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At least 127 records · Page 7Linked to original sources

The influence of novel bioactive glasses on in vitro osteoblast behavior.

Implant success requires a direct bond between bone and implant surface. Bioinert implants, such as titanium alloys, are commonly plasma-spray-coated with a bone-bonding, bioactive material such as hydroxyapatite. Such coatings tend to be chemically and topographically inhomogeneous without reproducible properties. A family of bioactive glasses that can be enameled and reliably adheres to titanium alloy has been developed. In this study the cytocompatibility of two of these glass compositions was tested in the as-cast condition. The effects of these glasses on the early and late events of osseous tissue formation in vitro were determined with MC3T3-E1.4 mouse osteoblast-like cells. MC3T3-E1.4 cells were cultured on glasses containing 55 and 50 wt % SiO(2), with titanium alloy (Ti6Al4V) and tissue culture polystyrene as controls. Cellular adhesion and proliferation, and alkaline phosphatase activity were studied over 5 to 15 days in culture. Qualitative and quantitative assays of mineralization were conducted. The osteoblast-like cells showed increased proliferation when grown on a bioactive glass containing 50 wt % silica. However, the adhesion, differentiation and mineralization behavior were similar on both glass compositions used in this study. These bioactive glasses proved to be cytocompatible substrata for osteoblast-like cell culture, and yielded higher cellular proliferation than titanium alloy.

Alkaline Phosphatase↗

Asymmetrically functional surface properties on biocompatible phospholipid polymer membrane for bioartificial kidney.

To obtain a bioartificial kidney composed of a porous polymer membrane and renal cells, a polysulfone (PSf) membrane (PSM) blended with 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer was prepared. The PSM flat membrane with a porous structure could be prepared from the polymer blend containing 1 wt % of the MPC polymer in PSf by the phase inversion technique in a dry-wet process. Asymmetrical surface properties were observed on both sides of the membrane surfaces. That is, the sponge layer formed at the substrate-contacting surface of the membrane had 10-20 microm pores, but the pores in the micrometer range could not be observed for a skin layer formed at the air-contacting surface of the membrane. At the sponge layer surface, the MPC unit composition was 7 times larger than that at the skin layer surface. The amount of proteins adsorbed on the surface corresponded to the MPC unit composition. On the skin layer, a small amount of adsorbed proteins and platelet adhesion could be suppressed compared with those on the sponge layer. However, the skin layer had a moderate protein adsorption, so it showed a sufficient cytocompatibility to enable renal tubule epithelial cells to adhere and proliferate in the membrane. Thus, it functioned well as a renal tubule. Therefore, because of both its hemocompatibility and cytocompatibility, we could conclude that the PSM membrane is useful for as a renal tubule device for a bioartificial kidney.

Adsorption↗

Increased osteoblast adhesion on nanograined hydroxyapatite and partially stabilized zirconia composites.

To improve the mechanical properties of hydroxyapatite (HA, Ca(10)(PO(4))(6)(OH)(2)) for orthopedic applications, numerous investigators have proposed combining HA with high strength materials, specifically zirconia. Despite the fact that, compared to pure HA, it is now well-established that zirconia and HA composites have improved mechanical properties, the cytocompatibility properties of this composite remain largely uninvestigated. For these reasons, the objective of the present in vitro study was to synthesize HA and partially stabilized zirconia composites for osteoblast (bone-forming cell) adhesion assays. Various sintering temperatures and amounts of zirconia in HA composites were used in order to ascertain their influence on osteoblast adhesion. Results demonstrated increased interactions between HA and partially stabilized zirconia, when either higher sintering temperatures (between 900 and 1,300 degrees C for 1 h) or higher zirconia contents (between 10 and 40 wt %) were used during material synthesis. More importantly, greater osteoblast adhesion was measured on HA-zirconia composites sintered either at lower temperatures (specifically, 900 degrees C) or with lower amounts of zirconia added to HA composites (specifically, 10 wt %). Results further indicated that when sintered at lower temperatures the composites possessed smaller nanometer grain sizes with increased surface roughness and a more stable HA phase. For these reasons, this study suggests that to optimize osteoblast adhesion on HA and partially stabilized zirconia composites for orthopedic applications, low sintering temperatures and low amounts of zirconia should be used. This suggests that a delicate balance must be reached between increasing mechanical properties of HA without decreasing osteoblast cytocompatibility properties through zirconia addition.

Animals↗

Enzymatic stabilization of gelatin-based scaffolds.

The definitive goal of this research is to develop protein-based scaffolds for use in soft tissue regeneration, particularly in the field of dermal healing. The premise of this investigation was to characterize the mechanical properties of gelatin cross-linked with microbial transglutaminase (mTGase) and to investigate the cytocompatibility of mTGase cross-linked gelatin. Dynamic rheological analysis revealed a significant increase in the storage modulus and thermal stability of gelatin after cross-linking with mTGase. Static, unconfined compression tests showed an increase in Young's modulus of gelatin gels after mTGase cross-linking. A comparable increase in gel strength was observed with 0.03% mTGase and 0.25% glutaraldehyde cross-linked gelatin gels. In vitro studies using 3T3 fibroblasts indicated cytotoxicity at a concentration of 0.05% mTGase after 72 h. However, no significant inhibition of cell proliferation was seen with cells grown on lower concentrations of mTGase cross-linked gelatin substrates. The mechanical improvement and cytocompatibility of mTGase cross-linked gelatin suggests mTGase has potential for use in stabilizing gelatin gels for tissue-engineering applications.

Animals↗

Porous akermanite scaffolds for bone tissue engineering: preparation, characterization, and in vitro studies.

The aim of this study was to develop a bioactive, degradable, and cytocompatible akermanite (Ca2MgSi2O7) scaffold with high porosity and pore interconnectivity. In brief, porous akermanite scaffolds were prepared using polymer sponge method. The porosity and corresponding compressive strength were evaluated. The in vitro degradability was investigated by soaking the scaffolds in Ringer's solution. Hydroxyapatite (HAp)-formation ability of akermantite scaffolds in simulated body fluid (SBF) and the effect of ionic products from the scaffolds dissolution on osteoblasts were investigated. In addition, bone marrow stromal cells (BMSC) adhesion and proliferation on the scaffolds were evaluated. Differentiation of the cells was assessed by measuring alkaline phosphatase (ALP) activity. The results showed that akermanite scaffolds possessed 63.5-90.3% of porosity, with a corresponding compressive strength between 1130 and 530 kPa. The weight loss of the scaffolds and ionic content of the Ringer's solution increased with the increase in soaking time, indicating the degradability of scaffolds. HAp was formed on the scaffolds in SBF and the ionic products from akermanite scaffolds dissolution stimulated osteoblasts proliferation, indicating good in vitro bioactivity. Furthermore, BMSC adhered and spread well on akermanite scaffolds and proliferated with the increase in the culture time, and the differentiation rate of osteoblasts on scaffolds was comparable to that on blank culture plate control. Our results suggested that akermanite scaffolds were bioactive, degradable, and cytocompatible, and might be used as bone tissue engineering materials.

Animals↗

Biological responses to multilayered DNA-coatings.

This study was performed to evaluate the basic biological response to deoxyribonucleic acid (DNA)-based coatings for soft tissue implants. To that end, in vitro experiments were used to study their cytocompatibility, and in vivo subcutaneous implantation studies with transponders in a rat model were performed to evaluate their histocompatibility. The DNA-based coatings were fabricated using the electrostatic self-assembly technique using cationic poly-D-lysine or poly-allylamine hydrochloride and anionic DNA. Noncoated substrates served as controls. In vitro, the behavior of primary rat dermal fibroblasts was assessed in terms of cell proliferation and morphology. Both types of multilayered DNA-coatings significantly increased rat dermal fibroblast proliferation without altering the morphological appearance of the cells. The tissue response to multilayered DNA-coatings was assessed using an in vivo rat model, in which transponders were inserted subcutaneously for 4 and 12 weeks. No macroscopic signs of inflammation or adverse tissue reactions were observed at implant retrieval. Histological analyses demonstrated a uniform tissue response to all types of implants. All implants were encapsulated in a fibrous tissue capsule without intervening inflammatory cells at the implant surface. Histomorphometrically, multilayered DNA-coatings induced fibrous tissue capsules with similar quality and thickness compared to noncoated controls. In addition, all fibrous tissue capsules showed similar expression of alpha-smooth muscle actin. This study demonstrates that multilayered DNA-coatings are cytocompatible and histocompatible, and justifies further research on their functionalization with biologically active compounds to modulate tissue responses.

Actins↗

Temperature and pH sensitive hydrogels: an approach towards smart semen-triggered vaginal microbicidal vehicles.

Microbicides are drug delivery systems (DDSs) for the prevention of sexual transmission of HIV and other STDs. A topically applied vaginal microbicidal gel should provide uniform coating of vaginal tissue, retention of this gel layer prior to intercourse, and controlled release kinetics of antivirals to inactivate the viral load potentially introduced during sexual activity. Here, we describe the microbicide-oriented characterization of a DDS made with a dual pH sensitive and thermosensitive smart polymer gel composed of a random terpolymer of N-isopropyl acrylamide, butyl methacrylate, and acrylic acid. The system was engineered to coat vaginal tissue with a stable gel layer and to release entrapped model agents in a burst release profile in response to the presence of the infecting agent: semen. The gel rheology, layer erosion properties, model drug release kinetics, and cytocompatibility of the terpolymer system were studied. Negligible erosion of the gel in the presence of vaginal fluid simulant suggests prolonged retention. Burst release of molecular and macromolecular model compounds was observed when the system's pH changed from the vaginal pH to the pH of semen, and cytotoxicity studies showed that the terpolymer is equally cytocompatible as a commonly used polymeric vaginal carrier.

Administration, Intravaginal↗

Patterning of a random copolymer of poly[lactide-co-glycotide-co-(epsilon-caprolactone)] by UV embossing for tissue engineering.

The random copolymer, poly[lactide-co-glycotide-co-(epsilon-caprolactone)] (PLGACL) diacrylate was prepared by ring-opening polymerization of L-lactide, glycolide, and epsilon-caprolactone initiated with tetra(ethylene glycol). The diacrylated polymers were extensively characterized. With a UV embossing method, these copolymers were successfully fabricated into microchannels separated by microwalls with a high aspect (height/width) ratio. The PLGACL network films showed good cytocompatibility. Varieties of microstructures were fabricated, such as 10 x 40 x 60, 10 x 80 x 60, 25 x 40 x 60, or 25 x 80 x 60 microm(3) structures (microwall width x microchannel width x microwall height). The results demonstrated that smooth muscle cells (SMCs) can grow not only on the microchannel surfaces but also on the surfaces of the microwall and sidewall. The SMCs aligned along the 25 microm wide microwall with an elongated morphology and proliferated very slowly in comparison to those on the smooth surface with a longer cell-culture term. Few cells could attach and spread on the surface of the 40 microm wide microchannel, while the cells flourished on the 80 microm, or more than 80 microm, wide microchannel with a spindle morphology. The biophysical mechanism mediated by the micropattern geometry is discussed. Overall, the present micropattern, consisting of biodegradable and cytocompatible PLGACL, provides a promising scaffold for tissue engineering.

Caproates↗

Immunogold labelling of fibroblast focal adhesion sites visualised in fixed material using scanning electron microscopy, and living, using internal reflection microscopy.

A new immunogold labelling method for the visualisation of vinculin, an integral protein in focal adhesions of cells, is reported. Quantification of vinculin is indicative of substrate cytocompatibility (cytocompatibility is one aspect of biocompatibility; it is the cellular response to a biomaterial). For efficient labelling, most of the cell body above the cell-substrate interface was removed with detergent. The antigen blocking procedure, size of label (5 nm) and duration of silver-enhancement (6 min), for visualisation of the labelled sites on the whole cell by scanning electron microscopy (SEM), were determined. Imaging living cells with interference reflection light microscopy, followed by backscattered electron (BSE) imaging of the same fixed and immunolabelled cells confirmed the results. Collecting low voltage BSE images of embedded cells after the substrate had been removed provided 'sectional' views through the cell. This enabled visualisation of vinculin exclusively within the cell-substrate contact zone; the focal adhesions. The method could be of general use in the imaging of protein distribution at biological tissue/substrate interfaces.

Acrylic Resins↗

Compatibility of resorbable and nonresorbable guided tissue regeneration membranes in cultures of primary human periodontal ligament fibroblasts and human osteoblast-like cells.

The purpose of this study was (a) to evaluate the cytocompatibility of three resorbable and nonresorbable membranes in fibroblast and osteoblast-like cell cultures and (b) to observe the growth of those cells on the various barriers by scanning electron microscopy (SEM). Primary human periodontal ligament fibroblasts (HPLF) and human osteoblast-like cells (SAOS-2) were incubated with nonresorbable polytetrafluoroethylene (ePTFE) barriers and resorbable polylactic acid as well as collagen membranes. Cytotoxic effects were determined by XTT (mitochondrial metabolic activity) and sulforhodamine B assays (cellular protein content). In addition, HPLF and SAOS-2 grown for 21 days on the investigated barriers were evaluated by SEM. Data were analyzed statistically by ANOVA using the Wilcoxon-Mann-Whitney test (P < 0.05). No changes were established in the periodontal ligament fibroblasts and human osteoblast-like cells after incubation with the collagen membrane. Cytotoxic effects, however, were induced by the polylactic acid barrier which slightly inhibited cell metabolism of the periodontal fibroblasts (XTT: 90.1% +/- 3.6 of control value). Moderate cytotoxic reactions were caused by the nonresorbable ePTFE membrane in HPLF-cultures (XTT: 82.7% +/- 3.5) and osteoblast-like cell monolayers (XTT: 80.0% +/- 0.6%). Mitochondrial activity in both cell cultures was significantly reduced by ePTFE barriers in comparison to non-incubated control cells (P = 0.028). SEM analysis of cell behavior on barriers demonstrated the differences between these materials: collagen barriers were densely populated with HPLF and SAOS-2, whereas only few or no cells were seen to adhere to the ePTFE and polylactic acid membranes. Our findings indicate that the collagen barrier investigated is very cytocompatible and may be integrated into connective tissue well. On the contrary, the ePTFE and polylactic acid membranes induced slight to moderate cytotoxic reactions which may reduce cellular adhesion. Thus, gap formation between the barrier surface and the connective tissue may be promoted which may facilitate epithelial downgrowth and microbial accumulation. Consequently, these effects may reduce the potential gain in periodontal attachment.

Absorbable Implants↗

Biological aspects of root canal filling materials--histocompatibility,cytotoxicity, and mutagenicity.

In order to minimize the incidence of local and/or systemic side effects, the biocompatibility of all endodontic materials should be investigated by various in vitro and in vivo tests prior to clinical application. The battery of in vitro tests includes determinations of mutagenicity, cytotoxicity, and antibacterial effects. Several reports have shown that paraformaldehyde-containing ZnO-eugenol cements in particular, such as Endomethasone and N2, are antibacterial. On the other hand, it has been found that endodontic materials with strong antimicrobial activity are frequently mutagenic, i.e., primarily those which release formaldehyde. Cell culture tests clearly show significantly different cytocompatibility of the various types of endodontic sealers: in general, formaldehyde-containing ZnO-eugenol cements are classified as highly/extremely cytotoxic, whereas most Ca(OH)2-based sealers are rated as possessing good or excellent cytocompatibility. These results were confirmed by numerous histological studies in vivo. Sealers with inferior biocompatibility, such as formaldehyde-releasing materials, should no longer be applied in practice because safer alternatives are available.

Animals↗

Preparation and properties of ProNectin F-coated biodegradable hollow fibers.

ProNectin F-coated biodegradable hollow fibers were newly prepared and their cytocompatibility was evaluated in vitro. Although the coating efficiency onto poly(L-lactic acid) (PLLA) and poly(lactide-co-caprolactone) [p(LA/CL)] matrices was similar, the cell adhesion properties were greatly affected by the nature of the polymer substrate. ProNectin F-coated PLLA showed about seven times higher cytocompatibility than ProNectin F-coated p(LA/CL). The single-extruded melt spinning method and the core-sheath bicomponent melt spinning method were employed to prepare PLLA hollow fibers. The effect of the spinning conditions, such as the melt draw ratio, spinneret temperature, and take-up speed, on the diameter and wall thickness of the spun fibers was studied in detail. For single-extruded melt spinning, a segmented type of spinneret was used, and the effect of the flow rate of nitrogen, which was confined in the hollow part of fibers, was studied. X-ray photographs of the drawn hollow fibers, clarified the significant molecular orientation, which was much higher than that in drawn solid PLLA fiber under identical drawing conditions. The morphology and mechanical properties of hollow fibers demonstrated an increase in the tensile strength and a decrease in the thickness of the PLLA wall with increased nitrogen flow rates and melt draw ratios for single-extruded melt spinning. These results indicate the unique characteristics of ProNectin F-coated PLLA hollow fibers, which can be successfully utilized as a biodegradable substrate.

Absorbable Implants↗

Decreased functions of astrocytes on carbon nanofiber materials.

Carbon nanofibers possess excellent conductivity properties, which may be beneficial in the design of more effective neural prostheses; however, limited evidence on their cytocompatibility properties currently exists. The objective of the present in vitro study was to determine cytocompatibility properties of formulations containing carbon nanofibers pertinent to neural implant applications. Substrates were prepared from four different types of carbon fibers, two with nanoscale diameters (nanophase, or less than or equal to 100 nm) and two with conventional diameters (or greater than 100 nm). Within these two categories, both a high and a low surface energy fiber were investigated and tested. Carbon fibers were compacted in a manual hydraulic press via a uniaxial loading cycle. Astrocytes (glial scar tissue-forming cells) were seeded onto the substrates for adhesion, proliferation, and long-term function studies (such as total intracellular protein and alkaline phosphatase activity). Results provided the first evidence that astrocytes preferentially adhered and proliferated on carbon fibers that had the largest diameter and the lowest surface energy. Based on these results, composite substrates were also formed using different weight percentages (0-25 wt%) of the nanophase, high surface energy fibers in a polycarbonate urethane matrix. Results provided the first evidence of decreased adhesion of astrocytes with increasing weight percents of the high surface energy carbon nanofibers in the polymer composite; this further demonstrates that formulations containing carbon fibers in the nanometer regime may limit astrocyte functions leading to decreased glial scar tissue formation. Positive interactions with neurons, and, at the same time, limited astrocyte functions leading to decreased gliotic scar tissue formation are essential for increased neuronal implant efficacy.

Animals↗

Enhanced osteoblast adhesion on hydrothermally treated hydroxyapatite/titania/poly(lactide-co-glycolide) sol-gel titanium coatings.

Sol-gel processing was used to coat titanium substrates with hydroxyapatite (HA), TiO2, and poly(DL-lactic-glycolic acid). Coating surface characteristics were analyzed with XRD, EDS, AFM, SEM, and water contact angle measurements which indicated that the coatings had a high degree of crystallinity and good resistance to cracking. Coatings were also evaluated by cytocompatibility testing with osteoblast-like cells (or bone-forming cells). The cytocompatibility of the HA composite coatings prepared in the present in vitro study was compared to that of a traditional plasma-sprayed HA coating. Results showed that osteoblast-like cell adhesion was promoted on the novel HA sol-gel coating compared to the traditional plasma-sprayed HA coating. In addition, hydrothermal treatment of the sol-gel coating improved osteoblast-like cell adhesion. Since osteoblast adhesion is a necessary prerequisite for subsequent formation of bone, these results provided evidence that hydrothermally sol-gel processed HA may improve bonding of titanium implants to juxtaposed bone and, thus, warrants further investigation.

Cell Adhesion↗

Biocompatibility and stability of disulfide-crosslinked hyaluronan films.

Hyaluronan (HA) can be chemically modified to engineer robust materials with pre-selected mechanical properties and resorption rates that can be dictated by the intended clinical use. Disulfide-crosslinked HA films were prepared by air oxidation of thiol-modified HA, followed by treatment with 0.3% hydrogen peroxide. The degradation of the disulfide-crosslinked films in vitro was very slow (<10% in 7 days) in buffer alone and shorter (t1/2=3-5 days) in the presence of hyaluronidase (HAse). The cytocompatibility of the disulfide-crosslinked HA films was determined using two separate conditions: (i) in vitro culture of mouse fibroblasts in indirect contract with the films, and (ii) in vitro culture of fibroblasts directly on films coated with poly d-lysine. Excellent cytocompatibility was observed in murine fibroblasts that were cultured in indirect contact with thiolated HA films. Although cells were unable to attach and spread on thiolated HA films, pre-coating the thiolated HA films with poly D-lysine resulted in attachment and spreading equivalent to that observed on polystyrene. Rates of resorption in vivo were obtained by subcutaneous implantation of disulfide-crosslinked HA films into the backs of Wistar rats. Biocompatibility in vivo was determined in both subcutaneous flank and peritoneal cavity implantation of the films in Wistar rats. The disulfide-crosslinked HA films were less than 30% resorbed after 42 days in vivo, and histochemical and cytochemical analysis indicated that the films were well-tolerated with mild inflammatory response at both sites of implantation.

Absorbable Implants↗

Influence of the degree of acetylation on some biological properties of chitosan films.

In this study, we investigated in vitro the role of the degree of acetylation (DA) on some biological properties of chitosan films. We noticed that, whatever the DA, all chitosan films were cytocompatible towards keratinocytes and fibroblasts. We also demonstrated that the higher the DA of chitosan, the lower was the cell adhesion on the films. Fibroblasts appear to adhere twice as much as keratinocytes on these materials. We observed that keratinocyte proliferation increases when the DA of chitosan films decreases. Thus, DA influences the cell growth in the same way as cell adhesion. On the other hand, although they remain alive, fibroblasts do not proliferate on chitosan films. This behaviour is related to an extremely high adhesion on this kind of material, which certainly inhibits cell growth. In conclusion, DA plays a key role in cell adhesion and proliferation, but does not change the cytocompatibility of chitosan. In parallel, it is also important to notice the role played by the surface morphology of the material, a second major parameter which influences the mechanism of adhesion.

Acetylation↗

Evaluation of cell behaviour related to physico-chemical properties of polymeric membranes to be used in bioartificial organs.

In bioartificial organs using isolated cells, polymeric semipermeable membranes are used as immunoselective barriers as a means for cell oxygenation and also as substrata for adhesion of anchorage-dependent cells. The capacity of the membrane to perform its functions and to provide a cytocompatible support for cell culture depends in particular on its surface properties. In this study we investigated the physico-chemical aspects of the interaction between the membrane and mammalian cells in order to provide guidelines to the selection of cytocompatible membranes. We evaluated the adhesion and metabolic behaviour of isolated liver cells cultured on various polymeric membranes such as those modified by protein adsorption. The physico-chemical properties of the membranes were characterised by contact angle measurements. The different parameters such as acid (gamma+), base (gamma-) and Lifshitz-van der Waals (gammaLW) of the surface free energy were calculated according to Good-van Oss's model. The adsorption of protein modified markedly both contact angle and components of membrane surface tension. In particular, base parameter of surface tension decreased drastically with increased water contact angle. For each investigated membrane we observed that cell adhesion increased with increasing base parameter of membrane surface tension. The absolute value of cell adhesion is higher in the presence of serum proteins adsorbed on the membrane surface, which change the wettability by increasing the base parameter of surface tension. Also, the metabolic functions improve on hydrophilic membranes. Liver cells synthesised urea with a rate that increased with increasing base parameter value of membrane surface tension. The metabolic activity is particularly expressed at high levels when cells were cultured on polycarbonate and cellulose acetate membranes.

Animals↗

Assessment of the cytotoxicity of photocrosslinked dextran and hyaluronan-based hydrogels to vascular smooth muscle cells.

The cytotoxicity of polysaccharide-based hydrogels and solutions was studied in vitro after 48h of indirect exposure of the materials with vascular smooth muscle cells. Dextran and/or hyaluronan were derivatized using glycidyl methacrylate, and hydrogels were formed in the presence of photoinitiators and ultraviolet radiation in multiwell inserts to avoid direct contact with cell monolayers. Observation of cell morphology indicated that dextran hydrogels, a blend of non-derivatized hyaluronan into dextran hydrogel. and a hyaluronan solution were highly cytocompatible. However, hydrogels made of derivatized hyaluronan were cytotoxic when compared to unexposed sham controls that contained multiwell inserts but no hydrogels. Results from quantitative assays for proliferation and viability corroborated the qualitative observations, and scrape wound assays revealed a significant increase in smooth muscles cell migration/proliferation after indirect exposure to several of the polysaccharide-based materials. Results from this study demonstrate that hydrogels made of dextran and hyaluronan solution show good cytocompatibility in vitro. making these degradable matrices interesting candidates for drug delivery purposes.

Animals↗