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Estrogen reduces endothelial progenitor cell senescence through augmentation of telomerase activity
Imanishi, T., T. Hano, et al. (2005), J Hypertens 23(9): 1699-706.
Abstract: BACKGROUND: Recent studies have demonstrated that aging or senescence constitutes a potential limitation to the ability of endothelial progenitor cells (EPCs) to sustain ischemic tissue and repair. Conversely, estrogens have been shown to accelerate recovery of the endothelium after vascular injury. OBJECTIVE: To investigate whether estrogens are able to prevent senescence of EPCs. METHODS AND RESULTS: Human EPCs were isolated from peripheral blood and characterized. After ex-vivo cultivation, the cells became senescent as determined by acidic beta-galactosidase staining. 17beta-estradiol dose-dependently inhibited the onset of EPC senescence in culture. Because cellular senescence is critically influenced by telomerase, which elongates telomeres, we measured telomerase activity using a polymerase chain reaction (PCR)-enzyme-linked immunosorbent assay (ELISA) technique. 17beta-estradiol significantly increased telomerase activity. Interestingly, reverse transcriptase-PCR analysis demonstrated that 17beta-estradiol dose-dependently increased the catalytic subunit, telomerase reverse transcriptase (TERT) - an effect that was significantly inhibited by pharmacological phosphatidylinositol 3-kinase (PI3-K) blockers (either wortmannin or LY294002). Because the expression of TERT is regulated by the PI3-K/Akt pathway, we examined the effect of 17beta-estradiol on Akt activity in EPCs. Immunoblotting analysis revealed that 17beta-estradiol dose-dependently led to phosphorylation and, thus, to activation of Akt in EPCs. We also examined whether the protective effect of 17beta-estradiol on EPC senescence translates into the augmentation of mitogenic activity in EPCs. A [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenol)-2-(4-sulfophenyl) -2H-tetrazolium] (MTS) assay demonstrated that the mitogenic potential in EPCs treated with 17beta-estradiol exceeded that in untreated (control) EPCs (P < 0.01). In addition, EPCs released vascular endothelial growth factor (VEGF) protein--an effect that was significantly augmented by 17beta-estradiol. Finally, in a Matrigel assay, EPCs treated with both 17beta-estradiol and VEGF were shown to be more likely to integrate into the network formation than those treated with VEGF alone. CONCLUSION: The inhibition of EPC senescence by estrogen in vitro may improve the functional activity of EPCs in a way that is important for potential cell therapy.

Etiology of implant orthopedic infections: A survey on 1027 clinical isolates
Arciola, C. R., Y. H. An, et al. (2005), Int J Artif Organs 28(11): 1091-100.
Abstract: In spite of the recent achievements derived from modern protocols of prophylaxis, orthopedic surgical infections still remain unacceptably frequent, especially in light of the often devastating outcomes of septic complications. The spectrum and the prevalence of the bacteria most frequently involved in orthopedic infections are here explored, with particular reference to those infections associated to implant biomaterials, which were grouped based on device typology. During a 30 months period (from September 2000 to April 2003), 1027 microbial strains were consecutively isolated from 699 patients undergoing revision surgery at the Rizzoli Orthopedic Institute. 775 (75.5%) of all these microorganisms were identified as belonging to the Staphylococcus genus, 82 (8%) to the Enterobacteriaceae family, 75 (7.3%) to the Pseudomonas genus, 54 (5.3%) to the Enterococcus genus and 20 (1.9%) to the Streptococcus genus. While confirming the importance of staphylococci as the most diffuse cause of infection, our data indicate an unexpectedly high prevalence of S. epidermidis on infected hip and knee arthroprostheses, respectively of 42% and 44%. The spectrum of bacteria infecting either internal or external fracture fixation devices appears to differ from that of hip and knee arthroprostheses and more closely resembles that of infections non-associated to medical devices, being characterized by a relatively higher prevalence of Staphylococcus aureus (over 40%) and Pseudomonas aeruginosa. Enterobacteriaceae and members of the Streptococcus and Corynebacterium genera are frequently associated with implants in which surgical incisions were made near the perineum, determining a completely altered spectrum.

Evaluating protein attraction and adhesion to biomaterials with the atomic force microscope
Wang, M. S., L. B. Palmer, et al. (2004), Langmuir 20(18): 7753-9.
Abstract: Failure of implanted biomaterials is commonly due to nonspecific protein adsorption, which in turn causes adverse reactions such as the formation of fibrous capsules, blood clots, or bacterial biofilm infections. Current research efforts have focused on modifying the biomaterial interface to control protein reactions. Designing biomaterial interfaces at the molecular level, however, requires an experimental technique that provides detailed, dynamic information on the forces involved in protein adhesion. The goal of this study was to develop an atomic force microscope (AFM)-based technique to evaluate protein adhesion on biomaterial surfaces. In this study, the AFM was used to evaluate (i) protein-protein, (ii) protein-substrate, and (iii) protein-dextran interactions. The AFM was first used to measure the pull-off forces between bovine serum albumin (BSA) tips/BSA surfaces and BSA tips/anti-BSA surfaces. Results from these protein-protein studies were consistent with the literature. More importantly, the successful measurement of antibody-antigen binding interactions demonstrates that both the BSA and anti-BSA proteins retain their folded conformation and remain functional following our immobilization protocol. The AFM was also used to quantify the physiochemical interactions of proteins during adhesion to various self-assembled monolayers (SAMs) and dextran-coated substrates representative of potential biomaterial interface modifications. Dextran, which renders surfaces very hydrophilic, was the only surface coating that BSA protein did not adhere to. Hydrophobic interactions were not found to play a significant role in BSA adhesion. Therefore, the dextran molecules may resist protein adhesion by repulsive steric effects or hydration pressure. Moreover, the AFM-based methodology provides dynamic, quantitative information about protein adhesion at the nanoscale level.

Evaluating the in vitro adhesive strength of biomaterials. Biosimulator for selective leak closure
Carbon, R. T., S. Baar, et al. (2003), Biomaterials 24(8): 1469-75.
Abstract: AIMS: Surgical measures always involve problems of hemostasis and tissue management. Tissue sealing can be carried out in this case, with knowledge of the adhesive strength of liquid and fleece-bound sealing being of great clinical relevance. By employing a leak closure model, it has been possible to perform in vitro standardized technological-biomechanical experiments on a biomembrane mounted in a pressure chamber (biosimulator). FINDINGS: Liquid sealing (fibrin gluing: 4.1 hPa, photopolymerisate: 82.9 hPa) offers minimal and maximal adhesive strengths. Fleece-bound sealing does not depend on the carrier material and is always more stabile than pure liquid sealing, whereby systems that must be coated "on the spot" (prepare-to-use: 22.3-25.3 hPa) exhibit significantly lower adhesive strength (p < 0.0001) than a biodegradable collagen system, which is ready-to-use thanks to its fibrinogen-based coating (TachoComb H). Practicability, effectiveness and efficiency are further advantages of ready-to-use systems. CONCLUSIONS: The biosimulator, presented here, is flexible in terms of its application for tissue management and it enables reproducible, economic and ecological evaluation of repair systems, e.g. tissue sealings.

Evaluating the interaction of bacteria with biomaterials using atomic force microscopy
Razatos, A., Y. L. Ong, et al. (1998), J Biomater Sci Polym Ed 9(12): 1361-73.
Abstract: Bacterial infection of biomaterials represents one of the most important reasons for the failure of transdermal or implanted medical devices. The first and least understood step in biomaterial-associated infections is the initial interaction between bacteria and a surface. This initial interaction can be either attractive or repulsive depending on the physiochemical nature of the biological and synthetic surfaces, as well as the properties of the interstitial fluid. We have shown that atomic force microscopy (AFM) can be employed as an exquisitely sensitive and versatile tool for quantifying the interaction between bacteria and surfaces in physiological solutions. The forces of interaction between an AFM cantilever tip and a uniform lawn of bacteria immobilized on glass were determined. By comparing the interactions of cantilever tips with lawns of isogenic E. coli strains carrying genetic lesions that alter their cell surface composition, it was possible to evaluate the effect of macromolecules such as lipopolysaccharide and capsular polysaccharide on the adhesion process. Mutations that result in the synthesis of truncated lipopolysaccharide or in the overproduction of the negatively charged capsular polysaccharide colanic acid render the interaction of the bacteria with the AFM tip unfavorable due to increased electrostatic repulsion. Furthermore, AFM could be used to evaluate the adhesion of bacteria onto commercially relevant biomaterials. In one approach, micron-size polystyrene beads were attached to AFM tips which were then used to measure forces. Unfortunately, this approach is limited by the meager number of materials manufactured as beads of a size suitable for AFM measurements. As an alternative approach, AFM cantilever tips were coated with a confluent layer of bacteria and used to probe planar surfaces. In this configuration, AFM could be employed to measure the force of interaction between virtually any bacterium and surface of interest.

Evaluation of a new antithrombogenic polyurethane-polysiloxane complex biomaterial
Ohkubo, N., H. Hirose, et al. (1986), ASAIO Trans 32(1): 193-7.

Evaluation of a new approach to the safety assessment of biomaterials
Benson, W. H. and R. A. Stackhouse (1986), Drug Chem Toxicol 9(3-4): 275-83.
Abstract: The effectiveness of a bacterial luminescence inhibition assay in assessing the toxicity of compounds which are released from biomaterials was evaluated. Luminescence from a strain of bacteria most closely resembling Photobacterium phosphoreum was measured. The concentration that inhibited luminescence by 50% (EC50) was determined for selected plasticizers, monomers and additives. The intraperitoneal (i.p.-ALD) and intravenous (i.v.-ALD) approximate lethal doses were determined using mice. By ranking the reference compounds toxic/non-toxic, a 37.5% agreement was obtained for the i.p.-ALD and i.v.-ALD toxicity data. While there was only a 12.5% agreement for the i.p. ALD and EC50 values, there was a 75% agreement between the i.v.-ALD and EC50 values. Although additional validation is required, these results provide preliminary indications that the bacterial luminescence inhibition assay is a promising alternative approach to the safety assessment of biomaterials.

Evaluation of a novel biomaterial in the suprachoroidal space of the rabbit eye
Einmahl, S., M. Savoldelli, et al. (2002), Invest Ophthalmol Vis Sci 43(5): 1533-9.
Abstract: PURPOSE: Drug delivery to treat diseases of the posterior segment of the eye, such as choroidal neovascularization and its complications, is hampered by poor intraocular penetration and rapid elimination of the drug from the eye. The purpose of this study was to investigate the feasibility and tolerance of suprachoroidal injections of poly(ortho ester) (POE), a bioerodible and biocompatible polymer, as a biomaterial potentially useful for development of sustained drug delivery systems. METHODS: After tunnelization of the sclera, different formulations based on POE were injected (100 microL) into the suprachoroidal space of pigmented rabbits and compared with 1% sodium hyaluronate. Follow-up consisted of fundus observations, echography, fluorescein angiography, and histologic analysis over 3 weeks. RESULTS: After injection, POE spread in the suprachoroidal space at the posterior pole. It was well tolerated and progressively disappeared from the site of injection without sequelae. No bleeding or retinal detachment occurred. Echographic pictures showed that the material was present in the suprachoroidal space for 3 weeks. Angiography revealed minor pigment irregularities at the site of injection, but no retinal edema or necrosis. Histology showed that POE was well tolerated in the choroid. CONCLUSIONS: POE suprachoroidal injections, an easy, controllable, and reproducible procedure, were well tolerated in the rabbit eye. POE appears to be a promising biomaterial to deliver drugs focally to the choroid and the retina.

Evaluation of a poly(vinyl pyrollidone)-coated biomaterial for urological use
Tunney, M. M. and S. P. Gorman (2002), Biomaterials 23(23): 4601-8.
Abstract: The associated problems of bacterial biofilm formation and encrustation that may cause obstruction or blockage of urethral catheters and ureteral stents often hinders the effective use of biomaterials within the urinary tract. In this in vitro study, we have investigated the surface properties of a hydrophilic poly(vinyl pyrollidone) (PVP)-coating applied to polyurethane and determined its suitability for use as a urinary tract biomaterial by comparing its lubricity and ability to resist bacterial adherence and encrustation with that of uncoated polyurethane and silicone. The PVP-coated polyurethane was significantly more hydrophilic and more lubricious than either uncoated polyurethane or silicone. Adherence of a hydrophilic Escherichia coli isolate to PVP-coated polyurethane and uncoated polyurethane was similar but significantly less than adherence to silicone. Adherence of a hydrophobic Enterococcus faecalis isolate to PVP-coated polyurethane and silicone was similar but was significantly less than adherence to uncoated polyurethane. Struvite encrustation was similar on the PVP-coated polyurethane and silicone but significantly less than on uncoated polyurethane. Furthermore, hydroxyapatite encrustation was significantly less on the PVP-coated polyurethane than on either uncoated polyurethane or silicone. The results suggest that the PVP-coating could be useful in preventing complications caused by bacterial biofilm formation and the deposition of encrustation on biomaterials implanted in the urinary tract and, therefore, warrants further evaluation.

Evaluation of a series of tyrosine-derived polycarbonates as degradable biomaterials
Ertel, S. I. and J. Kohn (1994), J Biomed Mater Res 28(8): 919-30.
Abstract: A series of four polycarbonates derived from the ethyl, butyl, hexyl, and octyl esters of desaminotyrosyl-tyrosine was prepared by condensation polymerization. The resulting polymers had weight average molecular weights ranging from 120,000-450,000, and their chemical structure was confirmed by elemental analysis, nuclear magnetic resonance, and Fourier transform infrared spectroscopy. The polycarbonates were evaluated as degradable biomaterials. Their surface properties were determined by electron spectroscopy for chemical analysis, attenuated total reflectance-Fourier transformed infrared spectroscopy, and contact angle measurement. The degree of surface hydrophobicity was related to the length of the alkyl ester pendent chain. The tensile properties were dependent on the chemical structure of the polymers: For thin, solvent cast film specimens, the tensile modulus varied from 1.2-1.6 GPa, and the strength at break from 60-220 MPa. The degradation of polymeric films was followed in vitro by measuring changes in mechanical strength for up to 40 weeks, and the decrease in molecular weight and changes in surface chemistry for up to 80 weeks. The length of the pendent chain affected the degradation behavior and strength retention; the polymers with short pendent chains were more readily hydrolyzable. For sterilization, ethylene oxide treatment was less destructive, as judged by molecular weight retention, than gamma-irradiation. Spin-cast films of all tested polycarbonates were not cytotoxic toward cultured rat lung fibroblasts. The cell response was influenced by the chemical structure of the polymer. The least hydrophobic polycarbonate (having a short ethyl ester pendent chain) was a more stimulating substrate for cell growth than the more hydrophobic polymers (carrying longer alkyl ester pendent chains).

Evaluation of ameroid ring constrictors for treatment for single extrahepatic portosystemic shunts in dogs: 168 cases (1995-2001)
Mehl, M. L., A. E. Kyles, et al. (2005), J Am Vet Med Assoc 226(12): 2020-30.
Abstract: OBJECTIVES: To evaluate use of an ameroid ring constrictor (ARC) for treatment for single extrahepatic portosystemic shunts (PSSs) and identify factors associated with postoperative death, continued portosystemic shunting, and long-term outcome in dogs. DESIGN: Retrospective study. ANIMALS: 168 dogs with a single extrahepatic PSS. PROCEDURE: Medical records of dogs that had a single extrahepatic PSS and were treated with an ARC were reviewed. Signalment, history, clinical signs, results of preoperative blood analyses and portal pressure measurements, PSS location, ARC size, postoperative complications, and postoperative scintigraphy results were recorded. Owners were interviewed 6 months to 6 years after surgery. Results-Postoperative complications developed in 10% of dogs. Postoperative mortality rate was 7.1%. Predictive factors for postoperative death included high preoperative WBC count and postoperative complications. Twenty-one percent of dogs in which portal scintigraphy was performed 6 to 10 weeks after surgery had continued shunting. Predictive factors for persistent shunting included low preoperative plasma albumin concentration, high portal pressure after complete occlusion, and high portal pressure difference (postocclusion minus baseline). Clinical outcome in 108 dogs was classified as excellent (80%), good (14%), or poor (6%). Predictive factors for excellent long-term clinical outcome included high preoperative plasma albumin concentration, low preoperative leukocytosis, low portal pressure after complete occlusion, absence of postoperative seizures, and absence of continued shunting. CONCLUSIONS AND CLINICAL RELEVANCE: Use of an ARC for treatment for a single extrahepatic PSS resulted in low morbidity and mortality rates. Certain preoperative factors were associated with increased risk of postoperative death, continued portosystemic shunting, and long-term outcome.

Evaluation of an in situ forming hydrogel wound dressing based on oxidized alginate and gelatin
Balakrishnan, B., M. Mohanty, et al. (2005), Biomaterials 26(32): 6335-42.
Abstract: Wound dressings that can be formed in situ offer several advantages over the use of preformed dressings such as conformability without wrinkling or fluting in the wound bed, ease of application and improved patient compliance and comfort. Here we describe such an in situ forming hydrogel wound dressing from gelatin, oxidized alginate and borax. Periodate oxidized alginate rapidly cross-links proteins such as gelatin in the presence of borax to give in situ forming hydrogels that are both non-toxic and biodegradable. The composite matrix has the haemostatic effect of gelatin, the wound healing-promoting feature of alginate and the antiseptic property of borax to make it a potential wound dressing material. The hydrogel was found to have a fluid uptake of 90% of its weight which would prevent the wound bed from accumulation of exudates. The water vapour transmission rate (WVTR) of the hydrogel was found to be 2686+/-124 g/m2/day indicating that the hydrogel can maintain a moist environment over wound bed in moderate to heavily exuding wound which would enhance epithelial cell migration during the healing process. The wound healing efficacy of hydrogel was evaluated in experimental full thickness wounds using a rat model which demonstrated that within 2 weeks, the wound covered with gel was completely filled with new epithelium without any significant adverse reactions. These in situ forming hydrogels fulfil many critical elements desirable in a wound dressing material.

Evaluation of autologous bone marrow mesenchymal stem cell-calcium phosphate ceramic composite for lumbar fusion in rhesus monkey interbody fusion model
Wang, T., G. Dang, et al. (2005), Tissue Eng 11(7-8): 1159-67.
Abstract: Autologous bone marrow mesenchymal stem cell (BMSC)-calcium phosphate ceramic composites were constructed in vitro and implanted as a bone graft substitute for lumbar anterior interbody fusion in rhesus monkeys to determine the osteogenic capacity of the composites. Nine adult rhesus monkeys underwent lumbar L3-L4 and L5-L6 diskectomy and interbody fusion via an anterior retroperitoneal approach. Two fusion sites in each animal were randomly assigned to two of three treatments: autogenous tricortical iliac crest bone graft (autograft group), cell-free ceramic graft (ceramic group), or BMSC-ceramic composite graft (BMSC group). Autologous BMSCs were expanded in culture and stimulated with osteogenic supplement. The spinal fusion segments were evaluated by radiography, biomechanical testing, histologic analysis, and histomorphometric analysis 3 months postsurgery. The BMSC group achieved lumbar interbody fusion superior to that of the ceramic group, both biomechanically and histologically. The BMSC group and the autograft group showed equivalent biomechanical stiffness. Ceramic residues were significantly greater in the ceramic group versus the BMSC group. The results indicate that BMSC-ceramic composites can enhance bone regeneration and achieve osseous spinal fusion 3 months after implantation in the rhesus monkey interbody fusion model.

Evaluation of biodegradable synthetic scaffold coated on arterial prostheses implanted in rat subcutaneous tissue
Wang, Z., S. Wang, et al. (2005), Biomaterials 26(35): 7387-401.
Abstract: Polyester arterial prostheses impregnated with various synthetic biodegradable materials and with gelatin were implanted subcutaneously in rats for 3-180 days. The inflammation was assessed by quantifying the activity of alkaline phosphatase and by histology. The degradation of the scaffold materials was determined by scanning electron microscopy (SEM), size exclusion chromatography (SEC), and differential scanning calorimetry (DSC). The alkaline phosphatase activity induced by the polymer-impregnated grafts was similar to that induced by the non-impregnated controls during most of the post-implantation periods. Histological studies revealed that the acute inflammatory response was moderate to mild and was similar for all types of specimens, except for the gelatin-impregnated grafts that induced a severe acute inflammation during the first 2 weeks post-implantation. At 4 and 6 months, significant disintegration of the scaffold was observed, accompanied by enhanced tissue infiltration and a reactivation of the acute inflammatory phase. Linear and exponential degradation rates of the synthetic polymers were described. The relative degradation rates of the biodegradable polymers were ranked as following: PLLACL > PDLLA > PLLA > PCEL. In conclusion, biodegradable polymers may provide an option as sealant/scaffolding materials for vascular prosthesis. It is suggested that the degradation rate of the polymer scaffolding materials should be higher to achieve early healing while without inducing strong inflammation.

Evaluation of biological responses to polymeric biomaterials by RT-PCR analysis III: study of HSP 70, 90 and 47 mRNA expression
Kato, S., T. Akagi, et al. (1998), Biomaterials 19(7-9): 821-7.
Abstract: In this study, in order to analyze how cells recognize biomaterials, mRNA was evaluated on various substrates as shown in the attached HeLa S3 cells. The expressed Heat-Shock Protein (HSP) 70A, 70B, 90 and 47 mRNA were isolated and detected using the RT-PCR method. As a result, 70B, 90 and 47 mRNA expressions varied with differences in the hydrophilicity hydrophobicity of the substrates. HSP mRNAs outcome was induced significantly in the HeLa S3 cells that had adhered onto the hydrophilic surfaces. On the other hand, in the cells that had adhered to the hydrophobic surfaces, HSP mRNAs expressions were low. In the non-adhered HeLa S3 cells the tendency found in HSP 70B and HSP 47 mRNA's expressions was the same as that found in adhered cells, while there were no significant differences in the HSP 90 mRNA expression among either of the samples. We have concluded that HSP mRNAs expression is an important marker in the study of cell-polymer interactions.

Evaluation of biological responses to polymeric biomaterials by RT-PCR analysis IV: study of c-myc, c-fos and p53 mRNA expression
Kato, S., T. Akagi, et al. (2000), Biomaterials 21(5): 521-7.
Abstract: In order to investigate how cells recognize biomaterials, mRNA that was expressed in attached human fibroblasts on various substrates was evaluated. The expressed oncogenes (c-fos and c-myc) and tumor suppressor gene (p53) mRNA were then isolated and detected using the RT-PCR method. As a result, c-fos and c-myc mRNA expression varied with respect to differences in the hydrophilicity-hydrophobicity of the substrates. Both c-fos and c-myc mRNA expression were low in the fibroblasts that had adhered to hydrophilic surfaces. The tendency of c-fos mRNA expression was similar to the adhesion curve of the cells. c-myc mRNA was largely induced in fibroblasts that had adhered to hydrophobic surfaces. p53 mRNA were largely induced in fibroblasts that had adhered to hydrophilic surfaces, while in the cells that had adhered to hydrophobic surfaces, p53 mRNA expression was low. We concluded that the expression of oncogenes and p53 mRNA is a powerful method for studying cell-polymer interactions or the evaluation of the carcinogenic activity of biomaterials.

Evaluation of biological responses to polymeric biomaterials by RT-PCR analysis. I. Study of IL-1 beta mRNA expression
Kishida, A., S. Kato, et al. (1996), Biomaterials 17(13): 1301-5.
Abstract: In this study, we introduce a novel research methodology, the evaluation of mRNA expression of cells contacting with polymeric materials using reverse transcription-polymerase chain reaction (RT-PCR) analysis HL-60 was used as a model of the macrophages. The expression of interleukin-1 beta (IL-1 beta) mRNA, a cytokine secreted by macrophages, was selected to estimate the extent of inflammation. The expression of IL-1 beta mRNA in the HL60 cells cultured on various substrates and in various conditions was studied. Expression of IL-1 beta could be successfully determined by RT-PCR analysis. A 48 h incubation period was necessary to clarify the expression of IL-1 beta mRNA. It became clear that lipopolysaccharide stimulation was not necessary in this analysis because of the high sensitivity of RT-PCR analysis. It is concluded that RT-PCR analysis is a powerful tool for studying cell-polymer interaction, and is a complementary method for ELISA.

Evaluation of biological responses to polymeric biomaterials by RT-PCR analysis. II: Study of HSP 70 mRNA expression
Kato, S., T. Akagi, et al. (1997), J Biomater Sci Polym Ed 8(10): 809-14.
Abstract: In order to investigate how cells recognize biomaterials, mRNA that was expressed in attached HeLa S3 cells on various substrates was evaluated. As culture substrates, cellulose, ethylene-vinyl alcohol copolymer (EVAL), nylon, tissue culture polystyrene (TCPS), high-density polyethylene (PE), silicone rubber, and tetrafluoroethylene-hexafluoropropylene copolymer (6F) were used. HeLa S3 cells were cultured on these substrates for 24 h. The expressed HSP 70s mRNA was then isolated and detected using the RT-PCR method. As a result, the expression of HSP 70B mRNA was largely induced in cells that adhered to hydrophilic surfaces. On the other hand, on hydrophobic surfaces, the HSP 70B mRNA expression was low. It is concluded that HSP 70B mRNA expression is sensitive to differences in the hydrophilicity-hydrophobicity of the substrates.

Evaluation of cartilage repair tissue after biomaterial implantation in rat patella by using T2 mapping
Watrin-Pinzano, A., J. P. Ruaud, et al. (2004), Magma 17(3-6): 219-28.
Abstract: To evaluate the ability of MR T2 mapping (8.5 T) to characterize ex vivo longitudinally, morphologically and quantitatively, alginate-based tissue engineering in a rat model of patellar cartilage chondral focal defect. Calibrated rat patellar cartilage defects (1.3 mm) were created at day 0 (D0) and alginate sponge with (Sp/C+) or without (Sp/C-) autologous chondrocytes were implanted. Animals were sacrificed sequentially at D20, D40 and D60 after surgery and dissected patellae underwent MRI exploration (8.5 T). T2 values were calculated from eight SE images by using nonlinear least-squares curve fitting on a pixel-by-pixel basis (constant repetition time of 1.5 s, eight different echo times: 5.5, 7.5, 10.5, 12.5, 15.0, 20.0, 25.0 and 30.0 ms). On the T2 map, acquired in a transversal plane through the repair zone, global T2 values and zonal variation of T2 values of repair tissue were evaluated versus control group and compared with macroscopic score and histological studies (toluidine blue, sirius red and hematoxylin-eosin). "Partial", "total" and "hypertrophic" repair patterns were identified. At D40 and D60, Sp/C+ group was characterized by a higher proportion of "total" repair in comparison to Sp/C- group. At D60, the proportion of "hypertrophic" repair was two fold in Sp/C- group versus Sp/C+ group. As confirmed morphologically and histologically, the T2 map also permitted the distinction of three types of repair tissue: "total", "partial" and "hypertrophic". "Total" repair tissue was characterized by high T2 values versus normal cartilage (p<0.05). Zonal variation, reflecting the collagen network organization, appeared only at D60 for Sp/C+ group (p<0.05). "Hypertrophic" tissue, mainly observed at D60, presented high T2 global values without zonal variation with cartilage depth. These results confirm the potency of the MR T2 map (8.5 T) to characterize macroscopically and microscopically the patterns of the scaffold guided-tissue repair of a focal chondral lesion in the rat patella ("total", "partial" and "hypertrophic"). On T2 map, three parameters (i.e. MRI macroscopic pattern, T2 global values and zonal variation of T2 values) permit to characterize chondral repair tissue, as a virtual biopsy.

Evaluation of cell colonization on biomaterials: preventing cell attachment to plastic containers
Amedee, J., R. Bareille, et al. (1994), Biomaterials 15(12): 1029-31.
Abstract: In biocompatibility evaluation involving cell culture models, we use samples of biomaterials of different forms and sizes. During cell seeding onto biomaterials of an inadequate size to cover the bottom of the culture wells completely, cells have the opportunity to attach to the plastic. As described in this report with two culture models and two biomaterials, we use an agarose gel sublayer to prevent this phenomenon.

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Last Modified: 8 February 2006