Biomedical subjects

J A Jansen

Publications and source records attributed to J A Jansen.

At least 73 records · Page 4Linked to original sources

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

Release of bioactive transforming growth factor beta(3) from microtextured polymer surfaces in vitro and in vivo.

Transforming growth factor beta(3) (TGF-beta(3)) has been under investigation with the objective of improving wound healing. Yet, little experimental knowledge exists about applications of TGF-beta(3) in implantology and tissue engineering. The aims of this study were to determine the release kinetics and bioactivity of TGF-beta(3) released from microtextured silicone and poly-L-lactic acid (PLA) surfaces in vitro and in vivo. We loaded surfaces with 100 ng of TGF-beta(3). An in vitro assay showed that TGF-beta(3) was released in a burstlike manner. Released TGF-beta(3) was capable of inhibiting the proliferation of mink lung epithelial cells, indicating that released TGF-beta(3) had remained at least partly active. Subsequently, an in vivo experiment (1 h-3 days) was performed with implants loaded with TGF-beta(3). In cryosections, TGF-beta(3) activity was assessed by an in situ bioassay. We found that active TGF-beta(3) was released for up to 24 h. Furthermore, released TGF-beta(3) could be detected up to 320 microm from the implant. On the basis of these observations, we conclude that TGF-beta(3) loaded onto microtextured polymer membranes remains functional when released in vitro and in vivo and, therefore, may represent an alternative for introducing a growth factor into a wound to achieve long-term and long-range biological effects.

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↗

PubMed2001

Cell and tissue behavior on micro-grooved surfaces.

In this review, we discuss substrates and implant surfaces provided with micrometer-sized groove and ridge patterns. Such "microgrooves" influence cell behavior: the cells align themselves, and migrate guided by the surface grooves. This phenomenon is known as "contact guidance". First, cell structure and cell attachment behavior are described. Then techniques for the production of microgrooves are addressed, and a summary is given of a number of previous in-vitro and in-vivo experiments on this subject. Based on the knowledge of cell movement, we suggest a theory involving the dynamics of fibrous cellular components in the filopodium. Finally, future directions for this type of research, and implications for medical and dental implantology, are addressed.

Journal Article↗

PubMed2001

A percutaneous device as model to study the in vivo performance of implantable amperometric glucose sensors.

Glucose kinetics were investigated in subcutaneous tissue of rabbits, in which a percutaneous device was implanted. The device was used for collection of tissue fluid and as carrier of an amperometric glucose sensor. Changes in glycaemia were reflected in subcutaneous tissue fluid. However, a limited number of responses of the implanted sensors were observed. Histologic evaluation showed thin fibrous capsules surrounding the implants. Accumulations of inflammatory cells were observed inside the subcutaneous chamber. The experiments again showed that changes in blood glucose concentration can be measured in subcutaneous tissue fluid collected with a percutaneous device. Nevertheless, implanted glucose sensors could not reliably monitor these changes. Supported by our histological observations and sufficient in vitro performance, we suppose that the cellular reaction to the sensor plays an important role in this poor in vivo performance. In combination with adsorption of tissue fluid proteins, this results in a reversible deactivation of implanted sensors. The exact mechanisms involved in this process are currently unknown and need further investigation.

Journal Article↗

PubMed2001

Pain and instability during biting with mandibular implant-retained overdentures.

We tested in a randomized controlled clinical trial the effect of pain and instability of dentures on bite force with different degrees of mucosal support. The trial involved 3 groups who had received: 1) a new conventional denture (CD-group), 2) an implant-mucosa-borne overdenture on 2 IMZ implants (IMZ-group) or 3) a mainly implant-borne overdenture retained by a transmandibular implant (TMI-group). Fifty-three women and 15 men, mean age 59 years, participated in this study. Bite force measurements were made unilaterally with a transducer and bilaterally with a bite fork. After the measurements, subjects were asked whether or not biting had caused pain or tilting of one of the dentures. Significantly more complete-denture wearers reported pain. They reported more frequent pain in the mandible than in the maxilla (P < 0.001), whereas implant-groups seemed to experience more often pain in the maxilla. On the transducer, maxillary dentures of the CD-group tilted less (P < 0.01) and mandibular dentures more (P < 0.05) compared to the implant-groups. With the bite fork, tilting occurred more often in the incisal-cuspid area than in the molar region (P < 0.001). No effect of pain and tilting was observed on maximum bite force. It appears that oral implants used to stabilize mandibular dentures permit subjects to exert higher bite forces and reduce the pain as otherwise felt in the mandible during maximum biting. Due to this stabilization, pain and instability of the maxillary denture can become the limiting factor for a further increase in bite force.

Adult↗

PubMed2001

Analysis of integrin expression in U2OS cells cultured on various calcium phosphate ceramic substrates.

Earlier we observed that calcium phosphate (Ca-P)-coated implant substrates stimulated the differentiation of osteoblast-like cells compared to uncoated substrates. This suggests that this difference in osteogenic induction is due to the chemical composition of the substratum. We hypothesized that Ca-P coatings modulate integrin expression patterns, because those receptors are the sensors of the cell. Therefore, in the present study we quantitatively analyzed integrin expression of osteosarcoma cells and their proliferation behavior on various well-defined Ca-P substrates. For this study we used the osteosarcoma cell line U2OS. Five groups of substrates were used: thermanox (Th), uncoated titanium (Ti), dense sintered hydroxyapatite (HA), and two Ca-P-coated titanium discs (TiHA-O% and TiHA-5%). At day 5, cell numbers were significantly lower (p < 0.05) for both types of Ca-P-coated titanium substrates compared to the other substrates. There were no significant differences between HA and uncoated titanium. From day 5 to 8, accumulated cell number was ranking highest to lowest HA > Th = Ti > TiHA-0% > TiHA-5%. Integrin expression at day 5 and day 8 of incubation was analyzed by flow cytometry for integrin subunits beta 1, alpha 3, alpha 4, alpha 5, alpha 6, and alpha v. Fluorescence-activated cell sorting (FACS) analysis showed that the cells express high levels of beta 1, low levels of alpha 4, alpha 5, and alpha 6, and moderate levels of alpha 3 and alpha v integrin subunits on the various biomaterial substrates. Minor differences in integrin expression between the various substrates were seen. Therefore, the observed differences in proliferation between the coatings may reside in modulating the functional properties of integrins.

Alkaline Phosphatase↗

PubMed2001

Influence of rhBMP-2 on rat bone marrow stromal cells cultured on titanium fiber mesh.

Titanium (Ti) fiber mesh is a candidate scaffold material for the creation of bone graft substitutes (BGS). Two densities (3.54 x 10(4) cells/cm(2) [LD or low density] and 3.54 x 10(5) cells/cm(2) [HD or high density]) of rat bone marrow stromal cells were seeded on Ti-fiber mesh discs. Cells were cultured for up to 16 days, 7 days of which the cells were in the presence of various concentrations of rhBMP-2 (0, 10, 100, and 1,000 ng/mL) in order to evaluate osteogenic expression. Scanning electron microscopy (SEM), light microscopy (LM), energy dispersive spectroscopy (EDS), DNA and calcium (Ca) content measurements, and x-ray diffraction (XRD) analysis were performed. SEM and EDS evaluation showed that a confluent layer of cells was present on top of the meshes together with collagen bundles and calcified globular accretions. Light microscopical evaluation showed a densely stained layer in the upper part of the mesh. SEM and Ca content measurement showed that calcification starts at 8 days. In addition, it was demonstrated that DNA content peaked at 8 days. LM, SEM, and Ca content evaluation revealed positive effects of increasing the cell seeding density, the rhBMP-2 concentration and the culture time on mineralization. Increasing the cell seeding density also showed a positive effect on DNA content. No effects of rhBMP-2 concentration were seen on DNA content. Finally, XRD revealed that the deposited matrix contained a precipitate of a stable calcium phosphate phase. We conclude that (1) titanium fiber mesh sustains excellent osteogenic expression in vitro, (2) increasing the cell seeding density has a positive effect on osteogenic expression in titanium mesh in vitro, and (3) in high density specimens, rhBMP-2 concentrations of 100 ng/mL and 1,000 ng/mL stimulate extracellular matrix calcification in a dose-responsive manner.

Animals↗

PubMed2001

Ectopic bone formation in titanium mesh loaded with bone morphogenetic protein and coated with calcium phosphate.

The osteoinductive properties of porous titanium fiber mesh, with or without a calcium phosphate coating and loaded with recombinant human bone morphogenic protein-2 (rhBMP-2) or rhBMP-2 and native bovine BMP (S-300) were investigated in a rat ectopic assay model. A total of 112 calcium phosphate-coated and 112 noncoated porous titanium implants, either loaded with rhBMP-2 and S-300 or loaded with rhBMP-2 alone, were subcutaneously placed in 56 Wistar-King rats. The rats were killed 5, 10, 20, and 40 days postoperatively, and the implants were retrieved. Histologic analysis demonstrated that all growth factor and carrier combinations induced ectopic cartilage and bone formation at 5 and 10 days, respectively. At 20 days, bone formation increased and was characterized by trabecular bone and bone marrow-like tissue. At 40 days, more lamellar bone and hemopoietic bone marrow-like tissue were present. At both times, more bone had been formed in calcium phosphate-coated implants than in noncoated samples. Further, in rhBMP-2 and S-300-loaded specimens, bone formation was higher than in rhBMP-2 only-loaded specimens. In rhBMP-2 only-loaded specimens, bone formation was mainly localized inside the mesh material, whereas in specimens loaded with both rhBMP-2 and S-300, the bone was localized inside and surrounding the titanium mesh. The histological findings were confirmed by calcium content and alkaline phosphatase activity measurements. In addition, all specimens showed osteocalcin expression as early as 5 days postoperatively. Our results show that the combination of titanium mesh with BMPs can induce ectopic bone formation and that this bone formation seems to be similar to "enchondral" ossification. In addition, a thin calcium phosphate coating can have a beneficial effect on the bone-inducing properties of a scaffold material. Finally, rhBMP-2 and native BMP act synergistically in ectopic bone induction.

Alkaline Phosphatase↗