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At least 163 records · Page 9Linked to original sources
PubMed2004

[Study on the interface of human hepatocyte L-02 polypropylene:simple culture method of human hepatocyte with spheroidal aggregate culture].

OBJECTIVE: To found new interface of human hepatocyte/poly propylene with good cytocompatibility for made polypropylene hollow fibers bioreactor of bioartificial liver in future. METHODS: Using the macromolecular hydroperoxide groups on the polypropylene membrane surface as initiators, acrylamides were polymerized on the polypropylene membranes, under induction by both UV irradiation and Fe2+ reduction. Growth characteristics of human hepatocyte L-02 were detected when it was cultured on polystyrene, polypropylene and modified polypropylene membrane surface. RESULTS: Water contact angle measurement of the polypropylene and the modified polypropylene membranes decreased from (72 +/- 5) degrees to (30 +/- 4) degrees , which indicated that the hydrophilicity of the membrane was improved obviously after the grafting modification. Human hepatocyte L-02 could not adhere and spread on modified polypropylene membrane surface, and grown in spheroidal aggregate with higher density and higher proliferation ratio measured by MTT method. CONCLUSIONS: Acrylamide polymerized on the polypropylene membranes is a good method which not only improved human hepatocytes cytocompatibility but also found a new simple culture method with spheroidal aggregate culture of human hepatocyte.

Cell Culture Techniques↗
PubMed2004

[Study on the interface of human hepatocyte/micropore polypropylene ultrafiltration membrane].

OBJECTIVE: To found a new interface of human hepatocyte/micropore polypropylene ultrafiltration membrane (MPP) with good cytocompatibility so as to construct bioartificial bioreactor with polypropylene hollow fibers in future. METHODS: MPP ultrafiltration membrane underwent chemical grafting modification through ultraviolet irradiation and Fe(2+) reduction. The contact angles of MPP and the modified MPP membranes were measured. Human hepatic cells L-02 were cultured. MPP and modified MPP membranes were spread on the wells of culture plate and human hepatic cells and cytodex 3 were inoculated on them. Different kinds of microscopy were used to observe the morphology of these cells. RESULTS: The water contact angle of MPP and the modified MPP membranes decreased from 78 degrees +/- 5 degrees to 27 degrees +/- 4 degrees (P < 0.05), which indicated that the hydrophilicity of the membrane was improved obviously after the grafting modification. Human hepatocyte L-02 did not adhere to and spread on the modified MPP membrane surface, and only grew on the microcarrier cytodex 3 with higher density and higher proliferation ratio measured by MTT. CONCLUSION: Grafting modification of acrylamide on MPP membrane is a good method to improve the human hepatocyte cytocompatibility with MPP ultrafiltration membrane.

Bioartificial Organs↗
PubMed2005

[Experimental research on the effect of nanophase ceramics on osteoblasts functions].

In order to study the cytocompatibility of nanophase hydroxyapatite ceramic in vitro, we prepared hydroxyapatite by use of the wet chemistry techniques. The grain size of hydroxyapatite of interest to the present study was determined by scanning electron microscopy and atomic force microscopy with image analysis software. Primary culture of osteoblast from rat calvaria was established. Protein content, synthesis of alkaline phosphatase and deposition of calcium-containing mineral by osteoblasts cultured on nanophase hydroxyapatite ceramics and on conventional hydroxyapatite ceramics for 7, 14, 21 and 28 days were examined. The results showed that the average surface grain size of the nanophase and that of the conventional HA compact formulations was 55 (nanophase) and 780 (conventional) nm, respectively. More importantly, compared to the synthesis of alkaline phosphatase and deposition of calcium-containing mineral by osteoblasts cultured on nanophase was significantly greater than that on conventional ceramics after 21 and 28 days. The cytocompatibility was significantly greater on nanophase HA than on conventional formulations of the same ceramic.

Alkaline Phosphatase↗
PubMed1994

[Assays of cytotoxicity of the Nickel-Titanium shape memory alloy].

The equiatomic Nickel-Titanium (NiTi) alloy has exceptional mechanical properties such as shape memory and superelasticity. It already has applications in orthodontics and is a promising orthopaedic biomaterial. Cytocompatibility studies must therefore be undertaken. The objective of this study is to determine the biological response that NiTi elicits compared to other orthopaedic metals currently used in orthopaedic surgery. Cytotoxicity tests constitute an efficient first step in a biocompatibility study and contribute to reduce animal use in laboratory. Direct contact and agar diffusion cytotoxicity assays were performed following ASTM standards #F813-83 and #F895-84 respectively. Confluent L-929 fibroblasts culture plates were incubated (directly or under an agar bed) in presence of NiTi, titanium (Ti), vitallium (Co-Cr-Mo) and 316L stainless steel discs. Following exposition to specimens, a vital dye was added to the plates. All cultures were evaluated for cytotoxic reactions, under light microscopy. Direct contact and agar diffusion assays indicated that all metals tested induced a mild biological reaction. Specimens were ranked according to an index of biological response, they are enumerated here in decreasing order of cytotoxicity: NiTi approximately Co-Cr-Mo >> pure grade 4 Ti approximately pure grade 1 Ti approximately Ti 6A1 4V approximately 316L stainless steel. Furthermore, plasma surface modification increased the cytocompatibility of NiTi.

Biocompatible Materials↗
PubMed1997

[Cultures of corneal epithelium on hydrogel for synthetic epikeratoplasty].

PURPOSE: To improve the LLH480 hydrogel cytocompatibility and to assess the interactions between corneal epithelium and biomaterials. METHODS: Rabbit corneal epithelium organotypic cultures were carried out on hydrogel samples coated with fibronectin, laminin, type IV collagen, or heparan sulfate proteoglycan. The control group consisted of epithelial cultures carried out on hydrogel with no coating. Cellular migration was quantified, and statistically analyzed. 8-day cultures were processed for transmission electron microscopy. RESULTS: Epithelial migration on hydrogel was significantly increased (p < 0.001) in the heparan sulfate proteoglycan group, significantly decreased (p < 0.05 and p < 0.001) with laminin or type IV collagen coating, and showed no significant modifications in the fibronectin group. Transmission electronic microscopy showed a monolayer of epithelial cells adherant to the surface of the hydrogel. Deposit of extracellular matrix and sketch of adhesion focal points were also noted. CONCLUSION: LLH480 cytocompatibility can be improved by heparan sulfate proteoglycan coating.

Animals↗
PubMed1998

Evaluation of a novel alginate gel dressing: cytotoxicity to fibroblasts in vitro and foreign-body reaction in pig skin in vivo.

Calcium alginate dressings have beneficial effects on wound healing by providing a moist wound environment. However, cytotoxicity and the nonbiodegradable nature of calcium alginate dressings induce unresolved chronic foreign-body reaction. In this study, a novel freeze-dried alginate gel dressing (AGA-100) low in calcium ions was evaluated for cytotoxicity to L929 cells in vitro and in full-thickness pig wounds in vivo. Cytotoxicity testing on L929 cells showed the cytocompatibility of AGA-100 extracts, while extracts from Kaltostat, a well-established alginate dressing, induced cytopathic effects. In an in vivo study using pigskin, AGA-100, Kaltostat, and gauze were applied on 1-in-diameter circular full-thickness wounds on the back of pigs and the time course of wound closure was evaluated. Kaltostat and gauze dressings were used as controls. For histologic evaluation, wound tissue was harvested on day 18. AGA-100-treated wounds showed rapid wound closure compared to control wounds on day 15. Foreign-body reaction was marked in Kaltostat- and gauze-treated wounds, and differed significantly from AGA-100-treated wounds. Based on these data, AGA-100 could reduce the cytotoxicity to fibroblasts and foreign-body reaction that have been observed with currently available calcium alginate dressings; it was also found to be useful as an alginate dressing.

Alginates↗
PubMed1998

First use of cultured human urothelial cells for biocompatibility assessment: application to urinary catheters.

For several years, studies performed to estimate in vitro biocompatibility of urinary catheters have been carried out using permanent cell lines. But for a rational design of the testing procedure, the cell culture model should relate to the material application. This work presents the results of a probe study designed to obtain an in vitro model of normal human urothelial cells (HUC) and to test the relevance of this system in cytocompatibility experiments of urinary catheters currently used.A comparison is made with continuous cell lines, the use of which is recommended by normalization bodies. We exposed monolayers of HUC (well characterized for their proliferation, qualitative evaluation, and quantitative measurement of cytokeratins) and two continuous human cell lines to liquid extracts (either pure or diluted in the culture medium) of nine available catheters, including positive (latex) and negative controls, for a 24 h incubation. Then colorimetric assays (Neutral Red and MTT) were performed. The extracts of two polyurethanes provoked a significant toxic effect on HUC only, suggesting differences in sensitivity between the models used. This effect could be due to the presence of a great amount of barium (used as a radioopacifier) in extracts, as highlighted by results of absorption emission spectroscopy. A culture model of HUC may be of relevance for the screening of materials intended for urological practice.

Biocompatible Materials↗
PubMed1998

Evaluation of proliferation and protein expression of human bone marrow cells cultured on coral crystallized in the aragonite of calcite form.

The two crystalline forms of CaCO3, aragonite (from natural coral) and calcite (from natural limestone), have been used with success as bone graft substitutes. However, natural coral transformed into calcite by heating has never been tested. The objective of this work was to study the proliferation and alkaline phosphatase, osteonectin, and osteocalcin expression of human bone marrow cells cultured on CaCO3 crystallized both in the aragonite form (natural coral) and in the calcite form (natural coral modified by heating). The methods used to characterize calcite obtained from the coral were volumic porosimetry, scanning electron microscopy (SEM) and X-ray diffraction. Cell colonization of the material was assessed by SEM performed on days 1, 7, 20, and 30 and [3H]thymidine incorporation was performed on days 3, 7, 12, 18, 25, and 32. Phenotypic expression was assessed by using in situ cytochemistry (alkaline phosphatase), immunocytochemistry (osteonectin and osteocalcin), and hybridization (osteocalcin, beta-actin, and alkaline phosphatase mRNA). Results showed the transformation of aragonite into calcite after heating, the conservation of macroporosity, and a modification of the surface. Calcite appeared to have a smoother and more uniform surface than aragonite crystals. As for [3H]thymidine there was an increase incorporation from days 3 to 18, a stabilization from days 18 to 25, and a decrease from days 25 to 32. After 20 days of culture, immunological studies using monoclonal antibodies to osteocalcin, osteonectin, cytochemical analysis of alkaline phosphatase activity, and in situ hybridization using osteocalcin, beta-actin, and alkaline phosphatase cDNA indicated that the cells had not lost their osteoblastic phenotype. These experiments demonstrate that coral crystallized in the aragonite or calcite form present a similar degree of specific cytocompatibility.

Animals↗
PubMed2003

Photocrosslinked hyaluronic acid hydrogels: natural, biodegradable tissue engineering scaffolds.

Ideally, rationally designed tissue engineering scaffolds promote natural wound healing and regeneration. Therefore, we sought to synthesize a biomimetic hydrogel specifically designed to promote tissue repair and chose hyaluronic acid (HA; also called hyaluronan) as our initial material. Hyaluronic acid is a naturally occurring polymer associated with various cellular processes involved in wound healing, such as angiogenesis. Hyaluronic acid also presents unique advantages: it is easy to produce and modify, hydrophilic and nonadhesive, and naturally biodegradable. We prepared a range of glycidyl methacrylate-HA (GMHA) conjugates, which were subsequently photopolymerized to form crosslinked GMHA hydrogels. A range of hydrogel degradation rates was achieved as well as a corresponding, modest range of material properties (e.g., swelling, mesh size). Increased amounts of conjugated methacrylate groups corresponded with increased crosslink densities and decreased degradation rates and yet had an insignificant effect on human aortic endothelial cell cytocompatibility and proliferation. Rat subcutaneous implants of the GMHA hydrogels showed good biocompatibility, little inflammatory response, and similar levels of vascularization at the implant edge compared with those of fibrin positive controls. Therefore, these novel GMHA hydrogels are suitable for modification with adhesive peptide sequences (e.g., RGD) and use in a variety of wound-healing applications.

Absorbable Implants↗
PubMed2007

Fabrication of agar-gelatin hybrid scaffolds using a novel entrapment method for in vitro tissue engineering applications.

Scaffolds of agar and gelatin were developed using a novel entrapment method where agar and gelatin molecules mutually entrapped one another forming stable cell adhesive matrices. Glutaraldehyde was used as a crosslinking agent for gelatin. Three types of hybrid matrices were prepared using agar and gelatin in different proportions in the weight ratio of 1:1, 2:1, and 3:1. Surface characterization of dry scaffolds was carried out by scanning electron microscope. Swelling studies were carried out in phosphate buffer saline (PBS) at physiological pH 7.4. The integral stability of the scaffolds was evaluated by estimating the released disintegrated gelatin from them in PBS at pH 7.4. The attachment kinetics of the cells was evaluated by culturing mouse fibroblast cell line NIH 3T3 on films. The cytocompatibility of these matrices was determined by studying growth kinetics of NIH 3T3 cells on them and morphology of cells was observed through optical photographs taken at various days of culture. It was found that the matrices containing agar and gelatin in 2:1 weight ratio exhibited best growth kinetics. The results obtained from these studies have suggested that the above-described method is a cheap and easy way to fabricate agar-gelatin hybrid scaffolds to grow cells which can be used in various in vitro tissue engineering applications like screening of drugs.

Agar↗
PubMed2002

Hydroxylapatite with substituted magnesium, zinc, cadmium, and yttrium. II. Mechanisms of osteoblast adhesion.

The present in vitro study investigated osteoblast adhesion on hydroxylapatite (HA) doped with either cadmium (Cd), zinc (Zn), magnesium (Mg), or yttrium (Y). Compared with any other dopant tested in the present study, osteoblast adhesion was significantly (p < 0.05) greater on HA doped with Y after 4 h; in addition, osteoblast adhesion increased with concentration (2-7 mol%) of Y in HA. The findings that HA doped with greater amounts of Y adsorbed higher concentrations of calcium and, subsequently, of vitronectin and collagen (proteins known to mediate osteoblast adhesion), but not of albumin, laminin, and fibronectin, may explain the observed enhanced adhesion of osteoblasts on these substrates. Interactions (i.e., adsorption and configuration/bioactivity) of vitronectin and collagen may have been promoted by increased porosity of doped HA. Through doping with Y, the present study provided the first evidence that HA can be synthesized and processed with improved cytocompatibility properties for osteoblast adhesion, and thus offered essential information for the design of novel proactive bioceramics. Proactive bioceramics which elicit specific, timely, and desired responses from surrounding cells and tissues are necessary for improving bonding of orthopaedic/dental implants to juxtaposed bone; such osseointegration will, undoubtedly, enhance implant efficacy.

Adsorption↗
PubMed2002

Fabrication of poly(alpha-hydroxy acid) foam scaffolds using multiple solvent systems.

The present studies describe the fabrication and characterization of highly porous and interconnected poly(alpha-hydroxy acid) foam scaffolds produced using a phase separation multisolvent system, followed by a sublimation process. Fabrication parameters, including solvent composition, polymer concentration, freezing temperature, polymer type, and polymer molecular weight, were optimized to produce the desired foam microstructure. Analyses of selected samples with scanning electron microscopic images and mercury intrusion porosimetry indicated polymer foams with pore size ranges of 100-350 microm, a porosity >90%, and an interconnecting open-pore foam structure. Scaffold degradation profiles varied according to the type and molecular weight of the polymers. Cytocompatibility assays demonstrated that the preferred foam structures were nontoxic and osteoprecursor cells seeded into the scaffolds exhibited the ability to attach, propagate, and differentiate into a calcified structure.

Absorbable Implants↗
PubMed1990

Cytotoxicity testing of wound dressings using normal human keratinocytes in culture.

Comparative cytotoxicity testing of 16 wound dressings of different composition show that normal human keratinocytes (NHK) growing on a fibroblastic feeder layer are as sensitive to toxic materials by direct contact as the confluent MRC5 fibroblasts used for standard cell culture cytotoxicity testing, and slightly more sensitive when extracts of the dressings were tested. After direct contact with each of the cell types, we found effects due to 12 dressing samples (75%), but the extracts of only 6 of them induced changes in cell shape or cell death on NHK, and 4 of them on MRC5 cells. In order to assess the compatibility of these dressings with a pure population of epidermal cells, the cell type responsible for reepidermization of healing wounds, we then tested the sensitivity, both to dressing samples and extracts, of normal human keratinocytes (NHK) grown in chemically defined medium and without a feeder layer: The results show epidermal cytocompatibility of 10 dressing extracts, while 6 others induced cytopathic effects. Three of these extracts specifically damaged epidermal cells and inhibited their proliferation. When comparing the sensitivities of NHK (in defined medium) and MRC5 cells, we observed complete correlation for 75% of the dressings by extract testing and in 94% of the cases after direct contact.

Bandages↗
PubMed1993

Endothelial cell compatibility testing of three different Pellethanes.

There is a need for viable small diameter vascular grafts, the luminal surface of which could be seeded by endothelial cells (ECs) to prevent thrombosis. In order to select candidates for EC seeding before implantation, the in vitro cytocompatibility of three different Pellethanes (polyetherurethanes) using human ECs was investigated. The methodology included two stages depending on either direct contact between cells and materials or contact between cells and material extracts, obtained under standardized conditions. By the latter method, we observed a cytotoxic effect on cell growth with 2363-55 D Pellethane extract at a 50% (v/v) concentration in the nutrient medium, likely provoked by leachables and correlated with the lowest levels of tPA, PAI1, and vWF antigens in the supernatants. By the former method, we studied EC attachment and growth. Morphology was studied by classical means and completed by scintigraphy and microautoradiography after 111Indium-labeling of the EC monolayer. Differentiation was determined by the release of vWF antigen and measurement of vWF activity (multimeric organization) after human thrombin stimulation. Despite an inhibition of proliferation for both 55 D and 75 D types (compared to the control), a functional monolayer of ECs was obtained on 75 D. Pellethane 75 D could be the best support for in vitro endothelization.

Blood Vessel Prosthesis↗
PubMed2003

Biomimetic surface modification of poly (L-lactic acid) with gelatin and its effects on articular chondrocytes in vitro.

Our objective in this study was to investigate the efficiency of two treatments for poly (L-lactic acid) (PLLA) surface modification with gelatin, via entrapment and coupling, using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The properties of original PLLA, gelatin-entrapped, and coupled PLLA films were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). The water contact angle indicated that the incorporation of gelatin resulted in a change in hydrophilicity, and the ESCA data suggested that the modified PLLA films became enriched with nitrogen atoms. The cytocompatibility of modified PLLA films might be improved. Therefore, we examined the attachment and proliferation of bovine articular chondrocyte seeded on modified PLLA films and virgin films. A whole-cell enzyme-linked immunosorbent assay (cell ELISA) that detects 5-bromo-2'-deoxyuridine (BrdU) incorporation during DNA synthesis and collagen type II secretion was applied to evaluate the chondrocytes on different PLLA films and tissue culture plates (TCPS). Cell viability was estimated by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay, and cell function was assessed by measuring glycosaminoglycan (GAG) secreted by chondrocytes. These results implied that gelatin used to modify the PLLA surface through entrapment and coupling could enhance chondrocyte adhesion, proliferation, and function.

Animals↗
PubMed2003

In vitro evaluation of degradation and cytotoxicity of a novel composite as a bone substitute.

The purpose of this study was to prepare and evaluate in vitro the feasibility and cytocompatibility of a novel composite (GGT) as a large defect bone substitute. The composite is tricalcium phosphate ceramic particles combined with genipin crosslinked gelatin. After soaking the GGT composites in Ringer solutions at 37 degrees C for 7, 14, 28, 42, 56, and 84 days, the in vitro biologic degradation rate and biocompatibility were determined. Substances released from soaked GGT composites were analyzed with an ultraviolet visible light spectrophotometer. In addition, the solution soaking the GGT was co-cultured with osteoblasts to determine whether or not the released substances from GGT could facilitate the growth of bone cells. After they had been cultured for 2 days, the osteoblasts were tested for differentiation and proliferation by alkaline phosphatase (ALP) activity and a MTT assay. Results indicate that the concentration of the genipin solution is a critical factor in deciding the crosslinking degree of the GGT composite. Complete crosslinking reaction in the GGT composite occurred when 0.5 wt % of genipin had been added. Cytotoxic testing revealed that 80 ppm of the genipin in the culture medium served as the level over which cytotoxicity to osteoblasts could be produced. In addition, we found that gelatin and calcium continuously were released from the GGT composite in the soaking solution, which promoted differentiation and proliferation of the osteoblasts.

Alkaline Phosphatase↗
PubMed2004

Collagen-hydroxyapatite composite prepared by biomimetic process.

A novel bone graft substitute comprising a porous, collagenous scaffold was biomimetically coated with hydroxyapatite using a simulated body fluid solution chemistry method. The scaffold had a porosity of approximately 85%, with pore sizes between 30 microm and 100 microm. Glutaraldehyde vapor was used to stabilize the collagenous scaffold, giving a significantly increased thermal stability over an unstabilized scaffold, as shown by differential scanning calorimetry. A thin layer (<10 microm) of crystalline hydroxyapatite was deposited onto the stabilized collagenous scaffold by soaking the collagenous construct in simulated body fluid in the presence of calcium silicate glass. The presence of crystalline hydroxyapatite was confirmed by X-ray diffraction, energy-dispersive X-ray spectroscopy, and scanning electron microscopy. In vitro cytotoxicity testing of the composite construct using L-929 fibroblasts (ISO 10993-5) and rabbit periosteal cells revealed a cytocompatible material that supported cellular attachment and proliferation.

Animals↗
PubMed2003

Polymers with nano-dimensional surface features enhance bladder smooth muscle cell adhesion.

Previous studies investigating the design of synthetic bladder wall substitutes have involved polymers with micro dimensional structures. Since the body is made up of nano-structured components (e.g., extracellular matrix proteins), the focus of the present in vitro study was to design nano-structured polymers for use as synthetic bladder constructs that mimic the topography of natural bladder tissue. In order to complete this task, novel nano-structured biodegradable polymeric films of poly-lactic-co-glycolic-acid (PLGA), poly-ether-urethane (PU), and poly-caprolactone (PCL) were fabricated and separately treated with various concentrations of NaOH (for PLGA and PCL) and HNO(3) (for PU) for select time periods. These treatments reduced the polymer surface feature dimensions from conventional micron dimensions to biologically inspired nanometer dimensions. Select cytocompatibility properties of these biomaterials were tested in vitro. Results provide the first evidence that adhesion of bladder smooth muscle cells is enhanced as polymer surface feature dimensions are reduced into the nanometer range. In addition, surface analysis results reveal that the polymer nanometer surface roughness is the primary design parameter that increases bladder smooth muscle cell adhesion. For this reason, the "next generation" of tissue-engineered bladder constructs with increased efficacy should contain surfaces with nanometer (as opposed to micron) surface features.

Animals↗