Biomimetic Bacterial Capsule for Enhanced Aptamer Display and Cell Recognition.

Great effort has been made to encapsulate or coat living mammalian cells for a variety of applications ranging from diabetes treatment to three-dimensional printing. However, no study has reported the synthesis of a biomimetic bacterial capsule to display high-affinity aptamers on the cell surface for enhanced cell recognition. Therefore, we synthesized an ultrathin alginate-polylysine coating to display aptamers on the surface of living cells with natural killer (NK) cells as a model. The results show that this coating-mediated aptamer display is more stable than direct cholesterol insertion into the lipid bilayer. The half-life of the aptamer on the cell surface can be increased from less than 1.5 to over 20 h. NK cells coated with the biomimetic bacterial capsule exhibit a high efficiency in recognizing and killing target cells. Therefore, this work has demonstrated a promising cell coating method for the display of aptamers for enhanced cell recognition.

Nanoparticle-Decorated Biomimetic Extracellular Matrix for Cell Nanoencapsulation and Regulation.

Cell encapsulation has been studied for various applications ranging from cell transplantation to biological production. However, current encapsulation technologies focus on cell protection rather than cell regulation that is essential to most if not all cell-based applications. Here we report a method for cell nanoencapsulation and regulation using an ultrathin biomimetic extracellular matrix as a cell nanocapsule to carry nanoparticles (CN2 ). This method allows high-capacity nanoparticle retention at the vicinity of cell surfaces. The encapsulated cells maintain high viability and normal metabolism. When gold nanoparticles (AuNPs) are used as a model to decorate the nanocapsule, light irradiation transiently increases the temperature, leading to the activation of the heat shock protein 70 (HSP70) promoter and the regulation of reporter gene expression. As the biomimetic nanocapsule can be decorated with any or multiple NPs, CN2 is a promising platform for advancing cell-based applications.

Elastic Macroporous Matrix-Supported In Situ Formation of Injectable Extracellular Matrix-Like Hydrogel for Carrying Growth Factors and Living Cells.

Hydrogels loaded with biologics hold great potential for various biomedical applications such as regenerative medicine. However, biologics may lose bioactivity during hydrogel preparation, shipping, and storage. While many injectable hydrogels do not have this issue, they face a dilemma between fast gelation causing the difficulty of injection and slow gelation causing the escape of solutions from an injection site. The purpose of this study is to develop an affinity hydrogel by integrating a pre-formed elastic macroporous matrix and an injectable hydrogel. The data shows that the macroporous hydrogel matrix can hold a large volume of solutions for the formation of in situ injectable hydrogels loaded with growth factors or living cells. The cells can proliferate in the composite hydrogels. The growth factors can be stably sequestered and sustainably released due to the presence of aptamers. When both living cells and growth factors are loaded together into the hydrogels, cells can proliferate under culture conditions with a reduced serum level. Therefore, a macroporous and elastic matrix-supported formation of aptamer-functionalized injectable hydrogels is a promising method for developing the carriers of biologics.

Ushering in the era of regenerative surgery.

Tissue loss, irrespective of etiology, often requires extensive reconstruction. In many instances, the need exceeds what current treatments and technologies modern medicine can offer. Tissue engineering has made immense strides within the past few decades due to advances in biologics, biomaterials, and manufacturing. The convergence of these three domains has created limitless potential for future surgical care. Unfortunately, there still exists a disconnect on how to best implant these 'replacement parts' and care for the patient. It is therefore vital to develop paradigms for the integration of advanced surgical and tissue engineering technologies. This paper explores the convergence between tissue engineering and reconstructive surgery. We will describe the clinical problem of tissue loss, discuss currently available solutions, address limitations, and propose processes for integrating surgery and tissue engineering, thereby ushering in the era of regenerative surgery.

Performance of a Hydrogel Coated Nitinol with Oligonucleotide-Modified Nanoparticles Within Turbulent Conditions of Blood-Contacting Devices.

INTRODUCTION: Hydrogels offer a wide range of applications in the antithrombotic modification of biomedical devices. The functionalization of these hydrogels with potentially drug-laden nanoparticles in the context of deviceassociated turbulence is critically under-studied. Thus, the purpose of this study was to use a hydrogel-coating nitinol surface as a model to understand the functions of hydrogels and the capture of nanoparticles under clinically relevant flow conditions. METHODS: Nitinol was coated by an oligonucleotide (ON) functionalized hydrogel. Nanoparticles were functionalized with complementary oligonucleotides (CONs). The capture of CONfunctionalized nanoparticles by the ON-functionalized hydrogel surfaces was studied under both static and dynamic attachment conditions. Fluorescent-labelling of nanoparticles was utilized to assess capture efficacy and resistance to removal by device-relevant flow conditions. RESULTS: The specificity of the ON-CON bond was verified, exhibiting a dose-dependent attachment response. The hydrogel coating was resistant to stripping by flow, retaining >95% after exposure to one hour of turbulent flow. Attachment of nanoparticles to the hydrogel was higher in the static condition than under laminar flow (p < 0.01), but comparable to that of attachment under turbulent flow. Modified nitinol samples underwent one hour of flow treatment under both laminar and turbulent regimes and demonstrated decreased nanoparticle loss following static conjugation rather than turbulent conjugation (36.1% vs 53.8%, p < 0.05). There was no significant difference in nanoparticle functionalization by upstream injection between laminar and turbulent flow. CONCLUSION: The results demonstrate promising potential of hydrogelfunctionalized nitinol for capturing nanoparticles using nucleic acid hybridization. The hydrogel structure and ONCON bond integrity both demonstrated a resistance to mechanical damage and loss of biomolecular functionalization by exposure to turbulence. Further investigation is warranted to highlight drug delivery and antithrombogenic modification applications of nanoparticle-functionalized hydrogels.

High-affinity one-step aptamer selection using a non-fouling porous hydrogel.

Aptamers, commonly referred to as chemical antibodies, are used in a wide range of applications including drug delivery and biosensing. However, the process of aptamer selection poses a substantial challenge, as it requires numerous cycles of enrichment and involves issues with nonspecific binding. We present a simple, fast instrument-free method for aptamer enrichment and selection based on a diffusion-binding process in a three-dimensional non-fouling porous hydrogel with immobilized target proteins. Low-affinity aptamer candidates can be rapidly released from the hydrogel, whereas high-affinity candidates are restricted due to their strong binding to the immobilized protein targets. Consequently, a one-step enriched aptamer pool can strongly bind the protein targets. This enrichment is consistent across five proteins with isoelectric points in varying ranges. With thrombin as a representative model, the anti-thrombin aptamer identified from an enriched aptamer pool has been found to have a binding affinity that is comparable to those identified over ten cycles of selection using traditional methods.

Display of Polyvalent Hybrid Antibodies on the Cell Surface for Enhanced Cell Recognition.

Cell surface engineering with exogeneous receptors holds great promise for various applications. However, current biological methods face problems with safety, antigen escape, and receptor stoichiometry. The purpose of this study is to develop a biochemical method for displaying polyvalent antibodies (PAbs) on the cell surface. The PAbs are synthesized through the self-assembly of DNA-Ab conjugates under physiological conditions without the involvement of any factors harsh to cells. The data show that PAb-functionalized cells can recognize target cells much more effectively than monovalent controls. Moreover, dual Ab incorporation into the same PAb with a defined stoichiometric ratio leads to the formation of a polyvalent hybrid Ab (DPAb). DPAb-functionalized cells can effectively recognize target cell models with antigen escape, which cannot be achieved by PAbs with one type of Ab. Therefore, this work presents a novel biochemical method for Ab display on the cell surface for enhanced cell recognition.

Synthetic DNA for Cell Surface Engineering: Experimental Comparison between Click Conjugation and Lipid Insertion in Terms of Cell Viability, Engineering Efficiency, and Displaying Stability.

The cell surface can be engineered with synthetic DNA for various applications ranging from cancer immunotherapy to tissue engineering. However, while elegant methods such as click conjugation and lipid insertion have been developed to engineer the cell surface with DNA, little effort has been made to systematically evaluate and compare these methods. Resultantly, it is often challenging to choose a right method for a certain application or to interpret data from different studies. In this study, we systematically evaluated click conjugation and lipid insertion in terms of cell viability, engineering efficiency, and displaying stability. Cells engineered with both methods can maintain high viability when the concentration of modified DNA is less than 25-50 µM. However, lipid insertion is faster and more efficient in displaying DNA on the cell surface than click conjugation. The efficiency of displaying DNA with lipid insertion is 10-40 times higher than that with click conjugation for a large range of DNA concentration. However, the half-life of physically inserted DNA on the cell surface is 3-4 times lower than that of covalently conjugated DNA, which depends on the working temperature. While the half-life of physically inserted DNA molecules on the cell surface is shorter than that of DNA molecules clicked onto the cell surface, lipid insertion is more effective than click conjugation in the promotion of cell-cell interactions under the two different experimental settings. The data acquired in this work are expected to act as a guideline for choosing an approximate method for engineering the cell surface with synthetic DNA or even other biomolecules.

Feasibility of anaerobic digestion as a treatment for the aqueous pyrolysis condensate (APC) of birch bark.

Biooil produced via biomass pyrolysis includes an aqueous-acidic phase and a dense and rich organic phase. The aqueous phase has a low heating value and is considered a waste stream. In this study fractional condensation was employed to separate the liquid product of birch bark pyrolysis into an aqueous pyrolysis condensate (APC) and a dense biooil fraction. The APC contained high amounts (~100 g/kg) of acidic acid (AA) and was investigated for anaerobic digestion (AD). The AA in the APC could be converted to biogas, however, it contained elevated concentrations of microbial inhibitors (24 g/kg total phenolics). The inhibiting effect could be mitigated by acclimatization of the microbial population, which in turn converted some of the additional organics. The production of methane further improved with the addition of biochar to adsorb some of the inhibitors. The results imply that a waste product can be converted into a potential energy carrier.