Chronic pulmonary disorders, marked by excessive extracellular matrix deposition (ECM) or fibrosis, are the most resistant to present clinical therapies resulting in prognoses of 50% life expectancy three years from diagnosis. Inadequacies of current treatments may be attributable to limitations in non-invasive therapeutic administration modalities. However, with the use of polyketal microparticles (PKMs), a novel drug delivery vehicle, a myriad of therapeutic schemes may be explored. Polyketals are a new polymeric family characterized by tissue biocompatibility, rapid hydrolysis, and benign degradation byproducts making it attuned for pulmonary applications. Potential treatments such as siRNA, oligo nucleotides, enzymes and other biomolecules can be encapsulated within PKMs and administered non-invasively via inhalation. For this study, we selected a model therapeutic peptide, Ac-SDKP, with established anti-fibrotic properties as the load for PKMs. For lung dysfunctions accompanied by fibrotic scarring, Ac-SDKP possesses promise in restoring the normal ECM framework. To assess PKMs viability as a pulmonary drug delivery vehicle three objectives were initially defined: 1) Synthesize particles possessing aerodynamic properties conducive for aerosolization 2) Optimization of the therapeutic load, Ac-SDKP, in PKMs to levels that will translate to clinical dosing concentrations, and 3) Determine the biocompatibility of the PKMs in the lung. Optimization of the Ac-SKDP loading within PKMs and size analysis revealed that a solid in oil in water double emulsion particle synthesis technique produced the most ideal microspheres. Based on previous reports, the loading efficiency attained, when locally dispensed, should reach clinical dosing requirements. Synthesized particles were compatible with aerosolization criteria; i.e., diameters below 3 μm and low polydispersities. In addition, we evaluated PKM tissue biocompatibility using a murine lung model. Examination of bronchoalveolar lavage fluid demonstrated only a slight inflammatory response to intratracheal particle injections of PKMs whereas PLGA, a commonly used biomaterial, elicited a significantly higher response. Histological assessment of the lungs following particle injection verified PKMs biocompatibility superiority. In conclusion, small-diameter PKMs are a suitable delivery system for pulmonary drug delivery, capable of delivering small peptide therapeutics and evading the local inflammatory response. The present work will enable expansion of therapeutic avenues capable of combating chronic lung disease.

The delivery of growth factors has been attempted for a number of different therapies. The approach of delivering therapeutic growth factors in a safe and efficient manner is difficult and certain criteria should be met. These criteria include: binding the appropriate growth factors, maintaining their bioactivity, and delivering these proteins with controllable release kinetics for an extended period of time. These criteria encompass a set of guidelines that hope to mimic in vivo biological events such as neovascularization. The central goal of this thesis is to meet these criteria by introducing a novel delivery strategy for growth factors using a biocompatible polycation and heparin. It was hypothesized that a polycation could interact with heparin to form a complex with the potential to deliver bioactive growth factors with an adaptable release. This hypothesis was tested by examining the release kinetics of bFGF from the complex and investigating whether the released bFGF maintained its bioactivity. The [polycation:heparin:bFGF] complex was formed by mixing the components in water, resulting in a precipitate. This precipitate was able to deliver bFGF with controllable release kinetics and the bioactivity of the released bFGF was higher than bolus bFGF and comparable to heparin stabilized bFGF. This system is expected to have the ability to bind and deliver numerous heparin-binding growth factors. In conclusion, the delivery system developed in this research provides a novel mechanism for controlled release of growth factors. This delivery strategy has met the criteria listed earlier and this research has laid the foundation for a successful delivery vehicle. Further, a biocompatible polycation was synthesized, which is a critical component of the delivery system. This polycation exhibited in vitro and in vivo biocompatibility that was orders of magnitude higher than existing polycations and has the potential to be very useful in a variety of biomedical applications. This design principle is also expected to serve as a platform for the synthesis of other biocompatible polycations.

Ultrasound has been identified as a possible non-invasive drug delivery device that could avoid many of the problems found in traditional therapeutics. Studies have shown that ultrasound can deliver molecules into cells; however, the applicability of ultrasound has been limited due to uncontrollable cellular viability losses after sonication. In this study, we sought to quantify the heterogeneous bioeffects of ultrasound in order to gain more insight into how ultrasound affects cells. We were also concerned with identifying the causes of and preventing programmed cell death caused by ultrasound exposure. In order to accomplish these objectives, we used flow cytometry to group cells into quantifiable characteristic populations. This allowed us to identify the relative importance of different forms of rapid cell death. We found that up to 65% of cells (at the highest ultrasound pressure studied) can lose viability rapidly and, for the first time, quantified them among three distinct populations: (1) cells that retain normal size but lose plasma membrane integrity; (2) intact nuclei surrounded by plasma membrane remnants; (3) debris resulting from cellular lysis. Our analysis was supported by mechanical sorting of these populations and subsequent imaging using confocal microscopy. We then monitored the viable populations for 6 h after ultrasound exposure. Results indicated that up to 15% of viable cells (at the highest ultrasound pressure studied) underwent apoptosis, which we showed was associated with an influx of intracellular Ca2+; therefore, we developed a method of chelating intracellular Ca2+ after sonication in an effort to maintain viability of those cells. Using this technique, we showed for the first time that cells could be saved, and we were able to prevent apoptosis by 50%, thereby increasing the overall viability of cells exposed to ultrasound. We conclude that ultrasound is a useful method to deliver molecules into cells and that appropriate selection of sonication conditions can minimize cell death by rapid and apoptotic mechanisms.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2008. / Härtill 5 uppsatser.

Thesis (Ph.D.)--University of Wollongong, 2008. / Typescript. Includes bibliographical references: leaf 190-205.

Dissertation (Ph.D.)--Auburn University, / Abstract. Vita. Includes bibliographic references.

Thesis (Ph. D.)--University of Texas at Austin, 2006. / Vita. Includes bibliographical references.

Thesis (Ph. D.)--University of Texas at Austin, 2006. / Vita. Includes bibliographical references.

Dissertation (Ph.D.) -- University of Texas Southwestern Medical Center at Dallas, 2006. / Vita. Bibliography: p.210-222