Biocatalytic Polymer Coatings: On‐Demand Drug Synthesis and Localized Therapeutic Effect under Dynamic Cell Culture Conditions

Abstract

Biocatalytic surface coatings are prepared herein for localized synthesis of drugs and their on-demand, site-specific delivery to adhering cells. This novel approach is based on the incorporation of an enzyme into multilayered polymer coatings to accomplish enzyme-prodrug therapy (EPT). The build-up of enzyme-containing multilayered coatings is characterized and correlations are drawn between the multilayer film assembly conditions and the enzymatic activity of the resulting coatings. Therapeutic effect elicited by the substrate mediated EPT (SMEPT) strategy is investigated using a prodrug for an anticancer agent, SN-38. The performance of biocatalytic coatings under flow conditions is investigated and it is demonstrated that EPT allows synthesizing the drugs on-demand, at the time desired and in a controllable amount to suit particular applications. Finally, using cells cultured in sequentially connected flow chambers, it is demonstrated that SMEPT affords a site-specific drug delivery, that is, exerts a higher therapeutic effect in cells adhering directly to the biocatalytic coatings than in the cells cultured "downstream". Taken together, these data illustrate biomedical opportunities made possible by engineering tools of EPT into multilayered polymer coatings and present a novel, highly versatile tool for surface mediated drug delivery.

Liposomal Drug Deposits in Poly(Dopamine) Coatings: Effect of Their Composition, Cell Type, Uptake Pathway Considerations, and Shear Stress

“…Recently, we were the first to consider shear stress in the context of SMDD by analyzing biocatalytic films in the presence of shear stress using hepatocytes. [31] First, the goal was to compare the initial adhesion and spreading of endothelial cells on M n2 films considering shear stress. Microfluidic channels were coated with M n2 or PDA as control and the cells were allowed to adhere for 3 h before shear stress (t 4 ¼ 4 dyn cm À2 ) was applied for 3 h. The initial 3 h adhesion in the absence of shear stress was needed for the cells to adhere.…”

Enzyme Prodrug Therapy Engineered into Biomaterials

“…Novelty of this presentation lies in that to the best of our knowledge, we provide the fi rst example of drug eluting biomaterials with a facile external control over the nature of the drug to be released (anti-infl ammatory, cytotoxic), drug dosage, and time of release. [ 15 ] This experimental setup also revealed that a more pronounced therapeutic effect was achieved locally, at the site of drug synthesis, compared to that observed on cells proliferating "downstream" from the location of drug release. [19][20][21]23 ] In doing so, our results signifi cantly extend functionality of these and other retained their enzymatic activity over as long as 7 days.…”

“…[ 12,13 ] Towards this end, we used an enzyme with excellent prior characterization in EPT techniques, β-Glucuronidase (β-Glu), [ 1,12 ] functionalized hydrogel biomaterials [ 13 ] and multilayered polymer coatings [ 14,15 ] with this enzyme, and subsequently used the resulting biomaterials as substrates for mammalian cell culture. We proposed to engineer the tools of EPT into biomaterials to achieve "substrate mediated enzyme prodrug therapy", SMEPT.…”

“…[ 9,11,21,30 ] Tools of EPT are particularly important in this context [ 31 ] and, together with success of SMEPT in achieving localized synthesis and delivery of anti-proliferative drugs, [ 15 ] this justifi es broader investigation of SMEPT for these applications. Scope and utility of the latter in chemotherapy signifi cantly depends on the opportunities in targeted drug delivery.…”

Effects of geometrical confinement in membrane pores on enzyme-based layer-by-layer assemblies

“…In addition, the polyionic character of opposite charge could help to preserve the secondary structure when facing denaturating agents [4,15]. Enzymes in LbL films can increase their life-time and stability [16,17], which makes LbL an interesting option for biomedical applications such as enzyme prodrug therapy [16,18]. Since LbL has no restriction with respect to adsorbing substrate size and topology [19], protein multilayer films have been deposited on planar substrates, colloidal particles and membranes [4,20].…”