Nathan C. Gianneschi

Stimuli-responsive nanomaterials for biomedical applications

Nature employs a variety of tactics to precisely time and execute the processes and mechanics of life, relying on sequential sense and response cascades to transduce signaling events over multiple length and time scales. Many of these tactics, such as the activation of a zymogen, involve the direct manipulation of a material by a stimulus. Similarly, effective therapeutics and diagnostics require the selective and efficient homing of material to specific tissues and biomolecular targets with appropriate temporal resolution. These systems must also avoid undesirable or toxic side effects and evade unwanted removal by endogenous clearing mechanisms. Nanoscale delivery vehicles have been developed to package materials with the hope of delivering them to select locations with rates of accumulation and clearance governed by an interplay between the carrier and its cargo. Many modern approaches to drug delivery have taken inspiration from natural activatable materials like zymogens, membrane proteins, and metabolites, whereby stimuli initiate transformations that are required for cargo release, prodrug activation, or selective transport. This Perspective describes key advances in the field of stimuli-responsive nanomaterials while highlighting some of the many challenges faced and opportunities for development. Major hurdles include the increasing need for powerful new tools and strategies for characterizing the dynamics, morphology, and behavior of advanced delivery systems in situ and the perennial problem of identifying truly specific and useful physical or molecular biomarkers that allow a material to autonomously distinguish diseased from normal tissue.

Enzyme‐Responsive Nanoparticles for Targeted Accumulation and Prolonged Retention in Heart Tissue after Myocardial Infarction

A method for targeting to and retaining intravenously injected nanoparticles at the site of acute myocardial infarction in a rat model is described. Enzyme-responsive peptide-polymer amphiphiles are assembled as spherical micellar nanoparticles, and undergo a morphological transition from spherical-shaped, discrete materials to network-like assemblies when acted upon by matrix metalloproteinases (MMP-2 and MMP-9), which are up-regulated in heart tissue post-myocardial infarction.

Programmable Shape‐Shifting Micelles

Nanoscale particles that undergo reversible and defined changes in morphology in response to stimuli are expected to have broad utility in a range of applications, including targeted drug delivery, detection strategies, soft templates, and self-healing materials. To date, programmable materials with these properties have not been reported, despite the many elegant examples of stimuli-responsive soft nanoparticles and micelles.[1–11] Inspired by the utility of DNA as an informational molecule in nanotechnology,[12–20] we report herein DNA-encoded polymeric materials that are capable of in situ controlled, selective, reversible, and user-defined shifts in morphology. The design is based on polymeric micelles formed from a novel set of amphiphilic DNA-brush copolymers (Figure 1).[16,21] By utilizing the sequence-selective recognition properties of DNA,[22] and its performance as a substrate for selective enzymatic cleavage,[23,24] information stored in the micelle shell may be read and manipulated in several modes, causing dramatic changes in morphology and particle size.

Controlling and Switching the Morphology of Micellar Nanoparticles with Enzymes

Micelles were prepared from polymer-peptide block copolymer amphiphiles containing substrates for protein kinase A, protein phosphatase-1, and matrix metalloproteinases 2 and 9. We examine reversible switching of the morphology of these micelles through a phosphorylation-dephosphorylation cycle and study peptide-sequence directed changes in morphology in response to proteolysis. Furthermore, the exceptional uniformity of these polymer-peptide particles makes them amenable to cryo-TEM reconstruction techniques lending insight into their internal structure.

Calcium phosphate-bearing matrices induce osteogenic differentiation of stem cells through adenosine signaling

Significance A mechanistic understanding of how calcium phosphate (CaP) minerals contribute to osteogenic commitment of stem cells and bone tissue formation is a necessary requirement for developing efficient CaP-based synthetic matrices to treat bone defects. This study unravels a previously unknown mechanism, phosphate-ATP-adenosine metabolic signaling, by which the CaP-rich mineral environment in bone tissues promotes osteogenic differentiation of human mesenchymal stem cells. In addition to a mechanical perspective on how biomaterials can influence stem cell differentiation through metabolic pathways, this discovery opens up new avenues for treating critical bone defects and bone metabolic disorders. Synthetic matrices emulating the physicochemical properties of tissue-specific ECMs are being developed at a rapid pace to regulate stem cell fate. Biomaterials containing calcium phosphate (CaP) moieties have been shown to support osteogenic differentiation of stem and progenitor cells and bone tissue formation. By using a mineralized synthetic matrix mimicking a CaP-rich bone microenvironment, we examine a molecular mechanism through which CaP minerals induce osteogenesis of human mesenchymal stem cells with an emphasis on phosphate metabolism. Our studies show that extracellular phosphate uptake through solute carrier family 20 (phosphate transporter), member 1 (SLC20a1) supports osteogenic differentiation of human mesenchymal stem cells via adenosine, an ATP metabolite, which acts as an autocrine/paracrine signaling molecule through A2b adenosine receptor. Perturbation of SLC20a1 abrogates osteogenic differentiation by decreasing intramitochondrial phosphate and ATP synthesis. Collectively, this study offers the demonstration of a previously unknown mechanism for the beneficial role of CaP biomaterials in bone repair and the role of phosphate ions in bone physiology and regeneration. These findings also begin to shed light on the role of ATP metabolism in bone homeostasis, which may be exploited to treat bone metabolic diseases.

X‐Ray Computed Tomography Imaging of Breast Cancer by using Targeted Peptide‐Labeled Bismuth Sulfide Nanoparticles

Enhanced visualization of breast cancer using X-ray microComputed Tomography is achieved using 10nm-diameter Bi2S3 nanoparticles, modified to display a tumor homing peptide (LyP-1, CGNKRTRGC). Accumulation within the tumor was increased by 260% over non-labeled nanoparticles.

Bio-Inspired Structural Colors Produced via Self-Assembly of Synthetic Melanin Nanoparticles

Structural colors arising from interactions of light with submicron scale periodic structures have been found in many species across all taxa, serving multiple biological functions including sexual signaling, camouflage, and aposematism. Directly inspired by the extensive use of self-assembled melanosomes to produce colors in avian feathers, we set out to synthesize and assemble polydopamine-based synthetic melanin nanoparticles in an effort to fabricate colored films. We have quantitatively demonstrated that synthetic melanin nanoparticles have a high refractive index and broad absorption spanning across the UV-visible range, similar to natural melanins. Utilizing a thin-film interference model, we demonstrated the coloration mechanism of deposited films and showed that the unique optical properties of synthetic melanin nanoparticles provide advantages for structural colors over other polymeric nanoparticles (i.e., polystyrene colloidal particles).

Therapeutic Enzyme‐Responsive Nanoparticles for Targeted Delivery and Accumulation in Tumors

An enzyme-responsive, paclitaxel-loaded nanoparticle is described and assessed in vivo in a human fibrosarcoma murine xenograft. This work represents a proof-of-concept study demonstrating the utility of enzyme-responsive nanoscale drug carriers capable of targeted accumulation and retention in tumor tissue in response to overexpressed endogenous enzymes.

Enzyme-directed assembly and manipulation of organic nanomaterials

Enzymes are the prime protagonists in the chemistry of living organisms. As such, chemists and biologists have long been fascinated by the array of highly selective transformations possible under biological conditions that are facilitated by enzyme-catalyzed reactions. Moreover, enzymes are involved in replicating, repairing and transmitting information in a highly selective and organized fashion through detection and signal amplification cascades. Indeed, because of their selectivity and potential for use outside of biological systems, enzymes have found immense utility in various biochemical assays and are increasingly finding applications in the preparation of small molecules. By contrast, the use of enzymatic reactions to prepare and build supramolecular and nanoscale materials is relatively rare. In this article, we seek to highlight efforts over the past 10 years at taking advantage of enzymatic reactions to assemble and manipulate complex soft, organic materials on the nanoscale. It is tantalizing to think of these processes as mimics of natural systems where enzymes are used in the assembly and transformation of the most complex nanomaterials known, for example, virus capsid assemblies and the myriad array of nanoscale biomolecular machinery.

Biological stimuli and biomolecules in the assembly and manipulation of nanoscale polymeric particles

Living systems are replete with complex, stimuli-responsive nanoscale materials and molecular self-assemblies. There is an ever increasing and intense interest within the chemical sciences to understand, mimic and interface with these biological systems utilizing synthetic and/or semi-synthetic tools. Our aim in this review is to give perspective on this emerging field of research by highlighting examples of polymeric nanoparticles and micelles that are prepared utilizing biopolymers together with synthetic polymers for the purpose of developing nanomaterials capable of interacting and responding to biologically relevant stimuli. It is expected that with the merging of evolved biological molecules with synthetic materials, will come the ability to prepare complex, functional devices. A variety of applications will become accessible including self-healing materials, self-replicating systems, biodiagnostic tools, drug targeting materials and autonomous, adaptive sensors. Most importantly, the success of this type of strategy will impact how biomolecules are stabilized and incorporated into synthetic devices and at the same time, will influence how synthetic materials are utilized within biomedical applications.