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- Abteilung für Funktionswerkstoffe der Medizin und der Zahnheilkunde (37)
- Graduate School of Life Sciences (13)
- Fakultät für Chemie und Pharmazie (11)
- Institut für Funktionsmaterialien und Biofabrikation (8)
- Lehrstuhl für Tissue Engineering und Regenerative Medizin (6)
- Institut für Organische Chemie (2)
- Institut für Pharmazie und Lebensmittelchemie (2)
- Klinik und Poliklinik für Hals-, Nasen- und Ohrenkrankheiten, plastische und ästhetische Operationen (2)
- Klinik und Poliklinik für Mund-, Kiefer- und Plastische Gesichtschirurgie (2)
- Klinik und Polikliniken für Zahn-, Mund- und Kieferkrankheiten (2)
Sonstige beteiligte Institutionen
Aim of this thesis was the development of functionalizable hydrogel coatings for melt electrowritten PCL scaffolds and of bioprintable hydrogels for biofabrication.
Hydrogel coatings of melt electrowritten scaffolds enabled to control the surface hydrophilicity, thereby allowing cell-material interaction studies of biofunctionalized scaffolds in minimal protein adhesive environments. For this purpose, a hydrophilic star- shaped crosslinkable polymer was used and the coating conditions were optimized. Moreover, newly developed photosensitive scaffolds facilitated a time and pH independent biofunctionalization.
Bioprintable hydrogels for biofabrication were based on the allyl-functionalization of gelatin (GelAGE) and modified hyaluronic acid-products, to enable hydrogel crosslinking by means of the thiol-ene click chemistry. Optimization of GelAGE hydrogel properties was achieved through an in-depth analysis of the synthesis parameters, varying Ene:SH ratios, different crosslinking molecules and photoinitiators. Homogeneity of thiol-ene crosslinked networks was compared to free radical polymerized hydrogels and the applicability of GelAGE as bioink for extrusion-based bioprinting was investigated. Purely hyaluronic acid-based bioinks were hypothesized to maintain mechanical- and rheological properties, cell viabilities and the processability, upon further decreasing the overall hydrogel polymer and thiol content.
Hydrogel coatings: Highly structured PCL scaffolds were fabricated with MEW and subjected to coatings with six-armed star-shaped crosslinkable polymers (sP(EO-stat-PO)). Crosslinking results from the aqueous induced hydrolysis of reactive isocyanate groups (NCO) of sP(EO-stat-PO) and increased the surface hydrophilicity and provided a platform for biofunctionalizations in minimal protein adhesive environments. Not only the coating procedure was optimized with respect to sP(EO-stat-PO) concentrations and coating durations, instead scaffold pre-treatments were developed, which were fundamental to enhance the final hydrophilicity to completely avoid unspecific protein adsorption on sP(EO-stat-PO) coated scaffolds. The sP(EO-stat-PO) layer thickness of around 100 nm generally allows in vitro studies not only in dependence on the scaffold biofunctionalization but also on the scaffold architecture. The hydrogel coating extent was assessed via an indirect quantification of the NCO-hydrolysis products. Knowledge of NCO-hydrolysis kinetics enabled to achieve a balance of sufficiently coated scaffolds while maintaining the presence of NCO-groups that were exploited for subsequent biofunctionalizations. However, this time and pH dependent biofunctionalization was restricted to small biomolecules. In order to overcome this limitation and to couple high molecular weight biomolecules another reaction route was developed. This route was based on the photolysis of diazirine moieties and enabled a time and pH independent scaffold biofunctionalization with streptavidin and collagen type I. The fibril formation ability of collagen was used to obtain different collagen conformations on the scaffolds and a preliminary in vitro study demonstrated the applicability to investigate cell-material interactions.
The herein developed scaffolds could be applied to gain deeper insights into the fundamentals of cellular sensing. Especially the complexity by which cells sense e.g. collagen remain to be further elucidated. Therefore, different hierarchies of collagen-like conformations could be coupled to the scaffolds, e.g. gelatin or collagen-derived peptide sequences, and the activation of DDR receptors in dependence on the complexity of the coupled substances could be determined. Due to the strong streptavidin-biotin bond, streptavidin functionalized scaffolds could be applied as a versatile platform to allow immobilization of any biotinylated molecules.
Gelatin-based bioinks: First the GelAGE products were synthesized with respect to molecular weight distributions and amino acid composition integrity. A detailed study was conducted with varying molar ratios of reactants and synthesis durations and implied that gelatin degradation was most dominant for high alkaline synthesis conditions with long reaction times. Gelatin possesses multiple functionalizable groups and the predominant functionalization of amine groups was confirmed via different model substances and analyses. Polymer network homogeneity was proven for the GelAGE system compared to free radical polymerized hydrogels with GelMA. A detailed analysis of hydrogel compositions with varying functional group ratios and UV- or Vis-light photoinitiators was executed. The UV-initiator concentration is restricted due to cytotoxicity and potential cellular DNA damages upon UV-irradiation, whereas the more cytocompatible Vis- initiator system enabled mechanical stiffness tuning over a wide range by controlling the photoinitiator concentration at constant Ene:SH ratios and polymer weight percentages. Versatility of the GelAGE bioink for different AM techniques was proved by exploiting the thermo-gelling behavior of differently degraded GelAGE products for stereolithography and extrusion-based printing. Moreover, the viability of cell-laden GelAGE constructs was demonstrated for extrusion-based bioprinting. By applying different multifunctional thiol-macromolecular crosslinkers the mechanical and rheological properties improved concurrently to the processability. Importantly, lower thiol-crosslinker concentrations were required to yield superior mechanical strengths and physico-chemical properties of the hydrogels as compared to the small bis-thiol-crosslinker. Extrusion-based bioprinting with distinct encapsulated cells underlined the need for individual optimization of cell-laden hydrogel formulations.
Not only the viability of encapsulated cells in extrusion-based bioprinted constructs should be assessed, instead other parameters such as cell morphology or production of collagen or glycosaminoglycans should be considered as these represent some of the crucial prerequisites for cartilage Tissue Engineering applications. Moreover, these studies should be expanded to the stereolithographic approach and ultimately the versatility and cytocompatibility of formulations with macromolecular crosslinkers would be of interest. Macromolecular crosslinkers allowed reducing polymer weight percentages and amounts of thiol groups and are thus expected to contribute to increased cytocompatibility, especially in combination with the more cytocompatible Vis-initiator system, which remains to be elucidated.
Hyaluronic acid-based bioinks: Different molecular weight hyaluronic acid (HA) products were synthesized to bear ene- (HAPA) or thiol-functionalities (LHASH) to enable pure HA thiol-ene crosslinked hydrogels. Depending on the molecular weight of modified HA products, polymer weight percentages and Ene:SH ratios, a wide range of mechanical stiffness was covered. However, the manageability of high molecular weight HA (HHAPA) product solutions (HHAPA + LHASH) was restricted to 5.0 wt.-% as a consequence of the high viscosity. Based on the same HA thiol component (LHASH), hybrid hydrogels of HA with GelAGE were compared to pure HA hydrogels. Although the overall polymer weight percentage of HHAPA + LHASH hydrogels was significantly lowered compared to hybrid hydrogels (GelAGE + LHASH), similar mechanical and physico-chemical properties of pure HA hydrogels were determined with maintained Ene:SH ratios. Low viscous low molecular weight HA precursor solutions (LHAPA + LHASH) prevented the applicability for extrusion-based bioprinting, whereas the non-thermoresponsive HHAPA + LHASH system could be bioprinted with only one-fourth of the polymer content of hybrid formulations. The high viscous behavior of HHAPA + LHASH solutions, lower polymer weight percentages, decreased printing pressures and consequently declined shear stress during printing, were hypothesized to contribute to high cell viabilities in extrusion-based bioprinted constructs compared to the hybrid bioink.
The low molecular weight HA precursor formulation (LHAPA + LHASH) was not applicable for extrusion-based printing, but this system has potential for other AM techniques such as stereolithography. Similar to the GelAGE system a more detailed study on the functions of encapsulated cells would be useful to further develop this system. Moreover, the initiation with the Vis-initiator should be conducted.
Aufgrund der sich umkehrenden Alterspyramide in Deutschland leiden bereits jetzt immer mehr Menschen an Gelenkknorpelschäden. Doch nicht nur das Alter, sondern auch Unfälle und Sportverletzungen und Übergewicht können zu irreversiblen Knorpeldefekten führen. Obwohl es diverse Behandlungsmöglichkeiten gibt, können die bisherige Methoden nicht als dauerhafte Heilung betrachtet werden. Im Rahmen des internationalen Forschungsprojektes BIO-CHIP sollte eine vielsprechende Behandlungsmethode mit neuartigen Arzneimitteln untersucht werden.
Als Ausgangsmaterial des Arzneimittels, ein hergestelltes Knorpelimplantat, dienen patienteneigene Knorpelzellen aus der Nase. Diese werden isoliert, vermehrt und letztlich auf einer Matrix zu einem Knorpelimplantat kultiviert. Wesentliche Voraussetzung für die Implantatfreigabe stellt neben toxikologischen und biologischen Unbedenklichkeitstests die Beurteilung der Viabilität dar. Diese wurde bisher anhand von Histologieschnitten von der Pathologie durchgeführt.
Ziel der vorliegenden Arbeit war die Entwicklung und Validierung eines standardisierten und objektiven Viabilitätstests für die Chondrozyten innerhalb der Knorpelmatrix. Hierfür wurde die LDH als Marker für irreversibel geschädigte Zellen verwendet. Die LDH Konzentration konnte mit dem CyQuant LDH-Assay durch die Messung der Absorption gemessen werden. Es konnte nachgewiesen werden, dass LDH die erforderliche Stabilität und Nachweisbarkeit im Medium besitzt. Mithilfe der Lyse, analog zum Herstellungsprozess, gezüchteter Mini-Knorpelimplantate, konnten die maximal erreichbaren LDH Konzentrationen ermittelt werden. Mithilfe dieser Konzentrationen wurde eine Eichkurve generiert. Diese dient als Beurteilung der Viabilität zukünftig gemessener Absorptionen des Überstandmediums.
Das entwickelte Verfahren erfordert keine invasiven Eingriffe am Implantat und zeichnet sich durch eine einfache Durchführung aus, da nur der Überstand gemessen werden muss. Die durchgeführte Validierung der Methode bescheinigte eine hohe Robustheit, Linearität, Genauigkeit und Präzision.
The implantation of any foreign material into the body automatically starts an immune reaction that serves as the first, mandatory step to regenerate tissue. The course of this initial immune reaction decides on the fate of the implant: either the biomaterial will be integrated into the host tissue to subsequently fulfill its intended function (e.g., tissue regeneration), or it will be repelled by fibrous encapsulation that determines the implant failure. Especially neutrophils and macrophages play major roles during this inflammatory response and hence mainly decide on the biomaterial's fate. For clinically relevant tissue engineering approaches, biomaterials may be designed in shape and morphology as well as in their surface functionality to improve the healing outcome, but also to trigger stem cell responses during the subsequent tissue regeneration phase.
The main focus of this thesis was to unravel the influence of scaffold characteristics, including scaffold morphology and surface functionality, on primary human innate immune cells (neutrophils and macrophages) and human mesenchymal stromal cells (hMSCs) to assess their in vitro immune response and tissue regeneration capacity, respectively. The fiber-based constructs were produced either via melt electrowriting (MEW), when the precise control over scaffold morphology was required, or via solution electrospinning (ES), when the scaffold design could be neglected. All the fiber-based scaffolds used throughout this thesis were composed of the polymer poly(ε caprolactone) (PCL).
A novel strategy to model and alleviate the first direct cell contact of the immune system with a peptide-bioactived fibrous material was presented in chapter 3 by treating the material with human neutrophil elastase (HNE) to imitate the neutrophil attack. The main focus of this study was put on the effect of HNE towards an RGDS-based peptide that was immobilized on the surface of a fibrous material to improve subsequent L929 cell adhesion. The elastase efficiently degraded the peptide-functionality, as evidenced by a decreased L929 cell adhesion, since the peptide integrated a specific HNE-cleavage site (AAPV-motif). A sacrificial hydrogel coating based on primary oxidized hyaluronic acid (proxHA), which dissolved within a few days after the neutrophil attack, provided an optimal protection of the peptide-bioactivated fibrous mesh, i.e, the hydrogel alleviated the neutrophil attack and largely ensured the biomaterial's integrity. Thus, according to these results, a means to protect the biomaterial is required to overcome the neutrophil attack.
Chapter 4 was based on the advancement of melt electrowriting (MEW) to improve the printing resolution of MEW scaffolds in terms of minimal inter-fiber distances and a concomitant high stacking precision. Initially, to gain a better MEW understanding, the influence of several parameters, including spinneret diameter, applied pressure, and collector velocity on mechanical properties, crystallinity, fiber diameter and fiber surface morphology was analyzed. Afterward, innovative MEW designs (e.g., box-, triangle-, round , and wall-shaped scaffolds) have been established by pushing the printing parameters to their physical limits. Further, the inter-fiber distance within a standardized box-structured scaffold was successfully reduced to 40 µm, while simultaneously a high stacking precision was maintained. In collaboration with a co-worker of my department (Tina Tylek, who performed all cell-based experiments in this study), these novel MEW scaffolds have been proven to facilitate human monocyte-derived macrophage polarization towards the regenerative M2 type in an elongation-driven manner with a more pronounced effect with decreasing pore sizes.
Finally, a pro-adipogenic platform for hMSCs was developed in chapter 5 using MEW scaffolds with immobilized, complex ECM proteins (e.g., human decellularized adipose tissue (DAT), laminin (LN), and fibronectin (FN)) to test for the adipogenic differentiation potential in vitro. Within this thesis, a special short-term adipogenic induction regime enabled to more thoroughly assess the intrinsic pro-adipogenic capacity of the composite biomaterials and prevented any possible masking by the commonly used long-term application of adipogenic differentiation reagents. The scaffolds with incorporated DAT consistently showed the highest adipogenic outcome and hence provided an adipo-inductive microenvironment for hMSCs, which holds great promise for applications in soft tissue regeneration.
Future studies should combine all three addressed projects in a more in vivo-related manner, comprising a co-cultivation setup of neutrophils, macrophages, and MSCs. The MEW-scaffold, particularly due to its ability to combine surface functionality and adjustable morphology, has been proven to be a successful approach for wound healing and paves the way for subsequent tissue regeneration.
Calcium phosphate cements (CPC) represent valuable synthetic bone grafts, as they are self-setting, biocompatible, osteoconductive and in their composition similar to the inorganic phase of human bone. Due to their long shelf-life, neutral setting and since water is sufficient for setting, hydroxyapatite (HA) forming cements are processed in different paste formulations. Those comprise dual setting, Ca2+ binding and premixed cement systems. With dual setting formulations, both dissolution and precipitation of the cement raw powder occur simultaneously to the polymerization of water-soluble monomers to form a hydrogel. Chelating agents are able to form complexes with Ca2+ released from the raw powder. Premixed systems mostly contain the raw powder of the cement and a non-aqueous binder liquid which delays the setting reaction until application in the moist physiological environment. In the present work, two of those reaction mechanisms allowed the development of HA based cement applications.
Drillable cements are of high clinical interest, as the quality of screw and plate osteosynthesis techniques can be improved by cement augmentation. A drillable, dual setting composite from HA and a poly(2-hydroxyethyl methacrylate) hydrogel was analyzed with respect to the influence of monomer content and powder-to-liquid ratio on setting kinetics and mechanical outcome. While the conversion to HA and crystal growth were constantly confined with increased monomer amount, a minimum concentration of 50 % was required to see impressive ameliorations including a low bending modulus and high fracture energy at improved bending strength. Increasing the liquid amount enabled injection of the paste as well as drilling after 10 min of pre-setting.
While classic bone wax formulations have drawbacks such as infection, inflammation, hindered osteogenesis and a lack of biodegradability, the as-presented premixed formulation is believed to exhibit outmatching properties. It consisted of HA raw powders and a non-aqueous, but water-miscible carrier liquid from poly(ethylene glycol) (PEG). The bone wax was proved to be cohesive and malleable, it withstood blood pressure conditions and among deposition in an aqueous environment, PEG was exchanged such that porous, nanocrystalline HA was formed. Incorporation of a model antibiotic proved the suitability of the novel bone wax formulation for drug release purposes.
Prefabricated laminates from premixed carbonated apatite forming cement and poly(ε-caprolactone) fiber mats with defined pore architecture were presented as a potential approach for the treatment of 2-dimensional, curved cranial defects. They are flexible until application and were produced in a layer-by-layer approach from both components such that the polymer scaffold prevents the cement from flowing. It was demonstrated that solution electrospinning with a patterned collector for the fabrication of perforated fiber mats was suitable, as high fiber volume contents in combination with an appropriate interface enabled the successful fabrication of mechanically reinforced laminates. Mild immersion of the scaffolds under alkaline conditions additionally improved the interphase followed by an increase in bending-strength.
Since few years, magnesium phosphate cements (MPC) have attracted increasing attention for bone replacement. Compared to CPC, MPC exhibit a higher degradation potential and high early strength and they release biologically valuable Mg2+. However, common systems offer some challenges while using them in non-classic cement formulations such as the need for foreign ion supply, the potential acidity of the reaction or the fast setting kinetics. Here, it was possible to develop a chelate-setting MPC paste with a broad spectrum of potential applications.
The general mechanism of the novel setting principle was tested in a proof-of-principle manner. The cement paste consisted of farringtonite with differently concentrated phytic acid solution for chelate formation with Mg2+ from the raw powder. Adjusting the phytic acid content and adding a magnesium oxide as setting regulator to compensate its retarding effect resulted in drillable formulations. Additionally, there is a strong clinical demand for well working bone adhesives especially in a moist environment. Mostly the existing formulations are non-biodegradable. Ex vivo adhesion of the above presented MPC under wet conditions on bone demonstrated over a course of 7 d shear strengths of 0.8 MPa. Further, the hardened cement specimens showed a mass loss of 2 wt.% within 24 d in an aqueous environment and released about 0.17 mg/g of osteogenic Mg2+ per day. Together with the demonstrated cytocompatibility towards human fetal osteoblasts, this cement system showed promising characteristics in terms of degradable biocements with special application purposes.
The focus of this thesis was to investigate how PCL and PLGA react to the heat exposure that comes with the MEW process over a defined timespan.
To assess the thermal stability of PCL during MEW over 25 d, an automated collection of fibers has been used to determine the CTS on each day of heating for three different temperatures. PCL is exceptionally stable over 25 d at 75 °C, whereas for 85 °C and 95 °C a slight upward trend during the last 10 d could be observed, which is an indication for thermal degradation. Same trend could be observed for diameter of fibers produced at a fixed collector speed. For all temperatures, CTS during the first 5 d decreased due to inhomogeneities of the melt. Physical analysis of the fibers by XRD and mechanical testing showed no significant changes.
To investigate the chemical details of the thermal durability, PCL was artificially aged over 25 d at 75 °C, 85 °C and 95 °C. Data from GPC analysis and rheology revealed that PCL is degrading steadily at all three temperatures. Combined with GC-MS analysis, two different mechanisms for degradation could be observed: random chain scission and unzipping. Additional GPC experiment using a mixture of PCL and a fluorescence labelled PCL showed that PCL was undergoing ester interchange reactions, which could explain its thermal stability.
PLGA was established successfully as material for MEW. GPC results revealed that PLGA degraded heavily in the one-hour preheating period. To reduce the processing temperature, ATEC was blended with PLGA in three mixtures. This slowed down degradation and a processing window of 6 h could be established. Mechanical testing with fibers produced with PLGA and all three blends was performed. PLGA was very brittle, whereas the blends showed an elastic behavior. This could be explained by ester interchange reactions that formed a loosely crosslinked network with ATEC.
The aim of this thesis was the development of a multifunctional coating system for AuNPs based on thioether polymers, providing both excellent colloidal stability and a variable possibility to introduce functionalities for biological applications.
First, two thioether-polymer systems were synthesised as a systematic investigation into colloidal stabilisation efficacy. Besides commonly used monovalent poly(ethylene glycol) (PEG-SR), its structural analogue linear poly(glycidol) (PG-SR) bearing multiple statistically distributed thioether moieties along the backbone was synthesised. Additionally, respective thiol analogues (PEG-SH and PG-SH) were produced and applied as reference.
Successive modification of varyingly large AuNPs with aforementioned thiol- and thioether-polymers was performed via ligand exchange reaction on citrate stabilised AuNPs. An increased stabilisation efficacy of both thioether-polymers against biological and physiological conditions, as well as against freeze-drying compared to thiol analogues was determined.
Based on the excellent colloidal stabilisation efficacy and multi-functionalisability of thioether-PG, a plethora of functional groups, such as charged groups, hydrophilic/hydrophobic chains, as well as bio-active moieties namely diazirine and biotin was introduced to the AuNP surface. Moreover, the generic and covalent binding of diazirine-modified PG-SR with biomolecules including peptides and proteins was thoroughly demonstrated.
Lastly, diverse applicability and bioactivity of aforementioned modified particles in various studies was displayed, once more verifying the introduction of functionalities. On the one hand the electrostatic interaction of charged AuNPs with hydrogels based on hyaluronic acid was applied to tune the release kinetics of particles from three-dimensional scaffolds. On the other hand the strong complexation of siRNA onto two positively charged AuNPs was proven. The amount of siRNA payload was tuneable by varying the surface charge, ionic strength of the surrounding medium and the N/P ratio. Moreover, the biological activity and selectivity of the biotin-streptavidin conjugation was verified with respectively functionalised particles in controlled agglomeration test and in laser-triggered cell elimination experiments. In the latter, streptavidin-functionalised AuNPs resulted in excellent depletion of biotinylated cells whereas unfunctionalised control particles failed, excluding unspecific binding of these particles to the cell surface.
Die vorliegende Arbeit befasste sich mit der bisher relativ unbekannten Polymerklasse der Polypeptoide, die hinsichtlich ihrer Verwendung als Biomaterial näher untersucht werden sollte. Hierbei war die Untersuchung des Polymerisationssystems ein wesentlicher Schwerpunkt. Dies beinhaltete zum einen die Synthesen verschiedener Monomere sowie deren Polymerisationskinetiken und zum anderen Studien über die Stabilität des aktiven Kettenendes. Um mehr über die Polypeptoide zu erfahren, wurden die erhaltenen Homopolymere nach der Strukturanalyse hinsichtlich ihrer physikochemischen Eigen-schaften untersucht. Im Anschluss erfolgte die Synthese von (amphiphilen) Blockco-polypeptoiden, die sich in wässrigen Lösungen zu definierten Morphologien zusammen-lagern. Die resultierenden Morphologien, sowohl mizellare als auch vesikuläre Strukturen, wurden mit verschiedenen Methoden, wie z. B. der Pyren-Fluoreszenz-Spektroskpie und der dynamischen Lichtstreuung, untersucht. Erste Erkenntnisse über die Biokompatibilität der Polypeptoide sollte die Bestimmung der Zellviabilität in verschiedenen Polymerlösungen liefern.
Die verschiedenen Studien über die Polypeptoide zeigten, dass diese Polymerklasse über eine besonders lebende Polymerisation synthetisiert werden kann. Dabei resultieren Produkte, die sich durch eine Poisson-Verteilung und eine hohe Endgruppengenauigkeit auszeichnen. Zusätzlich bestehen Polypeptoide aus einem abbaubaren Rückgrat und, im Vergleich zu den Polypeptiden, besitzen sie eine erhöhte proteolytische Stabilität. Amphiphile Blockcopolypeptoide sind zudem in der Lage, sich in Lösung zu verschiedenen Morphologien anzuordnen. Durch die Variierung der Seitenkette und des f kann sowohl die Selbstorganisation als auch das Mikroumfeld der Aggregate abgestimmt werden. Darüber hinaus können die amphiphile Blockcopolymere, die sich zu Mizellen anordnen, hydrophobe Substanzen solubilisieren.
Polypeptoide liefern all die nötige chemische Vielseitigkeit und potentielle Biokompatibilität, um bestehende sowie neuartige Probleme in biomedizinischen Anwendungen zu bewältigen. Zukünftige in vivo und in vitro Test werden das Potential, aber auch die Grenzen dieser neuen Polymerklasse als Biomaterial zeigen.
In einigen Punkten konnte diese Arbeit die Ergebnisse früherer Studien bestätigen.
Einstellungs- und Nadelmodi (wenn auch mit unterschiedlicher Ausprägung)
haben einen Einfluss auf die verbesserte Sichtbarkeit der Regionalanästhesienadeln
im Ultraschall. Damit diese sonographisch besser erkannt werden, sollten sie
in einem flacheren Winkel (30-45°) eingestochen werden. Größere Nadellumina
haben einen Vorteil in der Sichtbarkeit im Ultraschall. Auch konnte gezeigt werden,
dass der Nadelschliff einen Einfluss darauf hat, wie gut die Anästhesiekanülenspitze
sonographisch gesehen wird. Allein die (in der Arbeit untersuchten) Nadelbeschichtungs-
Arten brachten keinen wesentlichen Effekt für die verbesserte
Erkennbarkeit.
Es war jedoch auffallend, dass die subjektiven und objektiven Resultate in einem
relativ hohen Maße nicht übereinstimmten. Gründe dafür sehe ich v.a. im gewählten
Verfahren der objektiven Untersuchung (mit der Bildbearbeitungssoftware
muss die region of interest zielgenauer erfasst werden, um störende Umgebungseinflüsse
auszuschließen).
Echogene Nadeloberflächen (auch wenn diese nicht Gegenstand der vorliegenden
Untersuchung waren), weisen eine verbesserte Ultraschall-Reflexion auf. Sie
werden dazu beitragen, dass die sichere Anwendung der USRA weiter zunimmt.
Verbesserte Sonographiegeräte, zusätzliche Hilfsmittel (GPS-Unterstützung) und
neueste Nadelentwicklungen versprechen einen wichtigen Fortschritt auf dem
Gebiet der ultraschallunterstützten Regionalanästhesie (USRA). Diese Weiterentwicklungen
werden der USRA den Stellenwert als Gold-Standard in der
Regionalanästhesie sichern.
Ziel der Arbeit ist es, verschiedene Knochenersatzmaterialien der Tympanoplastik Typ 3
(autogenes Gewebe, Titan, Ionomerzement) bezüglich ihres Langzeitverhaltens im Mittelohr zu vergleichen. Es werden zwischen dem 21.12.1995 und dem 30.04.2011 in der Hals-Nasen-Ohrenklinik des Städtischen Klinikums Solingen operierte Patienten nachuntersucht. Insgesamt handelt es sich um 957 mit einer Tympanoplastik Typ III versorgte Patienten, die in diesem Zeitraum insgesamt 1093mal operiert worden sind. 676mal ist die Kette mit einer Titanprothese rekonstruiert worden, davon 301mal mit einer PORP und 375mal mit einer TORP (davon 21 bei intakter Stapessuprastruktur). Zu Beginn des Beobachtungszeitraums sind 56 Ionomerzement-prothesen eingesetzt worden, so dass 40 Ionomerzement-PORP und 16 Ionomerzement-TORP mit berücksichtigt worden sind. In 19 Fällen sind „sonstige“ Methoden (z.B. Knorpelüberhöhung des Steigbügels) zur Gehörknöchelchenkettenrekonstruktion gewählt worden.
Die Untersuchung zeigt, dass zur Kettenrekonstruktion eine Transposition autogener Ossikel angestrebt werden sollte. Stehen diese nicht zur Verfügung, empfiehlt sich die Verwendung von Titan-Prothesen. Aufgrund ihres hervorragenden In-Situ-Verhaltens sowie der nachgewiesen guten audiologischen Resultate sind sie derzeit das Mittel der Wahl.
Diese Arbeit befasst sich mit der Untersuchung von aus Patientenisolaten gewonnenen S. aureus Kulturen und deren Biofilmbildung auf implantatähnlichen Titan-Oberflächen. Ziel war es, den zeitlichen Ablauf bakterieller periprothetischer Infektionen über einen Zeitraum von 21 Tagen zu beschreiben und besser zu verstehen. Dazu sollte überprüft werden, ob ein fluoreszenzspektrometrisch ausgewertetes LIVE/DEAD Assay eine zusätzliche Aussage zum Status der im Biofilm befindlichen Zellen liefern kann. Zudem wurde die Biofilmentwicklung anhand etablierter fluoreszenzspektrometrischer Methoden (Concanavalin-A-Markierung extrazellulärer Polymerer Substanzen, DNA-Markierung mit Hoechst 33342) untersucht. Es konnte ein reproduzierbarer Verlauf der Entwicklung des Biofilms, sowie der DNA-Menge aufgezeigt werden. Das LIVE/DEAD Assay lieferte keine signifikanten Ergebnisse in Bezug auf das Verhältnis lebender zu toter S. aureus Zellen im Biofilm.
Weiter wurde die Angreifbarkeit des frühen, am Titan adhärenten Biofilms (Alter 1-5 Tage) durch das in der Orthopädie gängig eingesetzte Antibiotikum Gentamicin untersucht. Die Wirksamkeit konnte zu jedem getesteten Zeitpunkt der ersten fünf Tage durch Anzucht von Kolonien bestätigt werden. Auch wurde die Wirksamkeit über das LIVE/DEAD Assay überprüft, jedoch konnten hier keine aussagekräftigen Daten gewonnen werden, die diese Methode zur Überprüfung der Antibiotikawirksamkeit empfehlen könnten.