Biomedical subjects

J A Jansen

Publications and source records attributed to J A Jansen.

At least 19 recordsLinked to original sources

PubMed2002

Initial interaction of U2OS cells with noncoated and calcium phosphate coated titanium substrates.

From previous studies, we know that calcium phosphate (CaP) coated implants stimulate bone formation compared to uncoated implants. Nevertheless, the mechanism by which substrate surface characteristics affect cell function is unclear. In this study, we examined the initial interaction (30 min to 24 h) of U2OS cells with titanium substrates with or without a CaP coating. The effect of substrate roughness was also studied. When cell attachment was studied, we found that cells attached more readily to rough than to smooth surfaces. Also, more cells attached to the uncoated than to the CaP coated surface. After 24 h, cell numbers were similar for all substrate surfaces. Further, cells spread to a larger area on noncoated titanium than on the CaP coated substrates. At 24 h, the sequence of cell size was smooth titanium > rough titanium > CaP coated titanium. Shape measurements showed differences in cell shape between the cells on the different materials only at 7 h, not at different culture times. Cells expressed alpha2, alpha3, alpha5, alpha6, alphav, and beta1 subunits. Expression of alpha1, alpha4, alphavbeta3, beta3, beta4, and beta7 was extremely low or was not found. The beta1 integrin expression was higher on the coated than on the noncoated titanium at 3 h, but not on the other studied times. Expression of alpha2, alpha5, alpha6, and alphav expression was found to be upregulated at 24 h compared to earlier culture times on coated titanium, but not on uncoated titanium substrates. From this we conclude that the surface characteristics of a material (roughness and composition) can affect the initial interaction of cells with the material.

Calcium Phosphates↗

PubMed2002

Histological characterization of the early stages of bone morphogenetic protein-induced osteogenesis.

On the basis of currently available knowledge, we hypothesize that the initial bone formation, as induced by bone morphogenetic protein (BMP), is influenced by the chemical composition and three-dimensional spatial configuration of the used carrier material. Therefore, in the current study, the osteoinductive properties of porous titanium (Ti) fiber mesh with a calcium phosphate (Ca-P) coating (Ti-CaP), insoluble bone matrix (IBM), fibrous glass membrane (FGM), and porous particles of hydroxy apatite (PPHAP) loaded with rhBMP-2 were compared in a rat ectopic assay model at short implantation periods. Twelve Ti-CaP, 12 IBM, 12 FGM, and 12 PPHAP implants, loaded with rhBMP-2, were subcutaneously placed in 16 Wistar King rats. The rats were sacrificed at 3, 5, 7, and 9 days post-operative, and the implants were retrieved. Histological analysis demonstrated that IBM and Ti-CaP had induced ectopic cartilage and bone formation by 5 and 7 days, respectively. However, in PPHAP, bone formation and cartilage formation were seen together at 7 days. At 9 days, in Ti-CaP, IBM, and PPHAP, cartilage was seen together with trabecular bone. At 9 days, in FGM, only cartilage was observed. Quantitative rating of the tissue response, using a scoring system, demonstrated that the observed differences were statistically significant (Wilcoxon rank sum test, p < 0.05). We conclude that IBM, CaP-coated Ti mesh, FGM, and PPHAP provided with rhBMP-2 can indeed induce ectopic bone formation with a cartilaginous phase in a rat model at short implantation periods. Considering the different chemical composition and three-dimensional spatial configuration of the carrier materials used, these findings even suggest that endochondral ossification is present in rhBMP-2-induced osteogenesis, even though the amount of cartilage may differ.

Animals↗

PubMed2002

[Subcutaneously implantable glucose sensors in patients with diabetes mellitus; still many problems].

Despite considerable scientific efforts, no clinical method is currently available for the continuous monitoring of glucose in subcutaneous tissue fluid. In general, good results were obtained during in-vitro experiments with various implantable glucose sensors. However, after implantation these devices exhibited a progressive loss of sensor function. It is evident that the tissue reaction to the implanted sensor, especially the interactions at the sensor-tissue interface, plays an important role in this loss of function. Adequate strategies to improve in-vivo sensor performance can only be developed if there is a better understanding of the processes involved in sensor inactivation.

Biosensing Techniques↗

PubMed2002

Transforming growth factor-beta 1, 2, and 3 can inhibit epithelial tissue outgrowth on smooth and microgrooved substrates.

In this study, we describe the influence of parallel surface microgrooves, and of TGF-beta, on the outgrowth of corneal epithelial tissue. Microgrooves (depth 1 microm, width 1-10 microm) were made in polystyrene culturing surfaces. These surfaces were left untreated, or loaded with TGF-beta 1, 2, or 3 (6.0 ng/cm(2)). Subsequently, epithelial explants from bovine corneas were placed on the experimental surfaces. After 9 days of culturing, tissue outgrowth was evaluated. Furthermore, the tissue cultures were analyzed histologically. It was shown that epithelial tissue grew from the explants over all experimental surfaces. On microgrooved surfaces outgrowth proceeded in the direction of the grooves, rather than perpendicular to the grooves. The addition of each type of TGF-beta resulted in a reduction of outgrowth. However, outgrowth remained directed by the grooves. Further, the explants had shrunk after TGF treatment. Histology showed that this shrinkage was not related to alpha-smooth muscle actin expression in the explants. We conclude that microgrooves can direct, and TGF-betas can inhibit the outgrowth of epithelial tissue. This finding could be useful in biomaterial applications where the growth of epithelial tissue needs to be discouraged.

Animals↗

PubMed2002

Trabecular bone response to injectable calcium phosphate (Ca-P) cement.

The aim of this study was to investigate the physicochemical, biological, and handling properties of a new developed calcium phosphate (Ca-P) cement when implanted in trabecular bone. Ca-P cement consisting of a powder and a liquid phase was implanted as a paste into femoral trabecular bone of goats for 3 days and 2, 8, 16, and 24 weeks. The cement was tested using three clinically relevant liquid-to-powder ratios. Polymethylmethacrylate bone cement, routinely used in orthopedics, was used as a control. The Ca-P cement was easy to handle and was fast setting with good cohesion when in contact with body fluids. X-ray diffraction at the different implantation periods showed that the cement had set as an apatite and remained stable over time. Histological evaluation after 2 weeks, performed on 10 microm un-decalcified sections, showed abundant bone apposition on the cement surface without any inflammatory reaction or fibrous encapsulation. At later time points, the Ca-P cement implants were totally covered by a thin layer of bone. Osteoclast-like cells, as present at the interface, had resorbed parts of the cement mass. At locations where Ca-P cement was resorbed, new bone was formed without loss of integrity between the bone bed and the cement. This demonstrated the osteotransductive property of the cement, i.e., resorption of the material by osteoclast-like cells, directly followed by the formation of new bone. Histological and histomorphometrical evaluation did not show any significant differences between the Ca-P cement implanted at the three different liquid/powder ratios. The results indicate that the investigated Ca-P cement is biocompatible, osteoconductive, as well as osteotransductive and is a candidate material for use as a bone substitute.

Animals↗

PubMed2002

Effect of calcium phosphate coating crystallinity and implant surface roughness on differentiation of rat bone marrow cells.

In this study, we examined the effect of calcium phosphate (Ca-P) coating crystallinity and of surface roughness on growth and differentiation of osteogenic cells. Grit-blasted titanium substrates were provided with Ca-P coatings of different crystallinities. Rat bone marrow (RBM) cells were cultured on these substrates and on noncoated rough and smooth titanium substrates. After specific culture times, expression of osteogenic markers by the cells was studied. Cells cultured on crystalline coatings and on titanium substrates proliferate, express alkaline phosphatase, osteocalcin (OC), and show mineralization of the extracellular matrix. Rough titanium substrates only express low OC levels. Significantly higher OC levels were expressed on smooth titanium, and even higher levels on the crystalline Ca-P coating. No difference was found in calcification between smooth and rough titanium. The crystalline coating showed more calcification than the titanium substrates. When substrates without cells were incubated in medium, precipitation of calcium was found. On the titanium substrates, this precipitate disappeared after prolonged incubation. The precipitate on the crystalline coating was stable and increased with longer incubation times. On the amorphous coatings, no proliferation and differentiation of RBM cells were found. After longer culture periods, substrates showed extensive dissolution. Cells on the amorphous coatings did express high levels of prostaglandin E2. In contrast, prostaglandin E2 expression was low for the other substrates. We conclude that crystalline Ca-P coatings stimulate differentiation of RBM cells, to a higher extent than titanium substrates. Surface roughness only has a limited effect on phenotype expression of the cells. In contrast, thin amorphous coatings show negative effects on the growth and differentiation of cultured RBM cells.

Alkaline Phosphatase↗

PubMed2002

Soft-tissue response to silicone and poly-L-lactic acid implants with a periodic or random surface micropattern.

Tissue reaction to biomaterials is dependent on properties such as surface topography. The aim of this study was to evaluate the tissue reaction around implants with different surface topographies. We made coin-shaped silicone and poly-L-lactic acid (PLLA) implants with double-sided parallel microgrooves (depth 1.0 microm; width 10.0 microm) and random roughness on a micrometer scale. The control implants were smooth. These implants were inserted into subcutaneous pockets created on the flanks of goats. After 1, 3, or 12 weeks, the goats were sacrificed and the implants retrieved and histologically processed. Light microscopic evaluation revealed the formation of fibrous tissue capsules around all implant materials. The PLLA did not visibly degrade during the study period. Histomorphometric analyses were performed on capsule thickness, capsule quality, and on the implant-tissue interface quality. Compared with the silicone implants, the capsules around the PLLA implants showed significantly better capsule quality. Compared to the smooth implants, the capsules around the microgrooved implants were thicker, but the capsules around the roughened implants were thinner. However, randomly roughened implant surfaces generally elicited a stronger and more prolonged inflammatory reaction compared to smooth and microgrooved implant surfaces. We conclude that the application of microgrooves or random surface roughness to polymer implants apparently does not have beneficial effects on peri-implant tissue healing.

Animals↗

PubMed2002

Soft tissue response to microtextured silicone and poly-L-lactic acid implants: fibronectin pre-coating vs. radio-frequency glow discharge treatment.

From in vitro studies it is known that a plasma-treatment can enhance cell spreading. Similar effects can be observed after pretreatment of the surface with a protein coating, to mediate cell adhesion. The aim of the current study was to evaluate the in vivo effects of these surface modifications, in a three-month experiment in a goat model. We made silicone and poly-L-lactic acid implants with double-sided parallel micro-grooves (depth 1.0 microm, width 10.0 microm), a random surface roughness, or a smooth surface. Implants either received a radio-frequency glow discharge (RFGD) treatment, a fibronectin (Fn) pre-coating, or no pre-treatment. Subsequently, they were inserted into subcutaneous pockets created on the flanks of goats for 1, 3 or 12 weeks. Histological analysis showed that a fibrous tissue capsule had formed around all implants. Histomorphometrical analysis was performed on capsule thickness, capsule quality and the implant-tissue interface quality. Fn-treated surfaces showed a considerable early inflammatory reaction. Besides this, RFGD treatment or Fn pre-coating did not further influence any of the measured parameters. In conclusion, pre-treatment of polymer implant surfaces with Fn or RFGD treatment did not significantly influence tissue reaction around implants with micro-grooved, roughened or smooth surfaces.

Animals↗

PubMed2002

Osteoblast differentiation of bone marrow stromal cells cultured on silica gel and sol-gel-derived titania.

Primary cultures of osteogenic precursor cells derived from rat bone marrow stroma were performed on commercially available pure titanium discs (Ti c.p.) and surface modified Ti c.p.using a sol-gel technique (Ti sol). In separate repeated experimental runs, cell behavior and in vitro mineralization were compared with cultures on silica gel bioactive glass discs (S53P4). All substrates were incubated in simulated body fluid prior to the experiment. Overall, variable effects between experimental runs were seen. Apparently, this was due to the heterogeneous nature of the used cell population. Therefore, only careful conclusions can be made. Initial cell adhesion and growth rates between 3 and 5 days of culture--analyzed by cell numbers--were in general comparable for the two titanium substrates, while initial growth up to day 3 is suggested to be higher in Ti c.p. compared to Ti sol. Although initial cell adhesion on the S53P4 glass discs was lower than the titanium substrates, cell growth rates appeared to be higher on the silica gel compared to the two titanium substrates. Further, there were some indications that the early and late osteoblast differentiation markers, alkaline phosphatase and osteocalcin, monitored up to day 24, were elevated in Ti c.p cultures compared to Ti sol cultures. There were no differences observed in in vitro mineralization between the titanium groups. S53P4 seemed to display a substantially higher differentiating capacity for both osteogenic cell markers as well as in vitro mineralization compared to the two titanium substrates.

Alkaline Phosphatase↗

PubMed2002

Initial interaction of rat bone marrow cells with non-coated and calcium phosphate coated titanium substrates.

Initial interactions of rat bone marrow (RBM) cells with smooth titanium, rough titanium or calcium phosphate coated substrates were tested. Cells were seeded onto the substrates, and attachment, integrin expression and spreading and morphology were studied. We found no difference in attachment of RBM cells to the different materials. We did find differences in the percentage of attached cells within a certain time between replicate runs of the experiments. RBM cells on all materials express alpha1, alpha3, alpha5, alpha6 and beta1 subunits. Again there was a large difference in expression patterns on RBM cells in different runs. No difference was found in expression on the various materials. For alpha1, alpha5, alpha6 and beta1, no difference was found in expression between attached and unattached cells. Expression of alpha3 was similar on attached and unattached cells during early culture. At the end of culture, alpha3 expression was downregulated for attached cells and not for unattached cells. This resulted in a higher expression of alpha3 for unattached cells compared to attached cells. Cells did spread on all materials, and reached a larger cell size on smooth titanium than on the rough materials. Morphology of the cells on the materials differed. On smooth titanium, cells usually showed a compact cell body with short cellular extensions. On the rough materials, cells often showed elongated shapes, with many thin cellular extensions. From this we conclude that the substrate surface characteristics of the materials we used do not influence attachment or integrin expression during the initial cell-material interactions. On the other hand, spreading behavior and cell morphology do depend on substrate surface characteristics.

Animals↗

PubMed2002

A new method to produce macropores in calcium phosphate cements.

A new way to create macropores in calcium phosphate cements has been developed. The method consists in adding NaHCO3 to the starting cement powder (Biocement D) and using two different liquids: first a basic liquid to form the paste and later an acid liquid to obtain CO2 bubbles. Mercury intrusion measurements showed a dramatic increase both in macropores with an average size of 100 m and in the total porosity (even higher than 50% with respect to the Biocement D). This method does not change in any significant way the final reaction products of the starting material after being soaked 3 days in Ringer solution. Only, due to the increase of the porosity. the compressive strength of the porous cement decreases significantly.

Biodegradation, Environmental↗

PubMed2002

The effect of bone anchoring and micro-grooves on the soft tissue reaction to implants.

Here we aimed to compare the tissue reaction to smooth and micro-grooved implants, at different implantation sites. We hypothesised that subperiosteally less mobility is to be expected between an implant and the surrounding tissue, which can lead to a more subdued tissue response. In addition, we hypothesised that a similar effect can be reached when substrata are equipped with micro-grooves. Poly-L-lactic acid smooth or micro-grooved surfaces (width 2 or 10 microm, depth 1 microm) were implanted subperiosteally on the frontal bone of the skull, or subcutaneously in the flanks of goats for 2, 4 and 12 weeks. After sacrifice, implants and surrounding tissue were histologically processed. Light microscopical and histomorphometrical evaluation of the histomorphometrical analyses, capsule thickness, capsule quality and implant-tissue interface was performed. In addition, we stained for alpha-smooth muscle actin. collagen and CD-68 expression. All implants were surrounded by a fibrous capsule. Capsules around subperiosteal implants were more matured than around subcutaneous implants. In time, capsule thickness significantly decreased around subperiosteal implants, but increased around subcutaneous implants. Also, nowhere differences were found in the presence of collagen or alpha-smooth muscle actin. The interfacial cells around all implants frequently showed staining for the monocyte-macrophage marker CD-68. We concluded that in this model, decreased mobility of an implant relative to the surrounding tissue did positively influence the peri-implant tissue response, but the applied surface topography did not.

Actins↗

PubMed2002

Mandibular reconstruction: a clinical and radiographic animal study on the use of autogenous scaffolds and platelet-rich plasma.

This paper reports on an experimental animal study evaluating a method of mandibular reconstruction. After a successful pilot study, 28 goats underwent a continuity resection of the mandibular angle. Primary reconstruction was carried out using specially designed osteosynthesis plates and screws. The defect was bridged by the original cortical scaffold, filled with an autogenous bone graft from the iliac crest. To accelerate bone healing, platelet-rich plasma (PRP) was mixed with the particulate bone graft in 14 goats. All goats had uneventful healing. The osteosynthesis plates and screws withstood immediate loading for periods varying from 3 weeks to 3 months. The use of PRP appeared to enhance bone healing considerably.

Animals↗

PubMed2002

In vitro osteogenic differentiation of rat bone marrow cells subcultured with and without dexamethasone.

The aim of our study was to investigate the osteogenic potential of subcultured rat bone marrow cells. Rat bone marrow (RBM) cells were cultured with or without dexamethasone. Subsequently, osteogenic differentiation and expression was studied. When cells were cultured continuously in the presence of dexamethasone, cultures initially showed high alkaline phosphatase expression and abundant mineralization. Expression of differentiation markers decreased with passaging. After cells were passaged three times, no alkaline phosphatase activity and calcification were found. Primary cells cultured without dexamethasone showed low alkaline phosphatase and no calcification, and remained fibroblast-like. When these cells were subcultured in the presence of dexamethasone, the cells did show osteogenic differentiation. Nevertheless, this occurred at a significant lower level than with cells continuously cultured with dexamethasone. In addition, no differentiation was found after second passage. Our results indicate that subcultured undifferentiated RBM cells show osteogenic differentiation after addition of dexamethasone. Expression of alkaline phosphatase and mineralization is higher in cells continuously supplemented with dexamethasone. Still, even when dexamethasone is added continuously, RBM cells loose their osteogenic potential after several passages. Therefore, we conclude that subculture of undifferentiated rat bone marrow cells results in the loss of osteogenic potential of these cells.

Alkaline Phosphatase↗

PubMed2002

Modulation of integrin expression on rat bone marrow cells by substrates with different surface characteristics.

Biomaterials have been shown to be able to influence the growth and differentiation of osteogenic cells cultured on the surface. Although the precise mechanisms by which the materials influence osteogenic cells are unclear, it is possible that the materials manipulate the expression of integrins by the cells. We therefore studied the expression of a number of integrins by rat bone marrow (RBM) cells, after culture on culture polystyrene, on machined and grit-blasted titanium, and on calcium phosphate-coated titanium. Integrin expression was studied by FACS analysis. We found a large variation in the expression of integrins by cells in replicate experiments. After culture on polystyrene for 7 days, cells expressed alpha1, alpha2, alpha3, alpha5, alpha6, beta1, and beta3, although some of the subunits were expressed only occasionally. The cells did not express the alpha4 subunit. After culture of RBM cells for 8 days on coated and noncoated titanium substrates, cells always expressed alpha3, alpha5, alpha6, and beta1. The alpha1 and beta3 subunits were only expressed in some of the experiments. Frequently, the expression of alpha5, alpha6, and beta1 was higher on the coated than on the noncoated titanium substrates. Based on our results, we conclude that the studied materials are capable of influencing the expression of integrins by RBM cells cultured on relevant implant materials.

Animals↗

PubMed2002

[Tissue engineering].

Tissue engineering is a fast developing multidisciplinary field. Both from a medical and social point of view, there is great need to safe tissues and organs to overcome the shortage of donor organs and to reduce the health care cost. At the same time, progress made in the field of molecular sciences as well as in biomaterials and chemical engineering, offers great possibilities to manipulate cells, growth factors and carrier materials in such a way that growth of tissues can be achieved.

Bone Regeneration↗

PubMed2001

Influence of inflammatory cells and serum on the performance of implantable glucose sensors.

The objective of this investigation was to evaluate the influence of polymorphonuclear granulocytes on the performance of uncoated and cellulose acetate/Nafion coated amperometric glucose sensors in vitro. The response of these sensors was also investigated in serum. Uncoated and coated sensors showed lower sensitivities to glucose, with a significant drift in sensor output upon exposure to serum or leukocytes. Although the use of a coating resulted in higher sensitivity, the progressive loss of output was not completely prevented. Stimulated granulocytes were shown to excrete components, probably catalase and myeloperoxidase, which consumed the hydrogen peroxide formed by the oxidation of glucose. In addition, adsorbed serum proteins formed a diffusional barrier for glucose. Furthermore, serum was found to contain low-molecular weight components that alone inhibited glucose oxidase activity. Based on preliminary electrochemical results, we postulate that rabbit serum contains oxidizing substrates that compete with molecular oxygen for the acceptance of electrons from the oxidized enzyme. Consequently, future efforts should be aimed at elucidating the mechanisms involved in the interference of unknown serum components with electron transfer. In addition, further investigations have to be performed to develop an outer membrane that minimizes protein adsorption as well as the actions of inflammatory cells.

Animals↗

PubMed2001

Modulation of epithelial tissue and cell migration by microgrooves.

We used a polystyrene substratum to study the response of migrating epithelium to 1- or 5-microm depth microgrooves with groove/ridge widths of 1, 2, 5, or 10 microm. The migration of a tissue sheet was enhanced along the microgrooves, while migration across the microgrooves was inhibited. Changing the depth of the microgrooves had a greater effect on migration than alteration of the groove/ridge width. The migration of epithelial cells from a confluent monolayer culture followed a similar pattern to that of intact epithelial tissue. Cellular extensions generally followed the microgroove direction by tracking along the top of the ridges or following the ridge walls, as revealed by scanning electron microscopy. Actin filaments within the basal cell layer of the tissue were aligned with the microgrooves, unlike filaments in the superficial layers that did not appear to be affected by the presence of underlying microgrooves. The basal cell layer of the tissue conformed to the contours of the microgroove following migration. However, the ultrastructure of the tissue above the ridges resembled that of tissue on a flat surface. We concluded that surface microgrooves have the potential to direct the migration of immediately adjacent epithelial tissue, the effect of which is to guide epithelial tissue on the surface of implanted biomaterials.

Actin Cytoskeleton↗