High Throughput Cell-Based Screening of Biodegradable Polyanhydride Libraries

Abstract

A parallel screening method has been developed to rapidly evaluate discrete library substrates of biomaterials using cell-based assays. The biomaterials used in these studies were surface-erodible polyanhydrides based on sebacic acid (SA), 1,6-bis(p-carboxyphenoxy)hexane (CPH), and 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) that have been previously studied as carriers for drugs, proteins, and vaccines. Linearly varying compositional libraries of 25 different polyanhydride random copolymers (based on CPH:SA and CPTEG:CPH) were designed, fabricated, and synthesized using discrete (organic solvent-resistant) multi-sample substrates created using a novel rapid prototyping method. The combinatorial libraries were characterized at high throughput using infrared microscopy and validated using 1H NMR and size exclusion chromatography. The discrete libraries were rapidly screened for biocompatibility using standard SP2/0 myeloma, CHO and L929 fibroblasts, and J774 macrophage cell lines. At a concentration of 2.8 mg/mL, there was no appreciable cytotoxic effect on any of the four cell lines evaluated by any of the CPH:SA or CPTEG:CPH compositions. Furthermore, the activation of J774 macrophages was evaluated by incubating the cells with the polyanhydride libraries and quantifying the secreted cytokines (IL-6, IL-10, IL-12, and TNFalpha). The results indicated that copolymer compositions containing at least 50% CPH induced elevated amounts of TNFalpha. In summary, the results indicated that the methodologies described herein are amenable to the high throughput analysis of synthesized biomaterials and will facilitate the rapid and rational design of materials for use in biomedical applications.

Neuronal protection against oxidative insult by polyanhydride nanoparticle-based mitochondria-targeted antioxidant therapy

“…This is important because it indicates that the drug release rate over time to neurons will be constant, potentially leading to increased bioavailability. Adding to the vast literature on the biocompatibility of polyanhydride particles, functionalized or otherwise (55–57), our work shows that these nanoparticles were non-toxic to dopaminergic or primary cortical neurons (Figures 2a and 3a). …”

Applications of polymeric adjuvants in studying autoimmune responses and vaccination against infectious diseases

“…In recent years, this polymer has been approved by the FDA for human use owing to its tissue compatibility and controlled biodegradation properties [93]. Degradation of this polymer occurs through surface erosion mechanisms, resulting in controlled release of antigens.…”

Amphiphilic Polyanhydride Films Promote Neural Stem Cell Adhesion and Differentiation

“…Polyanhydrides have been studied extensively as vehicles for protein and vaccine delivery over the past decade [9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27] ; however, little work to date has investigated their use in tissue engineering applications. These surface eroding polymers are biologically inert, nontoxic, and nonmutagenic and are capable of providing sustained release kinetics of encapsulated proteins.…”

“…11,[13][14][15]17,19,21,24,28 Specifically, polyanhydrides based upon 1,6-bis( p-carboxyphenoxy)hexane (CPH) and 1,8-bis( p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) have been studied extensively because of their amphiphilic properties, controlled protein release and pro-tein stabilization capabilities, immune modulation, and cellular compatibility. 9,22,23,[25][26][27] These polymers can also be functionalized to target-specific cellular receptors. 12 Upon degradation, CPTEG:CPH copolymers result in small changes in pH (7.6-7.1) unlike other systems (i.e., PLGA), which result in much greater decreases in pH to as low as 2.6.…”