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Informatics challenges in tissue engineering and biomaterials
Rekow, D. (2003), Adv Dent Res 17: 49-54.
Abstract: Both tissue engineering and biomaterials have made tremendous strides recently, yet major questions remain unanswered. Tissue-engineered products have come to the market; others are in development. A fundamental issue that informatics could address for tissue engineering is to describe and to predict the cascade of biochemical and cellular reactions that occur as a function of time and implant material: surface texture, microporosity; pore size, density, and connectivity; and three-dimensional configuration. Behavior of ceramics, a subset of tissue-engineering scaffold materials and a mainstay of dental restorations, has been studied extensively for very thin layers and for thicknesses greater than 2 mm. Until recently, little has been known about dentally relevant thickness of 1-2 mm. Results have been surprising and are continuing to develop. Still, at least one fundamental question remains that could be addressed by informatics techniques: Where, along the spectrum of flat-polished material to 10-year clinical in vivo study, can we test to predict clinical performance of all-ceramic crowns accurately?

Infrainguinal ePTFE vascular graft with bioactive surface heparin bonding. First clinical results
Walluscheck, K. P., S. Bierkandt, et al. (2005), J Cardiovasc Surg (Torino) 46(4): 425-30.
Abstract: AIM: The ultimate aim of improved expanded polytetrafluoroethylene (ePTFE) vascular graft design is to achieve patency rates in femoropopliteal bypass grafting comparable with autologous saphenous vein grafts. Enhanced thromboresistance of the ePTFE material by bioactive surface heparin bonding is one conceivable path toward this goal. This retrospective study was performed to collect the first clinical data for a new ePTFE graft with bioactive surface heparin immobilization. METHODS: Between March 2003 and February 2004, 43 femoropopliteal or femorocrural ePTFE vascular prostheses with bioactive end-point immobilized heparin (Gore-Tex Propaten Vascular Graft), using the Car-meda BioActive Surface technology, were implanted in 40 patients. Twelve prostheses were implanted in above-knee and 31 in below-knee position. The indication for bypass grafting was limb-threatening ischemia in 88% of the patients. The mean follow-up was 16.6 months. RESULTS: The primary 1-year patency was 91% for above-knee bypass grafts and 92% for below-knee bypass grafts. The 2-year primary patency rate for above-knee bypass grafts was 68% and 81% for below-knee bypass grafts. Limb salvage was achieved in 98%. The perioperative mortality was 0%, but during follow-up 22% of the patients died with patent bypass grafts. CONCLUSIONS: While conventional ePTFE grafts, particularly in the below-knee position, have shown poor results even in the short-term, the bioactive heparinized ePTFE graft evaluated in this study provides patency rates comparable with autologous vein grafts. Because the influence of luminal heparin bonding is not only limited to thromboresistance but has impact on, amongst other elements, protein adsorption (thereby improving hemocompatibility), a continuous effect for long-term patency could also be expected.

Infrared laser light reduces loading time of dental implants: a Raman spectroscopic study
Lopes, C. B., A. L. Pinheiro, et al. (2005), Photomed Laser Surg 23(1): 27-31.
Abstract: OBJECTIVE: The aim of this study was to assess, through near-infrared Raman spectroscopy (NIRS), the incorporation of hydroxyapatite of calcium (CHA; approximately 960 cm(1))--on the healing bone around dental implants submitted or not to low-level laser therapy (LLLT) (lambda830 nm). BACKGROUND DATA: The process of maturation of the bone is important for the success of dental implants, as it improves the fixation of the implant to the bone, allowing the wearing of a prosthesis. LLLT has been suggested as a mean of improving bone healing because of its biomodulatory capabilities. METHODS: Fourteen rabbits received a titanium implant on the tibia; eight of them were irradiated with lambda830-nm laser (seven sessions at 48-h intervals, 21.5 J/cm(2) per session, 10 mW, phi approximately 0.0028 cm(2), 85 J/cm(2) treatment dose), and six acted as control. The animals were sacrificed at 15, 30, and 45 days after surgery. Specimens were routinely prepared for Raman spectroscopy. Twelve readings were taken on the bone around the implant. RESULTS: The results showed significant differences in the concentration of CHA on irradiated and control specimens at both 30 and 45 days after surgery (p < 0.001). Conclusion: It is concluded that LLLT does improve bone healing, and this can be safely assessed by Raman spectroscopy.

Infrared optical properties and AFM of spin-cast chitosan films chemically modified with 1,2 Epoxy-3-phenoxy-propane
Nosal, W. H., D. W. Thompson, et al. (2005), Colloids Surf B Biointerfaces 46(1): 26-31.
Abstract: Chemical modification of spin-cast chitosan films has been performed. This modification involves the attachment of 1,2 Epoxy-3-phenoxy-propane, commonly known as glycidyl phenyl ether (GPE), to the amine group of the chitosan molecule. Optical properties of modified films have been determined in the infrared region of the spectrum using spectroscopic ellipsometry, and are reported in this paper. Special attention is paid to the infrared region where the index of refraction and extinction coefficients from 750 to 4000 cm(-1) were determined. Difference plots of IR optical data before and after chemical modification were generated to confirm that modification had occurred. Optical modeling of infrared spectroscopic ellipsometry (IRSE) data with respect to chemical bond vibrations has also been performed. This modeling involved curve fitting of resonant chemical bond absorptions using Lorentz oscillators. These oscillator models allow for comparison of modified chitosan to unmodified chitosan. The purpose of this research was to determine infrared optical constants of chemically modified chitosan films This work shows that surface chemistry of biomaterials can be studied quite sensitively with spectroscopy ellipsometry, detecting as little as 100 ng/cm(2) of GPE.

Inhibition assay of biomolecules based on fluorescence resonance energy transfer (FRET) between quantum dots and gold nanoparticles
Oh, E., M. Y. Hong, et al. (2005), J Am Chem Soc 127(10): 3270-1.

Inhibition by heparin and derivatized dextrans of Staphylococcus aureus adhesion to fibronectin-coated biomaterials
Vaudaux, P., T. Avramoglou, et al. (1992), J Biomater Sci Polym Ed 4(2): 89-97.
Abstract: Recent data on cardiovascular device-centered infections suggest that some plasma and extracellular matrix proteins contribute to bacterial adhesion and colonization on biomaterials. We previously developed an in vitro assay to study the Staphylococcus aureus adhesion-promoting effect of surface-adsorbed fibronectin on flat PMMA coverslips coated with a monolayer amount of fibronectin. We screened the potential anti-adhesive properties of a group of substituted dextrans, previously shown to exhibit potent anticoagulant and anticomplementary activities. In comparison to unsubstituted dextran which showed no significant (< 20%) adhesion inhibition at 1 mg/ml, dextrans increasingly substituted with carboxylic and benzylamide groups (CMBD) exhibited increasing anti-adhesive activities. Three CMBD derivatives showing an increasing proportion (5-14%) of benzylamide groups showed inhibition of bacterial adhesion increasing from 33 to 51% at 1 mg/ml. Another category of substituted dextrans having a variable proportion (2-26%) of sulfonated benzylamide groups (CMBDS) produced active inhibition of S. aureus adhesion. In comparison to these heparin-like dextran derivatives, native heparin produced inhibition values of S. aureus adhesion which were intermediate between those of CMBD and CMBDS compounds. Furthermore, the anti-adhesive activity was still expressed when substituted dextrans were preincubated with fibronectin-coated PMMA but washed away at the time when radiolabeled bacteria were added to the adhesion assay. This indicates that the anti-adhesive effects of CMBDS could be exerted at the level of the S. aureus binding site of fibronectin. In conclusion, S. aureus adhesion on fibronectin-coated biomaterials can be efficiently blocked in vitro by soluble compounds such as dextran derivatives.

Inhibition of complement activation by soluble recombinant CR1 under conditions resembling those in a cardiopulmonary circuit: reduced up-regulation of CD11b and complete abrogation of binding of PMNs to the biomaterial surface
Larsson, R., G. Elgue, et al. (1997), Immunopharmacology 38(1-2): 119-27.
Abstract: The influence of soluble recombinant CR1 (sCR1) on complement activation, and its indirect effects on the coagulation system and cellular responses were assessed in two models for the study of blood/surface and blood/air interactions, as are encountered in e.g. cardiopulmonary bypass circuits. The concentrations of C3a and sC5b-9 and the amount of bound C3/C3 fragments were analyzed as indicators of complement activation. Thrombin-antithrombin complexes, the platelet count, surface-ATP, beta-thromboglobulin, and the expression of CD11b on leukocytes were the parameters analyzed to reflect coagulation and cellular responses. In addition, immunochemical analyses of the phenotypes of surface-bound leukocytes and platelets were performed. Recombinant sCR1, at doses ranging between 0.1-0.25 mg/ml, was found to completely inhibit the generation of sC5b-9, and of C3a by two thirds; the binding of C3 and/or C3 fragments to the surface was almost entirely abolished. As a result of the inhibition of complement activation, the expression of CD11b on PMNs, and the binding of these cells to the biomaterial surface was almost completely lost. In contrast, the thrombin-antithrombin complexes, the platelet count, and the adherence of platelets to the surface, as reflected by the ATP binding and the release of beta-thromboglobulin, were not affected. These data show that complement activation, in association with extra-corporeal treatment, causes activation and binding of PMNs to the biomaterial and that these effects can be completely abolished by the addition of soluble recombinant sCR1.

Inhibition of compound action potentials following administration of dental self-etching primer/adhesives
Onur, M. A., Z. C. Cehreli, et al. (2005), J Biomater Sci Polym Ed 16(4): 411-9.
Abstract: Self-etching dentin adhesive systems, a new generation of dental-bonding agents, were investigated for their ability to block nerve conductance in the rat sciatic nerve model. Clearfil SE Bond, Mac Bond and FL Bond were used as test materials. Isolated rat sciatic nerves obtained from female albino rats were placed between two electrodes in a bath containing Tyrode solution. The bonding agents were brought into contact with the nerves and the evoked compound action potentials were recorded versus time before and after contact with the materials. Statistical analysis was performed using the Friedman and Wilcoxon Signed-Ranks tests (P = 0.01) to compare the mean percentage of inhibition between test materials versus time. All test materials were able to block nerve transmission rapidly (3.3 +/- 0.5, 5.1 +/- 2.3 and 8.0 +/- 1.9 min for SE Bond, Mac Bond and FL Bond, respectively). Significant differences were found between FL Bond and SE and Mac Bond (P < 0.01). These results indicated that the bonded systems tested, intended primarily as dentinal tubule occluders, may also exert some direct desensitizing effect on intratubular or pulpal nerve terminals in extremely deep cavities or pulpal microexposures, contributing to decreased postoperative pain.

Inhibition of gene expression in vivo using multiplex siRNA
Chae, S. S. and T. Hla (2005), Methods Mol Biol 309: 197-203.

Inhibition of implant-associated infections via nitric oxide release
Nablo, B. J., H. L. Prichard, et al. (2005), Biomaterials 26(34): 6984-90.
Abstract: The in vivo antibacterial activity of nitric oxide (NO)-releasing xerogel coatings was evaluated against an aggressive subcutaneous Staphylococcus aureus infection in a rat model. The NO-releasing implants were created by coating a medical-grade silicone elastomer with a sol-gel-derived (xerogel) film capable of storing NO. Four of the bare or xerogel-coated silicone materials were subcutaneously implanted into male rats. Ten rats were administered 10 microl of a 10(8) cfuml(-1)S. aureus colony directly into the subcutaneous pocket with the implant prior to wound closure. Infection was quantitatively and qualitatively evaluated after 8d of implantation with microbiological and histological methods, respectively. A 82% reduction in the number of infected implants was achieved with the NO-releasing coating. Histology revealed that the capsule formation around infected bare silicone rubber controls was immunoactive and that a biofilm may have formed. Capsule formation in response to NO-releasing implants had greater vascularity in comparison with uninoculated or untreated controls. These results suggest that NO-releasing coatings may dramatically reduce the incidence of biomaterial-associated infection.

Inhibition of matrix metalloproteinase-9 by interferons and TGF-beta1 through distinct signalings accounts for reduced monocyte invasiveness
Nguyen, J., P. Knapnougel, et al. (2005), FEBS Lett 579(25): 5487-93.
Abstract: Cytokines may provide signals for regulating human monocyte matrix metalloproteinase-9 (MMP-9) activity. In this study, we investigated the roles of interferons (IFN) type I/II and transforming growth factor-beta1 (TGF-beta1) in MMP-9-mediated invasiveness. MMP-9 antibody and inhibitor, IFNs and TGF-beta1 inhibited monocyte transmigration through Matrigel. IFNs and TGF-beta1 downregulated MMP-9 mRNA, protein and activity levels. The inhibitory action of IFNs was associated with the STAT1/IRF-1 pathway since the JAK inhibitor AG490 blocked STAT1 phosphorylation, IRF-1 synthesis and counteracted the blockade of MMP-9 release. TGF-beta1-mediated MMP-9 inhibition appeared STAT1/IRF-1-independent but reversed by the protein tyrosine kinase inhibitor tyrphostin 25. Our data point out the importance of IFNs and TGF-beta1 in the control of monocyte MMP-9-mediated extravasation.

Inhibition of MDA-MB-231 cell cycle progression and cell proliferation by C-2-substituted oestradiol mono- and bis-3-O-sulphamates
Raobaikady, B., M. J. Reed, et al. (2005), Int J Cancer 117(1): 150-9.
Abstract: A natural metabolite of oestradiol (E2), 2-methoxyoestradiol (2-MeOE2), exerts both antitumour and antiangiogenic effects. 2-MeOE2 is currently in clinical trials for the treatment of a variety of cancers. We have previously shown that a number of sulphamoylated analogues of 2-MeOE2 possess enhanced potency and bioavailability with respect to 2-MeOE2. In our study, the effects of C-2-substituted E2 derivatives, with sulphamoylation at the C-3 and/or C-17 position, on ERalpha -ve MDA-MB-231 breast cancer cells were evaluated. Sulphamoylated derivatives were potent inhibitors of cell proliferation, and these effects were irreversible when compared to growth inhibitory effects induced by 2-MeOE2. Cell cycle analysis suggested that these derivatives caused cells to arrest at the G2-M phase of the cell cycle. Sulphamoylated analogues suppressed the clonogenic potential of MDA-MB-231 cells and also their growth on Matrigel culture substratum. Immunofluorescence studies showed fragmented nuclear bodies and an abnormal microtubule cytoskeleton in cells exposed to one of the potent compounds, 2-MeOE2-bis-sulphamate. In addition, these analogues induced phosphorylation of BCL-2, a protein considered to be the guardian of microtubule integrity. In each of the assays, the sulphamoylated derivatives were at least 10-fold more potent than the parent compound 2-MeOE2. In view of the enhanced potencies associated with sulphamoylated E2 derivatives in ERalpha -ve cells, these analogues should hold considerable therapeutic potential for the treatment of hormone-independent breast cancers.

Inhibition of Naja naja venom hyaluronidase by plant-derived bioactive components and polysaccharides
Girish, K. S. and K. Kemparaju (2005), Biochemistry (Mosc) 70(8): 948-52.
Abstract: The inhibitory effect of several bioactive compounds on the activity of hyaluronidase enzyme purified from Naja naja venom was investigated in vitro. Compounds were found to inhibit the hyaluronidase activity dose dependently. Among glycosaminoglycans, heparin, heparan sulfate, and dermatan sulfate showed maximum inhibition compared to chondroitin sulfates. Different molecular forms of chitosan inhibit the enzyme, and inhibition appears to depend on the chain length. In addition, plant-derived bioactive compounds also inhibited the activity of hyaluronidase dose dependently. Among those tested, aristolochic acid, indomethacin, quercetin, curcumin, tannic acid, and flavone exhibited inhibition, with aristolochic acid and quercetin completely inhibiting the enzyme activity. It is concluded that the inhibitors of hyaluronidase could be used as potent first aid agents in snakebite therapy. Furthermore, these inhibitors not only reduce the local tissue damage but also retard the easy diffusion of systemic toxins and hence increase survival time.

Inhibition of Staphylococcus adherence to biomaterials by extracellular slime of S. epidermidis RP12
Giridhar, G., A. S. Kreger, et al. (1994), J Biomed Mater Res 28(11): 1289-94.
Abstract: Adherence of selected strains of coagulase-negative staphylococci to various biomaterials, and the inhibition of their adherence by extracellular slime obtained from the RP12 strain of Staphylococcus epidermidis were studied in vitro. S. epidermidis RP12 adhered considerably more to polymethylmethacrylate (PMMA) discs than did the SP2 strain of S. hominis and the SE-360 strain of S. hyicus. Strain RP12 was less adherent to titanium alloy, ultrahigh molecular weight polyethylene (UHMWPE), and Teflon discs than to PMMA discs. Exposure of PMMA discs to extracellular slime extracted from strain RP12 greatly reduced adherence of strain RP12, SP2, SE-360, and S. epidermidis RP-62A. The active component(s) was present in the > 10 kD mol wt fraction obtained by Amicon YM10 ultrafiltration of crude slime; heat treatment of the fraction did not affect its inhibitory activity. When the bacteria and RP12 slime fractions were added simultaneously to the PMMA discs, the > 10 kD mol wt fraction of slime competitively inhibited adherence of strain RP12 to PMMA discs; in contrast, the < 10 kD mol wt fraction enhanced adherence of strain RP12 to PMMA discs.

Inhibition of the phosphatidylinositol 3-kinase/Akt pathway by inositol pentakisphosphate results in antiangiogenic and antitumor effects
Maffucci, T., E. Piccolo, et al. (2005), Cancer Res 65(18): 8339-49.
Abstract: The purpose of this study was to investigate the antiangiogenic and in vivo properties of the recently identified phosphatidylinositol 3-kinase (PI3K)/Akt inhibitor Inositol(1,3,4,5,6) pentakisphosphate [Ins(1,3,4,5,6)P5]. Because activation of the PI3K/Akt pathway is a crucial step in some of the events leading to angiogenesis, the effect of Ins(1,3,4,5,6)P5 on basic fibroblast growth factor (FGF-2)-induced Akt phosphorylation, cell survival, motility, and tubulogenesis in vitro was tested in human umbilical vein endothelial cells (HUVEC). The effect of Ins(1,3,4,5,6)P5 on FGF-2-induced angiogenesis in vivo was evaluated using s.c. implanted Matrigel in mice. In addition, the effect of Ins(1,3,4,5,6)P5 on growth of ovarian carcinoma SKOV-3 xenograft was tested. Here, we show that FGF-2 induces Akt phosphorylation in HUVEC resulting in antiapoptotic effect in serum-deprived cells and increase in cellular motility. Ins(1,3,4,5,6)P5 blocks FGF-2-mediated Akt phosphorylation and inhibits both survival and migration in HUVEC. Moreover, Ins(1,3,4,5,6)P5 inhibits the FGF-2-mediated capillary tube formation of HUVEC plated on Matrigel and the FGF-2-induced angiogenic reaction in BALB/c mice. Finally, Ins(1,3,4,5,6)P5 blocks the s.c. growth of SKOV-3 xenografted in nude mice to the same extent than cisplatin and it completely inhibits Akt phosphorylation in vivo. These data definitively identify the Akt inhibitor Ins(1,3,4,5,6)P5 as a specific antiangiogenic and antitumor factor. Inappropriate activation of the PI3K/Akt pathway has been linked to the development of several diseases, including cancer, making this pathway an attractive target for therapeutic strategies. In this respect, Ins(1,3,4,5,6)P5, a water-soluble, natural compound with specific proapoptotic and antiangiogenic properties, might result in successful anticancer therapeutic strategies.

Inhibition of the tissue reaction to a biodegradable biomaterial by monoclonal antibodies to IFN-gamma
Khouw, I. M., P. B. van Wachem, et al. (1998), J Biomed Mater Res 41(2): 202-10.
Abstract: Biomaterials are increasingly used for clinical applications. However, loss of function may occur owing to tissue reactions, which are mainly caused by a variety of inflammatory reactions. Recently, we demonstrated that macrophages (MO) and T cells play key roles in these reactions. Since immunological studies showed that the T cell-derived cytokine interferon-gamma (IFN-gamma) activates MO, the aim of this study was to investigate the possibility of modulating tissue reactions to biodegradable biomaterials by inactivating IFN-gamma. Dermal sheep collagen (DSC) was used as a test biomaterial. DSC impregnated with anti-IFN-gamma or phosphate-buffered saline (control) was implanted in rats. The results showed that cellular ingrowth and formation and function of giant cells were strongly delayed by anti-IFN-gamma. Also, MHC class II expression was strongly inhibited. In the treated DSC, some huge giant cells were formed at the interface but association with the DSC bundles did not occur. Finally, in both the control and treated DSC, T cells and NK cells were rarely detected. This study demonstrates that IFN-gamma plays an important role in the inflammatory reaction to biomaterials. This reaction can be modulated by anti-IFN-gamma, which warrants further studies of anti-IFN-gamma for clinical application to prevent unwanted tissue reactions to biomaterials.

Inhibitor of complement, Compstatin, prevents polymer-mediated Mac-1 up-regulation of human neutrophils independent of biomaterial type tested
Schmidt, S., G. Haase, et al. (2003), J Biomed Mater Res A 66(3): 491-9.
Abstract: The inflammatory reaction after cell contact with polymer materials is primarily mediated by activated neutrophils and may, in some cases, lead to exhaustion of neutrophil cell function. A direct consequence of this can be impairment of local or even systemic host defense mechanisms, which in turn can result in foreign body infections. Neutrophil activation, as indicated by the up-regulation of the Mac-1 adhesion receptor, is a reliable parameter for estimating the inflammatory risk due to implanted biomaterials. Because at blood contact, biomaterials immediately acquire a material-specific layer of blood proteins on their surface, including fibrinogen, complement, and immunoglobulin G, it is generally believed that after biomaterial contact, neutrophil activation primarily occurs by interaction with this protein layer. In this study, using our recently established polymer bead in vitro assay, we investigated whether complement inhibition alone can reduce biomaterial-mediated neutrophil activation, independent of the type of polymer and, hence, also its surface chemistry. Complement inhibition was achieved by using Compstatin, a recently developed complement inhibitor that binds to the complement component C3 preventing C3 convertase formation. We revealed significantly reduced (p < or = 0.025) Mac-1 receptor expression levels after 45 min of blood contact with the following polymers (without and with Compstatin): 1. polyurethane, 98.3%, 13.6%; 2. polymethylmetacrylate, 88.5%, 11.0%; and poly-D,L-lactide, 71.8%, 8.4%. Although these three polymer types acquire material-specific protein layers because of their different surface chemistry, complement inhibition by Compstatin alone proved to be sufficient to reduce neutrophil activation after surface contact, thus reducing the risk of biomaterial-mediated inflammatory reaction.

Inhibitory effects on esterase enzymes buche and ache in rainbow trout (Oncorhyncus mykiss) produced by the slow release insecticide chitosan diethyl phosphate
Placencia, J., A. Rudolph, et al. (2005), J Environ Sci Health B 40(5): 761-8.
Abstract: The study on the toxicity of chitosan diethyl phosphate (ChDP), a controlled release insecticide, on the activities of butyrylcholinesterase (BuChE) and acetylcholinesterase (AChE) in rainbow trout exposed to this pesticide was carried out. It was found that ChDP reduced BuChE activity in O. mykiss by a factor of eight at 6 days, with high fluctuation to the end of the exposition time at 12 days. The in vitro analysis of brain AChE treated with ChDP and Phenamiphos showed that it was competitively inhibited by both organophosphates. The values obtained for Km and Vmax for the AChE-ChDP (Km: 21.23 microM; Vmax: 43.10 micromol/min/g) and AChE-Phenamiphos (Km: 38.62 microM; Vmax: 38.91 micromol/min/g) systems were relatively low compared to values of the AChE (control) system (Km: 62.99 microM; Vmax: 63.29 micromol/min/g). Results reported in this study confirmed that chitosan diethyl phosphate performs similarly to organophosphate pesticides, producing inhibition in cholinesterases in rainbow trout.

Initial spreading dynamics of supported lipid monolayers
Blake, T. D. and J. De Coninck (2004), Langmuir 20(7): 2977-8.

Injectable biodegradable hydrogels composed of hyaluronic acid-tyramine conjugates for drug delivery and tissue engineering
Kurisawa, M., J. E. Chung, et al. (2005), Chem Commun (Camb)(34): 4312-4.
Abstract: The sequential injection of hyaluronic acid-tyramine conjugates and enzymes forms biodegradable hydrogels in vivo by enzyme-induced oxidative coupling, offering high potential as a promising biomaterial for drug delivery and tissue engineering.


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