Baiping Ren

The energy dissipation and Mullins effect of tough polymer/graphene oxide hybrid nanocomposite hydrogels

Nanocomposite hydrogels (NC gels) are considered to belong to the class of high strength hydrogels. Graphene oxide (GO), owing to its amphiphilic, mechanical, and optical properties, is widely used as a filler incorporated into different hosting materials (elastomers, plastics, and hydrogels) to improve their mechanical properties. In this work, we used in situ free radical polymerization to synthesize polyacrylamide (PAAm)/GO hybrid NC gels in the presence of GO nanosheets and a very small amount of chemical cross-linkers (N,N′-methylenebisacrylamide, MBA < 0.1 mol%). By optimizing GO and MBA concentrations, the resulting PAAm/GO gels can achieve an elastic modulus of 66 kPa, a fracture stress of 0.27 MPa, a fracture strain of 13.76 mm mm−1, deformed energy of 2.52 MJ m−3, and tearing energy of 964 J m−2. Due to the presence of physical interactions between PAAm and GO nanosheets, PAAm/GO gels demonstrate λ-dependent energy dissipation and Mullins self-recovery behaviors. The gels can rapidly recover their stiffness and toughness by 76% and 60%, respectively, after 30 min of resting at room temperature. The possible toughening mechanisms and Mullins effects of PAAm/GO gels were proposed and compared with those of filler rubbers and other high strength hydrogels. This work provides new viewpoints to develop tough hydrogels by the introduction of GO into other hydrogels with a good mechanical balance between strong chemical bonding and reversible physical bonding.

Membrane Interactions of hIAPP Monomer and Oligomer with Lipid Membranes by Molecular Dynamics Simulations

Interaction of human islet amyloid polypeptide (hIAPP) peptides with cell membrane is crucial for the understanding of amyloid toxicity associated with Type II diabetes (T2D). While it is known that the hIAPP-membrane interactions are considered to promote hIAPP aggregation into fibrils and induce membrane disruption, the membrane-induced conformation, orientation, aggregation, and adsorption behaviors of hIAPP peptides have not been well understood at the atomic level. Herein, we perform all-atom explicit-water molecular dynamics (MD) simulations to study the adsorption, orientation, and surface interaction of hIAPP aggregates with different sizes (monomer to tetramer) and conformations (monomer with α-helix and tetramer with β-sheet-rich U-turn) upon adsorption on the lipid bilayers composed of both pure zwitterionic POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) and mixed anionic POPC/POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine) (3:1) lipids. MD simulation results show that hIAPP monomer with α-helical conformation and hIAPP pentamer with β-sheet conformation can adsorb on both POPC and POPC/POPE bilayers via a preferential orientation of N-terminal residues facing toward the bilayer surface. The hIAPP aggregates show stronger interactions with mixed POPC/POPE lipids than pure POPC lipids, consistent with experimental observation that hIAPP adsorption and fibrililation are enhanced on mixed lipid bilayers. While electrostatic interactions are main attractive forces to drive the hIAPP aggregates to adsorb on both bilayers, the introduction of the more hydrophilic head groups of POPE lipids further promote the formation of the interfacial hydrogen bonds. Complement to our previous studies of hIAPP aggregates in bulk solution, this computational work increases our knowledge about the mechanism of amyloid peptide-membrane interactions that is central to the understanding the progression of all amyloid diseases.

Molecular Understanding of Aβ-hIAPP Cross-Seeding Assemblies on Lipid Membranes

Amyloid-β (Aβ) and human islet polypeptide (hIAPP) are the causative agents responsible for Alzheimers disease (AD) and type II diabetes (T2D), respectively. While numerous studies have reported the cross-seeding behavior of Aβ and hIAPP in solution, little effort has been made to examine the cross-seeding of Aβ and hIAPP in the presence of cell membranes, which is more biologically relevant to the pathological link between AD and T2D. In this work, we computationally study the cross-seeding and adsorption behaviors of Aβ and hIAPP on zwitterionic POPC and anionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG) mixed bilayers using all-atom molecular dynamics (MD) simulations, particularly aiming to the effects of the initial orientation of the Aβ-hIAPP assembly and the lipid composition of cell membranes on mutual structural and interaction changes in both Aβ-hIAPP assembly and lipid bilayers at the atomic level. Aβ-hIAPP cross-seeding assembly always preferred to adopt a specific orientation and interface to associate with both lipid bilayers strongly via the N-terminal strands of Aβ. Such membrane-bound orientation explains experimental observation that hybrid Aβ-hIAPP fibrils on cell membranes showed similar morphologies to pure hIAPP fibrils. Moreover, Aβ-hIAPP assembly, regardless of its initial orientations, interacted more strongly with POPC/POPG bilayer than POPC bilayer, indicating that electrostatic interactions are the major forces governing peptide-lipid interactions. Strong electrostatic interactions were also attributed to the formation of Ca2+ bridges connecting both negatively charged Glu of Aβ and PO4 head groups of lipids, which facilitate the association of Aβ-hIAPP with the POPC/POPG bilayer. It was also found that the strong peptide-lipid binding reduced lipid fluidity. Both facts imply that Aβ-hIAPP assembly may induce cell damage by altering calcium homeostasis and cell membrane phase. This work provides a better fundamental understanding of cross-seeding of Aβ and hIAPP on cell membranes and a potential pathological link between AD and T2D.

Seed-Induced Heterogeneous Cross-Seeding Self-Assembly of Human and Rat Islet Polypeptides

Amyloid peptides can misfold and aggregate into amyloid oligomers and fibrils containing conformationally similar β-sheet structures, which are linked to the pathological hallmark of many neurodegenerative diseases. These β-sheet-rich amyloid aggregates provide common structural motifs to accelerate amyloid formation by acting as seeds. However, little is known about how one amyloid peptide aggregation will affect another one (namely, cross-seeding). In this work, we studied the cross-seeding possibility and efficiency between rat islet amyloid polypeptide (rIAPP) and human islet amyloid polypeptide (hIAPP) solution with preformed aggregates at different aggregation phases, using a combination of different biophysical techniques. hIAPP is a well-known peptide hormone that forms amyloid fibrils and induces cytotoxicity to β-cells in type 2 diabetes, whereas rIAPP is a nonaggregating and nontoxic peptide. Experimental results showed that all different preformed hIAPP aggregates can cross-seed rIAPP to promote the final fibril formation but exhibit different cross-seeding efficiencies. Evidently, hIAPP seeds preformed at a growth phase show the strongest cross-seeding potential to rIAPP, which accelerates the conformational transition from random structures to β-sheet and the aggregation process at the fibrillization stage. Homoseeding of hIAPP is more efficient in initiating and promoting aggregation than cross-seeding of hIAPP and rIAPP. Moreover, the cross-seeding of rIAPP with hIAPP at the lag phase also reduced cell viability, probably because of the formation of more toxic hybrid oligomers at the prolonged lag phase. The cross-seeding effects in this work may add new insights into the mechanistic understanding of the aggregation and coaggregation of amyloid peptides linked to different neurodegenerative diseases.

Identification of a New Function of Cardiovascular Disease Drug 3-Morpholinosydnonimine Hydrochloride as an Amyloid-β Aggregation Inhibitor

Cardiovascular disease (CVD) and Alzheimer’s disease (AD) have a mutual cause-and-effect relationship, and they share some common risk factors. Although numerous Food and Drug Administration (FDA)-approved drugs have been developed for CVD treatment, no drugs are clinically available for AD treatment. Given the common disease-causing factors and links between the two diseases and the well-demonstrated drugs for CVD, we propose to re-examine the new potential of the existing CVD drugs as amyloid-β (Aβ) inhibitors. 3-Morpholinosydnonimine hydrochloride (SIN-1) is an FDA-approved drug for inhibiting platelet aggregation in CVD. Herein, we examine the inhibition activity of SIN-1 on the aggregation and toxicity of Aβ1–42 using combined experimental and computational approaches. Collective experimental data from ThT, circular dichroism, and atomic force microscopy demonstrate that SIN-1 can effectively inhibit amyloid formation at every stage of Aβ aggregation by prolonging lag phase, slowing down aggregation rate, and reducing final fibril formation. The cell viability assay also shows that SIN-1 enables the protection of SH-SY5Y cells from Aβ-induced cell toxicity. Such an inhibition effect is attributed to interference with the structural transition of Aβ toward a β-sheet structure by SIN-1. Furthermore, molecular dynamic simulations confirm that SIN-1 preferentially binds to the C-terminal β-sheet grooves of an Aβ oligomer and consequently disrupts the β-sheet structure of Aβ and Aβ–Aβ association, explaining experimental observations. This work discovers a new function of SIN-1, making it a promising compound with dual protective roles in inhibiting both platelet and Aβ aggregations against CVD and AD.

Integration of antifouling and antibacterial properties in salt-responsive hydrogels with surface regeneration capacity

The development of new antimicrobial materials and strategies is of importance for many biomedical and industrial applications. In this work, we report a new strategy to integrate distinct antimicrobial, antifouling, and stimuli-responsive properties into a single hydrogel to realize bacteria resistance, killing, and releasing functions. To achieve this design, we conjugated salt-responsive anti-polyelectrolyte polyDVBAPS (poly(3-(dimethyl(4-vinylbenzyl)ammonio)propyl sulfonate)) with antifouling polyHEAA (poly(N-hydroxyethyl acrylamide)) and antimicrobial AgNPs (silver nanoparticles) to form a hybrid hydrogel of polyDVBAPS-g-polyHEAA@AgNPs, among which polyHEAA functions as a general antifouling background to prevent bacteria adsorption on the surface, AgNPs act as antimicrobial agents to kill bacteria on the surface, and polyDVBAPS uses its unique salt-responsive, anti-polyelectrolyte property to release adherent bacteria from the surface. In this design, polyDVBAPS-g-polyHEAA@AgNPs hydrogels not only effectively resist bacteria attachment and kill the adherent bacteria, but also regenerate the antifouling surface of the hydrogel by releasing the adhered bacteria to keep the surface free from bacteria. The polyDVBAPS-g-polyHEAA@AgNPs hydrogels exhibited high surface resistance to bacteria adsorption (<106 cells per cm2) for up to 4 days, high antibacterial activity by killing ∼99% of attached bacteria of both E. coli and S. aureus, and surface regeneration ability by releasing >96% of adherent live or dead bacteria from the surface upon a simple treatment of 1.0 M NaCl solution for 10 min. Upon the release of AgNPs, AgNPs were reloaded into the hydrogel again to achieve multiple antifouling, bactericidal, and regenerative properties. This work demonstrates a new design for a new multifunctional hydrogel to effectively achieve antimicrobial, antifouling, and surface regeneration properties, making this hydrogel very promising for antimicrobial applications.

Tanshinones inhibit hIAPP aggregation, disaggregate preformed hIAPP fibrils, and protect cultured cells

Misfolding and aggregation of amyloid peptides are the key pathological events in many neurodegenerative diseases. The development of effective inhibitors and drugs to prevent amyloid peptide aggregation is considered as an important therapeutic strategy for treating these diseases. We previously reported on tanshinones, ingredients from the Chinese herb Danshen (Salvia miltiorrhiza Bunge), as a potent inhibitor against amyloid-β1-42 (Aβ) aggregation and toxicity. Considering the common structural and aggregation features, and the correlation of type II diabetes (T2D) and Alzheimers disease (AD), herein we examine the inhibition activity of two tanshinone I (TS1) and IIA (TS2) components on the aggregation and toxicity of hIAPP1-37 using combined experimental and computational approaches. Collective experimental data from ThT, CD, and AFM confirm that both tanshinones show comparable inhibition ability to reduce hIAPP aggregates by inhibiting the fibrillation process and changing the fibrillogenesis pathway, leading to the formation of some amorphous aggregates. More importantly, both tanshinones are capable of disassembling preformed hIAPP fibrils, but TS1 shows better potency in fibril dissembling than TS2. MTT and LDH assays also show that the tanshinones at very low concentrations of 5 μM can reduce the hIAPP-induced cell toxicity. Molecular dynamics (MD) simulations further reveal that both tanshinones preferentially bind to β-sheets to prevent lateral association of hIAPP aggregates and thus to inhibit fibril growth, explaining experimental observations. This work discovers that tanshinones act as common inhibitors to inhibit the aggregation of both hIAPP and Aβ, disaggregate preformed hIAPP and Aβ amyloid fibrils, and protect cells from hIAPP- and Aβ-induced toxicity, making them very promising agents against AD, T2D, and probably other amyloid-misfolding diseases.

Molecular simulation aspects of amyloid peptides at membrane interface

The interactions of amyloid peptides with cell membranes play an important role in maintaining the integrity and functionality of cell membrane. A thorough molecular-level understanding of the structure, dynamics, and interactions between amyloid peptides and cell membranes is critical to amyloid aggregation and toxicity mechanisms for the bench-to-bedside applications. Here we review the most recent computational studies of amyloid peptides at model cell membranes. Different mechanisms of action of amyloid peptides on/in cell membranes, targeted by different computational techniques at different lengthscales and timescales, are rationally discussed. Finally, we have proposed some new insights into the remaining challenges and perspectives for future studies to improve our understanding of the activity of amyloid peptides associated with protein-misfolding diseases. This article is part of a Special Issue entitled: Protein Aggregation and Misfolding at the Cell Membrane Interface edited by Ayyalusamy Ramamoorthy.

Role of Protein Charge Density on Hepatitis B Virus Capsid Formation

The role of electrostatic interactions in the viral capsid assembly process was studied by comparing the assembly process of a truncated hepatitis B virus capsid protein Cp149 with its mutant protein D2N/D4N, which has the same conformational structure but four fewer charges per dimer. The capsid protein self-assembly was investigated under a wide range of protein surface charge densities by changing the protein concentration, buffer pH, and solution ionic strength. Lowering the protein charge density favored the capsid formation. However, lowering charge beyond a certain point resulted in capsid aggregation and precipitation. Interestingly, both the wild-type and D2N/D4N mutant displayed identical assembly profiles when their charge densities matched each other. These results indicated that the charge density was optimized by nature to ensure an efficient and effective capsid proliferation under the physiological pH and ionic strength.

Genistein: A Dual Inhibitor of Both Amyloid β and Human Islet Amylin Peptides

Abnormal misfolding and aggregation of amyloid peptides into amyloid fibrils are common and critical pathological events in many neurodegenerative diseases. Most inhibitors or drugs have been developed to prevent amyloid aggregation of a specific peptide, showing sequence-dependent inhibition mechanisms. It is more challenging to develop or discover inhibitors capable of preventing the aggregation of two or more different amyloid peptides. Genistein, a major phytoestrogen in soybean, has been widely used as an anti-inflammation and cerebrovascular drug due to its antioxidation and antiacetylcholinesterase effects. Herein, we examine the inhibitory effects of genistein on the aggregation of amyloid-β (Aβ, associated with Alzheimers disease) and human islet amylin (hIAPP, associated with type 2 diabetes) and Aβ- and hIAPP-induced neurotoxicity using a combination of experimental and computational approaches. Collective experimental results from thioflavin T (ThT), atomic force microscopy (AFM), and circular dichroism (CD) demonstrate that genistein shows strong inhibition ability to prevent the conformational transition of both Aβ and hIAPP monomers to β-sheet structures, thus reducing final amyloid fibrillization from Aβ and hIAPP monomer aggregation by 40-63%. Further 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH), and large unilamellar vesicle (LUV) assays show that genistein helps to increase cell viability, decrease cell apoptosis, and reduce cell membrane leakage, where the cell protection effect of genistein is likely correlated with its reduced membrane leakage. Comparative molecular dynamics (MD) simulations reveal that genistein prefers to bind the β-sheet groove, a common structural motif of amyloid fibrils, of both Aβ and hIAPP oligomers to interfere with their self-aggregation. This work for the first time demonstrates genistein as a dual inhibitor of Aβ and hIAPP aggregation. Further structural optimization and refinement of genistein may generate a series of effective sequence-independent inhibitors against the aggregation and toxicity of different amyloid peptides.