Covalently grafted human serum albumin coating mitigates the foreign body response against silicone implants in mice.

Implantable biomaterials and biosensors are integral components of modern medical systems but often encounter hindrances due to the foreign body response (FBR). Herein, we report an albumin coating strategy aimed at addressing this challenge. Using a facile and scalable silane coupling strategy, human serum albumin (HSA) is covalently grafted to the surface of polydimethylsiloxane (PDMS) implants. This covalently grafted albumin coating remains stable and resistant to displacement by other proteins. Notably, the PDMS with covalently grafted HSA strongly resists the fibrotic capsule formation following a 180-day subcutaneous implantation in C57BL/6 mice. Furthermore, the albumin coating led to reduced recruitment of macrophages and triggered a mild immune activation pattern. Exploration of albumin coatings sourced from various mammalian species has shown that only HSA exhibited a promising anti-FBR effect. The albumin coating method reported here holds the potential to improve and extend the function of silicone-based implants by mitigating the host responses to subcutaneously implanted biomaterials.

A paradigm for high-throughput screening of cell-selective surfaces coupling orthogonal gradients and machine learning-based cell recognition.

The combinational density of immobilized functional molecules on biomaterial surfaces directs cell behaviors. However, limited by the low efficiency of traditional low-throughput experimental methods, investigation and optimization of the combinational density remain daunting challenges. Herein, we report a high-throughput screening set-up to study biomaterial surface functionalization by integrating photo-controlled thiol-ene surface chemistry and machine learning-based label-free cell identification and statistics. Through such a strategy, a specific surface combinational density of polyethylene glycol (PEG) and arginine-glutamic acid-aspartic acid-valine peptide (REDV) leads to high endothelial cell (EC) selectivity against smooth muscle cell (SMC) was identified. The composition was translated as a coating formula to modify medical nickel-titanium alloy surfaces, which was then proved to improve EC competitiveness and induce endothelialization. This work provided a high-throughput method to investigate behaviors of co-cultured cells on biomaterial surfaces modified with combinatorial functional molecules.

miR-22 eluting cardiovascular stent based on a self-healable spongy coating inhibits in-stent restenosis.

The in-stent restenosis (IRS) after the percutaneous coronary intervention contributes to the major treatment failure of stent implantation. MicroRNAs have been revealed as powerful gene medicine to regulate endothelial cells (EC) and smooth muscle cells (SMC) in response to vascular injury, providing a promising therapeutic candidate to inhibit IRS. However, the controllable loading and eluting of hydrophilic bioactive microRNAs pose a challenge to current lipophilic stent coatings. Here, we developed a microRNA eluting cardiovascular stent via the self-healing encapsulation process based on an amphipathic poly(ε-caprolactone)-poly(ethylene glycol)-poly(ε-caprolactone) (PCL-PEG-PCL, PCEC) triblock copolymer spongy network. The miR-22 was used as a model microRNA to regulate SMC. The dynamic porous coating realized the uniform and controllable loading of miR-22, reaching the highest dosage of 133 pmol cm-2. We demonstrated that the sustained release of miR-22 dramatically enhanced the contractile phenotype of SMC without interfering with the proliferation of EC, thus leading to the EC dominating growth at an EC/SMC ratio of 5.4. More importantly, the PCEC@miR-22 coated stents showed reduced inflammation, low switching of SMC phenotype, and low secretion of extracellular matrix, which significantly inhibited IRS. This work provides a simple and robust coating platform for the delivery of microRNAs on cardiovascular stent, which may extend to other combination medical devices, and facilitate practical application of bioactive agents in clinics.

Gradient Porous Structure Templated by Breath Figure Method.

Surfaces with gradient topography are important in various fields but are difficult to fabricate. Herein, we report a facile and robust way to fabricate a surface with gradient topography of porous structure, in one direction, based on the breath figure (BF) method for the first time. The influencing factors including relative humidity (RH), sample immersion time, and solvent composition, affecting the speed, time, and model of the droplet growth, respectively, were investigated to control gradient BF pores with different ranges of pore sizes. Applying appropriate parameters, gradient BF pores with a diameter difference over 400% were prepared on one sample. The mechanism of gradient duration of solvent evaporation at different regions of a sample for fabricating gradient pores was proposed and experimentally verified with recording optical and thermographic changes of the sample in the BF procedure. This new method provides a novel site for gradient topography fabrication.

High-throughput hyaluronic acid hydrogel arrays for cell selective adhesion screening.

As a component of extracellular matrix (ECM), hyaluronic acid (HA) has plenty of applications in the biomedical field such as tissue engineering. Due to its non-adhesive nature, HA requires further grafting of functional molecules for cell related study. RGD and YIGSR are two kinds of cell adhesion peptides. YIGSR enhances endothelial cell (EC) adhesion, which is important for endothelialization after implantation of stents to prevent in-stent restenosis. However, the effect of combined densities of these peptides for EC and smooth muscle cell (SMC) adhesion has not been explored in a quantitative and high-throughput manner. In this work, single or orthogonal gradient densities of RGD and YIGSR were grafted onto the HA hydrogel array surfaces using thiol-norbornene click chemistry. Optimized peptide combinations for EC preponderant adhesion were found in hydrogel arrays and confirmed by scaling samples.

Rapid build-up of high-throughput screening microarrays with biochemistry gradients via light-induced thiol-ene "click" chemistry.

Microarrays have become extremely powerful experimental tools for high-throughput screening of cell behaviors in multivariate microenvironments. Herein, a microarray-based high-throughput platform with biochemistry gradients was developed using poly(limonene carbonate) (PLimC) as a substrate through thiol-ene click chemistry. ATR-IR, XPS, Raman spectrum, and water contact angle results demonstrated that the sulfhydryl molecules, including PEG (polyethylene glycol) and RGD (arginine-glycine-aspartic acid) peptide, could be grafted onto PLimC substrates, while the grafting density could be well controlled by regulating the intensity of UV irradiation. Then, microarrays with a gradient of RGD grafting density were fabricated by using UV irradiation patterned by a photomask and a gradient light filter. Adhesion experiments of smooth muscle cells and 3T3-L1 mouse embryonic fibroblast cells proved that the cell behaviors were highly determined by the RGD density. This platform puts forward a facile, high-throughput method to study the effect of biochemical signal density on cell behaviors.

Rapid Buildup Arrays with Orthogonal Biochemistry Gradients via Light-Induced Thiol-Ene "Click" Chemistry for High-Throughput Screening of Peptide Combinations.

The concept of high-throughput screening sheds new light on fabrication and analysis of materials. Herein, a combinatorial surface-modified platform with biochemical gradients was developed through thiol-ene "click" chemistry by adjusting the intensity of ultraviolet (UV) irradiation. Contact angle, X-ray photoelectron spectroscopy, and ellipsometry measurement results demonstrated that the sulfhydryl molecules including polyethylene glycol and RGD (arginine-glycine-aspartic acid) and REDV (arginine-glutamic acid-aspartic acid-valine) peptides can be directly attached onto alkene-modified substrates, in which the graft density can be well controlled by the intensity of UV irradiation. The multistep attachment of different molecules onto substrates is archived via the multistep UV-initiated thiol-ene "click" reaction. The high-throughput arrays with the gradient density of single ligand and the orthogonal gradient density of two ligands were rapidly fabricated via the one-step UV gradient irradiation and the two-step orthogonal UV gradient-initiated thiol-ene "click" reaction. Endothelial cells (ECs) and smooth muscle cells (SMCs) were cocultured on the array with the orthogonal gradient density of RGD and REDV to screen the peptide combination with high EC selectivity, which is essential for in situ endothelialization during stent implant. From 64, 8 × 8, combinations investigated, a special combinatorial surface representing the really high competitiveness of ECs over SMCs was screened. This platform puts forward a facile, high-throughput method to study the combinatorial variation of biochemical signals to cell behavior.

Nanofibrous scaffold from electrospinning biodegradable waterborne polyurethane/poly(vinyl alcohol) for tissue engineering application.

A series of nanofibrous scaffolds, free of organic solvents, are prepared by electrospinning biodegradable waterborne polyurethane (BWPU) emulsion blending with aqueous poly(vinyl alcohol)(PVA). Tuning the proration of BWPU to PVA, various nanofibers with diameter from 370 to 964 nm are obtained. Strong intermolecular interaction existing between them benefits to the electrospun of BWPU emulsion, which is demonstrated by dynamic thermomechanical analysis and Fourier transform infrared spectroscopy. The nontoxic nanofibrous scaffolds with porous structure, which is similar to the natural extracellular matrix, favor to the attachment and proliferation of the L929 fibroblasts. Thus, the scaffolds are promising to be used as biomaterials for many natural tissues repair.

Synthesis and characterization of biodegradable lysine-based waterborne polyurethane for soft tissue engineering applications.

Biomaterials for soft tissue engineering scaffolds require a combination of multiple properties including suitable mechanical properties, biodegradability, and biocompatibility. In this work, a series of light-crosslinking waterborne polyurethanes (LWPUs) were prepared using l-lysine ethyl ester diisocyanate (LDI), 1,3-propanediol (PDO) and l-lysine as hard segments and poly(ε-caprolactone) (PCL) and poly(ethylene glycol) (PEG) as soft segments. The obtained LWPUs exhibited appropriate stretchability with a break elongation of 1400-2500% and an excellent strength of 12-18 MPa, which could admirably meet the requirements for soft tissue engineering scaffolds. In addition, the hydrophilic surfaces of LWPUs could effectively reduce protein adsorption and platelet adhesion and favor cell proliferation compared with traditional biomedical polyurethanes. The ultimate degradation products of LWPUs were proven to be nontoxic in a cytotoxicity test. More interestingly, a cytokine release test of macrophages adherent to the LWPU film surfaces shows that these macrophages secreted less pro-inflammation cytokine TNF-α and more anti-inflammation cytokine IL-10 after 3 days' culture, indicating that LWPUs possess the potential ability to aid in the transition of macrophages toward a wound healing phenotype. Furthermore, the LWPU films could support the adhesion and proliferation of endothelial cells. Thus, the obtained LWPUs have great potential for applications in soft tissue engineering scaffolds for tissue repair and wound healing.

Preparation of hydrocarbon/fluorocarbon double-chain phospholipid polymer brusheson polyurethane films by ATRP.

To fabricate artificial biomembrane mimicking cell surfaces, hydrocarbon/fluorocarbon double-chain phospholipid macromonomer was grafted on polyurethane (PU) film surfaces by surface-initiated atom transfer radical polymerization (SI-ATRP). The surface structures of modified PU film surfaces were characterized by X-ray photoelectron spectroscopy (XPS) and water contact angle measurement. The results indicate that initiator densities on these polymer film surfaces have a significant impact on graft polymerization of this fluorocarbon phospholipid macromonomer. The phospholipid polymer brushes grafted on PU film surfaces could self-assemble into biomimetic membranes under water environment, as demonstrated by liquid/liquid static contact angle measurement, atomic force spectroscopy (AFM), and attenuated total reflectance Fourier transform infrared (ATR-FTIR). These biomimetic membranes could maintain water within them as the "surrounding" water. Such would be favorable condition for the preservation of native conformational state of proteins and cell membranes. This work provides a new approach to fabricate biomimetic membranes on biomaterials surfaces.