Effectiveness of cell adhesive additives in different supramolecular polymers.

Supramolecular motifs in elastomeric biomaterials facilitate the modular incorporation of additives with corresponding motifs. The influence of the elastomeric supramolecular base polymer on the presentation of additives has been sparsely examined, limiting the knowledge of transferability of effective functionalization between polymers. Here it was investigated if the polymer backbone and the additive influence biomaterial modification in two different types of hydrogen bonding supramolecular systems, that is, based on ureido-pyrimidinone or bis-urea units. Two different cell-adhesive additives, that is, catechol or cyclic RGD, were incorporated into different elastomeric polymers, that is, polycaprolactone, priplast or polycarbonate. The additive effectiveness was evaluated with three different cell types. AFM measurements showed modest alterations on nano-scale assembly in ureido-pyrimidinone materials modified with additives. On the contrary, additive addition was highly intrusive in bis-urea materials. Detailed cell adhesive studies revealed additive effectiveness varied between base polymers and the supramolecular platform, with bis-urea materials more potently affecting cell behavior. This research highlights that additive transposition might not always be as evident. Therefore, additive effectiveness requires re-evaluation in supramolecular biomaterials when altering the polymer backbone to suit the biomaterial application.

Biomaterial screening of protein coatings and peptide additives: towards a simple synthetic mimic of a complex natural coating for a bio-artificial kidney.

Bio-artificial kidneys require conveniently synthesized membranes providing signals that regulate renal epithelial cell function. Therefore, we aimed to find synthetic analogues for natural extracellular matrix (ECM) protein coatings traditionally used for epithelial cell culturing. Two biomaterial libraries, based on natural ECM-coatings and on synthetic supramolecular small molecule additives, were developed. The base material consisted of a bisurea (BU) containing polymer, providing supramolecular BU-additives to be incorporated via specific hydrogen bonding interactions. This system allows for a modular approach and therefore easy fractional factorial based screening. A natural coating on the BU-polymer material with basement membrane proteins, laminin and collagen IV, combined with catechols was shown to induce renal epithelial monolayer formation. Modification of the BU-polymer material with synthetic BU-modified ECM peptide additives did not result in monolayer formation. Unexpectedly, simple BU-catechol additives induced monolayer formation and presented similar levels of epithelial markers and apical transporter function as on the laminin, collagen IV and catechol natural coating. Importantly, when this BU-polymer material was processed into fibrous e-spun membranes the natural coating and the BU-catechol additive were shown to perfectly function. This study clearly indicates that complex natural ECM-coatings can be replaced by simple synthetic additives, and displays the potency of material libraries based on design of experiments in combination with modular, supramolecular chemistry.

Development of Poor Cell Adhesive Immersion Precipitation Membranes Based on Supramolecular Bis-Urea Polymers.

A variety of biomedical applications requires tailored membranes; fabrication through a mix-and-match approach is simple and desired. Polymers based on supramolecular bis-urea (BU) moieties are capable of modular integration through directed non-covalent stacking. Here, it is proposed that non-cell adhesive properties can be introduced in polycaprolactone-BU-based membranes by the addition of poly(ethylene glycol) (PEG)-BU during immersion precipitation membrane fabrication, while unmodified PEG is not retained in the membrane. PEG-BU addition results in denser membranes with a similar pore size compared to pristine membranes, while PEG addition induces defect formation. Infrared spectroscopy and surface hydrophobicity measurements indicate that PEG-BU is retained during membrane processing. Additionally, PEG-BU incorporation successfully leads to poor cell adhesive surfaces. No evidence is observed to indicate PEG retention. The results obtained indicate that the BU system enables intimate mixing of BU-modified polymers after processing. Collectively, the results provide the first steps toward BU-based immersion precipitated supramolecular membranes for biomedical applications.

Functional peptide presentation on different hydrogen bonding biomaterials using supramolecular additives.

Supramolecular biomaterials based on hydrogen bonding units can be conveniently functionalized in a mix-and-match approach using supramolecular additives. The presentation of bioactive additives has been sparsely investigated in supramolecular-based elastomeric biomaterials. Here it was investigated how cell adhesive peptides are presented and affect the surface in supramolecular biomaterials based either on ureido-pyrimidinone (UPy) or bisurea (BU) moieties. Polycaprolactone modified with UPy or BU moieties served as the base material. RGD or cyclic (c)RGD were conjugated to complementary supramolecular motifs, and were mixed with the corresponding base materials as supramolecular additives. Biomaterial surface morphology changed upon bioactivation, resulting in the formation of random aggregates on UPy-based materials, and fibrous aggregates on BU-materials. Moreover, peptide type affected aggregation morphology, in which RGD led to larger cluster formation than cRGD. Increased cRGD concentrations led to reduced focal adhesion size and cell migration velocity, and increased focal adhesion numbers in both systems, yet most prominent on functionalized BU-biomaterials. In conclusion, both systems exhibited distinct peptide presenting properties, of which the BU-system most strongly affected cellular adhesive behavior on the biomaterial. This research provided deeper insights in the differences between supramolecular elastomeric platforms, and the level of peptide introduction for biomaterial applications.

Renal Epithelial Monolayer Formation on Monomeric and Polymeric Catechol Functionalized Supramolecular Biomaterials.

Induction of a functional, tight monolayer of renal epithelial cells on a synthetic membrane to be applied in a bioartificial kidney device requires for bio-activation of the membrane. The current golden standard in bio-activation is the combination of a random polymeric catechol (L-DOPA) coating and collagen type IV (Col IV). Here the possibility of replacing this with defined monomeric catechol functionalization on a biomaterial surface using supramolecular ureido-pyrimidinone (UPy)-moieties is investigated. Monomeric catechols modified with a UPy-unit are successfully incorporated and presented in supramolecular UPy-polymer films and membranes. Unfortunately, these UPy-catechols are unable to improve epithelial cell monolayer formation over time, solely or in combination with Col IV. L-DOPA combined with Col IV is able to induce a tight monolayer capable of transport on electrospun supramolecular UPy-membranes. This study shows that a random polymeric catechol coating cannot be simply mimicked by defined monomeric catechols as supramolecular additives. There is still a long way to go in order to synthetically mimic simple natural structures.

Antimicrobial peptide modification of biomaterials using supramolecular additives.

Biomaterials based on non-active polymers functionalized with antimicrobial agents by covalent modification or mixing are currently regarded as high potential solutions to prevent biomaterial associated infections that are major causes of biomedical device failure. Herewith a strategy is proposed in which antimicrobial materials are prepared by simply mixing-and-matching of ureido-pyrimidinone (UPy) based supramolecular polymers with antimicrobial peptides (AMPs) modified with the same UPy-moiety. The N-terminus of the AMPs was coupled in solution to an UPy-carboxylic acid synthon resulting in formation of a new amidic bond. The UPy-functionalization of the AMPs did not affect their secondary structure, as proved by circular dichroism spectroscopy. The antimicrobial activity of the UPy-AMPs in solution was also retained. In addition, the incorporation of UPy-AMPs into an UPy-polymer was stable and the final material was biocompatible. The addition of 4 mol % of UPy-AMPs in the UPy-polymer material protected against colonization by Escherichia coli, and methicillin-sensitive and -resistant strains of Staphylococcus aureus. This modular approach enables a stable but dynamic incorporation of the antimicrobial agents, allowing at the same time for the possibility to change the nature of the polymer, as well as the use of AMPs with different activity spectra. © 2018 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2018, 56, 1926-1934.