Browsing by Subject "PLGA"

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Design and development of an injectable, polymer-based immune priming center
(2010-05) Singh, Ankur; Roy, Krishnendu; Kwak, Larry W; Peppas, Nicholas A; Schmidt, Christine E; Suggs, Laura J; Williams III, Robert O
Immunotherapy, as a strategy to trigger immunity and eradicate a variety of chronic infectious diseases and cancers, has been explored for several decades with significant success in animal models. However, effective translation of these strategies into human clinical settings has proven elusive. Several cell-based anti-tumor therapies have progressed to clinical trials where antigen presenting dendritic cells (DCs) are isolated from patients, loaded with viral or tumor antigens and infused back in the patients. These ex-vivo “trained” DCs then present antigens to naïve T cells (adoptive therapy). However there exist several major limitations to this approach, including morbidity associated with patient cell isolation, high cost of ex vivo cell manipulation, time lag in “training” the immune cells, regulatory concerns, as well as the fact that ~ 90% of transplanted DCs die before they even home to lymph nodes. On the other hand, current immunotherapy approaches using recombinant proteins, synthetic peptides or nucleic acids, which "train" the immune cells in vivo to mount an immune response, have failed to address the tremendous challenge in generating efficient, sustained and protective immunity. There are two major challenges that must be overcome, (a) there exist relatively fewer numbers of immune cells at the sites of vaccine administration and given that these antigens themselves are weakly immunogenic, vaccine formulations must be tailored to attract large number of DCs to the immunization site and (b) immunologically, conventional nucleic acid or protein/peptide based vaccines do not elicit the required T helper type (Th) immunity along with a strong Cytotoxic T Lymphocyte (CTL) response against viral or tumor antigens and therefore new formulations must be able to “direct” the immune response towards a specific Th-type. Our goal was to design polymer-based sustained release formulations to addresses these challenges. Specifically, we have designed and developed delivery systems that can carry multiple biomolecules (nucleic acids, proteins, peptides, and chemoattractants) in a single injectable formulation. The delivery system promoted efficient migration of a large number of DCs to the site of injection and successful delivery of antigen resulting in activation of DCs. The multi-modal delivery system has the ability to bias or switch the immune response to the desired phenotype (e.g. Th1 or Th2) in a controlled manner. Using an infectious disease model against hepatitis B we have shown that co-encapsulation of Interleukin-10 (IL10) cytokine targeted siRNA within polymeric, surface-functionalized microparticles can further enhance DC activation and T cell proliferation in vitro as well as switch the hepatitis-specific immune response towards a strong Th1 phenotype in vivo. Further, in a weakly immunogenic A20 B cell lymphoma mouse model, a combination of microparticles and chemokine releasing in situ crosslinkable hydrogel provided significant Th1 type cellular immune response and delayed the onset of tumor development. Thus, the in situ crosslinkable hydrogel co-delivering microparticles and DC attracting chemokines creates an immune priming center with broad applications in a variety of disease models.
Development of multifunctional electrospun wraps for bone healing
(2020-11-17) Buie, Taneidra Walker; Cosgriff-Hernandez, Elizabeth; Suggs, Laura; Zoldan, Janeta; Laverty, David
The Masquelet technique is a two-staged procedure that uses an induced biological membrane and bone graft to reconstruct critical-sized bone defects. However, unpredictable clinical outcomes result due to the variable durability and the transient vascular network of the induced membrane, as well as high incidences of osteomyelitis. To this end, we have engineered a resorbable multifunctional electrospun wrap that guides formation of the induced membrane with improved durability and enhanced angiogenesis while simultaneously preventing infection. We achieve this by developing and combining an antimicrobial poly(lactic-co-glycolic) acid (PLGA) mesh and an angiogenic crosslinked gelatin mesh. We first confirmed the ability of electrospun PLGA to provide sustained release of gentamicin sulfate or gallium maltolate above its minimum inhibitory concentration (MIC). Studies that evaluated antimicrobial activity indicated that osteomyelitis-derived bacteria was not susceptible to released gallium maltolate at the hypothesized MIC and further established the accurate gallium maltolate MIC. The inhibitory concentration of each antimicrobial on osteoblasts was compared to the respective MIC to determine if they were safe and effective at released concentrations. Results concluded that the gentamicin sulfate-loaded PLGA mesh is safer and more effective mesh. Next, the bioactivity retention of vascular endothelial growth factor (VEGF) released from electrospun photo-crosslinked gelatin-methacrylate was confirmed. Subcutaneous implantation of the VEGF-loaded mesh in a rat corroborated resorption and the capacity for sustained release. A multifunctional electrospun wrap was then engineered to prevent osteomyelitis and guide formation of the induced membrane by combining the antimicrobial and angiogenic platforms with co-electrospinning. The combination of the two fiber populations was confirmed microscopically and offered independently tuned bimodal release of gentamicin sulfate and VEGF. Overall, this work provides the fundamentals to advance the development of a multifunctional electrospun wrap that can guide formation of the induced membrane and prevent osteomyelitis for improved clinical outcomes with the Masquelet technique. This work offers a substrate that can recruit and support cellular adhesion, provide a template for matrix deposition and tissue remodeling, and enable bimodal release of bioactive agents. These studies also enhance the capacity of electrospun platforms to serve as stand-alone therapies or combinatorial therapies in various bone regeneration applications.
Dexamethasone intravitreal implants : characterization, manufacture, and elucidation of drug release mechanisms
(2023-08) Costello, Mark Allen; Zhang, Feng, Ph. D.; Lynd, Nathaniel A; Williams III, Robert O; Smyth, Hugh DC
Long-acting injectable products based on the biodegradable polymer poly(lactide-co-glycolide) (PLGA) have been available in the USA since 1989, with more than 20 PLGA–based formulations approved for use to date. Despite the clinical and commercial success of many of these drug products, no generic formulations have gained FDA approval at the time of this writing. The lack of PLGA–based generics can largely be attributed to the technical challenges associated with development of formulations with drug release profiles equivalent to the reference product. Despite extensive study of the drug release mechanisms of PLGA, the inherent complexity of the copolymer is largely to blame for the challenges associated with generic product development. In this work, the FDA–approved product Ozurdex (dexamethasone intravitreal implant) was used as a model system to help address the challenges associated with generic product development of PLGA–based solid implants. Ozurdex is a small, rod-shaped implant (0.46 mm diameter, 6 mm long) formulated to deliver the corticosteroid dexamethasone to the posterior segment of the eye for 3–6 months. Ozurdex was thoroughly characterized and shown to be a porous implant consisting of a two-phase system of dexamethasone crystals embedded within a PLGA matrix due to a limited drug-polymer interaction. Compositionally equivalent, reverse-engineered implants were produced using a continuous hot-melt extrusion process that required careful control to manufacture implants structurally equivalent to Ozurdex. The drug release mechanisms of the reverse-engineered implants were studied in detail using a variety of analytical techniques to examine the implant throughout the drug release period. This work also demonstrated how sourcing PLGAs with similar, but subtly different, physicochemical properties can affect the manufacture and drug release kinetics of dexamethasone intravitreal implants.
Directing neuronal behavior via polypyrrole-based conductive biomaterials
(2011-05) Forciniti, Leandro; Zaman, Muhammad H.; Schmidt, Christine E.; Sanchez, Isaac C.; Maynard, Jennifer A.; Bonnecaze, Roger T.
The objective of my thesis is to explore the use of the conducting polymer, polypyrrole, in neural applications. In addition a supplementary aspect of dissertation will involves understanding the effects of external stimuli on nervous system cells, with the ultimate goal of designing therapeutic systems for nerve regeneration. In normal development and peripheral nervous system repair, nerves encounter naturally occurring chemical, physical, and electrical stimuli. Polypyrrole (PPy) has attracted much attention for use in numerous biomedical applications as it presents chemical, physical and electrical stimuli. In addition, PPy is particularly exciting because the extent by which chemical, physical, and electrical cues are presented to the injured nerve can be easily tailored. Thus, conducting polymers are excellent scaffolds for the exploration of how the cellular components of the nervous system (i.e., Schwann cells and neurons) interact with chemical, topographical, and electrical stimuli. This dissertation covers three main objectives and is supplemented by two additional topics. The two additional topics explore the effect stimuli present on the conducting polymer PPy have on neural interfaces. These fundamental studies use computational modeling to gain a better understanding of cellular motility on substrates containing different stimuli. Both topics are covered in the appendices of this dissertation. With regards to the three main objectives, I first characterized and optimized the electrochemical synthesis of the conducting polymer, PPy, for Schwann cell biocompatibility. Next, I investigated the effect the application of electrical cues through PPy has on Schwann cell migration. In addition to investigating the effect of the direct electrical current on Schwann cells I also considered the effect that electrical stimulation provided by PPy has on protein adsorption. Finally, I developed a hybrid PPy material that will provide advantageous properties for neural interfaces. Specifically, I describe the development of a polypyrrole:poly-(lactic-co-glycolic) acid blend for neural applications. In summary the three specific objectives covered in my thesis are: Specific Aim 1: Characterize and optimize the electrochemical synthesis of the conducting polymer, polypyrrole, for Schwann cell biocompatibility Specific Aim 2: Determine the effect of electrical stimulation on Schwann cell migration Specific Aim 3: Develop polypyrrole:poly-(lactic-co-glyolic) acid blends for neural engineering applications.