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At least 199 records · Page 11Linked to original sources

The experimental study on mixed culture of osteoblasts and tricalcium phosphate ceramics in vitro.

To study the effects of tricalcium phosphate (TCP) ceramics on osteoblasts, the rat osteoblasts were cultured with the TCP ceramics in vitro. Scanning electron microscopy and the colorimetric methyl-thiazol-tetrazolium assay showed that the osteoblasts could adhere well to the surface of the ceramics and the culture dish, and the proliferation of the cells was not inhibited. The results demonstrated that TCP ceramics possessed an excellent cytocompatibility with the osteoblasts, and had some promoting effects on proliferation of osteoblasts.

Animals↗

[Type I collagen in xenogenic bone material regulates attachment and spreading of osteoblasts over the beta1 integrin subunit].

Xenogenic bone biomaterials have been proposed as an alternative to autografts or allografts in human bone restoring or in complement of prosthetic surgery. When appropriate treatments were applied, immunological, inflammatory, bacteriological or virological adverse responses can be prevented. However, these treatments may interact with type I collagen, the major component of the organic bone matrix. Type I collagen can bind osteoblasts via specific cell surface receptors, the integrins. In this work, two different xenogenic biomaterials were studied. Both biomaterials have a bovine bone origin. They displayed similar architectural organization with connected plates and rods and similar surface topography and roughness. They differed by the presence or not of collagen type I. The first one was characterized by preservation of the type I collagen matrix associated with spindle-shaped hydroxypatite crystals and the second was solely composed by heat-modified apatite crystals. Osteoblast-like cells (Saos-2) were cultured on both biomaterials and examined in scanning and transmission electron microscopy after 7 and 14 days. Both biomaterials were cytocompatible as demonstrated by good ultrastructural cell preservation. (1) At the surface of the collagen containing biomaterial, cells were elongated in shape and oriented according to the trabecular architecture and to the superficial collagen network. After 14 days of culture, cells were confluent and the biomaterial surface was hidden by the cell sheet. The beta 1 integrin subunit was detected by immunogold in transmission electron microscopy in close relationship with the superficial collagen fibres of the biomaterial and with the outer cell surface. When cultures were carried out in presence of anti beta 1 integrin subunit, cells were packed and piled up with lack of specific orientation. (2) At the surface of the deproteinized biomaterial, cells were globular without specific disposition and often partially attached to the surface. After 14 days of culture, large areas of the biomaterial surface remained uncovered. Anti beta 1 subunits conjugated with gold particles were detected around the cells but with no specific association with the deproteinized biomaterial. These results strongly suggest that presence of type I collagen fibres in the matrix of a bone biomaterial is of major interest to determine cell attachment, spreading and orientation via interaction between type I collagen and beta 1 integrin subunit of osteoblasts.

Animals↗

A novel antibacterial titania coating: metal ion toxicity and in vitro surface colonization.

Postoperative implant-associated infection is still an unresolved and serious complication in modern surgery. Antibacterial and biocompatible surfaces could both reduce infection rates and promote tissue integration. In this respect, a comparative study of the antibacterial as well as the biocompatible potential of different metal ions in vitro is presented. The assays used were growth inhibition tests with different metal salts carried out with tissue cells and bacteria under corresponding culture conditions. Additionally, in vitro tests in direct surface contact with tissue cells and bacteria onto a novel copper containing sol-gel derived titanium dioxide coating (Cu-TiO2) and a fourfold Cu-TiO2 coating were performed. The values were compared to a non-filled titanium dioxide coating and standard Ti6Al4V alloy. SEM-investigations were performed to approve the results of the in vitro tests. Among Ag+, Zn2+, Co2+, Al3+ and Hg2+, the growth inhibition tests revealed an outstanding position of copper ions as antibacterial but nevertheless bio-tolerant additive. These results were affirmed by the cell tests in direct surface contact and SEM-investigations, where best cell growth was found on the Cu-TiO2 coatings. Highest antibacterial properties with a tolerable cytocompatibility could be observed on the fourfold Cu-TiO2 coatings. Consequently, surfaces with custom-tailored antibacterial properties may be established and could be of particular interest in revision and tumor arthroplasty.

3T3 Cells↗

Microspheres leaching for scaffold porosity control.

Scaffold morphology plays a key role in the development of tissue engineering constructs. The control of pore size, shape and interconnection is needed to achieve adequate nutrient transport and cell ingrowth. Several techniques are available for scaffold manufacturing, but none allows easy control of morphology and is, at the same time, applicable to a wide variety of materials. To investigate the possibility of processing a wide range polymers by solvent casting/particulate leaching with accurate control of scaffold morphology, three different porogens (gelatin microspheres, paraffin microspheres and sodium chloride crystals) were used to fabricate scaffolds from commonly employed biodegradable polymers. The outcome of processing was evaluated in terms of scaffold morphology and structure/properties relationships. Highly porous scaffolds were obtained with all porogens and well defined spherical pores resulted from microspheres leaching. Furthermore, scaffolds with spherical pores showed better mechanical performance and lower flow resistance. Cytocompatibility tests performed showed no evidence of processing residuals released from the scaffolds. Solvent casting/microspheres leaching, particularly gelatin microspheres leaching, can be used to process a large number of polymers and enables to tailor scaffold pore size, shape and interconnection, thus providing a powerful tool for material selection and optimization of scaffold morphology.

Microscopy, Electron, Scanning↗

Development of a new calcium phosphate powder-binder system for the 3D printing of patient specific implants.

A key requirement for three-dimensional printing (3-DP) of medical implants is the availability of printable and biocompatible powder-binder systems. In this study we developed a powder mixture comprising tetracalcium phosphate (TTCP) as reactive component and beta-tricalcium phosphate (beta-TCP) or calcium sulfate as biodegradable fillers, which can be printed with an aqueous citric acid solution. The potential of this material combination was demonstrated printing various devices with intersecting channels and filigree structures. Two post-processing procedures, a sintering and a polymer infiltration process were established to substantially improve the mechanical properties of the printed devices. Preliminary examinations on relevant application properties including in vitro cytocompatibility testing indicate that the new powder-binder system represents an efficient approach to patient specific ceramic bone substitutes and scaffolds for bone tissue engineering.

3T3 Cells↗

A novel bioactive porous bredigite (Ca7MgSi4O16) scaffold with biomimetic apatite layer for bone tissue engineering.

The aim of this study was to develop a novel bioactive, degradable and cytocompatible bredigite (Ca(7)MgSi(4)O(16)) scaffold with biomimetic apatite layer for bone tissue engineering. Porous bredigite scaffolds were prepared using polymer sponge method. The bredigite scaffolds with biomimetic apatite layer (BTAP) were obtained by soaking bredigite scaffolds in simulated body fluid (SBF) for 10 days. The porosity and in vitro degradability of BTAP scaffolds were investigated. In addition, osteoblast-like cell morphology, proliferation and differentiation on BTAP scaffolds were evaluated and compared with beta-tricalcium phosphate (beta-TCP) scaffolds. The results showed that BTAP scaffolds possessed 90% of porosity. The degradation of BTAP scaffolds was comparable to that of beta-TCP scaffolds. Cells on BTAP scaffolds spread well and presented a higher proliferation rate and differentiation level as compared with those on beta-TCP scaffolds. Our results indicated that BTAP scaffolds were degradable and possessed the function to enhance cell proliferation and differentiation, and might be used as bone tissue engineering materials.

Absorbable Implants↗

Screening of in vitro cytotoxicity by the adhesive film test.

A novel procedure, the adhesive film test, is proposed as a screening method to predict potential cytotoxicity of biomaterials. This in vitro test utilizes a sterile strip of acrylate-based medical adhesive as an anchorage substrate in cytotoxicity studies. The adhesive film allows direct fixation of test samples to the base of the petri dish, ensuring close contact between sample and cells. The test is based on the principle that toxic components present in the test material will readily leach out into the culture medium and adversely affect the local cell population. The main advantage of the adhesive film test is that a viable cell population can be added directly to the test plate and after an incubation period of 24 h, the cellular response can be recorded as either cytotoxic or cytocompatible. Microscopic examination can be followed by quantifying the results using a micrometer to measure cellular attachment areas, migration distances and zones of inhibition. In addition, the adhesive film used to attach the test samples is shown to support extensive fibroblast growth and attachment to its surface and hence can also function as a negative nontoxic control in cytotoxicity studies.

Adhesives↗

In vitro biocompatibility evaluation of a heparinizable material (PUPA), based on polyurethane and poly(amido-amine) components.

The biocompatibility of a new heparinizable material based on polyurethane and poly(amido-amine) (PUPA) was evaluated both in the heparinized and non-heparinized forms. The quantity of heparin present on the material was measured using radiolabelled heparin and biological tests. Heparin release in plasma from heparinized PUPA was investigated using in vitro methods. The behaviour of PUPA towards cellular and plasmatic blood components was studied. The influence of sterilization on the cytocompatibility response of both heparinized and non-heparinized PUPA was investigated; gamma-rays were found to be a suitable method of sterilization as no toxic response was noticed.

Animals↗

Biocompatibility evaluation of glass ionomer cement using cell culture techniques.

Cell culture techniques have been employed to carry out a preliminary investigation into the biocompatibility of a glass ionomer cement (GIC). A modification of the commonly used Agar Overlay test and a rapid, colorimetric assay based on the reduction of tetrazolium salt were used. The GIC is shown to leach a cytotoxic agent, possibly fluoride. This agent is effectively removed by an extraction procedure, and re-examination of the GIC demonstrates its cytocompatibility.

Agar↗

Cell culture methods for testing biocompatibility.

Cell culture systems may be of value in testing the biocompatibility of prosthetic materials before they are introduced into clinical use. In recent years, in vitro methods for assaying biomaterials have gained in importance owing to the growing concern over the use of animals for biomaterials testing. Significant effort is therefore being focused toward developing predictive and quantitative, but also simple and reliable, methods of testing using cultured cells. At present, a number of methods for measuring both the cytotoxicity and the specific cytocompatibility of different materials are available. The usefulness of these systems is no longer confined to screening new materials; they can be used to study the mechanisms of action of various materials during tissue/material interaction. This paper reviews the published literature on the use of cell culture models in evaluating biocompatibility and reports on the personal experience of the authors, who have been using cell culture systems for many years and for different purposes.

Biocompatible Materials↗

Mineralisation of chitosan scaffolds with nano-apatite formation by double diffusion technique.

The study of inorganic crystal assembly in organic matrices has given rise to increasing interest in various fields of materials science to the natural process of biomineralisation. To mimic the formation of hydroxyapatite as natural bone, a double diffusion technique is utilised in this study to nucleate the hydroxyapatite crystals onto three-dimensional porous polymeric scaffolds. The porous polymer scaffolds were produced from chitosan by a thermally induced lyophilisation technique, which yields highly porous, well-controlled anisotropic open pore architecture. The nucleation of hydroxyapatite crystals was initiated at ambient conditions on the surface of the polymer scaffold, which was in contact with a calcium solution chamber, due to diffusion of phosphate ions through the scaffold. The morphology of the mineralised scaffold as analysed by scanning electron microscopy shows that apatite crystals were not only formed on the surface of the scaffold, but also in the pore channels and attached to the pore walls. The X-ray diffraction and Fourier transformed infrared analyses confirmed the phase purity of the formed apatite crystals. The transmission electron microscopy analysis reveals the microstructure of the entangled nano-apatite in the chitosan polymeric matrix. The in-vitro cytocompatibility tests with osteoblast-like cells (Saos-2) demonstrated that the biomineralised scaffold is a suitable substrate for cell attachment and migration in bone tissue engineering.

Apatites↗

Novel injectable calcium phosphate/chitosan composites for bone substitute materials.

In this study, a novel injectable bone substitute material was developed which consists of chitosan, citric acid and glucose solution as the liquid phase, and tricalcium phosphate powder as the solid phase. This material was moldable because of its paste consistency after mixing. We used four groups of cement to investigate the mechanical properties and biocompatibility of the new biomaterial in vitro, which were named group A (10% citric acid), B (15% citric acid), C (20% citric acid) and D (25% citric acid). The setting times of the cements were 5-30 min. X-ray diffraction analysis showed that the products were hydroxyapatite (HA) and dicalcium phosphate anhydrous. When the concentration of citric acid was increased, the compressive strength of specimen increased. Through the simulated body fluid test, we observed the material was bioactive. Group D could induce Ca and P ions to deposit the surface group D quickly. These results indicated that the concentration of citric acid in the liquid component affected the mechanical properties and bioactivity of cements. The cell cultivation test showed that the cytocompatibility of the new biomaterial was good. The method for preparing the novel bone substitute material is simple. The starting material is more readily available and cheaper than HA, poly(methyl methacrylate), and so on. The cement could have good prospects for medical application.

Animals↗

Natural polymers for gene delivery and tissue engineering.

Although the field of gene delivery is dominated by viral vectors and synthetic polymeric or lipid gene carriers, natural polymers offer distinct advantages and may help advance the field of non-viral gene therapy. Natural polymers, such as chitosan, have been successful in oral and nasal delivery due to their mucoadhesive properties. Collagen has broad utility as gene activated matrices, capable of delivering large quantities of DNA in a direct, localized manner. Most natural polymers contain reactive sites amenable for ligand conjugation, cross-linking, and other modifications that can render the polymer tailored for a range of clinical applications. Natural polymers also often possess good cytocompatibility, making them popular choices for tissue engineering scaffolding applications. The marriage of gene therapy and tissue engineering exploits the power of genetic cell engineering to provide the biochemical signals to influence proliferation or differentiation of cells. Natural polymers with their ability to serve as gene carriers and tissue engineering scaffolds are poised to play an important role in the field of regenerative medicine. This review highlights the past and present research on various applications of natural polymers as particulate and matrix delivery vehicles for gene delivery.

Animals↗

Novel chitosan/collagen scaffold containing transforming growth factor-beta1 DNA for periodontal tissue engineering.

The current rapid progression in tissue engineering and local gene delivery system has enhanced our applications to periodontal tissue engineering. In this study, porous chitosan/collagen scaffolds were prepared through a freeze-drying process, and loaded with plasmid and adenoviral vector encoding human transforming growth factor-beta1 (TGF-beta1). These scaffolds were evaluated in vitro by analysis of microscopic structure, porosity, and cytocompatibility. Human periodontal ligament cells (HPLCs) were seeded in this scaffold, and gene transfection could be traced by green fluorescent protein (GFP). The expression of type I and type III collagen was detected with RT-PCR, and then these scaffolds were implanted subcutaneously into athymic mice. Results indicated that the pore diameter of the gene-combined scaffolds was lower than that of pure chitosan/collagen scaffold. The scaffold containing Ad-TGF-beta1 exhibited the highest proliferation rate, and the expression of type I and type III collagen up-regulated in Ad-TGF-beta1 scaffold. After implanted in vivo, EGFP-transfected HPLCs not only proliferated but also recruited surrounding tissue to grow in the scaffold. This study demonstrated the potential of chitosan/collagen scaffold combined Ad-TGF-beta1 as a good substrate candidate in periodontal tissue engineering.

Adenoviridae↗

Hydrogen plasma surface activation of silicon for biomedical applications.

Silicon has gradually been recognized to be an essential trace element in the normal metabolism of higher animals, and the role of silicon in the human body has aroused interests in the biomedical community. In fact, the interactions between silicon-based devices and the human body such as biosensors and microelectromechanical systems (MEMS) often suffer from poor biocompatibility. In this work, hydrogen plasma immersion ion implantation (H-PIII) is conducted to improve the bioactivity or bone conductivity of silicon. In order to investigate the formation mechanism of bone-like apatite on the surface of the hydrogen implanted silicon wafer, two comparative experiments, hydrogenation and argon bombardment, are performed. The H-PIII sample exhibits an amorphous surface consisting of Si-H bonds. After immersion in simulated body fluids, a negatively charged surface containing the functional group ([triple bond]Si-O-) is produced and bone-like apatite is observed to nucleate and grow on the surface. The surface of the H-PIII silicon wafer favors the adhesion and growth of osteoblast cells and good cytocompatibility may be inferred.

Biomedical Engineering↗

In situ crosslinkable hyaluronan hydrogels for tissue engineering.

We describe the development of an injectable, cell-containing hydrogel that supports cell proliferation and growth to permit in vivo engineering of new tissues. Two thiolated hyaluronan (HA) derivatives were coupled to four alpha,beta-unsaturated ester and amide derivatives of poly(ethylene glycol) (PEG) 3400. The relative chemical reactivity with cysteine decreased in the order PEG-diacrylate (PEGDA)>>PEG-dimethacrylate>PEG-diacrylamide>PEG-dimethacrylamide. The 3-thiopropanoyl hydrazide derivative (HA-DTPH) was more reactive than the 4-thiobutanoyl hydrazide, HA-DTBH. The crosslinking of HA-DTPH with PEGDA in a molar ratio of 2:1 occurred in approximately 9 min, suitable for an in situ crosslinking applications. The in vitro cytocompatibility and in vivo biocompatibility were evaluated using T31 human tracheal scar fibroblasts, which were suspended in medium in HA-DTPH prior to addition of the PEGDA solution. The majority of cells survived crosslinking and the cell density increased tenfold during the 4-week culture period in vitro. Cell-loaded hydrogels were also implanted subcutaneously in the flanks of nude mice, and after immunohistochemistry showed that the encapsulated cells retained the fibroblast phenotype and secreted extracellular matrix in vivo. These results confirm the potential utility of the HA-DTPH-PEGDA hydrogel as an in situ crosslinkable, injectable material for tissue engineering.

Animals↗

Preparation of acrylic grafted chitin for wound dressing application.

Chitin grafted with poly(acrylic acid) (chitin-PAA) was prepared with the aim of obtaining a hydrogel characteristic for wound dressing application. The chitin-PAA films were synthesized at various acrylic acid feed contents to investigate its effect on water sorption ability. Acrylic acid (AA) was first linked to chitin, acting as the active grafting sites on the chain that was further polymerized to form a network structure. The evidences of grafting were found from FTIR and solid state 13C NMR spectra. The TGA results exhibited the high degradation temperature of the grafted product suggesting the formation of a network structure. The degree of swelling (DS) of chitin-PAA films was found in the range of 30-60 times of their original weights depending upon the monomer feed content. The chitin-PAA film with 1:4 weight ratio of chitin:AA, possessed optimal physical properties. The cytocompatibility of the film was investigated with a cell line of L929 mouse fibroblasts. The morphology and behavior of the cells on the chitin-PAA film were determined after different time periods of culture up to 14 days. The L929 cells proliferated and attached well onto the film. These results suggested that the 1:4 chitin-PAA has a potential to be used as a wound dressing.

Acrylic Resins↗

Electrospinning of silk fibroin nanofibers and its effect on the adhesion and spreading of normal human keratinocytes and fibroblasts in vitro.

An electrospinning method was used to fabricate silk fibroin (SF) nanofiber nonwovens for cell culture of normal human keratinocytes and fibroblasts. The electrospinning of regenerated SF was performed with formic acid as a spinning solvent. For insolubilization, as-spun SF nanofiber nonwovens were chemically treated with an aqueous methanol solution of 50%. Morphology and microstructure of as-spun and chemically treated SF nanofibers were investigated by scanning electron microscopy and mercury porosimetry. As-spun SF nanofibers exhibited a circular cross-section with a smooth surface. From the image analysis, they had an average diameter of 80 nm and their diameters ranged from 30 to 120 nm. During the chemical treatment for 60 min, porosity of nonwovens composed of SF nanofibers decreased from 76.1% up to 68.1%. To assay the cytocompatibility and cell behavior onto the electrospun SF nanofibers, cell attachment and spreading of normal human keratinocytes and fibroblasts seeded on the SF nanofibers and interaction between cells and SF nanofibers were studied. Cell morphology on SF nanofibers was examined by scanning electron microscopy. Our results indicate that the SF nanofibers may be a good candidate for the biomedical applications, such as wound dressing and scaffolds for tissue engineering.

Adolescent↗