Biomedical subjects

X F Walboomers

Publications and source records attributed to X F Walboomers.

16 recordsLinked to original sources

PubMed2003

Soft tissue reaction to microgrooved poly-L-lactic acid implants loaded with transforming growth factor beta(3).

In both normal and disturbed wound healing, the generation of large, contracting scars can raise serious functional and cosmetic problems. A possible strategy to minimize or avoid the generation of scar tissue surrounding an implant is to apply transforming growth factor-beta(3) (TGF-beta(3)) to the implant. TGF-beta(3) (0, 1, or 2.5 microg) was freeze-dried onto poly-L-lactic acid (PLA) microgrooved substrates (width, 10 microm; depth, 1 microm) and implanted subcutaneously on the backs of rats for 2 and 8 weeks. After sacrifice, implants and surrounding tissue were histologically processed. Light microscopic and histomorphometric evaluation of capsule thickness, capsule quality, and implant-tissue interface was performed. In addition, we stained for alpha-smooth muscle actin (SMA), collagen, and ED-1 (a monocyte-macrophage marker). All implants were surrounded by a fibrous capsule. Capsules of the implants loaded with 1 or 2.5 microg of TGF-beta(3) showed significantly higher capsule quality. This meant that capsules were more mature compared with implants without TGF-beta(3). However, no significant differences were found in terms of thickness of the capsules or quality of the interface. Finally, apparently significant differences were also found in the expression of alpha-SMA, when comparing the various growth factor concentrations at both implantation points. In conclusion, the use of microgrooved PLA substrates with TGF-beta(3) did not lead to an overall improvement of periimplant tissue healing.

Actins↗

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

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

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

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↗

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↗

PubMed2000

Early spreading events of fibroblasts on microgrooved substrates.

We investigated the "contact guidance" phenomenon, shortly after cell attachment. For this purpose polystyrene substrates were produced, either smooth, or equipped with micogrooves (depth 0.5 micrometer, width 1-10 micrometer). On these substrates, fibroblasts were cultured for 15, 30, 45, 60, 120, or 240 min. Subsequently, they were studied with light microscopy, scanning electron microscopy, confocal laser scanning microscopy, and digital image analysis. Up to 1 h, cell attachment on the grooved substrates was impaired. Further, cells oriented to the direction of the microgrooves. This orientation was established fastest on the narrow grooves. After 30 min, cells showed abundant membrane extensions in all directions. Well-formed actin filaments were not present in the cell body at timepoints before 4 h. Furthermore, cells on smooth surfaces exhibited less filaments. The addition of cytochalasin-B only caused a delay of cell attachment and spreading. From these experiments, we conclude that a well-formed cellular actin cytoskeleton is no prerequisite for the occurrence of contact guidance. Actin microfilaments in the lamellipodia, and the interplay between the lamellipodium and extracellular matrix molecules seem to be the determining factor in the establishment of contact guidance.

Actins↗

PubMed2000

Microgrooved silicone subcutaneous implants in guinea pigs.

Cell-substratum interactions are of fundamental importance for the reaction of body tissues to surgically implanted foreign materials. In our study we investigated the influence of 2 microm wide microgrooves, with various depths (0.5-6 microm), on capsule formation around subcutaneous silicone implants, in an animal experiment. Silicone sheets with microtexture were glued around silicone tubes. These implants were placed subcutaneously in eight guinea pigs for 10 weeks. The implanted tubes were removed including all surrounding tissues, and processed for light microscopy and subsequent histomorphometrical evaluation. All removed implants were surrounded by a thin fibrous capsule, and it was observed that this capsule was separated from the implants by a thin, single layer of mono- and multinucleated phagocytotic cells. In histomorphometry no significant differences were seen in relation to the reaction towards the various textures. We conclude that microtextures do not have an effect on the morphological characteristics of capsule formation around silicone implants in soft tissue.

Animals↗

PubMed2000

Influence of transforming growth factor-beta3 on fibrous capsule formation around microgrooved subcutaneous implants in vivo.

Previous studies have shown that addition of transforming growth factor-beta3 (TGF-beta3) is capable of reducing scar tissue formation in skin defects. Therefore, we examined whether TGF-beta3 can also influence the organization of a fibrous capsule around implants in vivo. For this reason, 24 silicone implants with microgrooves with a groove depth of 1.0 microm and a ridge and groove width of 10.0 microm were made and loaded with human recombinant TGF-beta3 (0, 5, 50, and 250 ng). An in vitro release enzyme-linked immunosorbent assay (ELISA) test was done with another 10 implants to estimate the amount of TGF released from the implants. The implants were inserted subcutaneously in the backs of 6 guinea pigs. Each animal received four implants, which were left in place for 10 weeks. At the end of the implantation time, the implants were retrieved, embedded, and processed for histology. Histomorphometrical measurements were done on the capsule formation and the implant-cell interface quality and quantity. The results showed a fibrous capsule of 15 microm up to 50 microm thickness around all implants. There were no significant differences between the TGF-beta3-loaded implants or the controls. Frequently, inflammatory cells were present in the capsule. The implant-tissue interface was on average between 5 and 15 microm thick and consisted mostly out of one or two layers of macrophages or foreign body giant cells. Statistical analysis again showed no significant differences between the various TGF-beta3-coated implants and controls. Finally, we concluded that a microtextured surface can indeed be used for the release of TGF-beta3. On the other hand, this did not result in major differences in wound healing between implants loaded with 5, 50, or 250 ng of TGF-beta3 and controls.

Animals↗

PubMed1999

Attachment of fibroblasts on smooth and microgrooved polystyrene.

In this study rat dermal fibroblasts (RDFs) were cultured on smooth or microgrooved (1-20 microm wide, 0.5-5.4 microm deep) substrates. Polystyrene microgrooved substrates were produced by solvent casting on molds that had been produced by photolithographic techniques. We investigated the attachment of RDFs with various analytical techniques. Light microscopy and image analysis showed that RDFs were oriented on most microgrooves. The rate of orientation effectively was increased by an increase of groove depth. An analysis of confluent layers of RDF showed that at confluency microgrooves were able to support greater numbers of cells. However, the largest numbers of cells were not found on the narrowest and deepest microgrooves even though such microgrooves have the largest total surface and induce the strongest alignment. Interference reflection microscopy (IRM) showed that the RDFs form focal adhesions where the cell membrane is only 10 nm from the substrate. IRM also showed that RDFs follow the contours of shallow and wide microgrooves but bridge the grooves on deeper and narrower ones. This could explain why such grooves are not able to increase the numerical cell adhesion to a greater degree. The absence of contact between cells and the bottom of the grooves is a very important factor in establishing contact guidance.

Algorithms↗

PubMed1999

Contact guidance of rat fibroblasts on various implant materials.

Providing a substrate surface with micrometer-sized parallel grooves influences the behavior of cells growing on such substrates in vitro. Cells elongate in the direction of the groove and migrate guided by the grooves. It has been suggested that cellular alignment on microgrooves is predominantly dependent on groove dimensions and that surface chemical variation of the substrate material has little effect. Therefore we seeded primary rat dermal fibroblasts (RDF) on smooth and microgrooved (groove width 1-10 microm, depth 0.5 microm) polystyrene (PS), poly-L-lactic acid (PLA), silicone (SIL), and titanium (Ti) substrates. The production process was found to be more accurate for PS and PLA than for SIL and Ti substrates. A proliferation study, scanning electron microscopy, confocal laser scanning microscopy, and transmission electron microscopy revealed differences between RDF behavior on the materials. Our conclusions are (1) the accuracy of microtexture production by casting depends greatly on the material used; (2) even if no sharp discontinuities are present, microtextures still are potent tools for inducing contact guidance; and (3) besides surface texture, surface chemistry has a definitive influence on cell morphology.

Animals↗

PubMed1999

The effect of poly-L-lactic acid with parallel surface micro groove on osteoblast-like cells in vitro.

In this study we evaluated the behavior of rat bone marrow (RBM) cells on microgrooved poly-L-lactic acid (PLA) and polystyrene (PS) surfaces. The applied groove depth was 0.5, 1.0 or 1.5 microns, with a groove and ridge width of 1, 2, 5 or 10 microns. Scanning electron microscopical examination showed that a collagen-rich mineralized layer of extracellular matrix (ECM) was deposited. Alignment of the cells and matrix to the surface grooves was observed as described before. Quantitative evaluation, using a tetracycline labeling assay, revealed that more mineralized ECM was formed on the PLA than on the PS. Further, PLA surfaces with a groove depth of 1.0 micron and groove widths of 1 and 2 microns induced most mineralized ECM. Finally, alkaline phosphatase activity was also higher on most microgrooved PLA surfaces, compared with the other materials. On the basis of these observations, we concluded that microtextured surfaces are able to influence the differentiation of osteoblast-like cells and the deposition of mineralized matrix. Probably, this phenomenon can be used to increase the bone regeneration around oral implants.

Alkaline Phosphatase↗

PubMed1999

Soft tissue and epithelial models.

The applicability of a biomaterial for the manufacturing of oral implants is determined by its physicochemical and geometric surface properties. Research, therefore, is concerned with the cellular reactions that occur when an implant material comes into contact with body tissues. For permucosal oral implants, this involves both the reaction of bone and gingival cells. In vitro cell culturing--including the use of various analytical techniques like light microscopy, scanning and transmission electron microscopy, confocal laser scanning microscopy, and digital image analysis--is a good tool whereby investigators can obtain more insight into the relevant components of implant-tissue adhesion. In the current overview, the role of cell models in oral implant research is discussed, specifically with reference to responses of epithelial cells and fibroblasts.

Alveolar Process↗

PubMed1998

Microgrooved subcutaneous implants in the goat.

We investigated the behavior of microgrooved implants in soft tissue using polystyrene implantable disks, either smooth or microgrooved (1-10 microm) on both sides. The implants were placed subcutaneously in a goat for 1, 4, or 12 weeks. Light and transmission electron microscopy showed that fibrous capsule formation around the implants was fairly uniform. After 1 week the implants were covered with a fibrous capsule about 80 microm thick. The collagen matrix was loose, and many inflammatory cells were present. After 4 weeks the matrix was more dense and contained many newly formed blood vessels. At the implant surface a layer of inflammatory cells about 10 microm thick had accumulated. Finally, after 12 weeks the matrix had densified. One cellular layer of inflammatory cells was present at the implant surface. We carried out histomorphometric measurements of capsule thickness, inflammatory layer thickness, and the number of blood vessels. Capsule thickness appeared not to decrease with time. Further, these measurements showed that there were no differences in tissue reaction between smooth and microgrooved implants. On the basis of our observations, we suggest that 1 microm deep and 1-10 microm wide microgrooves do not influence tissue response around polystyrene implants in soft tissue.

Animals↗

PubMed1998

Growth behavior of fibroblasts on microgrooved polystyrene.

We investigated the contact guidance phenomenon of rat dermal fibroblasts (RDF) on microgrooved polystyrene substrates. Grooves were 1 microm deep, and between 1 and 10 microm wide. Light microscopy and digital image analysis (DIA) showed that RDF were oriented on all microgrooved substrates. Scanning electron microscopy showed that RDF cultured on 1 or 2 microm wide grooves were positioned on top of the ridges. On the wider 5 and 10 microm grooves, the cells were able to descend into the grooves. In confocal laser scanning microscopy, focal adhesions were lying in the same direction as the actin filament where they attached to. DIA confirmed an orientational behavior of focal adhesions and actin filaments on microgrooves. There were no differences in the measured orientation between the different grooves. Besides, no obvious preference was found for focal adhesions to lie along edges of the surface ridges. Transmission electron microscopy showed that focal adhesions were able to bend along the edges of ridges. On the basis of our observations, we suggest that the breakdown and formation of fibrous cellular components, especially in the filopodium, is influenced by the microgrooves. The microgrooves create a pattern of mechanical stress, which influences cell spreading and cause the cell to be aligned with surface microgrooves.

Animals↗