In Vivo Biodegradation of bisGMA and Urethane-Modified bisGMA-Based Resin Composite Materials.

The aim of this study was to compare the levels of in vivo chemical degradation sustained by bisphenyl-glycidyl-dimethacrylate (bisGMA)-based and urethane-modified bisGMA-based resin composites. A cohort of 58 patients was recruited for the study. Human salivary esterase activity (HSDE) was measured for each patient prior to restoration placement. Class V or III composite restorations without occlusal contacts were placed in adult patients using a 3-step adhesive (Scotchbond MP, 3M) and 1 of 2 resin composites: a traditional bisGMA-based (Z250; 3M) ( n = 28) or a urethane-modified bisGMA-based composite (TPH Spectra, Dentsply) ( n = 30). Patients followed a 2-min rinse (saline containing 20% ethanol) protocol before, immediately after, and 7 days after restoration placement. The rinse samples were analyzed for the presence of bisphenol A (BPA) and bishydroxypropoxyphenylpropane (bisHPPP), a bisGMA breakdown product, using high-performance liquid chromatography in combination with mass spectrometry. The overall mean ± standard error (SE) HSDE activity was 23.4 ± 1.9 U/mL, with no statistical difference between the Z250 (22.6 ± 2.8 U/mL) and TPH (24.1 ± 2.1 U/mL) groups ( P = 0.69). BPA was not detected from any rinse samples. BisHPPP was detected from both composites only in rinse samples immediately after resin composite placement (0.59 µg/mm2 ± 0.16 and 0.68 µg/mm2 ± 0.16 for Z250 and TPH, respectively, P = 0.767). There was no statistically significant correlation between HSDE and amount of bisHPPP obtained from the saliva for the Z250 group ( r = 0.071, P = 0.723), TPH group ( r = 0.266, P = 0.155), and both groups combined ( r = 0.080, P = 0.549). Conventional commercial resin composite materials used in the current study did not release any detectable amount of BPA and only showed detectable levels of bisHPPP for a short term after placement, suggesting that hydrolytic consumption of any available resin substrate is fast and the generated products are rapidly diluted below the detection level limit (<20 ppb) in the oral cavity. This short-term release of bisHPPP was not significantly affected by material type or esterase level in the saliva. Knowledge Transfer Statement: This clinical study demonstrated that the duration and degree of biodegradation of 2 representative formulations of resin composites was limited in both duration and amounts of detectable matrix derived degradation products. No significant level of potential biohazards was released following the application of the resin composites. The results of this study can help oral care professionals address concerns from their patients about possible health issues regarding the application of resin composite restorative materials.

Interaction of a block-co-polymeric biomaterial with immunoglobulin G modulates human monocytes towards a non-inflammatory phenotype.

Monocyte interactions with implanted biomaterials can contribute significantly to the ability of a biomaterial to support tissue integration and wound healing, as opposed to a chronic pro-inflammatory foreign body reaction, provided the materials are designed to do so. However, there are few biomaterials available designed to regulate immune cell response with the intention of reducing the pro-inflammatory activation state. Material chemistry is a powerful tool for regulating protein and cell interactions that can be incorporated into surfaces while maintaining desired mechanical properties. The aspects of material chemistry that can support monocyte activation away from a pro-inflammatory state are still poorly understood. Protein adsorption is a key initial event that transforms the surface of a biomedical device into a biological substrate that will govern subsequent cellular interactions. In this study, the chemistry of degradable block polyurethanes, termed degradable polar hydrophobic ionic (D-PHI) polyurethanes, were studied for their unique interactions with bound immunoglobulin G (IgG), a pro-inflammatory protein that supports monocyte-biomaterial interactions. The specific immunological active sites of the polyurethane-adsorbed protein were compared with IgG's adsorbed state on a homopolymeric material with surface chemistry conducive to cell interactions, e.g. tissue culture polystyrene (TCPS). IgG-coated TCPS supported sustained monocyte adhesion and enhanced monocyte spreading, effects not observed with IgG-coated PU. The degradable PU was subsequently shown to reduce the number of exposed IgG-Fab sites following pre-adsorption vs. IgG adsorbed to TCPS, with antibody inhibition experiments demonstrating that Fab-site exposure appears to dominate monocyte-biomaterial interactions. Minor changes in chemical segments within the PU molecular chains were subsequently investigated for their influence on directing IgG interactions towards reducing pro-inflammatory activity. A reduction in chemical heterogeneity within the PU, without significant differences in other material properties known to regulate monocyte response, was shown to increase Fab exposure and subsequently led to monocyte interactions similar to those observed for IgG-coated TCPS. These results infer that reduced IgG-Fab site exposure can be directed by material chemistry to attenuate pro-inflammatory monocyte interactions with biomaterial surfaces, and identify the chemical features of polymeric biomaterial design responsible for this process. STATEMENT OF SIGNIFICANCE: There is currently limited understanding of material design features that can regulate protein-material interactions in order to prevent adverse inflammatory responses to implanted biomaterials. In this paper, monocyte interactions with biomaterials (specifically a block co-polymeric degradable polyurethane [D-PHI] and tissue culture polystyrene [TCPS]) were investigated as a function of their interactions with adsorbed immunoglobulin G (IgG). D-PHI was shown to attenuate IgG-induced monocyte retention and spreading by reducing IgG-Fab site exposure upon adsorption relative to TCPS. Aspects of D-PHI chemistry important in regulating Fab site exposure were determined. This study thus identifies features of biomaterials, using D-PHI as a case study, which can contribute to the development of new immunomodulatory biomaterial design.

Immunomodulatory polymeric scaffold enhances extracellular matrix production in cell co-cultures under dynamic mechanical stimulation.

Despite the importance of immune cells in regulating the wound healing process following injury, there are few examples of synthetic biomaterials that have the capacity to push the body's immune cells toward pro-regeneration phenotypes, and fewer still that are designed with the intention of achieving this immunomodulatory character. While monocytes and their derived macrophages have been recognized as important contributors to tissue remodeling in vivo, this is primarily believed to be due to their ability to regulate other cell types. The ability of monocytes and macrophages to generate tissue products themselves, however, is currently not well appreciated within the field of tissue regeneration. Furthermore, while monocytes/macrophages are found in remodeling tissue that is subjected to mechanical loading, the effect this biomechanical strain on monocytes/macrophages and their ability to regulate tissue-specific cellular activity has not been understood due to the complexity of the many factors involved in the in vivo setting, hence necessitating the use of controlled in vitro culture platforms to investigate this phenomenon. In this study, human monocytes were co-cultured with human coronary artery smooth muscle cells (VSMCs) on a tubular (3mm ID) degradable polyurethane scaffold, with a unique combination of non-ionic polar, hydrophobic and ionic chemistry (D-PHI). The goal was to determine if such a synthetic matrix could be used in a co-culture system along with dynamic biomechanical stimulus (10% circumferential strain, 1Hz) conditions in order to direct monocytes to enhance tissue generation, and to better comprehend the different ways in which monocytes/macrophages may contribute to new tissue production. Mechanical strain and monocyte co-culture had a complementary and non-mitigating effect on VSMC growth. Co-culture samples demonstrated increased deposition of sulphated glycosaminoglycans (GAGs) and elastin, as well as increases in the release of FGF-2, a growth factor that can stimulate VSMC growth, while dynamic culture supported increases in collagen I and III as well as increased mechanical properties (elastic modulus, tensile strength) vs. static controls. Macrophage polarization toward an M1 state was not promoted by the biomaterial or culture conditions tested. Monocytes/macrophages cultured on D-PHI were also shown to produce vascular extracellular matrix components, including collagen I, collagen III, elastin, and GAGs. This study highlights the use of synthetic biomaterials having immunomodulatory character in order to promote cell and tissue growth when used in tissue engineering strategies, and identifies ECM deposition by monocytes/macrophages as an unexpected source of this new tissue. STATEMENT OF SIGNIFICANCE: The ability of biomaterials to regulate macrophage activation towards a wound healing phenotype has recently been shown to support positive tissue regeneration. However, the ability of immunomodulatory biomaterials to harness monocyte/macrophage activity to support tissue engineering strategies in vitro holds enormous potential that has yet to be investigated. This study used a monocyte co-culture on a degradable polyurethane (D-PHI) to regulate the response of VSMCs in combination with biomechanical strain in a vascular tissue engineering context. Results demonstrate that immunomodulatory biomaterials, such as D-PHI, that support a desirable macrophage activation state can be combined with biomechanical strain to augment vascular tissue production in vitro, in part due to the novel and unexpected contribution of monocytes/macrophages themselves producing vascular ECM proteins.

Monocyte/macrophage cytokine activity regulates vascular smooth muscle cell function within a degradable polyurethane scaffold.

Tissue engineering strategies rely on the ability to promote cell proliferation and migration into porous biomaterial constructs, as well as to support specific phenotypic states of the cells in vitro. The present study investigated the use of released factors from monocytes and their derived macrophages (MDM) and the mechanism by which they regulate vascular smooth muscle cell (VSMC) response in a VSMC-monocyte co-culture system within a porous degradable polyurethane (D-PHI) scaffold. VSMCs cultured in monocyte/MDM-conditioned medium (MCM), generated from the culture of monocytes/MDM on D-PHI scaffolds for up to 28 days, similarly affected VSMC contractile marker expression, growth and three-dimensional migration when compared to direct VSMC-monocyte co-culture. Monocyte chemotactic protein-1 (MCP-1) and interleukin-6 (IL-6) were identified as two cytokines present in MCM, at concentrations that have previously been shown to influence VSMC phenotype. VSMCs cultured alone on D-PHI scaffolds and exposed to MCP-1 (5 ng ml(-1)) or IL-6 (1 ng ml(-1)) for 7 days experienced a suppression in contractile marker expression (with MCP-1 or IL-6) and increased growth (with MCP-1) compared to no cytokine medium supplementation. These effects were also observed in VSMC-monocyte co-culture on D-PHI. Neutralization of IL-6, but not MCP-1, was subsequently shown to decrease VSMC growth and enhance calponin expression for VSMC-monocyte co-cultures on D-PHI scaffolds for 7 days, implying that IL-6 mediates VSMC response in monocyte-VSMC co-cultures. This study highlights the use of monocytes and their derived macrophages in conjunction with immunomodulatory biomaterials, such as D-PHI, as agents for regulating VSMC response, and demonstrates the importance of monocyte/MDM-released factors, such as IL-6 in particular, in this process.

Cariogenic bacteria degrade dental resin composites and adhesives.

A major reason for dental resin composite restoration replacement is related to secondary caries promoted by acid production from bacteria including Streptococcus mutans (S. mutans). We hypothesized that S. mutans has esterase activities that degrade dental resin composites and adhesives. Standardized specimens of resin composite (Z250), total-etch (Scotchbond Multipurpose, SB), and self-etch (Easybond, EB) adhesives were incubated with S. mutans UA159 or uninoculated culture medium (control) for up to 30 days. Quantification of the BisGMA-derived biodegradation by-product, bishydroxy-propoxy-phenyl-propane (BisHPPP), was performed by high-performance liquid chromatography. Surface analysis of the specimens was performed by scanning electron microscopy (SEM). S. mutans was shown to have esterase activities in levels comparable with those found in human saliva. A trend of increasing BisHPPP release throughout the incubation period was observed for all materials and was more elevated in the presence of bacteria vs. control medium for EB and Z250, but not for SB (p < .05). SEM confirmed the increased degradation of all materials with S. mutans UA159 vs. control. S. mutans has esterase activities at levels that degrade resin composites and adhesives; degree of degradation was dependent on the material's chemical formulation. This finding suggests that the resin-dentin interface could be compromised by oral bacteria that contribute to the progression of secondary caries.

Characterization of the annulus fibrosus-vertebral body interface: identification of new structural features.

Current surgical treatments for degenerative intervertebral disc disease do not restore full normal spinal movement. Tissue engineering a functional disc replacement may be one way to circumvent this limitation, but will require an integration of the different tissues making up the disc for this approach to be successful. Hence, an in-depth characterization of the native tissue interfaces, including annulus insertion into bone is necessary, as knowledge of this interface is limited. The objective of this study was to characterize the annulus fibrosus-vertebral bone (AF-VB) interface in immature (6-9 months old) and mature (18-24 months old) bovine discs, as well as to define these structures for normal adult human (22 and 45 years old) discs. Histological assessment showed that collagen fibers in the inner annulus, which are predominantly type II collagen, all appear to insert into the mineralized endplate zone. In contrast, some of the collagen fibers of the outer annulus, predominantly type I collagen, insert into this endplate, while other fibers curve laterally, at an ∼ 90° angle, to the outer aspect of the bone, and merge with the periosteum. This is seen in both human and bovine discs. Where the AF inserts into the calcified zone of the AF-VB interface, it passes through a chondroid region, rich in type II collagen and proteoglycans. Annulus cells (elongated cells that are not surrounded by proteoglycans) are present at this interface. This cartilage zone is evident in both human and bovine discs. Type X collagen and alkaline phosphatase are localized to the interface region. Age-associated differences in bovine spines are observed when examining the interface thickness and the matrix composition of the cartilaginous endplate, as well as the thickness of the mineralized endplate. These findings will assist with the design of the AF-VB interface in the tissue engineered disc.

Platelet Adhesion and Fibrinogen Accretion on a Family of Elastin-Like Polypeptides.

Previous work in our laboratory showed the potential of using a human recombinant elastin-like polypeptide (ELP) as a thromboresistant coating. In this work we investigate the use of three particular ELPs (ELP1, ELP2 and ELP4), that differ by molecular weight and number of repeating hydrophobic and cross-linking domains, as coatings to improve blood-contacting properties. All three ELPs were passively adsorbed on Mylar surfaces. Differences in water contact angle and surface concentration were found among the three ELP coatings, with the shortest polypeptide, ELP1, being the most hydrophilic and abundant on the surface (55°, 0.76 µg/cm(2)), followed by ELP2 (55°, 0.35 µg/cm(2)) and ELP4, the longest of the three (66°, 0.25 µg/cm(2)), respectively. The blood interactions of the ELP coatings were investigated by measuring fibrinogen adsorption and platelet adhesion in whole blood under laminar flow in a cone and plate viscometer configuration. In general, platelet adhesion to the ELP-coated surfaces was found to correlate with fibrinogen adsorption. Decreases in fibrinogen accretion and platelet adhesion were observed for ELP-coated compared to uncoated surfaces. The magnitude of the decreases was found to depend on the ELP sequence length, with ELP4 exhibiting the lowest levels of fibrinogen adsorption and platelet adhesion at 43 ± 24 ng/cm(2) and 113 ± 77 platelets/mm(2), respectively.

Biodegradation of resin-dentin interfaces increases bacterial microleakage.

Bis-GMA-containing resin composites and adhesives undergo biodegradation by human-saliva-derived esterases, yielding Bis-hydroxy-propoxy-phenyl-propane (Bis-HPPP). The hypothesis of this study is that the exposure of dental restorations to saliva-like esterase activities accelerates marginal bacterial microleakage. Resin composites (Scotchbond, Z250, 3M) bonded to human dentin were incubated in either buffer or dual-esterase media (pseudocholinesterase/cholesterol-esterase; PCE+CE), with activity levels simulating those of human saliva, for up to 90 days. Incubation solutions were analyzed for Bis-HPPP by high-performance liquid chromatography. Post-incubation, specimens were suspended in a chemostat-based biofilm fermentor cultivating Streptococcus mutans NG8, a primary species associated with dental caries, for 7 days. Bacterial microleakage was assessed by confocal laser scanning microscopy. Bis-HPPP production and depth and spatial volume of bacterial cell penetration within the interface increased with incubation time and were higher for 30- and 90-day PCE+CE vs. buffer-incubated groups, suggesting that biodegradation can contribute to the formation of recurrent decay.

Effect of phorbol esters on the macrophage-mediated biodegradation of polyurethanes via protein kinase C activation and other pathways.

It was previously found that re-seeding monocyte-derived macrophages (MDM) on polycarbonate-based polyurethanes (PCNUs) in the presence of the protein kinase C (PKC) activator phorbol myristate acetate (PMA) inhibited MDM-mediated degradation of PCNUs synthesized with 1,6-hexane diisocyanate (HDI), as well as esterase activity and monocyte-specific esterase (MSE) protein. However, no effect on the degradation of a 4,4'-methylene bisphenyl (MDI)-derived PCNU (MDI321) occurred. This finding suggested that oxidation, a process linked to the PKC pathway, was not activated in the same manner for all PCNUs. In the current study MDM were re-seeded onto the above PCNU surfaces with PMA, PKC-inactive 4alphaPMA and the PKC inhibitor bisindolylmaleimide I hydrochloride (BIM) for 48 h before assaying for PCNU degradation, esterase activity, MSE protein, DNA, cell viability and cell morphology. 4alphaPMA did not alter MDM-mediated HDI PCNU degradation but MDI321 degradation increased in this condition. BIM alone had no effect on any parameter; however, when BIM and PMA were added together, the PMA inhibition of biodegradation, esterase activity and MSE protein was partially reversed for MDM on HDI PCNUs only. Adding PMA to MDM on HDI PCNUs increased intercellular connections, whereas 4alphaPMA or BIM+PMA increased cell size. Although this study demonstrated a role for oxidation via a PKC-activated pathway in MDM-mediated PCNU degradation, phorbol esters appear to also activate non-PKC pathways that have roles in biodegradation. Moreover, the sensitivity to material surface chemistry in the MDM response to each PCNU dictates a multi-factorial degradative process involving alternate material specific oxidative and hydrolytic mechanisms.

Fibrinogen adsorption and platelet lysis characterization of fluorinated surface-modified polyetherurethanes.

A polyetherurethane (PU) was modified using fluorinated surface-modifying macromolecules (SMMs). A double radiolabel method was used simultaneously to measure the number of adhered platelets ((51)Cr) and the quantity of adsorbed Fg ((125)I), in a cone-and-plate instrument. The objectives were to determine if adsorbed Fg levels correlated to platelet adhesion on the surfaces, and to assess if any reductions in platelet adhesion for the SMM-treated surfaces resulted from surface-induced platelet lysis, rather than changes directly related to lower platelet activation and attachment on the novel surfaces. Platelet lysis was determined from lactate dehydrogenase (LDH) and unbound (51)Cr released into plasma isolated from whole blood exposed to test materials. The corresponding Fg adsorption, evaluated under the same platelet adhesion conditions, did not account for the reduced platelet adhesion on the treated surfaces. LDH and (51)Cr platelet release were very low and indicated no statistically significant differences between the materials. It was therefore concluded that platelet lysis did not contribute to the reduction in platelet adhesion characteristic observed on the SMM-treated surfaces. More importantly, the work emphasizes that the platelet activation cannot be inferred to by assessing the quantity of fibrinogen as is commonly done in the literature. The finding suggests a much more complex mechanism of action for the SMM surface modifiers. On-going work is investigating other Fg parameters such as protein binding affinity and protein conformational state in order to establish the mechanism by which the fluorinated surface modifiers may be reducing platelet adhesion via intermediary changes in initial protein adsorption.

Influence of silanated filler content on the biodegradation of bisGMA/TEGDMA dental composite resins.

It has been shown that an increase in the content of nonsilanated submicron colloidal silica filler particles within dental composites resulted in the release of more bis-phenol-A diglycidyl dimethacrylate (bisGMA)-derived product, bis-hydroxy-propoxyphenyl propane, following incubation with cholesterol esterase (CE). This work further investigates the enzyme-catalyzed biodegradation of fine composite resin systems, containing silanated micron-size irregular glass fillers, commonly used in clinical restorations. Model composite resin samples (10 or 60% weight fraction silanated barium glass filler, 1 mum average particle size) based on bisGMA/triethylene glycol dimethacrylate (TEGDMA) were incubated in buffer or buffer with CE (pH = 7.0, 37 degrees C) solutions for 32 days. The incubation solutions were analyzed using high-performance liquid chromatography, UV spectroscopy, and mass spectrometry. Both groups were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and X-ray photoelectron spectroscopy. In contrast with previous findings for nonsilanated submicron filler systems, the higher filler containing composite showed an increase in its stability with time, following exposure to esterase and when compared to the lower filler content material. As well, the 60% filler composite leached less unreacted monomer TEGDMA. Since the model composite resins studied here were identical and only the filler content varied, the differences in biostability could be specifically associated with the relative amount of resin/filler distribution. The clinical use of different materials in varied dental applications (ranging from fissure sealant to tooth-colored highly filled materials) must consider the potential for different degradation profiles to occur as a function of filler content.

Fibrinogen surface distribution correlates to platelet adhesion pattern on fluorinated surface-modified polyetherurethane.

In previous work, it had been shown that platelet adhesion could be reduced by fluorinating surfaces with oligomeric fluoropolymers, referred to as surface-modifying macromolecules (SMMs). In the current study, two in vitro blood-contacting experiments were carried out on a polyetherurethane modified with three different SMMs in order to determine if altered platelet adhesion levels could be related to the pattern of adsorbed protein and more specifically to the manner in which fibrinogen (Fg) distribution occurs at the surface. In the first experiment, the materials were placed in whole human blood and the adherent platelets were viewed with high-resolution scanning electron microscopy (SEM). In a second experiment, the materials were incubated with human plasma with the absence of platelets. The plasma contained 5% fluorescent-Fg. The materials were then viewed with a fluorescence microscope and images were collected to define the distribution of high-density fluorescent-Fg areas. The SEM and fluorescent-Fg images were imported to Image Pro Plus imaging software to measure the area, length and circularity and a bivariate correlation test was conducted between the two sets of data. For area and length morphology parameters, there were high and significant correlations (r > 0.9, p < 0.05) between the platelets and Fg aggregates. The data suggest that the Fg distribution may serve as a predictor of platelet morphology/activation and provides insight into the non-thrombogenic character of biomaterials containing the fluorinated SMMs.

Understanding the biodegradation of polyurethanes: from classical implants to tissue engineering materials.

After almost half a century of use in the health field, polyurethanes (PUs) remain one of the most popular group of biomaterials applied for medical devices. Their popularity has been sustained as a direct result of their segmented block copolymeric character, which endows them with a wide range of versatility in terms of tailoring their physical properties, blood and tissue compatibility, and more recently their biodegradation character. While they became recognized in the 1970s and 1980s as the blood contacting material of choice in a wide range of cardiovascular devices their application in long-term implants fell under scrutiny with the failure of pacemaker leads and breast implant coatings containing PUs in the late 1980s. During the next decade PUs became extensively researched for their relative sensitivity to biodegradation and the desire to further understand the biological mechanisms for in vivo biodegradation. The advent of molecular biology into mainstream biomedical engineering permitted the probing of molecular pathways leading to the biodegradation of these materials. Knowledge gained throughout the 1990s has not only yielded novel PUs that contribute to the enhancement of biostability for in vivo long-term applications, but has also been translated to form a new class of bioresorbable materials with all the versatility of PUs in terms of physical properties but now with a more integrative nature in terms of biocompatibility. The current review will briefly survey the literature, which initially identified the problem of PU degradation in vivo and the subsequent studies that have led to the field's further understanding of the biological processes mediating the breakdown. An overview of research emerging on PUs sought for use in combination (drug + polymer) products and tissue regeneration applications will then be presented.

Effect of composite resin biodegradation products on oral streptococcal growth.

Hydrolytic degradation by-products associated with the constitutive monomers 2,2-bis [4-(2-hydroxy-3-methacryloxypropoxy) phenyl] propane (bis-GMA), bisphenol A polyethylene glycol diether dimethacrylate (bis-EMA), and triethylene glycol dimethacrylate (TEDGMA) used in dental restorative composites include bis-hydroxy-propoxyphenyl propane (bis-HPPP), ethoxylated bisphenol A (E-bisPA), methacrylic acid (MA), and triethylene glycol (TEG). These products are generated from the interaction of human salivary esterases with the composites. Recent findings have indicated that TEGDMA has the ability to modulate oral bacteria but it is unclear which components of TEGDMA are related to the observed effects. The objective of the current study was to investigate the influence of TEGDMA derived degradation products MA and TEG on the growth of three strains of oral bacteria: S. mutans strains NG8 and JH1005, and S. salivarius AT2. Bacterial growth rates were measured at 37 degrees C, and pH values of 5.5 (representative of cariogenic state) or 7.0 at concentrations of 0-50mmol/l for MA (Sigma, US) and 0-100mmol/l for TEG (Sigma, US). It was found that at pH 5.5 TEG significantly stimulated the growth of both S. mutans strains ( p<0.05 ) in the concentration range of 0.5-10.0mmol/l and stimulated the growth of S. salivarius AT2 for the entire concentration range tested (p<0.05). TEG (above 50mmol/1) did not significantly affect the doubling times of S. salivarius at pH of 7.0 and it inhibited the growth of both S. mutans strains above 50mmol/l at the same pH value. At pH 5.5 MA inhibited the growth of all three strains with increasing concentration. At neutral pH, the growth of S. mutans NG8 strain was significantly reduced by MA ( p<0.05 ) above 10mmol/l. In summary, these results indicate that TEG and MA modulate the growth rate of important oral bacteria in a concentration and pH dependent manner.

The influence of resin chemistry on a dental composite's biodegradation.

Previous work reported that commercial dental composite resins containing a urethane-modified bisGMA (bisphenylglycidyl dimethacrylate)/TEGDMA (triethylene glycol dimethacrylate) (ubis) based monomer system showed a 10-fold reduction in the release of a bisGMA-derived product, bishydroxypropoxyphenyl propane (bisHPPP), as compared with that found for bisGMA/TEGDMA (bis) based composites after incubation with cholesterol esterase (CE). Unfortunately, these materials also differed substantially in filler type and content, making it impossible to directly relate any specific parameter to the differences in biodegradation levels. By controlling for filler content and type, the current study will seek to probe the biomolecular interactions between composite resin chemistry and esterase activity in order to help explain the observed differences in biodegradation levels between the ubis and bis resin systems. After 32 days of incubation, buffer and CE solutions were analyzed for degradation products using high-performance liquid chromatography, UV spectroscopy, and mass spectrometry. Both materials were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and X-ray photoelectron spectroscopy. In the CE groups, the ubis system showed a 2.6- to 86-fold reduction (dependent on the product) in the amount of isolated products relative to the bis system (p < 0.01). Scanning electron microscopy data also demonstrated the relative stability of the ubis system and X-ray photoelectron spectroscopy analysis showed a higher content of the ester bond at the surface of the bis samples. Fourier transform infrared data showed that both resins had similar conversions. Because both systems were identical except for their monomer systems, it was concluded that changes in biostability were associated with chemistry. Crosslinking, hydrophobicity, and solubility all relate to ubis's pro-stability.

Mutual influence of cholesterol esterase and pseudocholinesterase on the biodegradation of dental composites.

It has been demonstrated that human saliva contains cholesterol esterase (CE)- and pseudocholinesterase (PCE)-like hydrolase activities. While PCE has been shown to preferentially degrade triethylene glycol dimethacrylate (TEGDMA) and its derivatives, CE has a greater catalytic effect on the breakdown of bis-phenol-A-diglycidyl dimethacrylate (bisGMA) components in composite dental resins. The current study seeks to determine if there is a mutual influence between the different esterases with respect to the biodegradation of resin composite. Photopolymerized model composite resin samples (containing 60% by weight fraction of silanated barium glass filler) based on bisGMA/TEGDMA (bis) or urethane-modified bisGMA/TEGDMA/bisEMA (ubis) monomers were incubated in buffer, CE and/or PCE solutions (pH=7.0, 37 degrees C) for 8 and 16 days. The incubation solutions were analyzed for degradation products using high-performance liquid chromatography, UV spectroscopy and mass spectrometry. In the bis system, higher amounts (p<0.05) of a bisGMA derived product, bishydroxy-propoxyphenyl-propane (bisHPPP), were detected in the combined enzyme group as compared to the sum of the two individual enzyme groups. In the ubis system, similar comparisons showed that higher levels (p<0.05) of bisHPPP were detected in the combined group at 8 days while higher amounts (p<0.05) of a bisEMA derived product, ethoxylated bis-phenol A, were detected in the combined group at 16 days. The study concluded that CE and PCE act synergistically to increase the biodegradation of both composite resin materials.

Modulation of collagen proteolysis by chemical modification of amino acid side-chains in acellularized arteries.

In this study, we have examined the effects of specific chemical modifications of amino acid side-chains on the in vitro degradation of "native" collagen obtained from acellular matrix (ACM)-processed arteries. Two monofunctional epoxides of different size and chemistry were used to modify lysine, with or without methylglyoxal modification of arginine. Biochemical, thermomechanical, tensile mechanical, and multi-enzymatic (collagenase, cathepsin B, acetyltrypsin, and trypsin) degradation analyses were used to determine the effects of modifications.Collagen solubilization by enzymes was found to depend upon the size and chemistry of epoxides used to modify lysine residues. In general, the solubilization of native ACM collagen by collagenase, cathepsin B, trypsin, and acetyltrypsin was either unaltered or decreased after modification with glycidol. In contrast, n-butylglycidylether (n-B) treatment increased solubilization by all enzymes. Subsequent arginine modification significantly reduced collagen solubilization by acetyltrypsin for glycidol-treated ACM arteries, whereas increased collagen solubilization was observed for n-B-treated ACM arteries with all enzymes. Gel chromatographic analyses of collagen fragments solubilized by trypsin revealed that both the amount and sites of cleavage were altered after lysine and arginine modification. The ability to modulate the enzymatic degradation of tissue-derived materials as demonstrated in this study may facilitate the design of novel engineering scaffolds for tissue regeneration or collagen-based drug delivery systems.

Salivary esterase activity and its association with the biodegradation of dental composites.

Pseudocholinesterase (PCE) and cholesterol esterase (CE) can hydrolyze bisphenylglycidyl dimethacrylate (bisGMA) and triethylene glycol dimethacrylate (TEGDMA) monomers. This study will test the hypothesis that enzyme activities showing CE and PCE character are found in human saliva at levels sufficient to hydrolyze ester-containing composites important to restorative denstistry. The study also seeks to ask if the active sites of CE and PCE with respect to composite could be inhibited. Photo-polymerized model composite resin was incubated in PCE and CE solutions, in the presence and absence of a specific esterase inhibitor, phenylmethylsulfonyl fluoride (PMSF). Incubation solutions were analyzed for resin degradation products by high-performance liquid chromatography (HPLC), UV spectroscopy, and mass spectrometry. Saliva was found to contain both hydrolase activities at levels that could degrade composite resins. PMSF inhibited the composite degradation, indicating a material hydrolysis mechanism similar to the enzymes' common function.

Biodegradation of a dental composite by esterases: dependence on enzyme concentration and specificity.

Studies have shown that inflammatory (cholesterol esterase, CE) and salivary (pseudo-cholinesterase, PCE) enzymes can cause the breakdown of bisphenol-A diglycidyl dimethacrylate (bisGMA) and triethylene glycol dimethacrylate (TEGDMA) components from composite resins. Based on the above consideration, it was desired to show how CE- and PCE-catalyzed hydrolysis of resin components was dependent on the enzymes' concentration and to determine their distinct specificities (if any) towards resin components. Photopolymerized model composite resin samples (60% weight fraction silanated barium glass filler) based on bisGMA and TEGDMA monomers (55/45 weight ratio of the matrix, respectively) were incubated with PBS and either 0.01, 0.05, 0.1 or 1 unit/ml of CE or PCE for 16 days (pH 7.0, 37 degrees C). Incubation solutions were analyzed by high-performance liquid chromatography (HPLC), UV spectroscopy and mass spectrometry. The composite samples were characterized by scanning electron microscopy (SEM). Degradation rates of bisGMA and TEGDMA monomers were assessed. The results showed that CE had a greater specificity towards cleaving bisGMA while PCE showed a greater specificity towards TEGDMA. A strong enzyme concentration dependence was observed which suggests that the level of degradation products generated for a material will depend on the esterase make-up of an individual's saliva in combination with the specific formulation of monomer components used.

Isolation of methylene dianiline and aqueous-soluble biodegradation products from polycarbonate-polyurethanes.

Polycarbonate-polyurethanes (PCNUs) have provided the medical device industry with practical alternatives to oxidation-sensitive polyether-urethanes (PEUs). To date, many studies have focused on PCNUs synthesized with 4,4'-methylene diphenyl-diisocyanate (MDI). The relative hydrolytic stability of this class of polyurethanes is actually quite surprising given the inherent hydrolytic potential of the aliphatic carbonate group. Yet, there has been little information reporting on the rationale for the material's demonstrated hydrolytic stability. Recent work has shown that PCNU materials have a strong sensitivity towards hydrolysis when changes are made to their hard segment content and/or chemistry. However, knowledge is specifically lacking in regards of the identification of cleavage sites and the specific nature of the biodegradation products. Using high-performance liquid chromatography, radiolabel tracers and mass spectrometry, the current study provides insight into the distribution of biodegradation products from the enzyme-catalyzed hydrolysis of five different PCNUs. The hydrolytic sensitivity of the materials is shown to be related to the distribution of products, which itself is a direct consequence of unique micro-structures formed within the different materials. While an MDI-based polymer was shown to be the most hydrolytically stable material, it was the only PCNU that produced its diamine analog, in this case 4,4'-methylene dianiline (MDA), as a degradation product. Given the concern over aromatic diamine toxicity, this finding is important and highlights the fact that relative biostability is a distinct issue from that of degradation product toxicity, and that both must be considered separately when assessing the impact of biodegradation on biomaterial in vivo compatibility.