Lutz Scheideler

A Review on the Wettability of Dental Implant Surfaces II: Biological and Clinical Aspects

Dental and orthopedic implants have been under continuous advancement to improve their interactions with bone and ensure a successful outcome for patients. Surface characteristics such as surface topography and surface chemistry can serve as design tools to enhance the biological response around the implant, with in vitro, in vivo and clinical studies confirming their effects. However, the comprehensive design of implants to promote early and long-term osseointegration requires a better understanding of the role of surface wettability and the mechanisms by which it affects the surrounding biological environment. This review provides a general overview of the available information about the contact angle values of experimental and of marketed implant surfaces, some of the techniques used to modify surface wettability of implants, and results from in vitro and clinical studies. We aim to expand the current understanding on the role of wettability of metallic implants at their interface with blood and the biological milieu, as well as with bacteria, and hard and soft tissues.

Cellular Reactions of Osteoblasts to Micron- and Submicron-Scale Porous Structures of Titanium Surfaces

Osteoblast reactions to topographic structures of titanium play a key role in host tissue responses and the final osseointegration. Since it is difficult to fabricate micro- and nano-scale structures on titanium surfaces, little is known about the mechanism whereby the topography of titanium surfaces exerts its effects on cell behavior at the cellular level. In the present study, the titanium surface was structured in micron- and submicron-scale ranges by anodic oxidation in either 0.2 M H3PO4 or 0.03 M calcium glycerophosphate with 0.15 calcium acetate. The average dimensions of pores in the structured surface were about 0.5 and 2 µm in diameter, with roughness averaging at 0.2 and 0.4 µm, respectively. Enhanced attachment of cells (SaOS-2) was shown on micron- and submicron-scale structures. Initial cell reactions to different titanium surfaces, e.g. the development of the actin-containing structures, are determined by the different morphology of the surfaces. It is demonstrated that on either micron- or submicron-structured surfaces, many well-developed filopodia were observed to be primary adhesion structures in cell-substrate interactions, and some of them entered pores using their distinct tips or points along their length for initial attachment. Therefore, porous structures at either micro- or submicrometre scale supply positive guidance cues for anchorage-dependent cells to attach, leading to enhanced cell attachment. In contrast, the cells attached to a smooth titanium surface by focal contacts around their periphery as predominant adhesion structures, since repulsive signals from the environment led to retraction of the filopodia back to the cell bodies. These cells showed well-organized stress fibres, which exert tension across the cell body, resulting in flattened cells.

A review on the wettability of dental implant surfaces I: Theoretical and experimental aspects

The surface wettability of biomaterials determines the biological cascade of events at the biomaterial/host interface. Wettability is modulated by surface characteristics, such as surface chemistry and surface topography. However, the design of current implant surfaces focuses mainly on specific micro- and nanotopographical features, and is still far from predicting the concomitant wetting behavior. There is an increasing interest in understanding the wetting mechanisms of implant surfaces and the role of wettability in the biological response at the implant/bone or implant/soft tissue interface. Fundamental knowledge related to the influence of surface roughness (i.e. a quantification of surface topography) on titanium and titanium alloy surface wettability, and the different associated wetting regimes, can improve our understanding of the role of wettability of rough implant surfaces on the biological outcome. Such an approach has been applied to biomaterial surfaces only in a limited way. Focusing on titanium dental and orthopaedic implants, the present study reviews the current knowledge on the wettability of biomaterial surfaces, encompassing basic and applied aspects that include measurement techniques, thermodynamic aspects of wetting and models predicting topographical and roughness effects on the wetting behavior.

Aptamer-based capture molecules as a novel coating strategy to promote cell adhesion

The improvement of the cytocompatibility of medical implants is a major goal in biomaterials research. During the last years many researchers worked on the fascinating approach to seed the respective cell types on various artificial substrates before implantation. For instance, cell‐seeded implants are supposed to be better candidates for transplantable bone substitutes than conventional artificial bone grafts. To improve cell seeding efficiency and cytocompatibility, we designed a new coating material for medical implants. We used aptamers, highly specific cell binding nucleic acids generated by combinatorial chemistry with an in vitro selection called systematic evolution of exponential enrichment (SELEX). Aptamers do have high binding affinity and selectivity to their target. In our study, human osteoblasts from osteosarcoma tissue were used as a target to create the aptamer. Single aptamer mediated cell sorting assays showed the binding affinity with osteoblasts. Additionally, the aptamers immobilized on tissue culture plates could capture osteoblasts directly and rapidly from the cell solution. This model proves that aptamer coated artificial surfaces can greatly enhance cell adhesion. We assume that this strategy is capable to improve the clinical application of tissue engineered implants

Investigation of cell reactions to microstructured implant surfaces

Abstract Surface topography is one of the key parameters influencing cellular reactions towards artificial materials. Surfaces with defined microstructures may be useful for enhancement of the stable anchorage of transcutaneous implants in connective tissue or for prevention of epithelial downgrowth and subsequent exfoliation. Cell reactions of keratinocytes and fibroblasts were investigated on microstructured titanium experimental surfaces with alternating grooves and ridges in the range between 1–20 μm width and 0.4–2.0 μm depth. While fibroblasts displayed oriented cell growth on the structured surfaces, human keratinocytes failed to show orientation or enhanced number of focal contacts on structures in the 2–10 μm width range. In that respect, an influence of surface structure on initial cell adhesion could not be proven. The influence of a “bioactive” fibronectin (Fn) coating on adhesion and spreading of fibroblasts was tested on smooth and structured titanium model implant surfaces. Cell spreading was enhanced significantly by the fibronectin coating. Under mechanical shear stress conditions which simulated stresses during insertion of dental implants, the stimulating effects of Fn were lost on smooth surfaces due to abrasion of the coating, while complete abrasion was prevented by microstructured surfaces. The combination of microstructures with “bioactive” coatings may be used to trigger specific cell responses in areas of the implant surface with different functionality. Adhesion and growth of different cell types on microstructured surfaces was investigated by a modified technique at the electron microscopy (EM) level. The approach allows the detection of adhesion molecules in the different membrane domains by immunocytochemical gold labelling techniques. Preliminary results with this new technique suggest that vinculin is localized in the grooves rather than on the ridges in our model system.

Structural characterization of nanocrystalline hydroxyapatite and adhesion of pre-osteoblast cells

Nanocrystalline hydroxyapatite (Nano HA), a prototype of minerals of bones and teeth, attracts increasing interest in medicine and dentistry. Different parameters for synthesis and post-treatment were investigated to determine their effects on crystallinity of nano HA, and in vitro cell responses to nano HA were studied. XRD and TEM analyses indicate that the crystallinity of nano HA synthesized by a chemical method was within the range of 15–50 nm, which is adapted to natural minerals of hard tissues. Increasing the ageing temperature significantly increased the crystallinity of nano HA, while lengthening the ageing time or varying the post-ageing drying process did not have any influence on its crystallinity. Nano HA annealed between 300 and 900 °C showed a small increase in crystallinity with increasing annealing temperature due to the long-range ordering effect. Cell attachment and spreading on nano HA were lower than those on pure titanium, and decreased as the crystallinity of nano HA increased. However, cells on nano HA demonstrated well-developed filopodia and lamelliopodia, which facilitate migration of the cells on it. This may benefit osteogenesis at the interface between bone and nano HA in vivo.

Comparison of different in vitro tests for biocompatibility screening of Mg alloys.

Standard cell culture tests according to ISO 10993 have only limited value for the biocompatibility screening of degradable biomaterials such as Mg alloys. The correlation between in vitro and in vivo results is poor. Standard cytotoxicity tests mimic the clinical situation to only a limited extent, since in vivo proteins and macromolecules in the blood and interstitial liquid will influence the corrosion behaviour and, hence, biocompatibility of Mg alloys to a significant extent. We therefore developed a modified cytotoxicity test simulating the in vivo conditions by use of bovine serum as the extraction vehicle instead of the cell culture medium routinely used in standard cytotoxicity testing according to ISO 10993-5. The modified extraction test was applied to eight experimental Mg alloys. Cytotoxicity was assayed by inhibition of cell metabolic activity (XTT test). When extraction of the alloy samples was performed in serum instead of cell culture medium the metabolic activity was significantly less inhibited for six of the eight alloys. The reduction in apparent cytotoxicity under serum extraction conditions was most pronounced for MgZn1 (109% relative metabolic activity with serum extracts vs. 26% in Dulbeccos modified Eagles medium (DMEM)), for MgY4 (103% in serum vs. 32% in DMEM) and for MgAl3Zn1 (84% vs. 17%), resulting in a completely different cytotoxicity ranking of the tested materials when serum extraction was used. We suppose that this test system has the potential to enhance the predictability of in vivo corrosion behaviour and biocompatibility of Mg-based materials for biodegradable medical devices.

Effect of Heterogenic Surfaces on Contact Angle Hysteresis: Dynamic Contact Angle Analysis in Material Sciences

Recently, the author successfully applied the classic Wilhelmy balance method and the dynamic contact angle analysis (DCA) on initial interfacial reactions of surface-modified biomaterials. In this study, the authors present further results which underline the potential of these methods to yield time-resolved data of ionic and protein interfacial reactions. In contrast to many spectroscopic methods, an on-line method, which works time-resolved and without disturbing the interface, would be important in the process-engineered quality control. This is underlined by the fact, that many biomedical material surfaces currently are pre-biofunctionalized before their application in order to increase their biocompatibility and bifunctionality. The above-outlined statement of the problems involves many disciplines. In this approach, it is highlighted and discussed on the background of current research on biomaterials.

Cell reactions to microstructured implant surfaces

Cells are capable of reacting to a variety of differently patterned substrates. A number of hypotheses have already emerged to explain cell orientation and directed migration. It seems likely that no single hypothesis is able to explain in detail the behavior of cells that produce contact guidance. Understanding the mechanisms that influence the behavior of cells on microstructured surfaces could help to optimize the surface of future implants. A processing scheme has been developed for fabricating accurately structured Araldite replicas with titanium coating serving as implant imitates suitable for cross-sectioning. Using electron microscopy and immunogold labeling it could be shown that focal contacts of human gingival fibroblasts are formed at the walls and even at the bottom of grooves as narrow as 2 µm and as deep as 2.5 µm.

UV-A and UV-C light induced hydrophilization of dental implants.

OBJECTIVES Wettability is increasingly considered to be an important factor determining biological responses to implant materials. In this context, the purpose of this study was to compare the dynamic wettability of dental implants made from different bulk materials and modified by different surface modifications, and to analyze the respective changes of wettability upon irradiating these implants by UV-A or UV-C light. METHODS Four original screw-type implants were investigated: One grit-blasted/acid-etched and one anodically oxidized titanium, one zirconia and one polyetheretherketone implant. Additionally, experimental, screwless, machined titanium cylinders were included in the study. Part of that cylinders and of blasted/etched implants were further modified by a magnetron-sputtered photocatalytic anatase thin film. Scanning electron microscopy was used to investigate the surface micro- and nanostructures. Samples were treated by UV-A (382nm, 25mWcm(-2)) and UV-C (260nm, 15mWcm(-2)) for entire 40min, respectively, and their wettability was quantified by dynamic contact angle (CA) analysis from multi-loop Wilhelmy experiments. RESULTS All implants are characterized by submicron- and nanosized surface features. Unexposed implants were hydrophobic (CA>90°). Upon UV-A, solely the implants with anatase coating became superhydrophilic (CA<5°). Upon UV-C, the blasted/etched implants turned superhydrophilic, the anodized titanium and the zirconia implants were considerably (CA=34° and 27°, respectively) and the PEEK implants slightly (CA=79°) hydrophilized. SIGNIFICANCE The wettability of implant surfaces can be improved by UV irradiation. The efficiency of UV-A and UV-C irradiation to lower the CA by photocatalysis or photolysis, however, is strongly dependent on the specific material and surface. Thus, attempts to photofunctionalize these surfaces by irradiation is expected to result in a different pattern of bioresponses.