Despite being the most prevalent presentation of Age-Related Macular Degeneration (AMD), dry AMD (dAMD) lacks a therapeutic treatment. Retinal pigment epithelium (RPE) dysfunction preceding the onset of dAMD has inspired interest in regenerative medicine approaches seeking to replenish the RPE and preserve visual acuity. Cell delivery to the subretinal space however has been met with challenges surrounding ease of access and invasive surgical implantation. Two-dimensional scaffolds have made use of natural and polymeric materials to act as carriers for RPE cells and various progenitor lines. These substrates mitigate issues surrounding the handling of delicate cell sheets harvested for transplant. As well, they are often successful in preserving RPE phenotype, supporting growth, and can be fine tuned to possess morphologies comparable to native extracellular matrix (ECM). Despite aiming to act as replacement Bruch’s membrane on which RPE resides, two-dimensional substrates are often notably bulky and require traumatic surgery for implantation. As a result, the use of injectable methods of cell delivery has gained appeal. Bolus injections, despite improved methods of administration, are correlated with issues of inadequate cell localization. In response, three-dimensional hydrogel carriers for retinal applications aim to encapsulate cells, allowing for better cell distribution as these materials spread throughout the subretinal space. Increased viscosity of hydrogels as compared to saline injections, is hypothesized to improve cell loss and reduce aggregation. Of particular interest are in situ gelling systems, which undergo physical changes upon injection. Gelation upon delivery works to further assist in maintaining the cells within their target site. Purity and reproducibility concerns associated with the use of natural materials in the development of hydrogel cell carriers, have inspired the use of synthetic thermoresponsive poly-N-isopropylacrylamide (pNIPAAm). pNIPAAm undergoes a liquid to gel transition at a lower critical solution temperature (LCST) of 32°C. Copolymerization with various hydrophobic and hydrophilic groups can be used to adjust gel properties such as increasing or decreasing LCST, allowing for degradation, and improving water retention. In the work described herein, two NIPAAm-based thermoresponsive hydrogels intended for use as subretinal cell carriers are proposed. / Thesis / Master of Applied Science (MASc)

京都大学 / 新制・課程博士 / 博士(医学) / 甲第24794号 / 医博第4986号 / 新制||医||1066(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 寺田 智祐, 教授 武藤 学, 教授 上杉 志成 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Articular cartilage damage caused by sports accidents, trauma or gradual wear and tear can lead to degeneration and the development of osteoarthritis because cartilage tissue has only limited capacity for intrinsic healing. Osteoarthritis causes reduction of mobility and chronic pain and is one of the leading causes of disability in the elderly population. Current clinical treatment options can reduce pain and restore mobility for some time, but the formed repair tissue has mostly inferior functionality compared to healthy articular cartilage and does not last long-term. Articular cartilage tissue engineering is a promising approach for the improvement of the quality of cartilage repair tissue and regeneration. In this thesis, a promising new cell type for articular cartilage tissue engineering, the so-called articular cartilage progenitor cell (ACPC), was investigated for the first time in the two different hydrogels agarose and HA-SH/P(AGE-co-G) in comparison to mesenchymal stromal cells (MSCs). In agarose, ACPCs´ and MSCs´ chondrogenic capacity was investigated under normoxic (21 % oxygen) and hypoxic (2 % oxygen) conditions in monoculture constructs and in zonally layered co-culture constructs with ACPCs in the upper layer and MSCs in the lower layer. In the newly developed hyaluronic acid (HA)-based hydrogel HA-SH/P(AGE-co-G), chondrogenesis of ACPCs and MSCs was also evaluated in monoculture constructs and in zonally layered co-culture constructs like in agarose hydrogel. Additionally, the contribution of the bioactive molecule hyaluronic acid to chondrogenic gene expression of MSCs was investigated in 2D monolayer, 3D pellet and HA-SH hydrogel culture. It was shown that both ACPCs and MSCs could chondrogenically differentiate in agarose and HA-SH/P(AGE-co-G) hydrogels. In agarose hydrogel, ACPCs produced a more articular cartilage-like tissue than MSCs that contained more glycosaminoglycan (GAG), less type I collagen and only little alkaline phosphatase (ALP) activity. Hypoxic conditions did not increase extracellular matrix (ECM) production of ACPCs and MSCs significantly but improved the quality of the neo-cartilage tissue produced by MSCs. The creation of zonal agarose constructs with ACPCs in the upper layer and MSCs in the lower layer led to an ECM production in zonal hydrogels that lay in general in between the ECM production of non-zonal ACPC and MSC hydrogels. Even though zonal co-culture of ACPCs and MSCs did not increase ECM production, the two cell types influenced each other and, for example, modulated the staining intensities of type II and type I collagen in comparison to non-zonal constructs under normoxic and hypoxic conditions. In HA-SH/P(AGE-co-G) hydrogel, MSCs produced more ECM than ACPCs, but the ECM was limited to the pericellular region for both cell types. Zonal HASH/P(AGE-co-G) hydrogels resulted in a native-like zonal distribution of ECM as MSCs in the lower zone produced more ECM than ACPCs in the upper zone. It appeared that chondrogenesis of ACPCs was supported by hydrogels without biological attachment sites such as agarose, and that chondrogenesis of MSCs benefited from hydrogels with biological cues like HA. As HA is an attractive material for cartilage tissue engineering, and the HA-based hydrogel HA-SH/P(AGE-co-G) appeared to be beneficial for MSC chondrogenic differentiation, the contribution of HA to chondrogenic gene expression of MSCs was investigated. An upregulation of chondrogenic gene expression was found in 2D monolayer and 3D pellet culture of MSCs in response to HA supplementation, while gene expression of osteogenic and adipogenic transcription factors was not upregulated. MSCs, encapsulated in a HA-based hydrogel, showed upregulation of gene expression for chondrogenic, osteogenic and adipogenic differentiation markers as well as for stemness markers. In a 3D bioprinting process, using the HA-based hydrogel, gene expression levels of MSCs mostly did not change. Nevertheless, expression of three tested genes (COL2A1, SOX2, CD168) was downregulated in printed in comparison to cast constructs, underscoring the importance of closely monitoring cellular behaviour during and after the printing process. In summary, it was confirmed that ACPCs are a promising cell source for articular cartilage engineering with advantages over MSCs when they were cultured in a suitable hydrogel like agarose. The performance of the cells was strongly dependent on the hydrogel environment they were cultured in. The different chondrogenic performance of ACPCs and MSCs in agarose and HA-SH/P(AGE-co-G) hydrogels highlighted the importance of choosing suitable hydrogels for the different cell types used in articular cartilage tissue engineering. Hydrogels with high polymer content, such as the investigated HA-SH/P(AGE-co-G) hydrogels, can limit ECM distribution to the pericellular area and should be developed further towards less polymer content, leading to more homogenous ECM distribution of the cultured cells. The influence of HA on chondrogenic gene expression and on the balance between differentiation and maintenance of stemness in MSCs was demonstrated. More studies should be performed in the future to further elucidate the signalling functions of HA and the effects of 3D bioprinting in HA-based hydrogels. Taken together, the results of this thesis expand the knowledge in the area of articular cartilage engineering with regard to the rational combination of cell types and hydrogel materials and open up new possible approaches to the regeneration of articular cartilage tissue. / Gelenkknorpeldefekte, die durch Sportverletzungen, Unfälle oder graduelle Abnutzung ent-stehen, können zu Degeneration des Gewebes und zur Entstehung von Arthrose führen, da Knorpelgewebe nur über eine eingeschränkte Fähigkeit zur Selbstheilung verfügt. Arthrose reduziert die Beweglichkeit und verursacht chronische Schmerzen. Sie ist vor allem bei älte-ren Menschen einer der häufigsten Gründe für körperliche Behinderung. Die zurzeit verfüg-baren operativen Behandlungsmöglichkeiten können die Symptome meist für einige Zeit lindern, aber das dabei gebildete Ersatzgewebe zeigt meistens nur eingeschränkte Funktiona-lität im Vergleich zu natürlichem gesunden Knorpelgewebe und bleibt nur für eine begrenzte Zeit stabil. Tissue Engineering von Gelenkknorpelgewebe ist ein vielversprechender Ansatz, um die Qualität des Ersatzgewebes und der Knorpelregeneration zu verbessern. Diese Arbeit untersuchte einen neuen vielversprechenden Zelltyp für das Tissue Engineering von Knorpelgewebe, sogenannte Gelenkknorpel-Vorläuferzellen (ACPCs). Diese Zellen wurden erstmals in zwei verschiedenen Hydrogelen, Agarose und HA-SH/P(AGE-co-G), mit mesenchymalen Stromazellen (MSCs) verglichen. Die chondrogene Kapazität von ACPCs und MSCs in Agarose wurde unter normoxischen (21 % Sauerstoff) und hypoxischen (2 % Sauerstoff) Bedingungen in Monokultur und zonal geschichteter Kokultur untersucht. In den zonalen Kokulturen befanden sich ACPCs in einer oberen Schicht und MSCs in einer unte-ren Schicht. In dem neu entwickelten Hyaluronsäure (HA)-basierten Hydrogel HA-SH/P(AGE-co-G) wurde die chondrogene Differenzierung von ACPCs und MSCs ebenfalls in Monokultur und in zonal geschichteter Kokultur, wie im Agarose-Hydrogel, analysiert. Außerdem wurde der Beitrag des biologisch aktiven Moleküls Hyaluronsäure zur chondro-genen Genexpression von MSCs in 2D-, 3D-Pellet- und HA-SH-Hydrogel-Kulturen unter-sucht. Diese Arbeit zeigte, dass sowohl ACPCs als auch MSCs in Agarose- und HA-SH/P(AGE-co-G)-Hydrogelen chondrogen differenzieren konnten. ACPCs produzierten im Agarose-Hydrogel ein Gewebe, das dem Gelenkknorpel ähnlicher war als das von MSCs produzierte Gewebe, da es mehr Glykosaminoglykane (GAG), weniger Typ I Kollagen und nur geringe Aktivität der Alkalinen Phosphatase (ALP) aufwies. Hypoxische Bedingungen konnten die Produktion von extrazellulärer Matrix (ECM) durch ACPCs und MSCs nicht erhöhen, aber sie verbesserten die Qualität des von MSCs produzierten Gewebes. Die Herstellung von zon-alen Agarose-Konstrukten mit ACPCs in der oberen Schicht und MSCs in der unteren Schicht führte zu einer ECM-Produktion in zonalen Hydrogelen, die im Allgemeinen zwi-schen der ECM-Produktion der ACPC-Monokultur und der MSC-Monokultur lag. Zonale Kokultur von ACPCs und MSCs führte zwar nicht zu einer erhöhten ECM-Produktion, al-lerdings beeinflussten die beiden Zelltypen sich gegenseitig und modulierten zum Beispiel die Intensitäten der Typ II und Typ I Kollagen Färbungen im Vergleich zu Monokulturen unter normoxischen und hypoxischen Bedingungen. Im HA-SH/P(AGE-co-G)-Hydrogel produzierten die MSCs mehr ECM als die ACPCs, allerdings war die Verteilung der gebilde-ten ECM bei beiden Zelltypen auf den perizellulären Bereich beschränkt. Zonale HA-SH/P(AGE-co-G)-Hydrogele führten zu einer zonalen Verteilung von ECM, die der natürli-chen Struktur von Gelenkknorpel ähnlich war, da die MSCs in der unteren Schicht mehr ECM produzierten als die ACPCs in der oberen Schicht. Anscheinend wurde die chondroge-ne Differenzierung von ACPCs von Hydrogelen unterstützt, die, so wie Agarose, keine bio-logischen Bindestellen aufwiesen, und die Chondrogenese von MSCs profitierte von Hydro-gelen mit biologischen Signalen wie HA. Da HA ein attraktives Material für Tissue Engineering von Knorpel darstellt und das HA-basierte Hydrogel HA-SH/P(AGE-co-G) anscheinend die chondrogene Differenzierung von MSCs begünstigte, wurde der Beitrag von HA zur chondrogenen Genexpression in MSCs untersucht. Eine Hochregulation der chondrogenen Genexpression ließ sich in 2D- und 3D-Pellet-Kulturen von MSCs als Reaktion auf HA beobachten, während die Genexpression von osteogenen oder adipogenen Transkriptionsfaktoren nicht hochreguliert wurde. Der Ein-schluss von MSCs in einem HA-basierten Hydrogel führte zu einer Erhöhung der Genex-pression von chondrogenen, osteogenen, adipogenen und Stemness-Markern. Ein 3D-Druck-Prozess mit dem HA-basierten Hydrogel veränderte die Genexpression von MSCs in den meisten Fällen nicht. Dennoch wurde die Expression von drei getesteten Genen (COL2A1, SOX2, CD168) in gedruckten im Vergleich zu gegossenen Konstrukten herunterreguliert. Dies unterstrich die Wichtigkeit einer genauen Kontrolle des Verhaltens der Zellen während und nach dem Druck-Prozess. Zusammenfassend ließen sich ACPCs als vielversprechender neuer Zelltyp für das Tissue Engineering von Gelenkknorpelgewebe bestätigen. ACPCs haben Vorteile gegenüber MSCs, vor allem, wenn sie in einem passenden Hydrogel wie Agarose kultiviert werden. Die Leis-tung der Zellen war stark von den verschiedenen Hydrogelen und der Umgebung beeinflusst, die diese den Zellen darboten. Die unterschiedliche chondrogene Leistung von ACPCs und MSCs in Agarose- und HA-SH/P(AGE-co-G)-Hydrogelen zeigte deutlich die übergeordnete Relevanz der Auswahl von passenden Hydrogelen für die verschiedenen Zelltypen, die im Tissue Engineering von Gelenkknorpel Verwendung finden. Hydrogele mit einem hohen Polymergehalt, wie das eingesetzte HA-SH/P(AGE-co-G)-Hydrogel, können die Verteilung der gebildeten ECM auf den perizellulären Bereich beschränken und sollten weiterentwickelt werden, um einen niedrigeren Polymergehalt und damit eine homogenere ECM-Verteilung durch die kultivierten Zellen zu erreichen. Der Einfluss von HA auf die chondrogene Gen-expression und auf die Balance zwischen Differenzierung und Erhaltung der Stemness in MSCs ließ sich aufzeigen. In Zukunft sollten weitere Studien die Signalfunktionen von HA und den Einfluss des 3D-Drucks in HA-basierten Hydrogelen genauer zu untersuchen. Zusammengenommen erweitern die Ergebnisse dieser Arbeit das Wissen im Bereich des Tissue Engineerings von Gelenkknorpelgewebe, vor allem in Bezug auf eine rationale Kom-bination von Zelltypen und Hydrogel-Materialien, und eröffnen neue Ansätze zur Knorpel-regeneration.

No description available.

Despite the development of therapeutic interventions to prevent mortality following myocardial infarction (MI), there is a significant long-term risk of developing heart failure (HF). Injectable collagen hydrogels have demonstrated considerable promise as a therapeutic solution to reduce adverse ventricle remodeling associated with the development of HF post-MI. Matrix metalloproteinases (MMPs) are proteolytic enzymes involved in the degradation of the structural components of the extracellular matrix (ECM). The activation of MMPs following MI is an essential step in the cardiac repair process. However, uncontrolled enzymatic activity during this time has been associated with the formation of adverse fibrosis. Given the role of the proteases in tissue remodeling, MMPs may be a potential biomarker to predict the development of HF. This thesis work seeks to examine the effect of a novel hydrogel matrix therapy on cardiac tissue post-MI using broad-spectrum MMP-targeted radiotracer, [18F]BR3531. In Study 1, serial positron emission tomography (PET) imaging was performed to elucidate the spatial and temporal binding of [18F]BR351 post-MI using a murine model. Imaging was performed by administering [18F]BR351 at time points corresponding with periods of peak MMP activation post-MI. In vivo PET imaging and in vitro autoradiography demonstrated decreased [18F]BR351 binding in the infarct region. In Study 2, the model was used to evaluate the efficacy of a therapeutic collagen hydrogel to attenuate tissue remodeling. The groups that received the matrix treatment exhibited improved [18F]BR351 uptake in the infarct region. However, conflicting results between in vivo imaging and in vitro autoradiography, and immunohistochemistry using MMP2 and MMP9 indicate that [18F]BR351 may not be suited for MMP imaging in mouse models of MI.

Having access to clean water is not a certainty for every human being. Today, there are major problems with polluted water that not just affect us humans, but also the ecosystem around us. In recent years, research into making aerogels from protein nanofibrils (PNF) has increased. What is interesting about these gels is their properties of adsorbing contaminants in the water, such as organic molecules and metal ions. In this report, hydrogels and non-dry aerogels (called “non-dry” due to there being one additional heat-treatment step that can be performed to “dry” the aerogel to make it sturdier and more water-proof), have been produced by whey protein isolate (WPI), which is extracted from the dairy industry. This is a sustainable, cheap, and renewable raw material. The goals are thus: (1) to examine if there is a difference in adsorption capacity between hydrogels and non-dry aerogels in a static setup and (2) to understand and examine which product parameters give the gels the best adsorption efficiency. To achieve these goals, eight different gels were synthesized with eight different contents. The parameters have varied from the following; dialyzed or non-dialyzed WPI solution, straight or curly seeds and whether or not salt had been added to the gels. The samples’ adsorption efficiency was analyzed by micro equilibrium dialysis (MED) and UV-VIS spectroscopy. For hydrogels, the dialyzed solution with salt and curly seeds was the variant with the highest binding capacity for ThT at 34% of the initial ThT concentration. For non-dry aerogels the dialyzed solution with curly seeds and no added salt was the best variant for binding ThT, at 84% of initial ThT content bound. It was also seen that the gels that have non-dialyzed WPI solution or that the solutions contained salt increased the adsorption capacity. The conclusions drawn were that salt has a positive effect on the adsorption capacity of the gels that have formed a matrix, and a negative effect on the adsorption of loose fibrils as well as that the solutions may not need to be dialyzed. The type of seeds or fibrils effect on adsorption capacity was inconclusive. The adsorption capacity for non-dry aerogels was three times higher compared to hydrogels. / Att ha tillgång till rent vatten är inte en självklarhet för varje människa. Idag är det stora problem med förorenat vatten som inte bara påverkar oss människor utan också ekosystemet. På senare år har forskning inom att tillverka aerogeler av protein nanofibriller (PNF) ökat. Det som är intressant med dessa geler är deras egenskaper att adsorbera föroreningar i vattnet, som till exempel organiska molekyler och metalljoner. I detta arbete har icke-torra aerogels (kallas "icke-torra" på grund av att det finns ytterligare ett värmebehandlingssteg som kan utföras för att "torka" aerogelen för att göra dem robustare och mer vattentålig) och hydrogels producerats av vassleproteinisolat WPI, som utvinns från mjölkindustrin. Detta protein är ett hållbart, billigt och förnybart råmaterial. Målet med denna rapport är att (1) undersöka om det finns skillnader i adsorptionskapaciteterna mellan hydrogels och icke-torra aerogels i ett statiskt medium (2) förstå och undersöka vilka produktionsparametrar som ger den bästa adsorptionskapaciteten. För att uppnå dessa mål har åtta olika geler syntetiserats med åtta olika innehåll. De varierande parametrarna var följande; dialyserad eller icke dialyserad WPI-lösning, raka eller krulliga frön och om salt har tillsatts eller inte. Proverna analyserades med mikro-jämvikts-dialys och UV-VIS spektroskopi. För hydrogeler var den dialyserade lösningen med salt och krulliga seeds den variant som hade högst bindningskapacitet för ThT, vilket låg på 34 % av den orginellaThT-koncentrationen. För icke-tora aerogels var lösningen som var dialyserad med krulliga frön och inget tillsatt salt den variant som var bäst på att binda ThT, med en kapacitet på 84% av den orginella koncentrationen. Vi såg även att gelerna som hade odialyserad WPI-lösning alternativt att lösningarna som innehöll salt ökade adsorptionskapaciteten. Slutsatserna som dragits är att salt har en positiv påverkan på gelernas adsorptionsförmåga när de format en fibermatris och en negativ påverkan när de bara består av fria fibriller, samt att lösningarna kanske inte behöver dialyseras. Typ av frön eller fibrillers påverkan på adsorptionsförmåga kunde inte påvisas. Icke-torra aerogels hade en adsorptionsförmåga på ThT som är tre gånger högre jämfört med hydrogels.

No description available.

Soil moisture deficits often hinder rangeland revegetation efforts. Due to sheer numbers and germination timing, invasive annual species can easily outcompete those of desired seeded species for resources, including water. One way to increase seedling establishment may be to increase the amount of water available to seeded species. Superabsorbent polymers (SAPs) can absorb and release hundreds of times their weight in water and slowly release it. Concentrated SAP bands at or near seedling rooting depth may act as a soil water reservoir for germinating seedlings and increase rangeland seedling establishment. The objective of these studies was to evaluate the effect of banded SAP on soil moisture and seedling establishment through a series of eight glasshouse and field trials with different combinations of: species, SAP band rate, SAP band placement depth, seeding rate, fertilizer application, and seedling distance from the SAP band. Two grasses, bottlebrush squirreltail (Elymus elymoides) or Siberian wheatgrass (Agropyron fragile) were used as test species in all studies but one. In that study six species commonly used in rangeland restoration [bottlebrush squirreltail, crested wheatgrass (Agropyron cristatum (L.)), Lewis flax (Linum lewisii Pursh), yellow sweet clover (Melilotus officinalis), Wyoming big sagebrush (Artemisia tridentata Nuttall ssp. wyomingensis Beetle Young), and forage kochia (Kochia prostrata (L.) Schrad.)] were used. Concentrated band of SAP at rates up to 6000 kg ha-1 were placed below the seeds at depths ranging from the soil surface to 15 cm deep or mixed into the top 8 or 15 cm of soil. Measurements varied with trial. Soil moisture measurements included gravimetric water content, volumetric water content, and/or soil matric potential. Seedling growth parameters included seedling time to emerge, number emerged, longevity, percent of seedlings alive, shoot length, blade count, root length, root biomass, shoot biomass, and/or root:shoot ratio. Banded SAP at high rates increased usable soil moisture up to 49 days in the glasshouse and in spring of the first year after installation in the field. Rates ? 1500 kg ha-1 increased seedling longevity up to 12 days in the glasshouse but resulted in raised and cracked soil. This negatively impacted seedling survival in the field but not the glasshouse and may also reduce the amount of available soil moisture over time. Banded SAP at rates < 1500 kg ha-1, intended to reduce soil cracking, had no impact on soil moisture or live seedlings in the field. Further work is needed to investigate alternative application methods, such as blending the SAP into the soil.

Osteoarthritis (OA) is a progressive cartilage degeneration disease with a complex pathologic mechanism. Although OA has devastating effects on patient quality of life and places a significant burden on the healthcare system, no disease-modifying drugs have been found, and surgical treatment options are often unsustainable. 3D bioprinting is a novel field within tissue engineering that focuses on developing biocompatible constructs that can be implanted to replace an organ or tissue. Such constructs have a great potential to become treatments for OA. Understanding cell mobility within hydrogels could play a vital role in advancing the development of biocompatible constructs. However, due to the novelty of bioprinting, limited research on cell mobility within hydrogels is available. Therefore, this project aims to fill the gap in existing research regarding cell mobility within bioprinted constructs with varying mechanical properties. To achieve this goal, green fluorescent protein-tagged mesenchymal stromal cells (MSCs) were developed to assess progenitor cell mobility in bioprinted hydrogel constructs. Constructs were printed with three zones: hydrogel with embedded chondrocytes or MSCs; hydrogel spacer; and chemoattractant. Designed constructs were bioprinted (BioAssemblyBot, Advanced Solutions) using GelMA:HAMA bioinks containing photoinitiator with varying bioink percentages. Cell viability and directional mobility within constructs were assessed by fluorescence viability assay and time-lapse fluorescence microscopy. The protocol to evaluate cell mobility in bioprinted constructs and optimized bioprinting settings for GelMA:HAMA bioinks were gained through this project. Overall, this project allowed us to fill the gap in existing knowledge regarding MSC and chondrocyte mobility in hydrogels and contribute to developing a novel treatment method for OA.

Silicone Hydrogel (SiHy) contact lenses are highly successful compared to previous soft lenses; they were developed to provide superior oxygen permeability. However, the hydrophobic natures of the silicone segments enhance lipid sorption which may diminish the lens surface wettability, clarity and comfort. While lens and lens care product are designed to remove lipid deposition, there is lack of experimental evidence to evaluate the actual performances with respect to lipid removal. An in vitro model using an artificial tear fluid containing radiolabeled lipids was employed in this thesis research to evaluate the efficacy of different multi-purpose lens care solutions in removing lipids from SiHy contact lenses. Additional rubbing with the lens care solution is often encouraged by professionals. Part of this research evaluated the effect of additional rubbing process on lipid removal. Overall, a multi-purpose solution (MPS) for lens care, Opti-Free PureMoist®, removed the most lipid deposition from lenses (senofilcon A, comfilcon A, and balafilcon A and one conventional hydrogel lens polymacon). The overall removal percentages were approximately 55% of DPPC and 28% of cholesterol from a conventional hydrogel. However, the MPSs did not remove lipids effectively from SiHy lenses. The highest percentages of removal were 3.08% of DPPC and 0.76% of cholesterol from SiHy lotrafilcon B lenses with Opti-Free PureMoist. The rubbing process increased the amount of removal in some MPSs, but the effects were small. The lack of removal of lipid suggests that the surfactants in the MPSs are not hydrophobic enough to remove lipids from SiHy lenses. Apparently a majority of deposited lipids absorbed into the lens matrix as rubbing did not enhance removal significantly. Future study on determining the concentration profile of lipid sorption throughout the lens thickness is encouraged. Another topic in this research thesis is the use of hydrogel lenses to deliver comfort agents or lubricating molecules from lenses. A screening study was performed in this research to select possible agents to be loaded into several SiHy macromer formulations. Experiments showed that comfort agents PNVP and Kollidon were the best candidates for such a procedure.