Biomedical subjects

X F Walboomers

Publications and source records attributed to X F Walboomers.

At least 19 recordsLinked to original sources

PubMed2007

Macrophage behavior on multilayered DNA-coatings in vitro.

A pivotal factor to consider in the development of biomaterials and biomaterial coatings is the inflammatory response to these materials. The insertion of implants is followed by protein adsorption and subsequent interactions with cellular components of the biological surroundings, in which macrophages play a dominant role through the production of a myriad of signaling molecules. In view of this, the aims of the present study were to evaluate (i) gross protein adsorption to, and (ii) in vitro behavior of macrophages on novel biomaterial coatings, composed of poly-D-lysine (PDL) or poly(allylamine hydrochloride) (PAH) and DNA, and to compare these coatings with negative (noncoated glass) and positive controls (noncoated glass + LPS-stimulation). The results demonstrate that multilayered DNA-coatings do not affect gross protein adsorption compared to noncoated controls. Cell culture experiments showed that the attachment to, and viability and morphology of two types of macrophages cultured on multilayered DNA-coatings is comparable to noncoated controls. Still, macrophages repeatedly showed decreased secretion levels of the proinflammatory cytokine TNF-alpha on multilayered DNA-coatings, whereas no differences were observed in the secretion of IL-1beta, IL-10, and TGF-beta1. Appropriate animal studies are required to elucidate if these in vitro indications have clinical effects on the inflammatory and wound healing processes around implants.

Adsorption↗

PubMed2007

Biological responses to multilayered DNA-coatings.

This study was performed to evaluate the basic biological response to deoxyribonucleic acid (DNA)-based coatings for soft tissue implants. To that end, in vitro experiments were used to study their cytocompatibility, and in vivo subcutaneous implantation studies with transponders in a rat model were performed to evaluate their histocompatibility. The DNA-based coatings were fabricated using the electrostatic self-assembly technique using cationic poly-D-lysine or poly-allylamine hydrochloride and anionic DNA. Noncoated substrates served as controls. In vitro, the behavior of primary rat dermal fibroblasts was assessed in terms of cell proliferation and morphology. Both types of multilayered DNA-coatings significantly increased rat dermal fibroblast proliferation without altering the morphological appearance of the cells. The tissue response to multilayered DNA-coatings was assessed using an in vivo rat model, in which transponders were inserted subcutaneously for 4 and 12 weeks. No macroscopic signs of inflammation or adverse tissue reactions were observed at implant retrieval. Histological analyses demonstrated a uniform tissue response to all types of implants. All implants were encapsulated in a fibrous tissue capsule without intervening inflammatory cells at the implant surface. Histomorphometrically, multilayered DNA-coatings induced fibrous tissue capsules with similar quality and thickness compared to noncoated controls. In addition, all fibrous tissue capsules showed similar expression of alpha-smooth muscle actin. This study demonstrates that multilayered DNA-coatings are cytocompatible and histocompatible, and justifies further research on their functionalization with biologically active compounds to modulate tissue responses.

Actins↗

PubMed2006

Functionalization of multilayered DNA-coatings with bone morphogenetic protein 2.

The focus of the present study was to functionalize multilayered DNA-coatings with the osteoinductive factor bone morphogenetic protein 2 (BMP-2) using different loading modalities. The multilayered DNA-coatings were built up from either poly-d-lysine (PDL) or poly(allylamine hydrochloride) (PAH) and DNA using electrostatic self-assembly (ESA). The amounts of BMP-2 loaded into the multilayered DNA-coatings and its subsequent release characteristics were determined using radiolabeled BMP-2. Additionally, the effect of BMP-2 functionalized multilayered DNA-coatings on the in vitro behavior of bone marrow-derived osteoblast-like cells was evaluated in terms of proliferation, differentiation, mineralization, and cell morphology. The results demonstrate the feasibility of multilayered DNA-coatings to be functionalized by embedding BMP-2 according to three different loading modalities: superficial (s), deep (d), and double-layer (dl). BMP-2 was incorporated proportionally into the multilayered DNA-coatings as: s+(4*d)=dl. All differently loaded multilayered DNA-coatings showed an initial burst release followed by an incremental sustained release of the remaining BMP-2. In vitro experiments demonstrated that the loaded factor remained biologically active, as an accelerated calcium deposition was observed on s- and dl-loaded multilayered DNA-coatings, without affecting cell proliferation. In contrast, d-loaded multilayered DNA-coatings influenced osteoblast-like cell behavior by decreasing the deposition of calcium.

Alkaline Phosphatase↗

PubMed2006

The effect of combined hypergravity and micro-grooved surface topography on the behaviour of fibroblasts.

This study evaluated in vitro the differences in morphological behaviour between fibroblast cultured on smooth and micro-grooved substrata (groove depth: 1 mum, width: 1, 2, 5, 10 microm), which undergo artificial hypergravity by centrifugation (10, 24 and 50 g; or 1 g control). The aim of the study was to clarify which of these parameters was more important to determine cell behaviour. Morphological characteristics were investigated using scanning electron microscopy and fluorescence microscopy in order to obtain qualitative information on cell spreading and alignment. Confocal laser scanning microscopy visualised distribution of actin filaments and vinculin anchoring points through immunostaining. Finally, expression of collagen type I, fibronectin, and alpha(1)- and beta(1)-integrin were investigated by PCR. Microscopy and image analysis showed that the fibroblasts aligned along the groove direction on all textured surfaces. On the smooth substrata (control), cells spread out in a random fashion. The alignment of cells cultured on grooved surfaces increased with higher g-forces until a peak value at 25 g. An ANOVA was performed on the data, for all main parameters: topography, gravity force, and time. In this analysis, all parameters proved significant. In addition, most gene levels were reduced by hypergravity. Still, collagen type 1 and fibronectin are seemingly unaffected by time or force. From our data it is concluded that the fibroblasts primarily adjust their shape according to morphological environmental cues like substratum surface whilst a secondary, but significant, role is played by hypergravity forces.

Actin Cytoskeleton↗

PubMed2006

Hydrodynamic compression of young and adult rat osteoblast-like cells on titanium fiber mesh.

Living bone cells are responsive to mechanical loading. Consequently, numerous in vitro models have been developed to examine the application of loading to cells. However, not all systems are suitable for the fibrous and porous three-dimensional materials, which are preferable for tissue repair purposes, or for the production of tissue engineering scaffolds. For three-dimensional applications, mechanical loading of cells with either fluid flow systems or hydrodynamic pressure systems has to be considered. Here, we aimed to evaluate the response of osteoblast-like cells to hydrodynamic compression, while growing in a three-dimensional titanium fiber mesh scaffolding material. For this purpose, a custom hydrodynamic compression chamber was built. Bone marrow cells were obtained from the femora of young (12-day-old) or old (1-year-old) rats, and precultured in the presence of dexamethasone and beta-glycerophosphate to achieve an osteoblast-like phenotype. Subsequently, cells were seeded onto the titanium mesh scaffolds, and subjected to hydrodynamic pressure, alternating between 0.3 to 5.0 MPa at 1 Hz, at 15-min intervals for a total of 60 min per day for up to 3 days. After pressurization, cell viability was checked. Afterward, DNA levels, alkaline phosphatase (ALP) activity, and extracellular calcium content were measured. Finally, all specimens were observed with scanning electron microscopy. Cell viability studies showed that the applied pressure was not harmful to the cells. Furthermore, we found that cells were able to detect the compression forces, because we did see evident effects on the cell numbers of the cells derived from old animals. However, there were no other changes in the cells under pressure. Finally, it was also noticeable that cells from old animals did not express ALP activity, but did show similar calcified extracellular matrix formation to the cells from young animals. In conclusion, the difference in DNA levels as reaction toward pressure, and the difference in ALP levels, suggest that the osteogenic properties of bone marrow-derived osteoblast-like cells are different with respect to the age of the donor.

Aging↗

PubMed2006

Cyto- and histocompatibility of multilayered DNA-coatings on titanium.

DNA-containing biomaterial coatings offer potential beneficial effects for both soft and hard tissue implants because of the structural properties of DNA. In the current study, the aim was to assess the in vitro cyto- and in vivo histocompatibility of multilayered DNA-coatings generated using the electrostatic self-assembly technique, with poly-D-lysine or poly(allylamine hydrochloride) as the cationic counterparts of anionic DNA. Multilayered DNA-coatings were fabricated on titanium substrates. Noncoated titanium substrates served as controls. In vitro experiments with rat primary dermal fibroblasts (RDF) assessing their viability were performed using a Live/Dead assay and an MTT-based assay. The presence of multilayered DNA-coatings did not affect RDF cell viability. On the other hand, an increased proliferation was demonstrated on both types of multilayered DNA-coatings. An in vivo rat model was used to study the soft tissue histocompatibility of subcutaneously inserted implants during implantation periods of 4 and 12 weeks. Light microscopic analysis revealed that all implants were surrounded by a fibrous capsule containing alpha-smooth muscle actin, and that the presence of a multilayered DNA-coating did not induce any adverse effects in terms of inflammation and wound healing. Histomorphometrically, no significant differences in capsule quality or thickness were observed dependent on multilayered DNA-coating or implantation period. The cyto- and histocompatibility of multilayered DNA-coatings demonstrated in this study allows their use and functionalization with appropriate compounds to modulate cell and tissue responses in dental and medical implantology.

Animals↗

PubMed2006

The influence of the crystallinity of electrostatic spray deposition-derived coatings on osteoblast-like cell behavior, in vitro.

This article describes the influence of the crystallinity of carbonate apatite (CA) coatings on osteoblast-like cell behavior. Porous CA coatings were produced with electrostatic spray deposition (ESD), and subsequently, received heat treatments of 400, 500, or 700 degrees C to induce various coating crystallinities. As a result, an amorphous calcium phosphate (ACP), a crystalline CA (CCA), and a crystalline carbonated hydroxyapatite (CHA) structure were formed, respectively. Uncoated titanium substrates served as the control group. After seeding rat osteoblast-like cells, the initial cell attachment was similar between the groups, and approached 100% after 6 h. Between the various coatings, no differences were observed for proliferation, differentiation, or mineralization. However, proliferation of the osteoblast-like cells was lower on all coated substrates after longer culture periods, compared to the uncoated substrates, while at the same time differentiation was stimulated. Furthermore, after 8 and 16 days of incubation, scanning electron microscopy showed more signs of mineralization on coated substrates, compared to the uncoated substrates. In conclusion, porous ESD-derived CA coatings have a positive effect on the in vitro differentiation of osteoblast-like cells, compared to uncoated, as-machined titanium. However, this effect is not further enhanced by the degree of crystallinity of the ESD-derived CA coatings.

Alkaline Phosphatase↗

PubMed2005

The effect of combined cyclic mechanical stretching and microgrooved surface topography on the behavior of fibroblasts.

Under the influence of mechanical stress, cultured fibroblasts have a tendency to orient themselves perpendicular to the stress direction. Similar cell alignment can be induced by guiding cells along topographical clues, like microgrooves. The aim of this study was to evaluate cell behavior on microgrooved substrates, exposed to cyclic stretching. We hypothesized that cellular shape is mainly determined by topographical clues. On basis of earlier studies, a 10-microm wide square groove, and a 40-microm wide V-shaped groove pattern were used. Smooth substrates served as controls. Onto all substrates fibroblasts were cultured and 1-Hz cyclic stretching was applied (0, 4, or 8%) for 3-24 h. Cells were prepared for scanning electron microscopy, immunostaining of filamentous actin, alignment measurements, and PCR (collagen-I, fibronectin, alpha1- and beta1-integrins). Results showed that cells aligned on all grooved surfaces, and fluorescence microscopy showed similar orientation of intracellular actin filaments. After 3 h of stretch, cellular orientation started to commence, and after 24 h the cells had aligned themselves almost entirely. Image analysis showed better orientation with increasing groove depth. Statistical testing proved that the parameters groove type, groove orientation, and time all were significant, but the variation of stretch force was not. Substrates with microgrooves perpendicular to the stretch direction elicit a better cell alignment. The expression of beta1-integrin and collagen-I was higher in the stretched samples. In conclusion, we can maintain our hypothesis, as microgrooved topography was most effective in applying strains relative to the long axis of the cell, and only secondary effects of stretch force were present.

Animals↗

PubMed2005

Osteoclastic resorption of calcium phosphate coatings applied with electrostatic spray deposition (ESD), in vitro.

Calcium phosphate (CaP) coatings have been applied on titanium implants to improve the bioactivity in order to favor the initial bone healing response. Recently, a new technique has been developed to apply CaP coatings: electrostatic spray deposition (ESD). Although ESD-derived coatings have several benefits, it is not known whether they are degradable. This study was designed to examine the cell-mediated degradation of two ESD-derived coatings with different chemical compositions, that is, beta-tricalcium phosphate (beta-TCP) and carbonate apatite (CA). First, coatings were deposited and analyzed physiochemically. Subsequently, rat bone marrow-derived osteoclastlike cells were seeded on the coatings, and analyzed with osteoclast-specific markers, scanning electron microscopy, and transmission electron microscopy. Results showed that both coatings exhibited porous morphologies, with an average pore size of less than 1 microm (beta-TCP), or larger than 1 microm (CA). After heat treatment, both coatings were crystalline in structure. The Ca/P ratios were 1.4 to 1.5 for the beta-TCP coating, and 1.8 to 2.0 for the CA coating. After 8 and 12 days of culture, multinucleated osteoclastlike cells were observed on both coatings. The osteoclast phenotype was confirmed by tartrate resistant acid phosphatase (TRAP) staining, and immunostaining against the calcitonin receptor. Using scanning electron microscopy, numerous resorption lacunae were observed in both coatings. Finally, transmission electron microscopy of TRAP-positive cells confirmed the osteoclastlike aspect of the cells revealing multiple nuclei and a ruffled border. In conclusion, CaP coatings produced with the ESD process can be degraded by osteoclasts.

Absorbable Implants↗

PubMed2005

Effect of microtextured surfaces on the performance of percutaneous devices.

Along the percutaneous part of implantable devices, like (semi-)permanent catheters, epithelial downgrowth can occur. This process can eventually lead to implant loss. Various treatments for the catheter surface have been proposed, to improve their performance. The purpose of the current study was to investigate the effect of a microgroove pattern on the tube surface, on epithelial downgrowth. Catheterlike implants were made of silicone tubes, with anchoring cuffs made of titanium-fiber mesh. A thin sheet of silicone with microgrooves was applied on the tubes. Two types of texturing were used, a square groove of 10 microm wide and 1 microm deep; or a V-shaped groove of 40 microm wide. The grooves were directed either along the long axis of the catheter tube (grooves perpendicular to the skin surface) or circling around the catheter (grooves directed parallel to the skin surface). As controls, catheters with a smooth outer surface were used. Implants were placed in 30 rats, with a follow-up period of 9 weeks. During this time, animals were inspected biweekly, and catheter exit sites were evaluated using a scoring system. At the end of the 9-week period the implants and surrounding tissues were processed for histological evaluation. For the clinical evaluation of the exit sites, no statistical differences were found between the study groups. Histologically, epithelial downgrowth was observed for all samples. The histomorphometrical measurements showed that there were no differences in downgrowth between the smooth and parallel-grooved catheters. However, there was increased epithelial downgrowth along the catheters with grooves perpendicular to the skin. In conclusion, a grooved microtexture can direct epithelial tissue ingrowth, but this study found no beneficial effects of the guidance phenomenon.

Animals↗

PubMed2005

Integrins as linker proteins between osteoblasts and bone replacing materials. A critical review.

The adhesion of osteoblasts to substrates is mediated through proteins that have adsorbed to the substrate, providing integrins on the cell membrane with ligands to connect to. The integrins regulate cell behavior through bi-directional signaling pathways. This critical review has the purpose to consider the research that has been performed with osteoblasts, integrins, and bone replacing materials. Until now, most research has been done to investigate the integrin expression of osteoblasts in culture during cellular adhesion. However, it remains difficult to draw general conclusions from this research. Nevertheless, it can be concluded that the used substrates and protein or peptide coatings can influence the integrin expression and cellular behavior. Additional research has to be done to fully understand all the parameters involved in integrin expression, the adhesion of cells to substrates, and the subsequent cellular behavior. For this purpose, model substrates are under development. The signaling pathway is receiving more and more attention, but for biomaterial purposes, too little consideration is paid to the translation of the in vitro results to the in vivo situation, and to practical applications.

Animals↗

PubMed2005

The response of healing corneal epithelium to grooved polymer surfaces.

Corneal epithelial wounds heal rapidly by the inwards growth of tissue with a contracting wound front. A synthetic polymer lens to correct refractive error (an implantable contact lens) could be incorporated into the cornea using this wound healing process. Topographical cues on the polymer surface may facilitate epithelial tissue migration over the anterior device surface. Here, silicone discs with a defined surface geometry of parallel grooves (groove and ridge widths of 1, 2, 5 and 10 microm; groove depths of 1 and 5 microm) were implanted into corneas and maintained in organ culture. The nature and rate of epithelial tissue migration over the test surfaces was monitored for 8 days and evaluated using microscopy and histology. Irrespective of the pitch, deep groove geometries directed tissue migration laterally along the grooves but this prevented contraction of the wound front and retarded migration rates. No guidance occurred on any of the shallow groove geometries but these allowed inwards radial migration with a contracting wound front and supported migration rates equivalent to a flat surface. None of the geometries tested promoted tissue migration above a flat polymer surface and data suggested that parallel grooves may not be optimal for this application.

Animals↗

PubMed2004

Transforming growth factor-beta3-loaded microtextured membranes for skin regeneration in dermal wounds.

Adverse effects of wound healing, such as excessive scar tissue formation, wound contraction, or nonhealing wounds represent a major clinical issue in today's healthcare. Transforming growth factor (TGF)-beta3 has specifically been implicated in wound healing. Our hypothesis was that local administration of TGF-beta3 to excisional dermal wounds would diminish wound contraction and scar formation. Microtextured wound covers, containing different concentrations of TGF-beta3, were placed onto full-thickness excisional skin wounds in guinea pigs. Tattooed reference marks were used to quantify wound contraction. Sixty-four male guinea pigs in four study groups (5 ng TGF-beta3, 50 ng TGF-beta3, no growth factor, sham wound) were followed for up to 6 weeks. We analyzed 19 different parameters of wound healing. Results showed that, in some instances, the 50-ng TGF-beta3 group gave less contraction, whereas the 5-ng TGF-beta3 group gave more contraction. These differences confirm that TGF-beta3 has an optimum working concentration, and suggest this concentration to be closer to 50 ng than to 5 ng TGF-beta3. However, only very few significant differences occurred, and thus we conclude that the clinical relevance of our findings is negligible. Earlier studies, reporting clinically improved wound healing by TGF-beta3, could therefore not be confirmed by this study.

Actins↗

PubMed2004

Stretch-mediated responses of osteoblast-like cells cultured on titanium-coated substrates in vitro.

Cyclic stretching experiments on osteoblast-like cells have proven to be a useful tool in understanding the underlying mechanisms of load transduction at the bone-implant surface. However, most experimental setups use silicone rubber substrates, which are atypical for orthopedic and dental implant materials. Therefore, we investigated the responses of osteoblast-like cells to loading on titanium (Ti)-coated versus plain silicone substrates. Ti-coated substrates were made by a radio-frequency magnetron sputtering process, and characterized using Rutherford backscattering spectrometry, X-ray photoelectron spectroscopy, and contact-angle measurements. Osteoblast-like cells cultured from rat bone marrow were seeded on both types of substrates and stretched for 1 h continuously. Subsequently, cell proliferation, alkaline phosphatase activity, and calcium content were measured for up to 24 days after seeding. In addition light-, scanning electron-, and confocal laser scanning micrographs were made. The results showed that our Ti coating had a thickness of 50 nm and contained Ti/oxygen as 1:1. However, further characterization proved that the silicone material had a tendency to resurface through the coating. Osteoblast-like cells proliferated faster on the Ti-coated substrates, but differentiation was slower compared with the silicone substrates. It was concluded that that there was a definitive influence of the substrate material in mechanical stress models. Therefore, extrapolation of results obtained using silicone substrates cannot be translated directly toward the situation of metallic implant materials.

Alkaline Phosphatase↗

PubMed2004

Electrostatic spray deposition (ESD) of calcium phosphate coatings, an in vitro study with osteoblast-like cells.

Electrostatic spray deposition (ESD) is a recently developed technique to deposit a calcium phosphate (CaP) coating upon substrates. With this technique, an organic solvent containing calcium and phosphate is pumped through a nozzle. Between the nozzle and substrate a high voltage is applied. As a consequence, droplets coming out the nozzle disperse into a spray, and this spray is deposited upon the substrate. When the solvent has evaporated, a coating is formed on the substrate. ESD allows for a variation in coating composition and morphology. Titanium alloy (TiAl6V4) substrates were coated with a CaP layer using two different methods; radio frequency magnetron sputtering, and ESD. These surfaces were characterized with X-ray diffraction, Fourier transform infrared spectroscopy, an universal surface tester, scanning electron microscopy, and energy dispersive spectrometry. Subsequently, bone marrow cells were isolated from rat femora and cultured 1, 4, 8, 14 and 16 days. Cell proliferation, alkaline phosphatase activity, and osteocalcin concentration were assayed. RT-PCR was done for collagen type I and osteocalcin. SEM was also performed to observe cellular behaviour during culture. Two separate runs of the experiment were performed. In the first run, osteoblast-like cells on both CaP coatings showed similar results in all assays. In the second run, proliferation and osteogenic expression had increased on ESD coatings. On basis of these results, we conclude that the novel ESD coating behaved similar to, or even better than the known RF magnetron sputter coating. Thus, ESD could be a valid addition to already existing CaP coating processes.

Adsorption↗

PubMed2004

The influence of bone formation on anchoring percutaneous devices with titanium fibre mesh flanges.

For man-made percutaneous devices (PD), it is known that anchoring will improve the clinical success. Previously, our Department has designed PDs that use a sheet of titanium (Ti) fibre mesh for anchoring. In nature, a very successful natural PD occurs, namely the tooth. Teeth are anchored in the alveolar bone. In the current study, we evaluated whether a sheet of (ectopic) bone can be made, and be used to anchor a skin-penetrating device. Using available tissue engineering techniques, sheets of Ti fibre mesh were loaded with osteoblast-like cells. These sheets, and non-loaded controls, were placed subcutaneously in 20 syngeneic rats. After four weeks 10 rats were sacrificed, and tissues were prepared for histology. On the other 10 rats, a percutaneous Ti bar was screwed. These rats were evaluated clinically up to eight weeks. Finally, also their tissues were prepared for histology. The results showed, that bone formation was only established in one cell-loaded implant, of the four-week group. Clinical evaluation, and the histomorphometrical data, showed no differences between cell-loaded and unloaded samples. We concluded that the combination of Ti mesh with rat bone marrow cells was not able to generate bone formation after subcutaneous implantation predictably. Thus, our original aim could not be met. On the other hand, our results did confirm the biocompatible behaviour of a PD equipped with a Ti fibre mesh anchoring flange.

Animals↗

PubMed2003

Intermittent versus continuous stretching effects on osteoblast-like cells in vitro.

The objective of this study was to quantify and compare stretch-mediated responses of primary rat osteoblast-like cells to uniform cyclic strain applied intermittently or continuously. Primary rat osteoblast-like cells were seeded and cultured in silicone rubber dishes for 2 days. They were then subjected to 1000 microstrains at 1 Hz for periods of 60 consecutive minutes or to a series of 15-min stretch followed by 15-min rest, until a total stretch of 60 min. After stretching, cells were incubated and assayed on days 4, 8, 16, and 24 for DNA content, alkaline phosphatase (ALP) activity, and calcium (Ca) content. Additionally, qualitative information was obtained via scanning electron and confocal laser scanning micrographs. Significant increases in DNA were observed for cells stretched intermittently versus cells stretched continuously and versus controls. Results showed significant decreases (p < 0.05) in ALP for cells between stretched groups and between both stretched groups versus controls. Additionally, Ca content was greater in cells stretched intermittently versus controls on days 4 and 8 and versus cells stretched continuously on day 24. In conclusion, intermittently strained cells demonstrated significant decreases in ALP and increases in DNA and Ca versus cells strained continuously. This supports the theory that cells respond to mechanical loading in a "trigger-like" response.

Alkaline Phosphatase↗

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↗