Effect of resin hydrophilicity and temperature on water sorption of dental adhesive resins

This study examined the effects of copolymer hydrophilicity and temperature on water sorption and solubility characteristics of five copolymer blends of increasing degree of hydrophilicity using gravimetric measurements. Six resin disks (15 mm in diameter x 1 mm in thickness) were prepared from each copolymer blend and were stored in deionised water at 23, 37 and 55 degrees C. Water sorption and solubility of the resin disks were measured before and after water immersion and desiccation. Multiple regression analysis of water sorption was performed on two independent variables, copolymer hydrophilicity and temperature. Maximum water sorption increased significantly with Hoy's total cohesive energy density (delta(t)), Hoy's solubility parameter for polar forces (delta(p)) and hydrogen bonding (delta(h)), but was not influenced by temperature. However, a significant positive relationship was observed between diffusion coefficients (obtained using Fick's law of diffusion) and temperature. The water absorption activation energy was 10 kJ/mol for the most hydrophilic copolymer blend R5 and 35-51 kJ/mol for copolymer blends R1-R4. The positive relationship between maximum water uptake and copolymer hydrophilicity suggests that water molecules diffuse through the polymer matrices by binding successively to the polar sites via hydrogen bonding. Such water sorption may determine the durability of resin-dentine bonds.     

Effects of resin hydrophilicity on water sorption and changes in modulus of elasticity

As acidic monomers of self-etching adhesives are incorporated into dental adhesives at high concentrations, the adhesive becomes more hydrophilic. Water sorption by polymers causes plasticization and lowers mechanical properties. The purpose of this study was to compare the water sorption and modulus of elasticity (E) of five experimental neat resins (EX) of increasing hydrophilicity, as ranked by their Hoy's solubility parameters and five commercial resins. METHODS: After measuring the initial modulus of all resin disks by biaxial flexure, half the specimens were stored in hexadecane and the rest were stored in water. Repeated measurements of stiffness were made for 3 days. Water sorption and solubility measurements were made in a parallel experiment. RESULTS: None of the specimens stored in oil showed any significant decrease in modulus. All resins stored in water exhibited a time-dependent decrease in modulus that was proportional to their degree of water sorption. Water sorption of EX was proportional to Hoy's solubility parameter for polar forces (delta(p)) with increasing polarity resulting in higher sorption. The least hydrophilic resin absorbed 0.55 wt% water and showed a 15% decrease in modulus after 3 days. The most hydrophilic experimental resin absorbed 12.8 wt% water and showed a 73% modulus decrease during the same period. The commercial resins absorbed between 5% and 12% water that was associated with a 19-42% reduction in modulus over 3 days.     

Effect of chain modifications on the physicomechanical properties of silsesquioxane-based dental nanocomposites

The objective of this study was to evaluate the physicomechanical properties of a series of polyhedral silsesquioxane (SSQ) methacrylate monomers developed for dental applications. The effect of chain modifications on the properties of the SSQ-based monomers was also evaluated. Physicomechanical properties that are investigated include polymerization shrinkage, degree of conversion, hardness, and modulus. Results obtained were compared with unfilled 1:1 (control) bis-GMA/TEGDMA materials (typical monomers used in dental composites). All samples investigated were cured using 400-500 nm light at 500 mW/cm(2) for 40 s. Shrinkage associated with curing and post-gel reactions for all synthesized SSQ compounds were found to range from (0.04 +/- 0.01)% to (0.33 +/- 0.03)% with degree of conversion ranging from (56.68 +/- 2.81)% to (84.53 +/- 2.62)%. At all time intervals, post-gel shrinkage associated with control was found to be significantly greater than all SSQ compounds. No significant difference in degree of conversion was observed for control, and all SSQ compounds except for SSQ attached with eight equivalents of short-chain methacrylate. Mechanical properties associated with SSQ compounds were found to be significantly lower than control. However, through chain modifications, mechanical properties of SSQ compounds can be improved by approximately 50%.     

Effects of filler composition on flexibility of microfilled resin composite

The effects of the filler composition on physical and mechanical properties of microfilled composites was investigated by measuring water absorption, solubility, compressive, flexural, and impact strength. A series of experimental composites, consisting of UDMA/TEGDMA comonomer matrix and prepolymerized fillers, was fabricated. The prepolymerized fillers were composed of hydrophobic colloidal silica and two monomers in varying ratios, trimethylolpropanetrimethacrylate (TMPT), and polyesterdiacrylate (PEDA). TMPT/PEDA ratios were 100:0, 64:36, 46:54, 18:82, and 0:100%. There were no significant differences in water sorption and solubility, regardless of the amount of PEDA monomer. Young's modulus and modulus of resilience increased with decreasing PEDA ratio. Fracture energy exhibited drastic changes (30.1 x 10(-5) J to 93.4 x 10(-5) J). The highest value of flexural strength (96.0 +/- 3.5 MPa) was obtained when the TMPT-PEDA filler was 46:54. The impact strengths of composites fabricated with TMPT-PEDA filler of 46:54 (11.2 +/- 1.4 kJ/m(2)), 18:82 (10.6 +/- 3.2 kJ/m(2)), and 0:100 (13.1 +/- 3.8 kJ/m(2)) were significantly higher than those with 100:0 (6.0 +/- 1.8 kJ/m(2)) or 64:36 (7.1 +/- 2.4 kJ/m(2)). Based upon the results, it was concluded that the mechanical properties of microfilled composites were improved by the modification of prepolymerized filler composition.     

Trifunctional methacrylate monomers and their photocured composites with reduced curing shrinkage,water sorption,and water solubility

Novel trifunctional methacrylates, 1,1,1-tris[4-(2'-acetoxy-3'-methacryloyloxypropoxy)phenyl]ethane (Ac-THMPE) and 1,1,1-tris[4-(2'-acetoxy-3'-methacryloyloxypropoxy)phenyl]methane (Ac-THMPM), have been prepared by acetylation of the hydrophilic hydroxyl groups of 1,1,1-tris[4-(2'-hydroxy-3'-methacryloyloxypropoxy)phenyl]ethane (THMPE) and 1,1,1-tris[4-(2'-hydroxy-3'-methacryloyloxypropoxy)phenyl]methane (THMPM), respectively, for use as dental monomers. Decrease in monomer viscosity resulted from the acetylation. Unfilled resins and composites based on the four trimethacrylates were evaluated for photopolymerization conversion, water contact angle, and curing shrinkage. Water sorption, water solubility, and flexural strength of the composites prepared from the trimethacrylate were measured. Those data obtained for the trimethacrylate-containing materials were compared with control 2,2-bis[4-(2'-hydroxy-3'-methacryloyloxypropoxy)phenyl]propane (bis-GMA)-based materials in order to evaluate the applicability of the trimethacrylates as dental monomers. The acetylation of hydroxyl groups appeared to be an effective method to decrease curing shrinkage, water sorption, and water solubility of the dental composites. When compared with the bis-GMA composite, the composites based on Ac-THMPM and Ac-THMPM showed much lower curing shrinkage, water sorption, and water solubility, along with approximately equal conversion and flexural strength.     

Synthesis and characterization of antibacterial dental monomers and composites

The objective of this study is to synthesize antibacterial methacrylate and methacrylamide monomers and formulate antibacterial fluoride-releasing dental composites. Three antibacterial methacrylate or methacrylamide monomers containing long-chain quaternary ammonium fluoride, 1,2-methacrylamido-N,N,N-trimethyldodecan-1-aminium fluoride (monomer I), N-benzyl-11-(methacryloyloxy)-N,N-dimethylundecan-1-aminium fluoride (monomer II), and methacryloxyldecylpyridinium fluoride (monomer III) have been synthesized and analyzed by nuclear magnetic resonance (NMR) and mass spectrometry (MS). The cytotoxicity test and bactericidal test against Streptococcus mutans indicate that antibacterial monomer II is superior to monomers I and III. A series of dental composites containing 0-6% of antibacterial monomer II have been formulated and tested for degree of conversion (DC), flexure strength, water sorption, solubility, and inhibition of S. mutans biofilms. An antibacterial fluoride-releasing dental composite has also been formulated and tested for flexure strength and fluoride release. The dental composite containing 3% of monomer II has a significant effect against S. mutans biofilm formation without major adverse effects on its physical and mechanical properties. The new antibacterial monomers can be used together with the fluoride-releasing monomers containing a ternary zirconiun-fluoride chelate to formulate a new antibacterial fluoride-releasing dental composite. Such a new dental composite is expected to have higher anticaries efficacy and longer service life.     

Novel nano-particles as fillers for an experimental resin-based restorative material

The purpose of this study is to compare the properties of two experimental materials, nano-material (Nano) and Microhybrid, and two trade products, Clearfil AP-X and Filtek Supreme XT. The flexural strength and modulus after 24h water storage and 5000 thermocycles, water sorption, solubility and X-ray opacity were determined according to ISO 4049. The volumetric behavior (DeltaV) after curing and after water storage was investigated with the Archimedes principle. ANOVA was calculated with p<0.05. Clearfil AP-X showed the highest flexural strength (154+/-14 MPa) and flexural modulus (11,600+/-550 MPa) prior to and after thermocycling (117+/-14 MPa and 13,000+/-300 MPa). The flexural strength of all materials decreased after thermocycling, but the flexural modulus decreased only for Filtek Supreme XT. After thermocycling, there were no significant differences in flexural strength and modulus between Filtek Supreme XT, Microhybrid and Nano. Clearfil AP-X had the lowest water sorption (22+/-1.1 microg mm(-3)) and Nano had the highest water sorption (82+/-2.6 microg mm(-3)) and solubility (27+/-2.9 microg mm(-3)) of all the materials. No significant differences occurred between the solubility of Clearfil AP-X, Filtek Supreme XT and Microhybrid. Microhybrid and Nano provided the highest X-ray opacity. Owing to the lower filler content, Nano showed higher shrinkage than the commercial materials. Nano had the highest expansion after water storage. After thermocycling, Nano performed as well as Filtek Supreme XT for flexural strength, even better for X-ray opacity but significantly worse for flexural modulus, water sorption and solubility. The performances of microhybrids were superior to those of the nano-materials.     

Secretory component is cleaved by neutrophil serine proteinases but its epithelial production is increased by neutrophils through NF-kappa B- and p38 mitogen-activated protein kinase-dependent mechanisms

We previously showed that expression of polymeric immunoglobulin receptor (pIgR)/secretory component (SC), the epithelial receptor assuming transport of polymeric IgA in mucosal secretions, is strongly decreased in severe chronic obstructive pulmonary disease. Here, we evaluated in vitro the effects of polymorphonuclear neutrophil (PMN) mediators on pIgR/SC. On polyacrylamide gel electrophoresis analysis, soluble SC was rapidly cleaved by supernatants from phorbol-myristate-acetate-activated PMN, through a serine proteinase activity. Moreover, purified PMN serine proteinases also cleaved SC. Similarly, polymeric IgA was rapidly cleaved in monomers by neutrophil elastase, whereas secretory immunoglobulin A was relatively resistant to neutrophil elastase. Surface pIgR on human bronchial epithelial cells was also cleaved by serine proteinases, as shown by immunofluorescence. In contrast, pIgR/SC production by cultured epithelial cells (quantified by enzyme-linked immunosorbent assay) was significantly increased by supernatants from interleukin-8/formylmethionylleucylphenylalanine-activated PMN (122.6 +/- 17.3 versus 70.9 +/- 9 ng/mg protein, P < 0.01). Upregulation of pIgR/SC production by bronchial epithelial cells was abolished by nuclear factor kappa B- and p38 mitogen-activated protein kinase (MAPK) inhibitors. Moreover, supernatants from interleukin-8/formylmethionylleucylphenylalanine-activated PMN induced the phosphorylation of I kappa B-alpha and p38 MAPK in epithelial cells, independently of serine proteinases. Thus, PMN serine proteinases cleave pIgR/SC, whereas activated PMN induce an increased pIgR/SC expression through epithelial activation of nuclear factor kappa B and p38 MAPK pathways.     

Dendrimer/methyl methacrylate co-polymers: residual methyl methacrylate and degree of conversion

Dendrimer/methyl methacrylate co-polymers were studied for use in dental composites. The aim was to determine the effects of methyl methacrylate concentration in the resin mixture and polymerization method on the degree of conversion and residual monomer content of the copolymers. Two dendrimers were studied, D12 with 12 reactive methacrylate groups and D24 with 24 reactive groups. The concentration of methyl methacrylate varied from 20 wt% to 50 wt% of monomers. Camphorquinone (CQ) was used as the light-activation initiator and 2-(N,N-dimethylamino)ethyl methacrylate (DMAEMA) as the activator, both in the quantity of 3.0 wt%. Three polymerization methods were used: photo-polymerization, photo-polymerized immediately followed by post-polymerization at 120 degrees C for 15 min, and photo-polymerization followed by postpolymerization after 7 days. The degree of conversion was determined using FT-IR. Residual monomers were extracted with tetrahydrofuran and methanol and analyzed with HPLC. The highest degrees of conversion were 65 and 62%, and the lowest residual monomer contents 1.0 and 1.5% for D12 and D24, respectively. These were measured after heat-induced post-polymerization. For D12, increasing the proportion of methyl methacrylate decreased the degree of conversion and increased the residual monomer content after photo-polymerization. Post-polymerization enhanced the polymerization of the dendrimer co-polymers in respect of degree of conversion and residual monomer content. The present study suggested that the tested dendrimer/methyl methacrylate copolymers require heat-induced polymerization to reach the generally accepted levels of degree of conversion and residual monomers.     

Apparent conversion of adhesive resin in the hybrid layer, Part II: In situ studies of the resin-dentin bond

Penetration and conversion of adhesives into the hybrid layer (HL) is important to the quality and longevity of the adhesive resin (AR)-dentin bond. In this study, a methodology is developed to examine the degree of conversion and relative HEMA concentration with respect to Bis-GMA using Raman spectroscopy. This methodology will be used in the future reports related to this topic. Conversion in the AR of water-stored resin-dentin samples (84% +/- 3%) agreed well with that measured in commercial adhesive (Comm Adh) resin samples after 24-h water storage (80% +/- 2% from Part 1) and was significantly higher than Comm Adh without water storage (58% +/- 3% from Part 1) (p = 0.0005). Adhesive conversion was not significantly different (p = 0.5036) through the middle of the HL, with a mean of 83% +/- 6%. HEMA mole fraction, relative to Bis-GMA, was significantly higher (p = 0.0028) in the top half of the HL (0.67 +/- 0.03), when compared to HEMA in the AR (0.60 +/- 0.01). HEMA and EDMAB were identified through GC/MS as leachable components in the aqueous 24-h storage media. The effect of this elution explains the change in conversion measurements observed between dry and water-stored conditions, which is more appropriately described as the "apparent" conversion.     

Blood plasma and saliva levels of magnesium and other bivalent cations in patients with parotid gland tumors.

The plasma and saliva cations in parotid malignant tumors of stages II-III were studied in 31 patients before surgical therapy and in 27 control group volunteers. The magnesium (t-Mg), calcium (t-Ca), copper (t-Cu) and zinc (t-Zn) concentrations in plasma were determined, and t-Mg and t-Ca in saliva. Our results showed that salivary and plasma t-Mg concentrations were significantly higher in patients with parotid malignant tumors in comparison to control group (saliva: 0.25 +/- 0.04 mmol/L versus 0.14 +/- 0.03/L, p < 0.01; plasma: 1.05 +/- 0.06 mmol/L versus 0.86 +/- 0.05 mmol/L, p < 0.05). The t-Ca plasma concentrations were lower for patients with parotid malignant tumors by 20-22% in comparison to the control group (p < 0.05). Plasma and salivary t-Mg/t-Ca molar ratios are respectively 0.38 and 0.12 for control group, and respectively 0.61 and 0.31 for patients with parotid gland tumors. The t-Zn plasma concentration for patients with parotid malignant tumors (0.017 +/- 0.010 mmol/L) was significantly lower (p < 0.05) in comparison to control group (0.024 +/- 0.011 mmol/L). Plasma t-Cu/t-Zn molar ratio is respectively 0.68 for control group and 1.12 for patients with parotid gland tumors. The mechanism responsible for the increase of salivary magnesium as a consequence of the development of tumoral tissue needs to be clarified.     

The effect of inhaled hexamethonium bromide and atropine sulphate on airway responsiveness to histamine

The degree of protection against inhaled histamine achieved by inhalation of the ganglion blocker hexamethonium bromide plus placebo, hexamethonium plus atropine sulphate, and placebo plus placebo was examined in six atopic subjects, four of whom had current asthma. Hexamethonium was administered until there was systemic evidence of ganglionic blockade with a postural drop in blood pressure of 31 +/- 7.5 mm Hg (mean +/- SD) (p = 0.01) and an increase in heart rate of 30 +/- 3.1 bpm (mean +/- SD) (p = 0.01). Atropine was inhaled in a dose (18 mg nebulized during tidal breathing) known to produce systemic inhibition of cardiac and salivary cholinergic (muscarinic) receptors. The airway effects were measured by FEV1. Hexamethonium caused bronchoconstriction in all four subjects with asthma, which was reversed by atropine. The mean provocation concentration of histamine to provoke a 20% fall in FEV1 was 2.97 mg/ml after premedication with placebo, it was not different at 2.84 mg/ml after hexamethonium alone, and it increased slightly to 5.31 mg/ml after both hexamethonium and atropine (p = 0.06). The results suggest that the main effect of inhaled histamine is not by reflex bronchoconstriction but rather through stimulation of H1-receptors on airway smooth muscle. Therefore, histamine hyperresponsiveness in asthma is not primarily caused by a defect in the parasympathetic nervous supply to the airway.     

Kinetics and hydrolysis mechanism of polymeric prodrugs containing ibuprofen,ketoprofen, and naproxen as pendent agents

Polymeric prodrugs were prepared using methacrylic acid (MA) copolymerization with 2-hydroxyethyl methacrylate (HEMA), covalently linked with ibuprofen (HI), ketoprofen (HK), or naproxen (HN). It was previously shown that the acceptable composition of drug-linked monomer in polymeric prodrugs to prevent gastric mucosa irritation and maintain water solubility was in the range of 20-40 mol%. To investigate the applicability of these polymeric prodrugs, hydrolysis rates of HK-25, HN-29, and HI-30 (the number indicates the mole percent of the drug-linked monomers in the polymeric prodrugs), were studied in vitro with or without esterase. The polymeric prodrugs released a major fraction of the parent drugs and a fraction of the hydroxyethyl ester drug derivatives (drug-EtOH). The calculated hydrolysis rate constants and results correlated to the drug structural solubility and steric hindrance are discussed. The anti-inflammatory properties of these polymeric prodrugs were evaluated using carrageenan-induced edema test. The results indicate that HK-25 and HN-29 display greater potency to inhibit acute inflammatory processes than the free drugs over long periods. HI-30, however, retains a potency comparable to that of free ibuprofen.     

Water sorption,solubility and dimensional changes of denture base polymers reinforced with short glass fibers

The aim of this study was to determine water sorption, solubility and dimensional stability of injection and compression-molded polymethyl methacrylate based denture base polymer that was reinforced with various concentrations and lengths of E-glass fibers. For water sorption and solubility, 20 test groups with different fiber contents and lengths of fibers were prepared. Test specimens without fibers were used as a control. The water sorption and solubility was measured after 90 days water storage. For dimensional stability, rhombic test specimens were prepared and the dimensional changes were measured after processing, drying and storing in water for 4 days and 30 days and were compared with those on the brass model. The water sorption and solubility of injection-molded denture base polymer was lower compared to compression-molded specimens (p < 0.05). The dimensional accuracy of denture base polymer was not affected with fiber reinforcement (p > 0.05).     

HIV antibodies in whole saliva detected by ELISA and western blot assays

Paired serum and saliva samples were tested by enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) for the presence of human immunodeficiency virus (HIV) antibodies. The study group included 36 individuals known to be HIV seropositive and 14 healthy, seronegative controls. HIV antibodies were detected in all but one of the saliva samples of the seropositive subjects. In this particular patient, seroconversion was documented 1 week earlier by sequential testings. A further saliva sample obtained 2 months later was ELISA positive for salivary HIV antibodies. Antibodies against HIV proteins gp120 and gp160 were detected by Western blot assay in all saliva specimens taken from HIV seropositive subjects (including the ELISA-negative patient who seroconverted. Antibodies against other viral proteins (p65, p55, p51, gp41, p35, p24 p18) were found in saliva haphazardly without any clear-cut correlation with the clinical stage of the disease. Pretreatment of the saliva with protease inhibitor was essential for the diagnostic use of saliva for the detection of HIV antibodies by Western blot assay. Calculation of the ratio of titres in serum to those in saliva showed the highest ratios in symptomless subjects (mean +/- SD; 1844 +/- 1412) and the lowest in patients with acquired immune deficiency syndrome (AIDS) (mean +/- SD; 811 +/- 445). The ratio of serum to saliva by ELISA showed a positive correlation with salivary flow rate, indicating a dilution of salivary HIV antibodies with increasing salivary flow rate. The gingival bleeding index was negatively correlated with the ratio, supporting the concept that salivary HIV antibodies transudate from blood to saliva via gingival fluid.     

Mechanical properties of polymeric membranes obtained by radiation cast-polymerization of hydroxyalkyl and hydroxypolyethyleneglycol methacrylate monomers

Polymeric membranes have been prepared by radiation cast-polymerization of hydroxyalkyl and hydroxypolyethyleneglycol methacrylate monomers, and the mechanical properties of the membranes before and after swelling have been studied as a function of the molecular structure of the monomers. The degree of hydration of the polymers from hydroxypolyethyleneglycol methacrylate monomers increased with increasing the number of ethyleneglycol units in the monomers, and that from hydroxyalkyl methacrylate monomers decreased with an increase in the number of methylene units. The mechanical properties such as tensile strength, tear strength, Young's modulus, and elongation at break varied with the length of the methylene and ethyleneglycol units in the monomers. It was found that flexible polymeric membranes having various properties for medical applications could be obtained by radiation cast-polymerization of hydroxyalkyl and hydroxypolyethyleneglycol methacrylate monomers.     

NIR-spectroscopic investigation of water sorption characteristics of dental resins and composites.

A near infrared (NIR) method using the 5200 cm(-1) absorption of water has been employed to examine water absorbed in photopolymerized dental resins and composites in the form of 0.01-cm- to 0.15-cm-thick specimens. The concentration, c [mol L(-1)], of absorbed water in specimens of thickness t [cm] was calculated by means of Beer's law, A = e ct. A is the NIR absorbance and e is the absorptivity of absorbed water. e depends on the environment of the water molecule and it is necessary to estimate e for water in each material. Water sorption was determined gravimetrically and correlated to the absorbance in the NIR spectrum. Once the relationship between e and water content was known for a material, water sorption was determined rapidly on very thin specimens for faster equilibration. Where dissolution of the specimen occurred, the solubility behavior of the specimen was evaluated from a comparison of NIR and gravimetric measurements. The NIR absorptivity, e, of water absorbed in a polymeric medium was found to be inversely related to the degree of hydrophilicity and hydrogen bonding capability of the polymer. The presence of water clusters in a polyethylene oxide methacrylate polymer was inferred from convex-up curvature in the plot of e vs. water content.     

Water sorption, solubility and effect of post-curing of glass fibre reinforced polymers.

Different polymer matrices are used for dental glass fibre composites. The aims of this study were to determine water sorption and solubility of glass fibre composites with different polymer matrices. In addition, the effect of post-curing of matrix polymers with heat on the water sorption and solubility values was investigated. Commercial one-phase and two-phase (powder-liquid) monomer systems were used in polymer matrix of E-glass fibre composite. Rhombic unreinforced and fibre reinforced test specimens were polymerized by autopolymerization or by light only, or additionally post-cured with heat. Water sorption and solubility determination method was based on ISO/DIS 1567-1997 draft for international standard with 7 d immersion time. In addition, water sorption was measured at second time for 30 d immersion time to determine saturation time of test specimens by water. Five test specimens of unreinforced polymer and reinforced polymer were tested and the quantity of fibres was determined by combustion analysis. Water sorption values of different brands of polymer matrices ranged from 0.9 to 8.3 wt% (P < 0.001, ANOVA). High sorption values were explained by microscopic voids in the polymer matrix and by composition of polymer matrix. Solubility values ranged from 0.02 to 2.5 wt% (P < 0.001, ANOVA). Generally, fibre inclusion and post-curing of polymer matrix reduced water sorption and solubility. The results of this study suggest that the water sorption and solubility of fibre composites varies according to the brand of polymer matrix and homogenity of polymer matrix. Water sorption of polymer matrix might influence hydrolytic stability of polymer-glass fibre composite.     

Enhancing hepatocyte adhesion by pulsed plasma deposition and polyethylene glycol coupling

Decreased hepatocyte adhesion to polymeric constructs limits the function of tissue engineered hepatic assist devices. We grafted adhesion peptides (RGD and YIGSR) to polycaprolactone (PCL) and poly-L-lactic acid (PLLA) in order to mimic the in vivo extracellular matrix and thus enhance hepatocyte adhesion. Peptide grafting was done by a novel technique in which polyethylene glycol (PEG)-adhesion peptide was linked to allyl-amine coated on the surface of PCL and PLLA by pulsed plasma deposition (PPD). Peptide grafting density, quantified by radio-iodinated tyrosine in YIGSR, was 158 fmol/cm(2) on PLLA and 425 fmol/cm(2) on PCL surfaces. The adhesion of hepatocytes was determined by plating 250,000 hepatocytes/well (test substrates were coated on 12 well plates) and quantifying the percentage of adhered cells after 6 h by MTT assay. Adhesion on PCL surfaces was significantly enhanced (p < 0.05) by both YIGSR (percentage of adhered cells = 53 +/- 7%) and RGD (53 +/- 12%) when compared to control surfaces (31 +/- 8%). Hepatocyte adhesion on PLLA was significantly (p < 0.05) enhanced on PLLA-PEG-RGD surfaces (76 +/- 14%) compared to control surfaces (42 +/- 19%) and more (68 +/- 25%) but not statistically significant (p = 0.15) on PLLA-PEG-YIGSR surfaces compared to control surfaces. These results indicate that hepatocyte adhesion to PCL and PLLA based polymeric surfaces can be enhanced by a novel adhesion peptide grafting technique using pulsed plasma deposition and PEG cross-linking.     

Assays for total and antigen-specific polymeric IgA in serum based on binding to secretory component

Binding assays with secretory component (SC) were used to detect polymeric IgA antibody to E. coli lipopolysaccharide and to estimate total polymeric IgA in sera from 14 patients with alcoholic liver disease and eight normal controls. Radioiodinated human SC was shown to bind to polymeric IgA and IgM but not to monomeric IgA, secretory IgA or IgG. Serum aliquots (0.5 ml) were totally depleted of IgM using 2 ml anti-IgM affinity columns and the effluent sera were titrated in microtitre plates coated with lipopolysaccharide, the binding of polymeric IgA being detected by adding 10 ng radiolabelled SC. Total polymeric IgA was measured via its capacity to inhibit the binding of 5 ng labelled SC to IgM coated wells, quantitation being achieved by comparison with the inhibition produced by purified polymeric IgA. Total lipopolysaccharide-specific IgA antibody was detected by ELISA in sera from both patients and controls, 1185 +/- 793 and 56 +/- 19 U/100 microliters (mean +/- SD), respectively; but polymeric IgA antibody was detected only in patients' sera (131 +/- 214 U/100 microliters). The concentration of total polymeric IgA was higher in patients' sera than in control sera (488 +/- 333 and less than 120 micrograms/ml respectively).