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The delivery of hydrophilic drugs from hydrophobic polymers is a long-standing challenge in the biomaterials field due to the limited solubility of the therapeutic agent within the polymer matrix. In this work, we develop a drug delivery mechanism that enables the impregnation and subsequent elution of hydrophilic drugs from a hydrophobic polymer material. This was achieved by synthesizing core cross-linked star polymer amphiphiles with hydrophilic cores and hydrophobic coronas. While significant work has been done to create nanocarriers for hydrophilic drugs, this work is distinct from previous work in that it designs amphiphilic and core cross-linked particles for controlled release from hydrophobic matrices. Ultraviolet-mediated atom transfer radical polymerization was used to synthesize the poly(ethylene glycol) (PEG)-based hydrophilic cores of the star polymers, and hydrophobic coronas of poly(caprolactone) (PCL) were then built onto the stars using ring-opening polymerization. We illustrated the cytocompatibility of PCL loaded with these star polymers through human endothelial cell adhesion and proliferation for up to 7 days, with star loadings of up to 40 wt %. We demonstrated successful loading of the hydrophilic drug heparin into the star polymer core, achieving a loading efficiency and content of 50 and 5%, respectively. Finally, the heparin-loaded star polymers were incorporated into a PCL matrix and sustained release of heparin was illustrated for over 40 days. These results support the use of core cross-linked star polymer amphiphiles for the delivery of hydrophilic drugs from hydrophobic polymer matrices. These materials were developed for application as drug-eluting and biodegradable coronary artery stents, but this flexible drug delivery platform could have impact in a broad range of medical applications.

Antibiotic-resistant bacteria are a severe threat to human health. The World Health Organization's Global Antimicrobial Surveillance System has revealed widespread occurrence of antibiotic resistance among half a million patients across 22 countries, with Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae being the most common resistant species. Antimicrobial nanoparticles are emerging as a promising alternative to antibiotics in the fight against antimicrobial resistance. In this work, selenium nanoparticles coated with the antimicrobial polypeptide, e-poly-l-lysine, (Se NP-e-PL) were synthesized and their antibacterial activity and cytotoxicity were investigated. Se NP-e-PL exhibited significantly greater antibacterial activity against all eight bacterial species tested, including Gram-positive, Gram-negative, and drug-resistant strains, than their individual components, Se NP and e-PL. The nanoparticles showed no toxicity toward human dermal fibroblasts at the minimum inhibitory concentrations, demonstrating a therapeutic window. Furthermore, unlike the conventional antibiotic kanamycin, Se NP-e-PL did not readily induce resistance in E. coli or S. aureus. Specifically, S. aureus began to develop resistance to kanamycin from ∼44 generations, whereas it took ∼132 generations for resistance to develop to Se NP-e-PL. Startlingly, E. coli was not able to develop resistance to the nanoparticles over ∼300 generations. These results indicate that the multifunctional approach of combining Se NP with e-PL to form Se NP-e-PL is a highly efficacious new strategy with wide-spectrum antibacterial activity, low cytotoxicity, and significant delays in development of resistance.

We have engineered biomaterials that display nanoclusters of ligands that bind both integrin and syndecan-4 cell receptors. These surfaces regulate cell behaviors under static conditions including adhesion, spreading, actin stress fiber formation, and migration. The syndecan-4 receptors are also critical mediators of cellular mechanotransduction. In this contribution we assess whether this novel class of materials can regulate the response of cells to applied mechanical stimulation, using the shear stress imparted by laminar fluid flow as a model stimulus. Specifically, we assess endothelial cell detachment due to flow, cell alignment due to flow, and cell adhesion from the flowing fluid. A high degree of cell retention was observed on surfaces containing integrin-binding ligands or a mixed population of integrin- and syndecan-binding ligands. However, the presence of both ligand types was necessary for the cells to align in the direction of flow. These results imply that integrin engagement is necessary for adhesion strength, but engagement of both receptor types aids in appropriate mechanotransduction. Additionally, it was found that surfaces functionalized with both ligand types were able to scavenge a larger number of cells from flow, and to do so at a faster rate, compared to surfaces functionalized with only integrin- or syndecan-binding ligands. These results show that interfaces functionalized with both integrin- and syndecan-binding ligands regulate a significant range of biophysical cell behaviors in response to shear stress.

Purpose: Selenium nanoparticles (Se NPs) are promising antibacterial agents to tackle the growing problem of antimicrobial resistance. The aim of this study was to fabricate Se NPs with a net positive charge to enhance their antibacterial efficacy. Methods: Se NPs were coated with a positively charged protein--recombinant spider silk protein eADF4([kappa] 16)--to give them a net positive surface charge. Their cytotoxicity and antibacterial activity were investigated, with negatively charged polyvinyl alcohol coated Se NPs as a control. Besides, these eADF4([kappa] 16)-coated Se NPs were immobilized on the spider silk films, and the antibacterial activity of these films was investigated. Results: Compared to the negatively charged polyvinyl alcohol coated Se NPs, the positively charged eADF4([kappa] 16)-coated Se NPs demonstrated a much higher bactericidal efficacy against the Gram-negative bacteria E. coli, with a minimum bactericidal concentration (MBC) approximately 50 times lower than that of negatively charged Se NPs. Cytotoxicity testing showed that the eADF4([kappa] 16)-coated Se NPs are safe to both Balb/3T3 mouse embryo fibroblasts and HaCaT human skin keratinocytes up to 31 [micro]g/mL, which is much higher than the MBC of these particles against E. coli (8 [+ or -] 1 [micro]g/mL). In addition, antibacterial coatings were created by immobilising the eADF4([kappa] 16)-coated Se NPs on positively charged spider silk films and these were shown to retain good bactericidal efficacy and overcome the issue of low particle stability in culture broth. It was found that these Se NPs needed to be released from the film surface in order to exert their antibacterial effects and this release can be regulated by the surface charge of the film, such as the change of the spider silk protein used. Conclusion: Overall, eADF4([kappa] 16)-coated Se NPs are promising new antibacterial agents against life-threatening bacteria.

A dual-responsive, cell capture and release surface was prepared through the incorporation of phenylboronic acid (PBA) groups into an oxime-based polyethylene glycol (PEG) hydrogel. Owing to its PEG-like properties, the unfunctionalized hydrogel was nonfouling. The use of highly efficient oxime chemistry allows the incorporation of commercially available 3,5-diformylphenyl boronic acid into the hydrogel matrix. Thus, the surface properties of the hydrogel were modified to enable reversible cell capture and release. Boronic ester formation between PBA groups and cell surface carbohydrates enabled efficient cell capture at pH 6.8. An increase to pH 7.8 resulted in cell detachment. This capture-and-release procedure was performed on MCF-7 human breast cancer cells, NIH-3T3 fibroblast cells, and primary human umbilical vein endothelial cells (HUVECs) and could be cycled with negligible loss in activity. The facile preparation of PBA-functionalized surfaces presented here has applications in biomedical fields such as cell diagnostics and cell culture.

Blood contacting devices are commonly used in today's medical landscape. However, such devices (including small diameter vascular grafts) are limited by poor blood compatibility and may fail due to thrombosis. An attractive strategy for improving the blood compatibility of such devices is to generate biomaterials that foster a confluent and functioning endothelial cell layer. Synthesizing materials that display integrin-binding peptide ligands is a common way to promote endothelialization. However, in addition to integrin-ligand binding, integrin clustering is necessary to achieve intracellular signaling events that influence cellular phenotype. In this study, we explored the impact of nano-scale clustering of integrin-binding ligands on endothelial cell functions by designing novel materials that promote the clustering of integrin receptors. RGD-functionalized copolymers were prepared via reversible addition-fragmentation chain transfer (RAFT) polymerization and used for the preparation of random and nano-clustered surfaces spanning a range of global and local RGD densities (global densities 0.4-1.9 μg of peptide per mg of polymer and local densities of 1-2.4 ligands per nano-cluster). The adhesion and migration of endothelial cells was improved on nano-clustered surfaces compared to random surfaces. The highest adhesion and migration speed of endothelial cells and the rapid development of the endothelial monolayer were observed on surfaces with the highest local and global peptide density. These results indicate that the nano-clustering of peptide ligands is a promising strategy for next generation cardiovascular biomaterials, especially for small diameter vascular graft applications where the development of a confluent and functioning endothelium holds the potential to prevent device failure due to thrombosis.

Transparent, high water content (>65%), and cytocompatible hydrogels, which also possess super high refractive indices (RI > 1.5), are needed for ophthalmological applications. Most hydrogels can achieve either high RI or high water content but not both in the same system because water is a low RI material. Here, high water content/high RI hydrogels fabricated through elevated-temperature UV polymerization of an aqueous solution of acrylamide (AM) and methacrylamide (MAM) with tri(ethylene glycol) dimethacrylate (TEDA) crosslinker are reported. By varying the AM:MAM ratios (2:8 to 8:2) and crosslinker density (5 to 11 mol %), it is discovered that high water content (66%) AM:MAM copolymer hydrogels exhibiting anomalously high refractive indices (1.53); they are also colorless, transparent (99.4%), and cytocompatible with human keratinocytes.

A methacrylic terpolymer system with non-fouling interfacial properties was synthesized by the random copolymerization of hexyl methacrylate, methyl methacrylate and sulfobetaine methacrylate (a monomer bearing a zwitterionic pendant group). Polymers were synthesized from feeds containing 0-15 mol.% of the zwitterion-containing methacrylate. Proton nuclear magnetic resonance verified the incorporation of sulfobetaine methacrylate into the polymer structure. Water absorption studies illustrate that the hydrophilicity of the material increases with increasing zwitterion concentration. The biological properties of the polymer were probed by fibrinogen adsorption, human umbilical vein endothelial cell adhesion and growth, and platelet adhesion. Strong resistance to protein adsorption and cell and platelet attachment was observed on materials synthesized from 15 mol.% sulfobetaine methacrylate. Results were compared to the non-fouling behavior of a PEGylated terpolymer formulation and it was observed that the poly(ethylene glycol)-containing materials were slightly more effective at resisting human umbilical vein endothelial cell adhesion and growth over a 7 day incubation period, but the zwitterion-containing materials were equally effective at resisting fibrinogen adsorption and platelet adhesion. The zwitterion-containing materials were electrospun into three-dimensional random fiber scaffolds. Materials synthesized from 15 mol.% of the zwitterion-containing monomer retained their non-fouling character after fabrication into scaffolds.

Transparent, high water content (65%), and cytocompatible hydrogels, which also possess super high refractive indices (RI 1.5), are needed for ophthalmological applications. Most hydrogels can achieve either high RI or high water content but not both in the same system because water is a low RI material. Here, high water content/high RI hydrogels fabricated through elevated-temperature UV polymerization of an aqueous solution of acrylamide (AM) and methacrylamide (MAM) with tri(ethylene glycol) dimethacrylate (TEDA) crosslinker are reported. By varying the AM:MAM ratios (2:8 to 8:2) and crosslinker density (5 to 11 mol %), it is discovered that high water content (66%) AM:MAM copolymer hydrogels exhibiting anomalously high refractive indices (1.53); they are also colorless, transparent (99.4%), and cytocompatible with human keratinocytes.

In a previous report we screened a combinatorial peptide library to identify novel ligands that bind with high affinity and specificity to human blood outgrowth endothelial cells (HBOEC). In this study we demonstrate the use of the phage display-selected-HBOEC-specific peptides as a tool to direct and modulate endothelial cell (EC) behavior with a focus on designing functional biomaterials intended for use in cardiovascular applications. First, we ensured that our peptide ligands did not interfere with EC function as tested by proliferation, migration, tube formation, and response to vascular endothelial growth factor. Second, peptides that supported EC function were incorporated into methacrylic terpolymers via chain transfer free radical polymerization. The HBOEC-specific peptide, TPSLEQRTVYAK, when covalently coupled to a terpolymer matrix, retained binding affinity towards HBOEC in a serum-free medium. Under the same binding conditions, the attachment of human umbilical vein endothelial cells (HUVEC) was limited, thus establishing HBOEC specificity. To our knowledge, this is the first report demonstrating specificity in binding to peptide-modified biomaterials of mature EC, i.e., HUVEC, and EC of progenitor origin such as HBOEC. The findings from this work could facilitate the development of autologous cell therapies with which to treat cardiovascular disease.

Large-scale expansion of highly functional adult human mesenchymal stem cells (aMSCs) remains technologically challenging as aMSCs lose self renewal capacity and multipotency during traditional long-term culture and their quality/quantity declines with donor age and disease. Identification of culture conditions enabling prolonged expansion and rejuvenation would have dramatic impact in regenerative medicine. aMSC-derived decellularized extracellular matrix (ECM) has been shown to provide such microenvironment which promotes MSC self renewal and “stemness”. Since previous studies have demonstrated superior proliferation and osteogenic potential of human fetal MSCs (fMSCs), we hypothesize that their ECM may promote expansion of clinically relevant aMSCs. We demonstrated that aMSCs were more proliferative (∼1.6×) on fMSC-derived ECM than aMSC-derived ECMs and traditional tissue culture wares (TCPS). These aMSCs were smaller and more uniform in size (median ± interquartile range: 15.5 ± 4.1 μm versus 17.2 ± 5.0 μm and 15.5 ± 4.1 μm for aMSC ECM and TCPS respectively), exhibited the necessary biomarker signatures, and stained positive for osteogenic, adipogenic and chondrogenic expressions; indications that they maintained multipotency during culture. Furthermore, fMSC ECM improved the proliferation (∼2.2×), size (19.6 ± 11.9 μm vs 30.2 ± 14.5 μm) and differentiation potential in late-passaged aMSCs compared to TCPS. In conclusion, we have established fMSC ECM as a promising cell culture platform for ex vivo expansion of aMSCs., Singapore-MIT Alliance for Research and Technology