiO and ZrO coatings showed evidence of a minor inflammatory response and more complete endothelialization of the aorta than that seen around D coated and uncoated St stents. The resulting neointimal growth in the aorta with TiO, ZrO, and D coated and uncoated St stents, measured 8 weeks after stenting (the ratio of the neointima in the stented artery to the non-stented artery) was 1.03 + 0.28, 0.85 + 0.36, 1.78 + 1.26, and 1.15 + 0.56, accordingly. From the data obtained it could be concluded that the increased neointima measured around D-coated stents, may be due to both, the inferior haemocompatibility of the diamond-like carbon coating and mechanical instability of D coating observed in an in vivo environment.     

In vitro and in vivo reactivity of porous, electrosprayed calcium phosphate coatings

The dissolution and/or precipitation behaviour of porous calcium phosphate (CaP) coatings, deposited using electrostatic spray deposition (ESD), was investigated (a) in vitro after soaking in simulated body fluid (SBF) for several time periods (2, 4, 8, and 12 weeks), and (b) in vivo after subcutaneous implantation of CaP-coated implants in the back of goats for identical time periods. Physical and chemical properties of coatings were characterized before and after in vitro/vivo testing by means of scanning electron microscopy, X-ray diffraction, Fourier-transform infrared spectroscopy, and energy dispersive spectroscopy. Moreover, part of the explants was prepared for light microscopical evaluation of the tissue response. In vitro, all apatitic ESD-coatings induced the formation of homogeneous and adherent CaP precipitation layers. Amorphous CaP, however, displayed a delayed precipitation of poorly adherent CaP layers, whereas heterogeneous calcification was observed on top of beta-TCP-coated substrates, indicating that beta-TCP and amorphous CaP coatings exhibit a poor ability of inducing calcification in SBF as compared to crystalline apatitic coatings. In vivo, no adverse tissue reactions (toxic effects/inflammatory cells) were observed using light microscopy, and all coatings became surrounded by a dense, fibrous tissue capsule after implantation. All ESD-coatings degraded gradually at a dissolution rate depending on the chemical phase (order of relative solubility: amorphous CaP approximately carbonate apatite>beta-TCP>carbonated hydroxyapatite), thereby enabling synthesis of CaP coatings with a tailored degradation rate.     

In vitro antimicrobial activity of the new antibiotic vermisporin

The antimicrobial activity of vermisporin, a new antibiotic produced by fermentation of the fungusOphiobolus vermisporis, was tested in vitro. Vermisporin inhibited 90 % ofBacteroides fragilis and otherBacteroides spp. at 1 µg/ml (range 0.25–1 µg/ml).Clostridium perfringens were inhibited by 1 µg/ml (range 0.25–2 µg/ml). Vermisporin inhibited 90 % ofStaphylococcus aureus, including methicillin-resistantStaphylococcus aureus, at 0.5 µg/ml (range 0.12–0.5 µg/ml). Vermisporin MICs for group A, B, C, F and G streptococci were < 1 µg/ml when tested in Haemophilus Test Medium but 8 µg/ml in the presence of blood. Vermisporin MICs forEnterobacteriaceae, Pseudomonas aeruginosa andHaemophilus influenzae exceeded 64 µg/ml. Inhibited organisms had MBCs 16- to 32-fold above the MICs.     

In vitro activity of seventeen antimicrobial agents againstGardnerella vaginalis

The in vitro activity of 17 antimicrobial agents was tested against 25 clinical isolates ofGardnerella vaginalis. Minimum inhibitory concentrations were determined by agar dilution. The isolates were sensitive to penicillin, ampicillin, ticarcillin, piperacillin, cephalothin, cefoxitin, cefotaxime, cefoperazone, N-formimidoyl-thienamycin, chloramphenicol, clindamycin and erythromycin. MIC₉₀ for theβ-lactam antibiotics ranged from 0.12 mg/l for penicillin to 2 mg/l for ticarcillin. Cefoperazone was the most active cephalosporin, inhibiting all isolates at 1.0 mg/l. N-formimidoyl-thienamycin was the most active of the newerβ-lactam compounds inhibiting all isolates with a concentration of 0.5 mg/l. Clindamycin and erythromycin were highly active, inhibiting all isolates at 0.6 mg/l. Susceptibility to tetracycline, gentamicin, metronidazole and tinidazole varied between strains. All isolates were resistant to rosoxacin. The hydroxy-metabolites of metronidazole and tinidazole were more active than the parent compounds, inhibiting all isolates     

In vitro and in vivo anti-allergic activity of soy sauce

Soy sauce (Shoyu) is a traditional fermented seasoning of Japan and available throughout the world. Polysaccharides were obtained from dialysate of Shoyu, and these Shoyu polysaccharides (SPS) were examined for anti-allergic activity in vitro and in vivo. The SPS originated from partially-degraded polysaccharides of soybeans by mold enzymatic hydrolyses, and Shoyu contained about 1% (w/v) SPS. First, the inhibitory effects of SPS on hyaluronidase, which is known to be related to inflammation and allergic responses, were as potent as those of an anti-allergic medicine, disodium cromoglycate. Second, SPS significantly inhibited the release of histamine from rat basophilic leukemia (RBL-2H3) cells, which had been induced by the antigen. Third, orally administered SPS had a significant suppressive effect on passive cutaneous anaphylaxis induced in the ears of mice. These results suggest that SPS may have anti-allergic activities, and soy sauce is a potentially promising seasoning for the treatment of allergic diseases through food.     

注射用头孢拉宗钠的体内外抗菌活性研究

目的评价注射用头孢拉宗钠的体内外抗菌活性。方法选取663株临床分离致病菌为实验菌,以头孢美唑、头孢西丁、头孢替坦、头孢米诺、头孢唑啉、头孢呋辛、头孢哌酮、头孢吡肟为对照药物,分别采用平皿二倍稀释法测定药物最低抑菌浓度（MIC）、最低杀菌浓度（MBC）、绘制杀菌曲线（KCs）并行诱导耐药实验,观察头孢拉宗的体外抗菌作用;建立小鼠腹腔感染模型,评价头孢拉宗皮下给药对金黄色葡萄球菌、大肠埃希菌、肺炎克雷伯杆菌感染小鼠的体内疗效。结果头孢拉宗对多种革兰阴性菌具有较高的抗菌活性,尤以对大肠埃希菌和肺炎克雷伯杆菌的抗菌活性更为突出,其MIC50、MIC90分别为0.5、8μg/ml,对产超广谱β-内酰胺酶大肠埃希菌和肺炎克雷伯杆菌的抗菌活性也较强,其MIC50、MIC90分别为4、8μg/ml;头孢拉宗对革兰阴性菌的抗菌作用大多优于头孢西丁、头孢美唑、头孢唑啉、头孢哌酮、头孢替坦和头孢呋辛,而对革兰阳性菌的抗菌活性较弱,与头孢替坦和头孢米诺相近。MBC和KCs测定结果表明,头孢拉宗对金黄色葡萄球菌、大肠埃希菌、肺炎克雷伯杆菌均具有杀菌作用。以1/4MIC剂量诱导20d后,头孢拉宗对金黄色葡萄球菌、大肠埃希菌和肺炎克雷伯杆菌的抗菌活性无明显改变。头孢拉宗皮下给药对大肠埃希菌ATCC25922、大肠埃希菌1515、肺炎克雷伯杆菌7、肺炎克雷伯杆菌9613感染小鼠的体内疗效明显优于头孢美唑和头孢唑啉（均P〈0.01）;对金黄色葡萄球菌感染小鼠的体内疗效与头孢美唑相近（P〉0.05）,但弱于头孢唑啉（P〈0.01）。结论注射用头孢拉宗钠对多数革兰阴性菌,特别是大肠埃希菌和肺炎克雷伯杆菌具有较强的体外抗菌活性;同时对大肠埃希菌、肺炎克雷伯杆菌、金黄色葡萄球菌腹腔感染小鼠的体内疗效也较好。     

Preparation and antibacterial properties of gelatin grafted with an epoxy silicone quaternary ammonium salt

Gelatin (GE) was modified with epoxy silicone quaternary ammonium salt (EPSiQA) under alkaline conditions (pH 10–11). Silyl and quaternary ammonium groups were linked to gelatin skeleton simultaneously. It was illustrated by XRD and DSC that the short-range order of GE is destroyed and the glass transition temperature (Tg) of GE drops 10 °C after modification. The measured contact angles and surface free energy calculated by Owens–Wendt equation showed that the surface energy of modified gelatin EPSiQA-GE is mainly contributed by the dispersive component of non-polarity silicone groups, the hydrophobility of EPSiQA-GE increases with the increase of grafted silicone units in gelatin. The results of minimum inhibitory concentration indicated that EPSiQA-GE has bactericidal property against Gram-positive bacteria, Gram-negative bacteria and has no antibacterial effect on mold.     

In vivo andin vitro anti-tumour activity of dimaprit

Dimaprit, a histamine H₂-receptor agonist, injected daily i.p. to fibrosarcoma-bearing mice, induced a decrease in tumour growth and an increase in survival. Dimaprit, added to tumour cell cultures (10^–4 M), inhibited the incorporation of³H-thymidine while embryonic cell cultures were unaffected. This particular anti-tumour activity is probably H₂-independent as histamine and impromidine have no effect on tumour cell cultures.     

In vitro and in vivo biological responses of plasma-sprayed hydroxyapatite coatings with posthydrothermal treatment

This study was undertaken to evaluate the effect of post-hydrothermal treatment on the biological responses of the plasma-sprayed hydroxyapatite (HA)-coated Ti-6Al-4V implant system both in vitro and in vivo. After hydrothermal treatment, the HA coating (HAC) shows the high mechanical strength and indices-of-crystallinity, denser microstructure, lower concentrations of amorphous and impurity phases, when compared with the as-sprayed HAC. The in vitro cell-culture studies, using UMR106 osteoblast-like cell, demonstrated no signifiacnt cell growth on both surface of as-sprayed and hydrothermal-treated HACs during 10-day culture. The in vivo studies, using the transcortical implant model in the femora of goats, evaluated the histological responses of two coatings. After 6 week of implantation, using backscattered electron images, no substantial histological variations in the extents of new bone apposition and new bone healing between the two HACs were observed. However, the as-sprayed HAC, owing to the dissolution induced the granular particles dissociated from the HAC, showed the statically lower extent of new bone apposition than hydrothermal-treated HAC at 12 weeks. The results suggest that hydrothermal treatment could be used to improve the mechanical strength, crystallinity, and phase composition of HAC, which are important factors of long-term fixation and stability of implant. Besides, the treated HAC could also achieve the initial fixation of implant in clinical use.     

In vitro antimicrobial activity of diethyldithiocarbamate and dimethyldithiocarbamate against methicillin-resistant Staphylococcus

Staphylococcus aureus has appeared which is highly resistant to both methicillin and aminoglycosides. Current therapy involves long-term intravenous therapy of vancomycin. Since vancomycin is currently the only drug used to treat these patients, there is a need to develop additional antimicrobial therapy. The in vitro antimicrobial effect of the metal chelator, diethyldithiocarbamate (DDTC) and its structural analog dimethyldithiocarbamate (DMTC) were investigated. Both DDTC and DMTC were effective against S. aureus including methicillin-resistant S. aureus (MRSA). By agar diffusion, DDTC at 10 micrograms per disk produced zone sizes of 12 to 21 mm and at 100 micrograms per disk produced zone sizes of 26 to 34 mm against MRSA. The DMTC produced slightly greater zone sizes against MRSA of 16 to 24 mm and 24 to 37 mm for 10 micrograms per disk and 100 micrograms per disk, respectively. The minimum inhibitory concentration (MIC) for DMTC against MRSA was 6 micrograms per ml. Both DDTC and DMTC were also effective against enterococci, Proteus mirabilis, Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Enterobacter cloacae, Enterobacter aerogenes, Salmonella species, Serratia marcescens and Citrobacter freundii at 100 micrograms per disk. The MICs of DMTC for Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Salmonella and Citrobacter freundii were approximately 128 micrograms per ml while the MICs for Proteus vulgaris, Proteus mirabilis, Pseudomonas aeruginosa and Serratia marcescens was greater than or equal to 256 micrograms per ml. In addition, DMTC was synergistic with gentamicin against MRSA and coagulase-negative staphylococcus species, Enterobacter cloacae, Klebsiella pneumoniae and Pseudomonas aeruginosa. Additive and synergistic effects of DMTC were displayed with gentamicin against S. aureus including methicillin-resistant S. aureus.(ABSTRACT TRUNCATED AT 250 WORDS)