Zur Erhöhung der mechanischen Stabilität mineralischer Knochenzemente aus Calciumorthophosphaten (CPC) wurde in einem TTCP/DCPA-System das Zementedukt TTCP mit verschiedenen biokompatiblen Oxiden (SiO2, TiO2, ZrO2) während des Herstellungsprozesses dotiert. Dies führte zur Bildung von Calciummetallaten und einer Herabsetzung der Löslichkeit der TTCP-Komponente des Zements. Gegenüber einem oxidfreien Zement konnte die Druckfestigkeit von 65 MPa auf 80 MPa (SiO2) bzw. 100 MPa (TiO2) gesteigert werden. In einem zweiten Ansatz zur Verbesserung der Injizierbarkeit wurden die Wechselwirkungen der Partikeloberflächen mit der flüssigen Zementphase betrachtet. Durch biokompatible Additive sollte eine repulsive elektrostatische Wechselwirkung eingestellt werden, um Partikelagglomerate effektiv zu dispergieren und eine verflüssigende Wirkung zu erreichen. Die Injizierbarkeit eines TTCP/DCPA-Zements durch eine Kanüle mit 800 µm Durchmesser konnte durch die Verwendung von 500 mM tri-Natriumzitrat-Lösung aufgrund einer deutlichen Herabsetzung der Viskosität der Zementpaste signifikant gesteigert werden (>95%, P/L 3,3/1, Kraftaufwand 20 N). Abschließend wurde der Einfluss der Partikelgrößenverteilung auf die Festigkeit und Injizierbarkeit einer auf monomodaler Partikelgrößenverteilung basierten Zementmatrix untersucht. Hierzu wurden einem mechanisch aktivierten a-TCP-System unreaktive, feinkörnige Füllstoffpopulationen (TiO2, CaHPO4, CaCO3) zugesetzt und systematisch deren Effekt in Verbindung mit einer Partikelaufladung durch tri-Natriumzitrat auf die rheologischen und mechanischen Eigenschaften untersucht. Erst die Kombination einer bimodalen Partikelgrößenverteilung mit tri-Natriumzitrat-Lösung führte zu einer starken Erniedrigung der Viskosität, damit zur nahezu vollständigen Injizierbarkeit der Zemente und einer teilweise signifikanten Steigerung der mechanischen Festigkeiten (z.B. 72 MPa reiner a-TCP-Zement auf 142 MPa mit Zusatz von CaHPO4). / An improvement of the mechanical strength of calcium phosphate bone cements in combination with a lower cement paste viscosity for better injectability was investigated. A first approach to improve the mechanical stability consisted in adding several biocompatible oxides (SiO2, TiO2, ZrO2) during the fabrication process of tetracalcium phosphate (TTCP). The formation of calcium metallates led to a decrease of the solubility of the oxide doped tetracalcium phosphates. The cements based on tetracalcium phosphates mixed with SiO2 and TiO2 exhibit a significant increase of the compressive strengths (65 MPa of pure cement up to 80 MPa (SiO2) and 100 MPa (TiO2)). A second approach to decrease the viscosity of CPC pastes considered the interactions between the particle surfaces and the liquid cement phase. To disperse agglomerates and improve the injection properties of CPC, biocompatible components were added to the liquid phase. As a result the injection properties of the TTCP/DCPA cement improved significantly. The use of 500 mM trisodium citrate solution as liquid phase changed the viscosity of the cement paste to a point, where complete injectability (> 95%) through an 800 mm diameter hypodermic needle could be achieved at low loads (20 N). In order to transfer the results of a multi-constituent cement to a single reactive component cement, different fine-particle-sized fillers were added to mechanically activated, monomodal α-tricalcium phosphate (α-TCP) cement. The inert fillers dicalcium phosphate anhydrous (DCPA), titanium dioxide (TiO2) and calcium carbonate (CC) were added to the α-TCP-matrix in different concentrations. For an effective ionic modification a 500 mM trisodium citrate solution was used as liquid phase as in the aforementioned TTCP/DCPA cement system. Only the combination of a bimodal particle size distribution and trisodium citrate solution led to a nearly complete injectability and a significantly increase of the mechanical stability of CPC.

No description available.

No description available.

The outcome of the innate immune response to biomaterials mainly determines whether the material will be incorporated in the body to fulfill its desired function or, when it gets encapsulated, will be rejected in the worst case. Macrophages are key players in this process, and their polarization state with either pro- (M1), anti-inflammatory (M2), or intermediate characteristics is crucial for deciding on the biomaterial’s fate. While a transient initial pro-inflammatory state is helpful, a prolonged inflammation deteriorates the proper healing and subsequent regeneration. Therefore, biomaterial-based polarization may aid in driving macrophages in the desired direction. However, the in vivo process is highly complex, and a mono-culture of macrophages in vitro displays only one part of the cellular system, but, to this date, there is a lack of established co-cultures to assess the immune response to biomaterials. Thus, this thesis aimed to establish a functional co-culture system of human macrophages and human mesenchymal stromal cells (hMSCs) to improve the assessment of the immune response to biomaterials in vitro. Together with macrophages, hMSCs are involved in tissue regeneration and inflammatory reactions and can modulate the immune response. In particular, endogenously derived hMSCs considerably contribute to the successful engrafting of biomaterials. This thesis focused on poly(ε-caprolactone) (PCL) fiber-based scaffolds produced by the technique of melt electrowriting (MEW) as biomaterial constructs. Via this fabrication technique, uniform, precisely ordered scaffolds varying in geometry and pore size have been created in-house. To determine the impact of scaffold geometries and pore sizes on macrophages, mono-cultures incubated on scaffolds were conducted. As a pre-requisite to achieve a functional co-culture system on scaffolds, setups for direct and indirect systems in 2D have initially been established. These setups were analyzed for the capability of cell-cell communication. In parallel, a co-culture medium suitable for both cell types was defined, prior to the establishment of a step-by-step procedure for the co-cultivation of human macrophages and hMSCs on fiber-based scaffolds. Regarding the scaffold morphologies tested within this thesis to improve M2-like polarization, box-shaped scaffolds outperformed triangular-, round- or disordered-shaped ones. Upon further investigation of scaffolds with box-shaped pores and precise inter-fiber spacing from 100 µm down to only 40 µm, decreasing pore sizes facilitated primary human macrophage elongation accompanied by their differentiation towards the M2 type, which was most pronounced for the smallest pore size of 40 µm. To the best of my knowledge, this was the first time that the elongation of human macrophages in a 3D environment has been correlated to their M2-like polarization. Thus, these results may set the stage for the design, the assessment, and the selection of new biomaterials, which can positively affect the tissue regeneration. The cell communication of both cell types, detected via mitochondria exchange in direct and indirect co-cultures systems, took place in both directions, i.e., from hMSCs to macrophages and vice versa. Thereby, in direct co-culture, tunneling nanotubes enabled the transfer from one cell type to the respective other, while in indirect co-culture, a non-directional transfer through extracellular vesicles (EVs) released into the medium seemed likely. Moreover, the phagocytic activity of macrophages after 2D co-cultivation and hence immunomodulation by hMSCs increased with the highest phagocytic rate after 48 h being most pronounced in direct co-cultivation. As the commonly used serum supplements for macrophages and hMSCs, i.e., human serum (hS) and fetal calf serum (FCS), respectively, failed to support the respective other cell type during prolonged cultivation, these sera were replaced by human platelet lysate (hPL), which has been proven to be the optimal supplement for the co-cultivation of human macrophages with hMSCs within this thesis. Thereby, the phenotype of both cell types, the distribution of both cell populations, the phagocytic activity of macrophages, and the gene expression profiles were maintained and comparable to the respective standard mono-culture conditions. This was even true when hPL was applied without the anticoagulant heparin in all cultures with macrophages, and therefore, heparin was omitted for further experiments comprising hPL and macrophages. Accordingly, a step-by-step operating procedure for the co-cultivation on fiber-based scaffolds has been established comprising the setup for 3D cultivation as well as the description of methods for the analysis of phenotypical and molecular changes upon contact with the biomaterial. The evaluation of the macrophage response depending on the cultivation with or without hMSCs and either on scaffolds or on plastic surfaces has been successfully achieved and confirmed the functionality of the suggested procedures. In conclusion, the functional co-culture system of human macrophages and hMSCs established here can now be employed to assess biomaterials in terms of the immune response in a more in vivo-related way. Moreover, specifically designed scaffolds used within the present thesis showed auspicious design criteria positively influencing the macrophage polarization towards the anti-inflammatory, pro-healing type and might be adaptable to other biomaterials in future approaches. Hence, follow-up experiments should focus on the evaluation of the co-culture outcome on promising scaffolds, and the suggested operating procedures should be adjusted to further kinds of biomaterials, such as cements or hydrogels. / Der Verlauf der angeborene Immunantwort auf Biomaterialien bestimmt maßgebend, ob das Material vom Körper angenommen wird, um so seine gewünschte Funktion zu erfüllen, oder ob es zur Einkapselung und im schlimmsten Fall zur Abstoßung kommt. Makrophagen spielen in diesem Prozess eine Schlüsselrolle, und ihr Polarisationszustand, entweder pro (M1), antiinflammatorisch (M2) oder ein dazwischenliegender Subtyp, ist dabei von entscheidender Bedeutung. Während ein vorübergehender proinflammatorischer Anfangszustand hilfreich ist, verschlechtert eine anhaltende Entzündung eine zeitnahe Heilung und die anschließende Regeneration. Daher könnte eine durch Biomaterialien beeinflusste Polarisation hilfreich sein, um die Makrophagen in die gewünschte Richtung zu lenken. Die in vivo Reaktion ist jedoch äußerst komplex und die Kultivierung von Makrophagen in vitro stellt nur einen Teil des Prozesses dar. An etablierten Co-Kultursystem zur Untersuchung der immunmodulierenden Eigenschaften von Biomaterialien mangelt es jedoch. Daher war es Ziel dieser Arbeit ein funktionelles Co-Kultursystem von humanen Makrophagen und humanen mesenchymalen Stromazellen (hMSCs) zu etablieren um die in vitro Bewertung der Immunantwort nach Kontakt mit Biomaterialien zu verbessern. Von Interesse sind hMSCs hierbei, da sie zusammen mit Makrophagen an der Geweberegeneration und an Entzündungsreaktionen beteiligt sind. Zudem weisen MSCs immunmodulierende Eigenschaften in Hinblick auf Makrophagen auf und sind aktiv am Verlauf der Fremdkörperreaktion nach der Transplantation von Biomaterial beteiligt. Im Rahmen dieser Arbeit wurden Poly(ε-caprolactone) (PCL)-Scaffolds auf Faserbasis als Biomaterialkonstrukte verwendet, welche mit der Technik des Melt Electrowriting (MEW) hergestellt wurden. Mit dieser Technik kann sowohl die Form der Scaffolds als auch die Porengröße variiert werden. Um Unterschiede der Scaffoldgeometrien und Porengrößen in Hinblick auf die Makrophagenreaktion zu untersuchen, wurden zunächst Versuche mit Makrophagen-Monokulturen durchgeführt. Zur Etablierung eines funktionellen Co-Kultursystem, wurde zu Beginn ein Aufbau für ein direktes und indirektes System in 2D erstellt. Dieser Aufbau wurde anschließend auf die Möglichkeit der Zell-Zell-Kommunikation darin analysiert. Weiterhin wurde ein, für beide Zelltypen, geeignetes Kulturmedium definiert, gefolgt von der Etablierung eines Protokoll für die Co-Kultivierung beider Zelltypen auf faserbasierten Scaffolds. Im Bezug zu dieser Arbeit wurden Scaffolds mit unterschiedlicher Geometrie mittels der Technik des Melt Electrowriting hergestellt um die Veränderung der Makrophagenpolarisation zu untersuchen. Dabei zeigte sich eine verstärkte M2-Polarisation auf Scaffolds mit einer kastenförmigen Morphologie, verglichen mit dreieckigen, runden oder ungeordnet-strukturierten Scaffolds. Die weitere Untersuchung von Scaffolds mit kastenförmigen Poren und präzisen Faserabständen von 100 µm bis zu 40 µm zeigte das kleinere Porengrößen die Elongation primärer menschlicher Makrophagen förderten. Begleitet wurde die verstärkte Elongation mit einer gesteigerten Polarisation in Richtung des M2 Typs. Dieser Effekt war nach Kultivierung von Makrophagen auf Scaffolds mit 40 µm Poren am stärksten ausgeprägt. Im Rahmen dieser Arbeit konnte damit erstmals eine länglichen Morphologie humaner Makrophagen mit einer Polarisierung in den M2 Typ korreliert werden. Diese Ergebnisse könnten daher für das Design neuer Biomaterialien, welche sich positiv auf die Geweberegeneration auswirken sollen, von Bedeutung sein. Die Zellkommunikation beider Zelltypen, welche über Mitochondrienaustausch im direkten und indirekten Co-Kultur-System nachgewiesen wurde, fand sowohl ausgehend von Makrophagen als auch von hMSCs statt. Dabei ermöglichten „Tunneling Nanotubes“ in der direkten Co-Kultur den Transfer von Mitochondrien von einem Zelltyp zum jeweils anderen, während in der indirekter Co-Kultur ein ungerichteter Transfer durch in das Medium freigesetzte extrazelluläre Vesikel (EVs) stattfand. Darüber hinaus wurde die phagozytotische Aktivität von Makrophagen nach Co-Kultivierung untersucht, um die immunmodulatorischen Eigenschaften von hMSCs nachzuweisen, wobei die höchste phagozytotische Aktivität nach 48 stündiger Co-Kultivierung festgestellt wurde. Da die üblicherweise verwendeten Serumzusätze für Makrophagen (humanes Serum (hS)) und hMSCs (fötales Kälberserum (FCS)) bei längerer Kultivierung den jeweils anderen Zelltyp nicht unterstützen konnten, wurden diese Seren durch humanes Thrombozytenlysat (hPL) ersetzt. Dieses erwies sich im Rahmen dieser Arbeit als optimale Ergänzung für die gemeinsame Kultivierung beider Zelltypen in der Co-Kultur. Dabei wurden der Phänotyp und die Populationsverteilung beider Zelltypen, sowie die phagozytotische Aktivität und die Veränderung des Genexpressionsprofils von Makrophagen untersucht und mit den jeweiligen Standard-Monokulturbedingungen verglichen. Des Weiteren konnte gezeigt werde, dass eine Zugabe von Heparin in Zellkulturen mit Makrophagen und hPL nicht nötig ist. Daher wurde auf den Zusatz von Heparin für alle weitere Experimente, die hPL und Makrophagen umfassten, verzichtet. Im letzten Teil der Arbeit wurde ein Protokoll für die Co-Kultivierung auf MEW Scaffolds erstellt. Neben der Etablierung eines Setups für die 3D-Kultivierung wurden sowohl Protokolle zur Bewertung phänotypischer als auch molekularer Veränderungen entwickelt. Durch Feststellung von Unterschieden in der Makrophagenreaktion in Abhängigkeit zu der Kultivierung mit / ohne hMSCs und entweder auf Scaffolds oder Plastik-Kulturschalen konnte die Funktionalität der Protokolle nachgewiesen werden. Mit dem in dieser Arbeit etabliertem funktionellen Co-Kultursystem von humanen Makrophagen und hMSCs können zukünftig Biomaterialien mit einem stärkeren in vivo -Bezug in Hinblick auf die Immunantwort bewertet werden. Darüber hinaus deuten Ergebnisse auf speziell konstruierte MEW-Scaffolds ein vielversprechendes Designkriterium für neu entwickelte Biomaterialien an, wobei die Polarisation der Makrophagen in Richtung des entzündungshemmenden, heilungsfördernden Typens durch eine gesteuerte Morphologieänderung beeinflusst werden kann. An diese Arbeit anschließende Experimente sollten sich auf die Untersuchung vielversprechender Scaffolds mittels Co-Kultivierung sowie auf die Anpassung der etablierten Protokolle an andere Biomaterialgruppen, wie beispielsweiße für Zemente oder Hydrogele, konzentrieren.

No description available.

Mycelium-composites has been emerging commercially as an environmentally sustainable alternative to conventional materials. By utilizing the fungal mycelium’s ability to bond with lignin and cellulose fibers of plants, mycelium-composites can be made that are recyclable and renewable. Mycelium therefore has the potential to contribute to creating more sustainable options in material selection for product designers. However, the visual appearance of mycelium materials might pose a challenge when it comes to their adaption into society. Case-studies have indicated a negative reaction to the mycelium-composites aesthetics which are characterized by imperfections and irregularities with an off-white coloring. Using theories surrounding aesthetics, materials experience and emotional design, the purpose of this study was to further develop the visual and/or aesthetical aspects of a mycelium-composite through a material driven design process The results of material development is presented in a product concept that displays the insight that can assist designers create mycelium-based products that are more accepted by a broader market.

Stabilization of highly sensitive noble metal nanoparticles is essential for their practical application. Bionanocomposites in which various types of noble metal nanoparticles, especially anisotropic noble metal nanoparticles, are immobilized into a macroscopic biomaterial membrane show promising applications in biomedical, catalytic, and environmental fields. This research focuses on developing two fabrication methods to generate novel bionanocomposite materials by immobilizing gold (Au) or silver (Ag) nanoparticles onto a "green" biomaterial, namely an eggshell membrane (ESM). Furthermore, the applications of the resulting bionanocomposite materials were demonstrated by studying their use as catalysts for environmental pollutant conversion and for the detection of two pollutant chemicals. The first fabrication method immobilizes ex situ synthesized nanoparticles onto a chemically modified ESM. Disulfide originating from the ESM was reduced by dithiothreitol into free thiol groups for binding to Au nanoparticles with different morphologies. The immobilization of Au nanoparticles greatly enhances their stability, making it possible to apply the resulting bionanocomposites for catalyzing the reduction reaction to convert pollutant p-nitrophenol (PNP) to p-aminophenol (PAP), with a great increase in their lifetime use from 2 to 10 reaction cycles. The second fabrication method utilizes the zwitterionic property of the protein based ESM for binding with Ag nanoparticles to form bionanocomposites. A seed mediated nanoparticle synthesis method originally performed in suspension was modified and adapted for the in situ synthesis of Ag nanodisks in this research. Ag nanoseeds were first immobilized onto an eggshell membrane using the static interaction between the nanoseeds and the membrane. Subsequently, Ag nanodisks were further grown directly on the Ag nanoseeds on the ESM. The final distribution density of Ag nanodisks can be adjusted by tuning the distribution density of Ag nanoseeds immobilized on the ESM. The performance of the resulting bionanocomposites were evaluated for both catalysis, and their application as substrates for surface enhanced Raman spectroscopy (SERS). The material performance was found to depend on the final distribution density of the Ag nanodisks on the ESM, offering the possibility to optimize bionanocomposite material performance by adjusting this density. A SERS based technique was further developed for detecting pollutant chemical species using the Ag nanodisks/ESM bionanocomposite material as a SERS substrate. Direct detection of thiram, a commonly used pesticide, was achieved at a concentration that is lower than that regulated by the US EPA. By using crystal violet as a SERS probe molecule, mercury, a heavy metal without an intrinsic Raman fingerprint, was indirectly detected not only at a limit of detection lower than most reported in the scientific literature, but also with a selectivity against a group of metal ions including Ba, Cu, Ca, Co, Mg, Mn, Ni, and Zn. It was also found that the detection sensitivity can be optimized by adjusting the Ag nanodisk distribution density on the ESM. The development of the fabrication approach and the use of ESM as a matrix material for immobilizing noble metal nanoparticles to form bionanocomposite materials demonstrates a novel strategy for meeting the needs of a variety of applications. The development of bionanocomposites for detecting pollutant species with different SERS activities by simply tuning the nanoparticle distribution density on the surface of the substrate, is a novel discovery, as it does not appear to have been previously reported in the literature. / Doctor of Philosophy / Noble metal nanoparticles exhibit special physical and chemical properties, which are totally different from the bulk material, making them promising candidates for use as novel materials in broad applications, such as catalysis, pollutant detection, antibacterial materials, etc. However, due to their high activity and poor colloidal stability (having high tendency to aggregate and lose activity), the nanoparticles require stabilization when being exploited for practical applications. A promising method to achieve this goal is to immobilize highly active noble metal nanoparticles onto a macroscopic membrane to form a nanocomposite. In this research, a "green" biomaterial, eggshell membrane (ESM), was utilized to immobilize noble metal nanoparticles. The resulting bionanocomposite materials were applied for catalyzing a reduction reaction to convert an environmental pollutant p-nitrophenol (PNP) to p-aminophenol (PAP) for environmental cleaning purposes, as well as detecting pollutant chemicals such as pesticide thiram and heavy metal mercury. General physical and chemical properties of the proteins in the ESM include rich chemical functional groups on the amino acid residue, and a zwitterionic property that allows the surface charge of the ESM to be changed under different pH levels. These properties, which have not been unleashed to immobilize noble metal nanoparticles in this field as of yet, were exploited in this research to create strong interactions between the noble metal nanoparticles and the ESM. This resulted in the formation of a bionanocomposite where the ESM served as a matrix for stably immobilizing the nanoparticles. Different bionanocomposites were fabricated using gold (Au) or silver (Ag) nanoparticles. The resulting bionanocomposite materials with gold nanoparticles were applied for catalyzing a reduction reaction for the conversion of p-nitrophenol, a commonly used chemical in the pharmaceutical photographic industries. The immobilized nanoparticles exhibited catalytic activity for ten reaction cycles and one hundred days after they were fabricated, while the colloidal nanoparticles (not immobilized nanoparticles) have catalytic activity for only two reaction cycles. For the chemical detection application, bionanocomposites with immobilized silver nanodisks were used as substrates for surface enhanced Raman spectroscopy. Different detection strategies were developed for detecting thiram with intrinsic Raman fingerprints and mercury without intrinsic Raman fingerprints. Outstanding detection sensitivities were achieved compared to those reported in the literature. For detection of mercury, a good selectivity was also obtained against a group of metal ions including Ba, Cu, Ca, Co, Mg, Mn, Ni, and Zn. The development of the fabrication approach and the use of ESM as a matrix material for immobilizing noble metal nanoparticles to form bionanocomposite materials demonstrates a good strategy for meeting the needs of a variety of applications

<p>In this thesis different aspects of calcium-aluminate (CA) as biomaterial are presented. Calcium aluminate is a chemically bonded ceramic with inherent properties making it suitable for use as biomaterial in some applications. In this thesis the emphasis is put on the basic chemical, physical and mechanical properties that may be achieved using the CA system as well as synthesis of the CA raw material. The basis for using CA in any application is the synthesis of the raw material. Different synthesis routes for producing CA are presented with focus on high temperature routes and the micro-structural and phase development during synthesis. As a base for further understanding of the CA properties a thorough outline of the reaction chemistry for CA is presented also including a description of how the reactions may be controlled and how formulations can be designed. The surface reactions of CA when subjected to simulated body fluid showed that CA is <i>in vitro</i> bioactive. An <i>in vivo</i> study in teeth also indicates that CA produces apatite at the tooth material interface. Dental materials are subjected to a harsh environment in the mouth with high mechanical forces, erosion and thermal changes. Also the demands on precise handling characteristics are high. For these reasons the <i>in vitro</i> evaluation of physical and mechanical properties are important. In this work several mechanical and physical properties of Ca-based formulations for dental applications has been tested using different methods. Some attention is also put on the specific characteristics of CA and the difficulties that arise when new material classes needs to be tested according to consensus standard methods. Finally studies on a CA-based formulation intended for Vertebroplasty is presented. The studies include basic mechanical properties as well as testing the material in an <i>in vitro</i> model utilising synthetic cancellous bone.</p>

Os estudos na área de engenharia de tecidos aplicada à reparação da cartilagem articular estão voltados ao desenvolvimento de uma matriz biocompatível que permita a diferenciação, proliferação e manutenção de células para produção de cartilagem hialina. A quitosana é um biomaterial e vem sendo estudada como suporte para condrócitos e para liberação controlada de substâncias. O objetivo deste trabalho foi estudar a biocompatibilidade do gel de quitosana associada ao fosfato de glicerol para reparação de defeitos osteocondrais induzidos experimentalmente na tróclea do talus de eqüinos. Foram utilizados cinco cavalos da raça Mangalarga, de três anos de idade, e por artroscopia foi criado um defeito osteocondral na tróclea lateral do talus de cada articulação. De forma aleatória um defeito foi escolhido para implante do gel de quitosana - fosfato de glicerol, e o defeito da articulação contralateral foi mantido vazio, servindo como controle. Para acompanhamento da evolução do processo de reparação da cartilagem articular foram realizados os exames físico, radiográfico e ultrassonográfico; análise do líquido sinovial (física, celularidade, quantificação de proteína, condroitim sulfato e ácido hialurônico); e análise da cartilagem articular (histológica e produção de proteoglicanos). Os resultados obtidos de todas as avaliações realizadas foram semelhantes entre os defeitos tratados e controle. O gel de quitosana fosfato de glicerol é biocompatível com o ambiente articular e pode ser indicado para futuras aplicações como suporte de células e para liberação controlada de medicamentos. / The tissue engineering studies applied to articular cartilage repair are focused on the development of scaffold biocompatibility allowing the differentiation, proliferation and cells maintenance providing production of the hyaline cartilage. Chitosan is a biomaterial that has been evaluated as a scaffold for chondrocyts implant and also as a drug-delivery control material. The aim of this work was to evaluate the chitosan glycerol phosphate gel biocompatibility in experimentally induced equine talus osteochondral defect. Five three years old Mangalarga breed horses were submitted to arthroscopy for osteochondral defect production on the lateral troclea of the talus in both tibiotarsal joints by arthroscopy. In a random form one defect was chosen for chitosan-glycerol phosphate gel implant, and the defect of the opposed joint was kept empty and used as a control. For the assessment of the articular cartilage repair process was performed the physic, radiographic and ultrassonographic exams; the synovial fluid analyze (physic, cellularity, protein quantification, chondroitin sulphate and hialuronan); and the articular cartilage analyze (hystologic and proteoglicans production). The results obtained in all evaluations performed were similar between the treated and control defects. The chitosan glycerol phosphate gel is biocompatible with the articular environment and can be indicate for future applications as an scaffold for cells support and drug-delivery control system.

A ocorrência de fraturas em animais e, especialmente nas espécies de grande porte, geram preocupação quanto ao custo do tratamento e a ocorrência de sequelas, pois há a possibilidade de complicações pela carga excessiva que esses animais exercem sobre os membros. Dessa forma é importante a imobilização imediata do membro e de maneira adequada, e avaliar se há necessidade de se realizar cirurgia para estabilização da fratura. Em algumas fraturas pode haver falhas ósseas, principalmente em fraturas cominutivas com extensa lesão de tecidos adjacentes, o que dificulta e prolonga o tempo de consolidação. Por essas razões vem sendo estudado, há alguns anos, o uso de substitutos ósseos na forma de cimentos ou mantas ósseas para reparar esses defeitos. O estudo com biomateriais como substitutos ósseos tem tido grande evolução nos últimos anos, e aqueles à base de quitosana, hidroxiapatita e colágeno apresentam grande vantagem, pois a quitosana estimula a regeneração óssea, a hidroxiapatita confere a dureza que o material exige, e o colágeno fornece maleabilidade. O presente projeto teve por objetivo avaliar o reparo ósseo após implante de compósito à base de quitosana, hidroxiapatita e colágeno, em falhas ósseas de tíbias de ovinos induzidas experimentalmente. Foram utilizados os métodos de imagem por densitometria ótica radiográfica e microtomografia computadorizada 3D para avaliar o grau e o tempo de reparo ósseo, assim como a densidade mineral óssea durante a cicatrização. As falhas ósseas foram confeccionadas nas tíbias de ovelhas, em ambos os membros, onde foi implantado o biomaterial em um deles, e o membro contralateral mantido como controle. Posteriormente, os animais foram submetidos a exames radiográficos, quinzenalmente, até 90 dias de pós operatório. Após este período, os animais foram submetidos à eutanásia e fragmentos das tíbias foram colhidos para avaliação por microtomografia computadorizada 3D. Observou-se, pelos resultados qualitativos das imagens radiográficas e dos dados estatísticos, que houve absorção gradativa do biomaterial, e que não houve diferença estatística em relação à densidade mineral óssea entre os dois grupos estudados, embora a DMO tenha sido maior nos membros com o biomaterial. Este método mostrou-se prático e eficaz, além de ser uma técnica não invasiva. A microtomografia computadorizada 3D revelou importantes informações quanto à porosidade óssea e volume ósseo entre os dois grupos. As imagens em 3D mostraram detalhes do osso que não eram possíveis de visualizar por meio das radiografias. Dessa forma é uma técnica bastante eficaz e detalhada, mas é necessária padronização de todas as amostras para poder quantificar os valores adequadamente. O estudo estatístico não mostrou diferenças entre os grupos com biomaterial e controle para os parâmetros de volume ósseo, grau de porosidade e porcentagem de volume ósseo. Porém, pelas imagens em 3D observa-se nitidamente maior quantidade de poros nas amostras controle em relação às amostras com biomaterial. Conclui-se com este estudo que as técnicas de densitometria óssea por raios-X e &micro;CT 3D são bastante eficazes para avaliar a cicatrização óssea em ovinos / The occurrence of fractures in animals and, especially, in large animals are conditions that generate concern about the cost of treatment and the occurrence of future complications because there is the possibility of complications due to excessive load that these animals receive in their limbs. Thus, it is important the limb immobilization immediately and appropriately and assess whether it is necessary to perform surgery to stabilize the fracture. In some kinds of fractures, it might have large bone defects, mainly in comminuted fractures with extensive damage to surrounding tissue, which makes it more difficult and prolongs the consolidation time. For these reasons, scientists have been studied the use of bone substitutes in the form of bone cements to repair these defects. The study of biomaterials as bone substitutes has had great progress in last years, and those based on chitosan, hydroxyapatite and collagen had great advantage because chitosan stimulates bone regeneration, hydroxyapatite confers the hardness that the material requires, and collagen provides flexibility to the material. This project aims to assess bone repair after deploying a composite of chitosan, collagen and hydroxyapatite in bone defects experimentally induced in sheep tibiae. For assessing the degree and the time of bone healing, it was used imaging techniques like radiographic bone densitometry and 3D micro-computed tomography. Bone defects were made in the sheep tibiae in both limbs where the biomaterial was deployed in one of them, and the control was the contralateral limb. After the surgeries, the animals were submitted to radiographic exams every two weeks until 90 days postoperatively. After this period, the animals were euthanized and harvested tibiae fragments for evaluation by 3D micro-computed tomography. Through the qualitative results of images and statistical data, it was observed, radiographically, that there was a gradual absorption of the biomaterial, and also that there was no statistical difference in relation to bone mineral density between the two groups. The method of determining the X-ray BMD proved to be practical and effective, and it is a noninvasive technique. 3D micro-computed tomography revealed important information about bone porosity and bone volume between the two groups. The 3D images showed bone details that were not possible to visualize through radiographs. Thus it is a highly effective and detailed technique, but it requires patterning of all the samples in order to quantify the amounts properly. Statistical analysis showed no differences between the groups for bone volume parameters, degree of porosity and percentage of bone volume. However in the 3D images, it can be observed clearly larger amount of pores in the control samples in comparison to those with biomaterial. It is concluded from this study that the techniques of bone densitometry X-ray and 3D &micro;CT are very effective to evaluate bone healing in sheep. Bone mineral density was higher in limbs with biomaterial and there was a higher bone volume and lower degree of porosity in the samples with biomaterial compared to the control group