Nature-Inspired Thylakoid-Based Photosynthetic Nanoarchitectures for Biomedical Applications.

"Drawing inspiration from nature" offers a wealth of creative possibilities for designing cutting-edge materials with improved properties and performance. Nature-inspired thylakoid-based nanoarchitectures, seamlessly integrate the inherent structures and functions of natural components with the diverse and controllable characteristics of nanotechnology. These innovative biomaterials have garnered significant attention for their potential in various biomedical applications. Thylakoids possess fundamental traits such as light harvesting, oxygen evolution, and photosynthesis. Through the integration of artificially fabricated nanostructures with distinct physical and chemical properties, novel photosynthetic nanoarchitectures can be catalytically generated, offering versatile functionalities for diverse biomedical applications. In this article, an overview of the properties and extraction methods of thylakoids are provided. Additionally, the recent advancements in the design, preparation, functions, and biomedical applications of a range of thylakoid-based photosynthetic nanoarchitectures are reviewed. Finally, the foreseeable challenges and future prospects in this field is discussed.

A chlorogenic acid functional strategy of anti-inflammation, anti-coagulation and promoted endothelial proliferation for bioprosthetic artificial heart valves.

Heart valve replacement has become an optimal choice for the treatment of severe heart valve disease. At present, most commercial bioprosthetic heart valves (BHVs) are made from porcine pericardium or bovine pericardium treated with glutaraldehyde. Nevertheless, due to the toxicity of residual aldehyde groups left after glutaraldehyde cross-linking, these commercial BHVs exhibit poor biocompatibility, calcification, risk of coagulation and endothelialization difficulty, which greatly affects the durability of the BHVs and shortens their service life. In this work, based on a chlorogenic acid functional anti-inflammation, anti-coagulation and endothelialization strategy and dual-functional non-glutaraldehyde cross-linking reagent OX-CO, a kind of functional BHV material OX-CA-PP has been developed from OX-CO cross-linked porcine pericardium (OX-CO-PP) followed by the convenient modification of chlorogenic acid through a reactive oxygen species (ROS) sensitive borate ester bond. The functionalization of chlorogenic acid can reduce the risk of valve leaf thrombosis and promote endothelial cell proliferation, which is beneficial to the formation of a long-term interface with good blood compatibility. Meanwhile, such a ROS responsive behavior can trigger intelligent release of chlorogenic acid on-demand to achieve the inhibition of acute inflammation at the early stage of implantation. The in vivo and in vitro experimental results show that the functional BHV material OX-CA-PP exhibits superior anti-inflammation, improved anti-coagulation, minimal calcification and promoted proliferation of endothelial cells, showing that this non-glutaraldehyde functional strategy has great potential for the application of BHVs and providing a promising reference for other implanted biomaterials.

A versatile modification strategy for functional non-glutaraldehyde cross-linked bioprosthetic heart valves with enhanced anticoagulant, anticalcification and endothelialization properties.

Valvular heart disease is a major threat to human health and transcatheter heart valve replacement (THVR) has emerged as the primary treatment option for severe heart valve disease. Bioprosthetic heart valves (BHVs) with superior hemodynamic performance and compressibility have become the first choice for THVR, and more BHVs have been requested for clinical use in recent years. However, several drawbacks remain for the commercial BHVs cross-linked by glutaraldehyde, including calcification, thrombin, poor biocompatibility and difficulty in endothelialization, which would further reduce the BHVs' lifetime. This study developed a dual-functional non-glutaraldehyde crosslinking reagent OX-VI, which can provide BHV materials with reactive double bonds (CC) for further bio-function modification in addition to the crosslinking function. BHV material PBAF@OX-PP was developed from OX-VI treated porcine pericardium (PP) after the polymerization with 4-vinylbenzene boronic acid and the subsequent modification of poly (vinyl alcohol) and fucoidan. Based on the functional anti-coagulation and endothelialization strategy and dual-functional crosslinking reagent, PBAF@OX-PP has better anti-coagulation and anti-calcification properties, higher biocompatibility, and improved endothelial cells proliferation when compared to Glut-treated PP, as well as the satisfactory mechanical properties and enhanced resistance effect to enzymatic degradation, making it a promising candidate in the clinical application of BHVs. STATEMENT OF SIGNIFICANCE: Transcatheter heart valve replacement (THVR) has become the main solution for severe valvular heart disease. However, bioprosthetic heart valves (BHVs) used in THVR exhibit fatal drawbacks such as calcification, thrombin and difficulty for endothelialization, which are due to the glutaraldehyde crosslinking, resulting in a limited lifetime to 10-15 years. A new non-glutaraldehyde cross-linker OX-VI has been designed, which can not only show great crosslinking ability but also offer the BHVs with reactive double bonds (CC) for further bio-function modification. Based on the dual-functional crosslinking reagent OX-VI, a versatile modification strategy was developed and the BHV material (PBAF@OX-PP) has been developed and shows significantly enhanced anticoagulant, anti-calcification and endothelialization properties, making it a promising candidate in the clinical application of BHVs.

N-Arylimidazoliums as Highly Selective Biomimetic Antimicrobial Agents.

Antibiotic resistance has become one of the greatest health threats in the world. In this study, a charge-dispersed dimerization strategy is described for the antimicrobial peptide (AMP) mimics via a tunable cationic charge to improve the selectivity between prokaryotic microbes and eukaryotic cells. This strategy is demonstrated with a series of charge-dispersed AMP mimics based on N-arylimidazolium skeletons. These N-arylimidazolium AMP mimics show potent antibacterial activity against strains along with a low rate of drug resistance, good hemocompatibility, and low cytotoxicity. In addition to the elimination of planktonic bacteria, N-arylimidazolium AMP mimics can also inhibit biofilm formation and destroy the established biofilm. More importantly, methicillin-resistant Staphylococcus aureus (MRSA)-induced lung-infected mice can be effectively treated by the intravenous administration of N-arylimidazolium AMP mimic, which enable the design of N-arylimidazolium AMP mimics to offer an alternative avenue to eradicate drug-resistant bacterial infection.

A bioprosthetic heart valve cross-linked by a non-glutaraldehyde reagent with improved biocompatibility, endothelialization, anti-coagulation and anti-calcification properties.

Valvular heart disease is an important disease that endangers human health and heart valve replacement has become one of the main treatments for patients with severe valvular heart disease. However, the traditional surgical valve replacement (SVR) suffers several drawbacks such as high risk, great trauma and long recovery time, and more than 30% of patients are intolerant to SVR, especially elderly patients. In recent years, with the development of minimally invasive technology, transcatheter heart valve replacement (THVR) as a method of implantation without thoracotomy has become an optimal treatment for severe valvular heart disease due to its advantages of minimal trauma, low risk and fast recovery. Meanwhile, the usage of bioprosthetic heart valves (BHVs) has been enlarged greatly with the rapid development of THVR and the aging population. Most BHVs in clinics are crosslinked by glutaraldehyde (Glut), which shows great mechanical properties and chemical stability. However, some problems such as poor biocompatibility, calcification, coagulation and endothelialization difficulty also need to be solved urgently for Glut-treated BHVs. In this work, a non-Glut treated BHV from 7a-ethyltetrahydro-oxazolo[3,4-c]oxazole (OX-Et) crosslinked porcine pericardium (PP) has been developed. Compared with glutaraldehyde-crosslinked porcine pericardium (Glut-PP), good physical and chemical properties similar to Glut-PP are shown for OX-Et treated porcine pericardium (OX-Et-PP). It is noteworthy that better biocompatibility, endothelialization performance, and anti-coagulant effect as well as the improved anti-calcification property can also be observed for OX-Et-PP in the in vitro and in vivo study, potentially making OX-Et-PP a good candidate in the application of BHVs.

Dual-responsive injectable hydrogels encapsulating drug-loaded micelles for on-demand antimicrobial activity and accelerated wound healing.

Disease microenvironment stimuli-responsive hydrogels are of special interests in enhancing the drug delivery specificity for biomedical applications. In order to achieve specific drug release characteristic at the inflammation region, a smart pH- and reactive oxygen species (ROS)-responsive injectable hydrogel with self-healing and remodeling capability was designed in our present work. By grafting phenylboronic acid to the side chain of the alginate polymer, a highly specific dual-responsive hydrogel with low pH and high ROS was obtained. Moreover, the hydrogel was endowed with antibacterial and anti-inflammatory properties respectively via the effective assembly of antibiotic amikacin (AM), and anti-inflammatory drug naproxen (Nap) which were preloaded into the micelles. Such hydrogel formulation not only preserved the structural integrity and excellent rheological properties of the hydrogel but also allowed for a controllable drug release rate at inflammation sites. The antibacterial experiment results in vitro demonstrated that the hydrogel could effectively kill bacteria by amikacin release, with the inhibitive rate reached to 90% for S. aureus and 98% for P.aeruginosa. Furthermore, the anti-inflammatory drug naproxen was also controllably released from the ROS-responsive micelles within 24 h under pH 5.0, and 10 mM H2O2in vitro. Most importantly, the smart hydrogels showed good biocompatibility, and greatly promoted the healing of infected wounds in vitro by cell scratch assay. In addition, it efficiently reduced the levels of TNF-α (the pro-inflammatory cytokine) by 2.80 times, and increased IL-10 (anti-inflammatory cytokine) by 2.41 times than the hydrogel control without antibiotic and anti-inflammatory drug. Within the dual-responsiveness of pH and ROS, the hydrogel reduced the inflammation response of the surrounding tissues significantly and accelerated the healing process of the infected area. Collectively, this smart hydrogel formula containing antibiotic and drug-loaded micelles is very promising to be applied topically against various microbial infections. We believe that this strategy can also be applied to various disease treatments.

Synergistic Chemical and Photodynamic Antimicrobial Therapy for Enhanced Wound Healing Mediated by Multifunctional Light-Responsive Nanoparticles.

Recently, rapid acquisition of antibiotic resistance, increased prevalence of antibiotic-resistant bacterial infections, and slow healing of infected wound have led to vast difficulties in developing innovative antimicrobial agents to obliterate pathogenic bacteria and simultaneously accelerate wound healing. To effectively solve this problem, we designed light-responsive multifunctional nanoparticles with conjugation of quaternary ammonium chitosan and photosensitizer chlorin e6 (Ce6) to merge chemical and photodynamic therapy to efficient antibacteria. The Mg/(-)-epigallocatechin-3-gallate (EGCG) complex rapidly responded to light irradiation under 660 nm with release of magnesium ions, which effectively accelerated wound healing without toxicity to mammalian cells. Notably, positively charged nanoparticles could efficiently adhere to the bacterial surface, and reactive oxygen species (ROS) produced under laser irradiation destroyed the membrane structure of the bacteria, which is irreversible, ultimately leading to bacteria death. Thus, multifunctional nanoparticles with a combination of chemical and photodynamic antimicrobial therapy would offer guidance to rational predicted and designed new effective antimicrobial nanomaterials. Most importantly, it may represent a promising class of antimicrobial strategy for potential clinical translation.

Peptide-/Drug-Directed Self-Assembly of Hybrid Polyurethane Hydrogels for Wound Healing.

Drug-loading hydrogels are promising candidates in the bioengineering research field; nevertheless, hydrophobic drug loading into a hydrophilic carrier system remains unsolved and is full of challenges. In this work, following the potential dual interactions between peptides and aromatic drugs, we developed a potent hybrid hydrogel formation method, namely, "peptide-/drug-directed self-assembly". The hybrid hydrogels were synthesized using polyethylene glycol (PEG)-based Fmoc-FF peptide hybrid polyurethane, in which curcumin could be encapsulated through self-assembly with Fmoc-FF peptide via π-π stacking. On the basis of this, curcumin loading capacity could be improved to as high as 3.3 wt % with sustained release. In addition, the curcumin loading enhanced the hydrogel mechanical properties from 4 kPa to over 10 kPa, similar to that of natural soft tissues. Furthermore, the hydrogels were injectable with self-healing properties since the Fmoc-FF peptide/curcumin coassembly was noncovalent and reversible. Spectroscopy results confirmed the existence of the coassembly of Fmoc-FF peptide/curcumin. Further in vivo experiments effectively demonstrated that the hydrogels could improve the cutaneous wound healing in a full-thickness skin defected model. This peptide-/drug-directed self-assembly of hybrid polyurethane hydrogel could be used as a promising platform for tissue-engineering scaffold and biomedical application.

Multifunctional Two-Photon AIE Luminogens for Highly Mitochondria-Specific Bioimaging and Efficient Photodynamic Therapy.

In recent years, photodynamic therapy (PDT) has drawn much attention as a noninvasive and safe cancer therapy method due to its fine controllability, good selectivity, low systemic toxicity, and minimal drug resistance in contrast to the conventional methods (for example, chemotherapy, radiotherapy, and surgery). However, some drawbacks still remain for the current organic photosensitizers such as low singlet oxygen (1O2) quantum yield, poor photostability, inability of absorption in the near-infrared (NIR) region, short excitation wavelength, and limited action radius of singlet oxygen, which will strongly limit the PDT treatment efficiency. As a consequence, the development of efficient photosensitizers with high singlet oxygen quantum yield, strong fluorescent emission in the aggregated state, excellent photostability, NIR excitation wavelength ranging in the biological transparency window, and highly specific targeting to mitochondria is still in great demand for the enhancement of PDT treatment efficiency. In this study, two new two-photon AIEgens TPPM and TTPM based on a rigid D-π-A skeleton have been designed and synthesized. Both AIEgens TPPM and TTPM show strong aggregation-induced emission (AIE) with the emission enhancement up to 290-folds, large two-photon absorption with the two-photon absorption cross section up to 477 MG, and highly specific targeting to mitochondria in living cells with good biocompatibility. They can serve as two-photon bioprobes for the cell and deep tissue bioimaging with a penetration depth up to 150 µm. Furthermore, high 1O2 generation efficiency with high 1O2 quantum yield under white light irradiation has been found for both TPPM and TTPM and high PDT efficiency to HeLa cells under white light irradiation has also been proven. To the best of our knowledge, AIEgens in this work constitute one of the strongest emission enhancements and one of the highest 1O2 generation efficiencies in the reported organic AIEgens so far. The great AIE feature, large two-photon absorption, high specificity to mitochondria in living cells, and high PDT efficiency to living cells as well as excellent photostability and biocompatibility of these novel AIEgens TPPM and TTPM reveal great potential in clinical applications of two-photon cell and tissue bioimaging and image-guided and mitochondria-targeted photodynamic cancer therapy.