L’architecture des biomatériaux de comblement osseux à un réel impact sur l’activité cellulaire, la vascularisation et la diffusion de facteurs de croissance. Dans un premier temps, différentes membranes de polystyrène (PS) composées de fibres alignées ou aléatoires ont été étudiées. Elles sont non biodégradables et pourraient servir de supports pour la régénération de larges défauts osseux. Nous avons montré la cytocompatibilité in vitro des membranes en analysant l’adhésion, la prolifération et la différenciation cellulaire. Ensuite, des membranes de PS ont été enrichies de grains de β-TCP ou de nanoparticules d’or. Nous les avons implantées dans un modèle de défaut crânien de taille critique chez la souris. L’addition de β-TCP a stimulé la repousse osseuse grâce à la grande bioactivité des céramiques. Les membranes ont été biotolérées, les fibres ont été encapsulées dans l’os néoformé mais également dans un tissu conjonctif dense. Les nanoparticules d’or immobilisées sur des fibres ont migré dans l’os ou ont été phagocytées. Dans un second temps, différentes formulations de granules poreux de β-TCP ont été analysées par nanotomographie aux rayons X. L’architecture macroporeuse des granules varie inversement avec la concentration en β-TCP. Les faces internes montrent une grande hétérogénéité de minéralisation. Pour mimer les conditions d’utilisation en chirurgie maxillo-faciale, les granules ont été empilés dans des tubes. Nous avons montré que l’architecture des empilements de granules dépendait de leur forme. L’empilement de granules commerciaux (contenant12,5g de β-TCP) mime l’architecture naturelle et les propriétés physiques de l’os. / The architecture of bone filling biomaterials has a real impact on cellular activity, vascularization and diffusion of growth factors. As a first step, different polystyrene (PS) scaffolds composed of aligned or random fibers were studied. They were non-biodegradable and could be used as supports for regeneration of large bone defects. We showed the in vitro cytocompatibility of the scaffolds by analyzing cell adhesion, proliferation and differentiation. Then, polystyrene scaffolds were enriched with β-TCP grains or gold nanoparticles. We implanted them in a model of critical size defect in mouse calvaria. Addition of β-TCP stimulated bone regrowth due to the high bioactivity of the ceramics. Scaffolds were biotolerated, fibers were encapsulated in the newly formed bone and also in a dense connective tissue. The gold nanoparticles immobilized on fibers migrated into the bone or were phagocytized. As a second step, different formulations of porous granules of β-TCP were analyzed by X-ray nanotomography. The macroporous architecture of granules varies inversely with the concentration of β-TCP. The internal faces howed a great heterogeneity of mineralization. Tomimic the conditions in maxillofacial surgery, granules were stacked in tubes. We have shown that the architecture of the granules depends on their shape.The stacks of commercial granules (containing 12.5 g ofβ-TCP) mimicked the natural architecture and physical properties of bone.

[ES] El desarrollo de la sociedad ha estado siempre ligado a los avances científicos y tecnológicos. Sin embargo, algunos de estos avances han supuesto la aparición de nuevos problemas, sobre todo de tipo ético, dando lugar a nuevos movimientos asociativos contrarios a ellos. Un ejemplo de ello es el uso de la experimentación animal, fundamentalmente en el campo de la medicina, y más concretamente en el ensayo de fármacos y dispositivos implantables en contacto con medios biológicos. En este sentido, desde el ámbito de los biomateriales y la ingeniería tisular se trabaja para buscar alternativas a la experimentación animal. Una de estas alternativas es el desarrollo de modelos in vitro a partir de soportes poliméricos para el crecimiento celular in vitro. Estas estructuras, además, podrían emplearse no sólo en ensayos de fármacos o investigación in vitro para reducir el uso de animales en experimentación, sino también para regeneración tisular, simulando desde tejidos simples en los que se tienen un único tipo celular, a tejidos más complejos a partir del co-cultivo celular. Así pues, en esta tesis se ha desarrollado un sistema tridimensional con estructura porosa estratificada que permite tanto el co-cultivo celular indirecto, como la realización de ensayos de liberación de fármacos. Para ello, se ha obtenido soportes porosos (scaffolds) por medio de la técnica de solvent-casting particle-leaching empleando como porógeno sal, cuyo tamaño de poro permite albergar células en su interior, sobre los que se han dispuesto, formando una estructura tipo sándwich, membranas electrohiladas. Estas membranas forman una estructura de fibras entrecruzadas, dejando entre ellas espacios de tamaño muy inferior al tamaño celular, de modo que permiten el paso de nutrientes y moléculas a través de ellas, pero actúan de barrera para las células impidiendo su migración a otras zonas del sistema tridimensional. Este tipo de estructuras permiten simular, por ejemplo, la arquitectura tubular renal, con la zona porosa intersticial central y las dos capas epitelial y endotelial externas, que estarían en contacto con la orina y la sangre, respectivamente. Para obtener estas estructuras, se ha optado por emplear polímeros de la familia de los poliésteres, en particular ácido poliláctico y poli(¿-caprolactona), así como su mezcla y copolímeros con ácido poliglicólico. Estas combinaciones permiten ajustar la hidrofilicidad, y por tanto la biodegradabilidad, la cinética de liberación de fármacos y el comportamiento biológico, según interese. Además, el uso de la técnica de electrospinning para el desarrollo de las membranas, permite obtener diferentes diámetros de fibra a partir de la modificación de los principales parámetros del electrohilado, permitiendo también regular las propiedades de estos materiales. Finalmente, para estudiar la liberación de fármacos desde el sistema anterior, las membranas electrohiladas se han cargado con curcumina mediante dos métodos diferentes: a través del método de electrohilado en disolución y con electrospinning coaxial, para obtener así diferentes perfiles de liberación. / [CA] El desenvolupament de la societat ha estat sempre lligat als avanços científics i tecnològics. Malauradament, alguns d'aquests avanços han suposat l'aparició de nous problemes, sobretot de tipus ètic, donant lloc a nous moviments associatius contraris a ells. Un exemple d'això és l'ús de l'experimentació animal, fonamentalment al camp de la medicina, i més concretament en el testatge de fàrmacs i dispositius implantables en contacte amb medis biològics. En aquest sentit, des de l'àmbit dels biomaterials i l'enginyeria tissular es treballa per a buscar alternatives a l'experimentació animal. Una d'aquestes alternatives és el desenvolupament de models in vitro a partir de suports polimèrics per al creixement cel·lular in vitro. Aquestes estructures, a més a més, podrien emprar-se no sols en testatge de fàrmacs o investigació in vitro per a reduir l'ús d'animals en experimentació, sinó també per a regeneració tissular, simulant des de teixits simples en què es té un únic tipus cel·lular, a teixits més complexos a partir del co-cultiu cel·lular. Així doncs, en aquesta tesi s'ha desenvolupat un sistema tridimensional amb estructura porosa estratificada que permet tant el co-cultiu cel·lular indirecte, com la realització d'assajos d'alliberació de fàrmacs. Per a això, s'ha obtingut suports porosos (scaffolds) mitjançant la tècnica solvent-casting particle-leaching emprant com a porògen sal, amb una grandària de porus que permet albergar cèl·lules en el seu interior, sobre els que s'han disposat, formant una estructura tipus sandvitx, membranes electrofilades. Aquestes membranes formen una estructura de fibres entrecreuades, deixant entre elles espais de grandària molt inferior a la grandària cel·lular, de mode que permeten el pas de nutrients i molècules a través d'elles, però actuen de barrera per a les cèl·lules impedint la seua migració a altres zones del sistema tridimensional. Aquest tipus d'estructures permeten simular, per exemple, l'arquitectura tubular renal, amb la zona porosa intersticial central i les dues capes epitelial i endotelial externes, que estarien en contacte amb l'orina i la sang, respectivament. Per a obtindre aquestes estructures, s'ha optat per emprar polímers de la família dels polièsters, en particular àcid polilàctic i poli(¿-caprolactona), així com la seua mescla i copolímers amb àcid poliglicòlic. Aquestes combinacions permeten ajustar la hidrofilicitat, i per tant la biodegradabilitat, la cinètica d'alliberació de fàrmacs i el comportament biològic, segons interesse. A més, l'ús de la tècnica d'electrospinning per al desenvolupament de les membranes, permet obtindre diferents diàmetres de fibra a partir de la modificació dels principals paràmetres de l'electrofilat, permetent també regular les propietats d'aquests materials. Finalment, per a estudiar l'alliberació de fàrmacs des del sistema anterior, les membranes electrofilades s'han carregat amb curcumina mitjançant dos mètodes diferents: a través del mètode d'emulsió i amb electrospinning coaxial, per a obtindre així diferents perfils d'alliberació. / [EN] The development of society has always been linked to scientific and technological advances. However, some of these advances have led to the appearance of new problems, especially of an ethical nature, giving rise to new associative movements opposed to them. An example of this is the use of animal experimentation, mainly in the field of medicine, and more specifically in the testing of drugs and implantable devices in contact with biological media. In this sense, the field of biomaterials and tissue engineering is working to find alternatives to animal experimentation. One of these alternatives is the development of in vitro models based on polymer supports for in vitro cell growth. These structures, moreover, could be used not only in drug testing or in vitro research to reduce the use of animals in experimentation, but also for tissue regeneration, simulating from simple tissues in which there is a single cell type, to more complex tissues from cell co-culture. Thus, in this thesis, a three-dimensional system with a stratified porous structure have been developed to allow both indirect cell co-culture and drug release assays. For this purpose, porous supports (scaffolds) have been obtained by means of the solvent-casting particle-leaching technique using salt as porogen, whose pore size allows cells to be housed in its interior, on which electrospun membranes have been arranged, forming a sandwich structure. These membranes form a structure of cross-linked fibers, leaving spaces between them that are much smaller than the cell size, so that they allow the passage of nutrients and molecules through them, but act as a barrier to the cells, preventing their migration to other areas of the three-dimensional system. This type of structure allows us to simulate, for example, the renal tubular architecture, with the central interstitial porous zone and the two outer epithelial and endothelial layers, which would be in contact with urine and blood, respectively. To obtain these structures, we have chosen to use polymers of the polyester family, in particular polylactic acid and poly(¿-caprolactone), as well as their blend and copolymers with polyglycolic acid. These combinations allow adjustment of hydrophilicity, and thus biodegradability, drug release kinetics and biological behavior, as desired. In addition, the use of the electrospinning technique for the development of membranes allows to obtain different fiber diameters by modifying the main electrospinning parameters, allowing also to regulate the properties of these materials. Finally, to study drug release from the previous system, the electrospun membranes have been loaded with curcumin by two different methods: through the emulsion method and with coaxial electrospinning, in order to obtain different release profiles. / Herrero Herrero, M. (2022). Desarrollo de andamiajes con porosidad estratificada basados en poliésteres como soportes en co-cultivo celular indirecto [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/190919

Thesis (MSc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: The need to analyse and detect volatile organic compounds (VOCs) at trace levels has led to the development of specialized sample preparation techniques. The requirement for trace analysis of VOCs stems from the negative effects they have on the environmental and human health. Methods for the analysis of non-polar VOCs commonly found as trace contaminants in water and analysis of more polar oxygenated compounds commonly found in zero-VOC water-based paints were developed. Solid phase micro extraction (SPME) was employed and extraction of the majority of the target analytes could be achieved at levels below 0.3 μg.l-1. In an attempt to further improve the detection of these two target analyte groups, novel materials based on poly(dimethyl siloxane) (PDMS) were investigated as possible extraction phases for VOCs, with the focus specifically on the analysis of the polar analytes in paint. Conventional free radical polymerization was used to synthesize poly(methyl methacrylate-graft-poly(dimethyl siloxane) (PMMA-g-PDMS), poly(methacrylic acid)-graft-poly(dimethyl siloxane) (PMAA-g-PDMS), polystyrene-graftpoly( dimethyl siloxane) (PSty-g-PDMS) and poly(butyl acrylate)-graft–poly(dimethyl siloxane) (PBA-g-PDMS). These polymers have a copolymer functionality which presents a series of different polarities. The MMA-g-PDMS and MAA-g-PDMS as well as the homopolymers were electrospun into nanofibers. The low glass transition temperature and molecular weight of the PBAg- PDMS meant that this polymer could not be electrospun. Scanning electron microscopy (SEM) was used to study the fiber morphology of the electrospun fibers and the non-beaded fibers were further investigated. Polyacrylonitrile-graft-poly(dimethyl siloxane) (PAN-g-PDMS) previously synthesized and electrospun by another member of the group were also investigated for use as possible extraction material in volatile analysis. The thermal stability of the nanofibers at 200°C was studied using thermal gravimetric analysis (TGA). This property is important since after the target analytes are extracted using the nanofibers, elevated temperatures are used to thermally desorp the volatile analytes from the extraction materials prior to GC analysis. The PAN-g-PDMS, MMA-g-PDMS and PMMA showed no significant weight loss during thermal evaluation, however, it was observed that the PMMA and PMMA-g-PDMS nanofibers looses their nanostructure and that the PAN-g-PDMS nanofibers changes colour from white to yellow to rust brown. The polymers based on MAA showed weight losses of more than 10% after one hour of exposure to the elevated temperatures, but the nanostructure remained intact. The PAN-g-PDMS, PMAA-g-PDMS and PMAA nanofibers were evaluated as possible extraction materials for VOC analysis. The nanofibers were evaluated using a similar approach to that of stir bar sorptive extraction (SBSE). Headspace sorptive extraction (HSSE) using a commercially available PDMS stir bar and the novel materials were used to evaluate the extraction efficiency of the different materials. The optimized extraction method developed using SPME were employed for the extraction using the nanofibers and PDMS stir bar. It was noted that the nanofibers lose their extraction capabilities during the first extraction/desorption cycle possibly due to thermal degradation therefore each of the materials can only be used in a single extraction. The majority of the non-polar analytes were extracted using the nanofibers at levels of 500 μg.l-1, however it was noted that the commercially available SPME extraction materials and the PDMS stir bar had superior extraction efficiencies for the specific target analytes. In the evaluation of the nanofibers for extraction of the more polar oxygenated analytes it was noted that 2-Ethylhexylacrylate was the only analyte to be extracted by all of the materials. The PAN-g-PDMS extracted three of the four analytes at levels of 100 μg.l-1. At lower analyte concentrations of 10 μg.l-1 only two of the four acrylate compounds were detected using the PAN-g-PDMS nanofibers. Ethyl acrylate was not extracted by any of the novel materials, whereas in SPME using the CAR/PDMS fiber, the LOD was determined to be below 1 μg.l-1. Although these materials were not superior to the commercially available phases, this is only the case for the specific target analytes analyzed. / AFRIKAANSE OPSOMMING: Die behoefte vir die analiese van vlugtige organiese verbindings (VOS) op spoorvlak, het gelei tot die ontwikkeling van gespesialiseerde monster voorbereidingstegnieke. Die vereiste vir die spoor analiese van die VOS het ontstaan uit die negatiewe uitwerking wat hierdie stowwe het op die omgewing en menslike gesondheid. Metodes vir die analiese van nie-polêre VOS wat algemeen voorkom as spoorkontaminante in water en polêre suurstofryke verbindings wat algemeen voorkom in nul-VOS water-gebaseerde verf was ontwikkel. Soliede fase mikro-ekstraksie (SFME) was gebruik, en die ekstraksie van die meerderheid van die teikenstowwe kon gedoen word op vlakke laer as 0,3 μg.l-1. In 'n poging om die opsporing van hierdie twee teiken analietgroepe verder te verbeter, is nuwe materiale gebaseer op polidimetielsiloksaan (PDMS), ondersoek as moontlik ekstraksiefases vir VOS, met die fokus spesifiek op die analiese van die polêre stowwe in verf. ’n Konvensionele vrye radikaal polimerisasieproses was gebruik om poli (metiel- metakrilaat)-entpoli( dimetielsiloksaan) (PMMA-g-PDMS), poli(metakrilaatsuur)-ent–poli (dimetielsiloksaan) (PMAA-g-PDMS), polistireen-ent-poli(dimetielsiloksaan) (PSty-g-PDMS) en poli(butielakrilaat)- ent-poli(dimetielsiloksaan) (PBA-g-PDMS) te sintetiseer. Hierdie ko-polimere het 'n kopolimeer funksionaliteit wat 'n reeks van verskillende polariteite bied. Die MMA-g-PDMS en MAA-g-PDMS sowel as die homopolimere was ge-elektrospin in orde om nanovesels te vorm. Die lae glasoorgangstemperatuur en molekulêre gewig van die PBA-g-PDMS het beteken dat hierdie polimeer nie elektrospin kon word nie. Skandeerelektronmikroskopie (SEM) was gebruik om die veselmorfologie van die ge-elektrospinde vesels te bestudeer en die nanovesels wat ’n eweredige oppervlak gehad het, was verder ondersoek. Poliakrilonitriel-ent-poli(dimetielsiloksaan) (PAN-g- PDMS) wat voorheen gesintetiseer en ge-elektrospin was deur 'n ander lid van die groep is ook ondersoek vir gebruik as moontlik ekstraksiemateriaal vir die analiese van vlugtige stowwe. Die termiese stabiliteit van die nanovesels was by 200°C bestudeer met behulp van ‘n termiese gravimetriese analiese (TGA) instrument. Hierdie eienskap is belangrik, aangesien die teikenstowwe by hoë temperature van die nanovesels gedesorbeer word voor die GC-analiese. Die PAN-g-PDMS, MMA-g-PDMS en PMMA het geen beduidende gewigsverlies tydens termiese evaluering gehad nie, alhoewel dit egter waargeneem was dat die PMMA en PMMA-g-PDMS nanovesels hulle nanostruktuur verloor en dat die PAN-g-PDMS nanovesels se kleur verander van wit na geel na roesbruin gedurende die termiese analiese. Die polimere wat gebaseer was op MAA het ’n gewigs-verlies van meer as 10% getoon na 'n uur van blootstelling aan die verhoogde temperature, maar die nanostruktuur het ongeskonde gebly. Die PAN-g-PDMS, PMAA-g-PDMS en PMAA nanovesels was geëvalueer as moontlike ekstraksiemateriale vir VOS-analiese. Die nanovesels was geëvalueer met 'n soortgelyke benadering tot dié van “stir bar“ sorpsie ekstraksie (SBSE). Bo-ruimte sorpsie ekstrasie is gebruik om die ekstraksie-doeltreffendheid van die verskillende materiale (kommersiële PDMS en nanovesels) te evalueer. Die geoptimaliseerde ekstraksiemetode ontwikkel in SFME was gebruik vir die ekstraksie van die VOS met die nanovesels en die PDMS “stir bar“. Dit was waargeneem dat die nanovesels hul ekstraksievermoë verloor tydens die eerste ekstraksie/desorpsie siklus, moontlik as gevolg van termiese degradasie dus, kon die materiale slegs ‘n enkele maal gebruik word vir die ekstraksie. Die meerderheid van die nie-polêre stowwe was ge-ëkstraeer deur gebruik te maak van die nanovesels op vlakke van 500 μg.l -1, maar die kommersieel beskikbare SFME ekstraksie materiale en die PDMS “stir bar“ se ekstraksie-doeltreffendheid vir die spesifieke stowwe was beter. In die evaluering van die nanovesels vir die ekstraksie van die meer polêre suurstofryke stowwe was daar waargeneem dat 2- etielheksielakrilaat die enigste analiet was wat ge-ëkstraeer was deur al die materiale. Die PAN-g- PDMS kon drie van die vier polêre stowwe op vlakke van 100 μg.l-1 opspoor. By laer analietkonsentrasies van 10 μg.l-1 kon slegs twee van die vier akrilaat verbindings opgespoor word deur gebruik te maak van hierdie nanovesels. Etielakrilaat was nie ge-ëkstraeer deur enige van die nuwe materiale nie, terwyl in SFME met die gebruik van die CAR/ PDMS vesel, die analiet op vlakke onder 1 μg.l-1 opgespoor kon word. Alhoewel hierdie nuwe materiale nie beter is as die kommersieel beskikbare ekstraksiemateriale nie is dit net die geval vir die spesifieke teiken analietgroepe wat ondersoek was in hierdie studie.

Electrospinning is a method of using high voltage electric fields to transform polymer solutions into nano-scale fibres. The field has seen significant work on processing different polymers and their resulting fibres but less work has focused the electrospinning process itself. The aim of this thesis is to present experimental observations of charge behaviour in the electrospinning process in the context of the underlying physics typically used to describe electrospinning. This thesis presents a review of existing methods of measuring aspects of the electrospinning process, and reviews published mathematical models of the process as representative examples of the current understanding of the underlying physics that drive the electrospinning phenomena. A novel measurement technique is introduced - high frequency data capture of the electric current flow simultaneously at the high voltage and collector electrode. This is used in three ways: to examine bulk charge density, to measure fibre flight time, and to quantify charge lost from the fibre in flight. Charge density is studied by comparing current and mass flow at the Taylor cone under a wide range of conditions. For 8% PVOH in water a constant bulk charge density was found of 7.7 C/kg. Flight time is studied by determining the time from the application of high voltage to the charged fibre first arriving at the collector electrode. It was found that for 8% PVOH the flight time depended strongly on applied voltage while electrode distance had a negligible effect. Charge loss was studied by comparing the magnitude of the simultaneous current flows in the quasi-steady state to determine if the charge flowing into the Taylor cone arrives with the fibre at the collector. For 8% PVOH, 8% PVOH with ionic salt, 9% PVOH in water and 18% PVB in ethanol, it was found that charge is always lost.

The objective of this research project was to evaluate the mechanical, morphological, and permeability properties of electrospun blends of polydioxanone and poly(lactic acid) for application as vascular grafts. Mechanical analysis was performed by uniaxial tensile testing to examine the peak load, peak stress, elastic modulus, and strain at break of the fibrous materials. The morphological characteristics of the polymer blends were analyzed using phase contrast microscopy, scanning electron microscopy, and image analysis software. Scanning electron microscopy and image analysis software were used to assess fiber diameter and pore size of electrospun scaffolds. Scaffold permeability measurements were also used to calculate fiber diameter and pore size, and the values were compared to those obtained using image analysis. The material property results acquired from the research suggest that the electrospun polymer blends have potential for use in vascular graft applications.

The overall goal of this study was to identify mineralized scaffolds which can serve as potential alternatives to bone graft substitutes intended for cleft palate repair. The aim of this preliminary study was to evaluate the role of fibrinogen (Fg) and nano-hydroxyapatite (nHA) in enhancing mineralization potential of polydioxanone (PDO) electrospun scaffolds. Scaffolds were fabricated by blending PDO:nHA:Fg in the following weight ratios: 100:0:0, 50:25:25, 50:50:0, 50:0:50, 0:0:100 and 0:50:50. Scaffolds were immersed in different simulated body fluids for 5 and 14 days to induce mineralization. The inclusion of fibrinogen induced sheet-like mineralization while individual fiber mineralization was noticed in its absence. Modified protocols of alizarin red staining and burn-out test were developed to quantify mineral content of scaffolds. After mineralization, 50:50:0 scaffolds were still porous and contained the most mineral. 50:25:25 scaffolds had the highest mineralization potential but lacked porosity. Therefore, it can be anticipated that these mineralized organic-inorganic electrospun scaffolds will induce bone formation.

This study examines the prospects of using the electrospinning process to fabricate tissue engineering scaffolds targeting a variety of regenerative applications, with a primary focus on the production of nerve guides for the treatment of long-defect nerve injuries in the peripheral nervous system. A basic overview of the conventional electrospinning process is provided, and the utility of this fabrication scheme in the production of collagen-based tissue engineering scaffolds is demonstrated. Next, a novel modification of the basic electrospinning process is presented. This process, called two pole air gap electrospinning, was developed to produce nerve guides that exhibit an anisotropic structure that mimics the extracellular matrix of native peripheral nerve tissue. This electrospinning process makes it possible to produce macroscopic nerve guides that are cylindrical in shape and composed of dense arrays of nano- to micron-scale diameter fibers. Unlike, conventional hollow core nerve guides, these electrospun constructs lack a central lumen, hence the designation 3D (for three-dimensional) nerve guide. The fibers are nearly exclusively arrayed in parallel with the long axis of the construct. This architectural feature provides thousands of individual channels, and aligned fibers that provide guidance cues that are designed to drive regenerating axons to grow in a highly directed fashion down the longitudinal axis of the guide. To supplement the structural cues provided by the fibrillar arrays of the electrospun 3D nerve guides, an alginate-based platform designed to deliver therapeutic reagents was developed and characterized. This platform makes it possible to fabricate gradients of therapeutic reagents within the fibrillar arrays of an electrospun nerve guide. Functional and structural analyses of these constructs supplemented with or without a gradient of NGF, in a long-defect nerve injury in the rodent sciatic nerve indicate that the 3D design is superior to the gold standard treatment, the autologous nerve graft. Animals treated with the 3D grafts recovered motor and sensory function faster and exhibited far higher nerve-to-nerve and nerve-to-muscle signal amplitudes in electrophysiological studies than animals treated with autologous grafts or conventional hollow core cylindrical grafts.

A superhydrophobic coating can be produced using a hydrophobic material textured with surface roughness on the micro-/nano-scale. Such a coating on the outside of a submersible body may result in reduced skin-friction drag due to a trapped layer of air in the coating. However, this layer may become unstable when subjected to elevated hydrostatic pressures, and a coating’s performance is compromised beyond a certain threshold (critical pressure). This thesis presents a numerical model for predicting the pressure tolerances of superhydrophobic coatings comprised of randomly deposited hydrophobic particles or fibers. We have also derived a set of force-balance-based analytical equations for predicting critical pressure in surfaces with ordered roughness, and compared our numerical model against it, observing reasonable agreement. The numerical model was then applied in a large parameter study, predicting critical pressure for coatings with a given set of microstructure properties.

Sterilization of tissue engineered scaffolds is an important regulatory issue and is at the heart of patient safety. With the introduction of new biomaterials and micro/nano structured scaffolds, it is critical that the mode of sterilization preserve these built-in features. Conventional sterilization methods are not optimal for engineered polymeric systems and hence alternate systems need to be identified and validated. PCL is polyester with a low melting point (heat labile), susceptible to hydrolysis and is popular in tissue engineering. Electrospinning generates some nanoscale features within the scaffold, the integrity of which can be affected by sterilization method. Chapter 1 explores the possibility of using Peracetic acid (PAA) to sterilize polymeric scaffolds that are sensitive to heat or moisture. PAA is a strong oxidizing agent that has been approved for sterilizing catheters and endoscopes. The ability of PAA to sterilize at room temperature, its breakdown into non-toxic end products and effectiveness at low concentrations are major advantages. Chapter 2 evaluates the ability of PAA-sterilized PCL scaffolds (PAA-PCL) to support survival and proliferation of mouse calvarial osteoblast cell line, MC3T3. While Ctrl-PCL scaffolds (ethanol-disinfected) showed robust cell survival, PAA-PCL scaffolds induced massive cell death. Following interrelated hypotheses are tested: the observed cytotoxicity was due adsorption of PAA and/or hydrogen peroxide onto PCL fibers during sterilization; and elimination of adsorbed residues will restore scaffold cytocompatibility. Neither extensive aeration nor chemical neutralization with sodium thiosulfate and catalase were effective in improving cell survival. However, quenching PAA-PCL scaffolds in 70% ethanol for 30 minutes effectively removed adsorbed PAA residues and completely restored cell viability and proliferation over a 7 day period. In order to test if PAA-induced toxicity was limited to electrospun PCL scaffolds, commercially available, porous polystyrene scaffolds (Alvetex®) was treated with PAA. Interestingly, a statistically significant increase in cell survival and proliferation resulted with PAA treatment and this was abolished by ethanol quenching. Combined, these results illustrate that PAA treatment can produce diametrically opposite effects on cell survival depending on substrate chemistry and that PAA can be used to effectively sterilize tissue engineering scaffolds without compromising cell viability.

Advanced biomaterials and sophisticated processing technologies aim to fabricate tissue-engineering scaffolds that can predictably interact within a biological environment at a cellular level. Sterilization of such scaffolds is at the core of patient safety and is an important regulatory issue that needs to be addressed prior to clinical translation. In addition, it is crucial that meticulously engineered micro- and nano- structures are preserved after sterilization. Conventional sterilization methods involving heat, steam and radiation are not compatible with engineered polymeric systems because of scaffold degradation and loss of architecture. Using electrospun scaffolds made from polycaprolactone (PCL), a low melting polymer, and employing spores of Bacillus atrophaeus as biological indicators, we compared ethylene oxide, autoclaving and 80% ethanol to a known chemical sterilant, peracetic acid (PAA), for their ability to sterilize as well as their effects on scaffold properties. PAA diluted in 20% ethanol to 1000 ppm or above, sterilized electrospun scaffolds in 15 min at room temperature while maintaining nano-architecture and mechanical properties. Scaffolds treated with PAA at 5000 ppm were rendered hydrophilic, with contact angles reduced to zero degrees. Therefore, PAA can provide economical, rapid and effective sterilization of heat-sensitive polymeric electrospun scaffolds used in tissue-engineering.