Single-Molecule Force Spectroscopy Reveals Multiple Binding Modes between DOPA and Different Rutile Surfaces.

Inspired by marine mussel adhesive systems, numerous 3,4-dihydroxyphenylalanine (DOPA)-containing surface coating materials have been recently designed. It is well known that DOPA has a strong adhesion ability to different kinds of wet surfaces. However, the molecular mechanism of DOPA adhesion remains elusive. Recent biophysical studies of DOPA adhesion by both surface force apparatus (SFA) and atomic force microscopy (AFM) suggest that DOPA can bind to a wide range of surfaces exhibiting diverse chemical properties through different binding mechanisms. Here, using AFM-based single-molecule force spectroscopy, we show that even for chemically well-defined crystal surfaces, DOPA can bind to them by multiple binding modes. The binding forces between DOPA and different rutile TiO2 surfaces can vary within a broad range from 40-800 pN at a pulling speed of 1000 nm s-1 and are largely dependent on the surface properties. Our findings indicate that the local chemical environment can greatly affect DOPA adhesion, and that single-molecule force spectroscopy is a unique tool to reveal the heterogeneity of DOPA adhesion to the same surface.

Mussel-Derived and Bioclickable Peptide Mimic for Enhanced Interfacial Osseointegration via Synergistic Immunomodulation and Vascularized Bone Regeneration.

Inadequate osseointegration at the interface is a key factor in orthopedic implant failure. Mechanistically, traditional orthopedic implant interfaces fail to precisely match natural bone regeneration processes in vivo. In this study, a novel biomimetic coating on titanium substrates (DPA-Co/GFO) through a mussel adhesion-mediated ion coordination and molecular clicking strategy is engineered. In vivo and in vitro results confirm that the coating exhibits excellent biocompatibility and effectively promotes angiogenesis and osteogenesis. Crucially, the biomimetic coating targets the integrin α2ß1 receptor to promote M2 macrophage polarization and achieves a synergistic effect between immunomodulation and vascularized bone regeneration, thereby maximizing osseointegration at the interface. Mechanical push-out tests reveal that the pull-out strength in the DPA-Co/GFO group is markedly greater than that in the control group (79.04 ± 3.20 N vs 31.47 ± 1.87 N, P < 0.01) and even surpasses that in the sham group (79.04 ± 3.20 N vs 63.09 ± 8.52 N, P < 0.01). In summary, the novel biomimetic coating developed in this study precisely matches the natural process of bone regeneration in vivo, enhancing interface-related osseointegration and showing considerable potential for clinical translation and applications.

Mussel-inspired resilient hydrogels with strong skin adhesion and high-sensitivity for wearable device.

Stretchable and self-adhesive conductive hydrogels hold significant importance across a wide spectrum of applications, including human-machine interfaces, wearable devices, and soft robotics. However, integrating multiple properties, such as high stretchability, strong interfacial adhesion, self-healing capability, and sensitivity, into a single material poses significant technical challenges. Herein, we present a multifunctional conductive hydrogel based on poly(acrylic acid) (PAA), dopamine-functionalized pectin (PT-DA), polydopamine-coated reduction graphene oxide (rGO-PDA), and Fe3+ as an ionic cross-linker. This hydrogel exhibits a combination of high stretchability (2000%), rapid self-healing (~ 94% recovery in 5 s), and robust self-adhesion to various substrates. Notably, the hydrogel demonstrates a remarkable skin adhesion strength of 85 kPa, surpassing previous skin adhesive hydrogels. Furthermore, incorporating rGO within the hydrogel network creates electric pathways, ensuring excellent conductivity (0.56 S m-1). Consequently, these conductive hydrogels exhibit strain-sensing properties with a significant increase in gauge factor (GF) of 14.6, covering an extensive detection range of ~ 1000%, fast response (198 ms) and exceptional cycle stability. These multifunctional hydrogels can be seamlessly integrated into motion detection sensors capable of distinguishing between various strong or subtle movements of the human body.

Efficient secretion of mussel adhesion proteins using a chaperone protein Spy as fusion tag in Bacillus subtilis.

BACKGROUND: Mussel foot proteins (Mfps) are considered as remarkable materials due to their extraordinary adhesive capability. Recombinant expression is an ideal way to synthesis these proteins at large scale. However, secretory expression of Mfps into culture medium has not been achieved in a heterologous host. METHODS AND RESULTS: Here, to realize the secretion of Mfp3 and Mfp5 in Bacillus subtilis, signal peptide screening was first performed. Minimal Mfp3-6×His was targeted into the growth medium with AmyE signal peptide. We found that a small chaperone protein Spy was secreted efficiently in B. subtilis, and the fusion proteins Spy-Mfp3-6×His and Spy-Mfp5-6×His could also be delivered into growth medium well. The yield of Spy-Mfp3-6×His and Spy-Mfp5-6×His reached 255 and 119 mg L-1 at shake flask conditions, respectively. Mfp3-6×His and Mfp5-6×His were finally purified via TEV protease cleavage and NTA affinity chromatography. CONCLUSION: Mfp3-6×His and Mfp5-6×His could be efficiently secreted using a chaperone protein Spy as fusion tag in B. subtilis.

Clinical application of a new hemostatic material using mussel-inspired catecholamine hemostat: A pilot study.

BACKGROUNDS/AIMS: This study aimed to evaluate clinical application of InnoSEAL Plus (a mussel-inspired catecholamine hemostat) as a new hemostatic material for humans. METHODS: Patients treated with topical hemostatic patches after liver resection were enrolled. They were divided into an experimental group (InnoSEAL Plus group) and two control groups (TachoSil® group and Surgicel Fibrillar® group) for efficacy evaluation. RESULTS: A total of 15 patients were enrolled. Each group had five patients. The 3-minute hemostasis success rate was 80.0% (4/5 patients) in the InnoSEAL Plus group, 80.0% (4/5 patients) in the TachoSil® group, and 40.0% (2/5 patients) in the Surgicel Fibrillar® group, showing no significant difference in the success rate among these groups (p > 0.05). All three groups exhibited 100% success rate for 10-minute hemostasis. Both InnoSEAL Plus and TachoSil® groups had one patient developing adverse events, which were treated easily with drug administrations. CONCLUSIONS: InnoSEAL Plus is expected to be functionally not inferior to other conventional hemostatic agents. However, it is necessary to confirm this through multicenter prospective studies in the future.

Rational integration of defense and repair synergy on PEEK osteoimplants via biomimetic peptide clicking strategy.

Polyetheretherketone (PEEK) has been widely used as orthopedic and dental materials due to excellent mechanical and physicochemical tolerance. However, its biological inertness, poor osteoinduction, and weak antibacterial activity make the clinical applications in a dilemma. Inspired by the mussel adhesion mechanism, here we reported a biomimetic surface strategy for rational integration and optimization of anti-infectivity and osteo-inductivity onto PEEK surfaces using a mussel foot proteins (Mfps)-mimic peptide with clickable azido terminal. The peptide enables mussel-like adhesion on PEEK biomaterial surfaces, leaving azido groups for the further steps of biofunctionalizations. In this study, antimicrobial peptide (AMP) and osteogenic growth peptide (OGP) were bioorthogonally clicked on the azido-modified PEEK biomaterials to obtain a dual-effect of host defense and tissue repair. Since bioorthogonal clicking allows precise collocation between AMP and OGP through changing their feeding molar ratios, an optimal PEEK surface was finally obtained in this research, which could long-term inhibit bacterial growth, stabilize bone homeostasis and facilitate interfacial bone regeneration. In a word, this upgraded mussel surface strategy proposed in this study is promising for the surface bioengineering of inert medical implants, in particular, achieving rational integration of multiple biofunctions to match clinical requirements.

A multicenter, prospective, randomized clinical trial of marine mussel-inspired adhesive hemostatic materials, InnoSEAL Plus.

PURPOSE: InnoSEAL Plus is an adhesive, coagulant-free hemostatic material that mimics the adhesion mechanism of marine mussels. This study reports on the safety and efficacy of InnoSEAL Plus for patients with hemorrhage after hepatectomy despite first-line hemostasis treatments. METHODS: This is a multicenter, prospective, single-blinded, randomized clinical trial involving 96 hepatectomy patients. TachoSil was used as a comparator group. Three-minute and 10-minute hemostatic success rates were monitored. Rebleeding rates were also observed. Safety was assessed by recording all novel undesirable symptoms. RESULTS: InnoSEAL Plus showed a 3-minute hemostasis rate of 100%, while TachoSil had a rate of 98.0% (48 of 49 patients), demonstrating that the 2 had similar hemostatic efficacies. The difference in efficacy between the test and comparator group was 2.04%, and the lower limit of the one-sided 97.5% confidence interval was -1.92%; as this is greater than the noninferiority limit of -23.9%, the 2 treatments were equivalent. Meanwhile, the 10-minute hemostatic success rate was the same in both groups (100%). No rebleeding occurred in either group. In the safety evaluation, 89 patients experienced adverse events (45 in the test group and 44 in the comparator group). The difference between the 2 groups was not significant. No death occurred after application of the test or comparator group product. CONCLUSION: Given that InnoSEAL Plus is a coagulation factor-free product, the hemostasis results are encouraging, especially considering that TachoSil contains a coagulation factor. InnoSEAL Plus was found to be a safe and effective hemostatic material for control of bleeding in hepatectomy patients.

Counterplotting the Mechanosensing-Based Fouling Mechanism of Mussels against Fouling.

Marine organisms react to various factors when building colonies for survival; however, severe accumulation of diverse organisms on artificial structures located close to water causes large industrial losses. Herein, we identify a concept in the development of antifouling surfaces based on understanding the surface stiffness recognition procedure of mussel adhesion at the genetic level. It was found that on a soft surface the combination of decreased adhesive plaque size, adhesion force, and plaque protein downregulation synergistically weakens mussel wet adhesion and sometimes prevents mussels from anchoring, mainly due to transcriptional changes within the mechanosensing pathway and the adhesive proteins in secretory glands. In addition, the use of soft substrates or antagonists of surface mechanosensing behavior suppresses mussel fouling significantly.

The position of lysine controls the catechol-mediated surface adhesion and cohesion in underwater mussel adhesion.

Intensive studies have found that 3,4-dihydroxyphenylalanine (Dopa) is one of the key molecules for underwater mussel adhesion. Although basic mechanisms of mussel adhesion have been elucidated, little is known about how mussels control the balance between surface adhesion and cohesion, which is critical for successful adhesion without peeling and/or tearing. In this work, we focused on lysine (Lys) molecules which are frequently flanked to Dopa residues in interfacial adhesive proteins, specifically their synergy and anti-synergy on surface adhesion and cohesion. Three model peptides were designed to characterize flanking Lys effects. Through nano-mechanistic analyses, we found that flanking Lys enhanced surface adhesion but disrupted Fe3+-mediated cohesion. Through nuclear magnetic resonance analyses and density functional theory calculations, we corroborated the synergetic effect on surface adhesion and anti-synergetic effect on cohesion. We also confirmed the consistency of flanking Lys effects in the actual protein system. Thus, we, for the first time, discovered that each Dopa molecule in interfacial adhesive proteins is participated in surface adhesion and cohesion differently through controlling the existence of flanking Lys. Our discovery enlightens how nature designs adhesive proteins through according roles of Dopa.

Metal-crosslinked ɛ-poly-L-lysine tissue adhesives with high adhesive performance: Inspiration from mussel adhesive environment.

Strong glue of mussels has long been considered as an ideal model to design synthetic bio-adhesives but the adhesive strength of metal-crosslinked mussel-inspired glues is not often satisfactory. Herein, inspired by the adhesive environment of mussels, we obtained metal-crosslinked ε-poly-L-lysine adhesives with high adhesive performance by introducing the elements of suitable adhesive environment (SAE) into the adhesives. The elements of SAE were clarified as weak alkaline conditions (pH ∼ 7.4) and low Fe3+ contents. The adhesive strength (∼105 kPa) of the metal-crosslinked adhesives endowed with the elements of SAE (PL-Cat/Fe-SAE) was about 8 times higher than that of fibrin glues. The high adhesive strength was found to originate from distinctive interfacial adhesion and cohesion strength of PL-Cat/Fe-SAE. PL-Cat/Fe-SAE showed strong interfacial adhesion capacity and nearly comparable cohesion strength to those PL-Cat/Fe adhesives with higher Fe3+ contents. The nearly comparable cohesion strength of PL-Cat/Fe-SAE was then found to be due to more amount of stable tris-complex existed in PL-Cat/Fe-SAE. In addition, PL-Cat/Fe-SAE was able to efficiently close the full thickness skin incisions. The study highlighted the importance of introducing SAE elements into the design of tissue adhesives and provided a facile and efficient strategy for constructing tissue adhesives with high adhesive performance.

Translational bioadhesion research: embracing biology without tokenism.

Bioadhesion has attracted a sizable research community of scientists and engineers that is striving increasingly for translational outcomes in anti-fouling and bioinspired adhesion initiatives. As bioadhesion is highly context-dependent, attempts to trivialize or gloss over the fundamental physical, chemical and biological sciences involved will compromise the relevance and durability of translation. This article is part of the theme issue 'Transdisciplinary approaches to the study of adhesion and adhesives in biological systems'.

Bio-/Nanoimmobilization Platform Based on Bioinspired Fibrin-Bone@Polydopamine-Shell Adhesive Composites for Biosensing.

Inspired by blood coagulation and mussel adhesion, we report novel adhesive fibrin-bone@polydopamine (PDA)-shell composite matrix as highly efficient immobilization platform for biomacromolecules and nanomaterials. Fibrin, as a bioglue, and PDA, as a chemical adhesive, are integrated in a one-pot simultaneous polymerization consisting of biopolymerization of fibrinogen and chemical polymerization of dopamine. Fibrin fibers act as adhesive bones to construct scaffold, while PDA coat on the scaffold to form adhesive shell, generating 3D porous composite matrix with unique bone@shell structure. Two types of enzymes (glucose oxidase and acetylcholinesterase) and Au nanoparticles were adopted as respective model biomolecules and nanomaterials to investigate the immobilization capability of the matrix. The bionanocomposites showed high efficiency in capturing nanoparticles and enzymes, as well as significant mass-transfer and biocatalysis efficiencies. Therefore, the bionanocomposites exhibited significant potential in biosensing of glucose and paraoxon with limits of detection down to 5.2 µM and 4 ppt, respectively. The biological-chemical-combined polymerization strategy and composite platform with high immobilization capacity and mass-transfer efficiency open up a novel way for the preparation of high-performance bionanocomposites for various applications, in particular, biosensing.

PYRROLO isolated from marine sponge associated bacterium Halobacillus kuroshimensis SNSAB01 - Antifouling study based on molecular docking, diatom adhesion and mussel byssal thread inhibition.

In the present study, an attempt has been made to explore the antifouling potential of bioactive compound isolated from sponge associated bacterium Halobacillus kuroshimensis SNSAB01. The crude extract of SNSAB01 strongly inhibited the growth of fouling bacterial strains with least minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The bioactive compound was characterized through FT-IR, HPLC, GCMS and NMR predicted as 'pyrrolo". From the mass spectral library, structure was elucidated as pyrrolo [1, 2-a] pyrazine-1, 4-dione, hexahydro. The in silico studies provided encouraging docking scores with two interactions by GLN 200 and GLU 304. The extract inhibited 89% diatom adhesion at 350 µg/ml concentration against Amphora sp. An EC50 value of 150 µg/ml for 50% inhibition of byssal thread of Perna viridis and LC50 was found to be 500 µg/ml. The LC50/EC50 ratio of 3.0 indicated nontoxic to nature. The result suggested that pyrrolo[1,2-a]pyrazine-1,4-dione can be used for antifouling coating.

Mussel-Inspired Injectable Hydrogel Adhesive Formed under Mild Conditions Features Near-Native Tissue Properties.

Injectable hydrogel adhesives, especially those that can strongly adhere to tissues and feature near-native tissue mechanical properties, are desirable biomaterials for tissue repair. Compared to nonadhesive injectable hydrogels for minimally invasive delivery of therapeutic agents, they can better retain the delivered agents at targeted tissue locations and provide additional local physical barriers. However, regardless of recent advances, an ideal injectable hydrogel adhesive with both proper adhesion and mechanical matching between hydrogels and tissues is yet to be demonstrated with cytocompatible and efficient in situ curing methods. Inspired by marine mussels, where different mussel foot proteins (Mfps) function cooperatively to achieve excellent wet adhesion, we herein report a dual-mode-mimicking strategy by modifying gelatin (Gel) biopolymers with a single-type thiourea-catechol (TU-Cat) functionality to mimic two types of Mfps and their mode of action. This strategy features a minor, yet impactful modification of biopolymers, which gives access to collective properties of an ideal injectable hydrogel adhesive. Specifically, with TU-Cat functionalization of only ∼0.4-1.2 mol % of total amino acid residues, the Mfp-mimetic gelatin biopolymer (Gel-TU-Cat) can be injected and cured rapidly under mild and cytocompatible conditions, giving rise to tissue adhesive hydrogels with excellent matrix ductility, proper wet adhesion, and native tissue-like stress relaxation behaviors. Such a set of properties originating from our novel dual-mode-mimicking strategy makes the injectable hydrogel adhesive a promising platform for cell delivery and tissue repair.

A cohort of new adhesive proteins identified from transcriptomic analysis of mussel foot glands.

The adaptive attachment of marine mussels to a wide range of substrates in a high-energy, saline environment has been explored for decades and is a significant driver of bioinspired wet adhesion research. Mussel attachment relies on a fibrous holdfast known as the byssus, which is made by a specialized appendage called the foot. Multiple adhesive and structural proteins are rapidly synthesized, secreted and moulded by the foot into holdfast threads. About 10 well-characterized proteins, namely the mussel foot proteins (Mfps), the preCols and the thread matrix proteins, are reported as representing the bulk of these structures. To explore how robust this proposition is, we sequenced the transcriptome of the glandular tissues that produce and secrete the various holdfast components using next-generation sequencing methods. Surprisingly, we found around 15 highly expressed genes that have not previously been characterized, but bear key similarities to the previously defined mussel foot proteins, suggesting additional contribution to byssal function. We verified the validity of these transcripts by polymerase chain reaction, cloning and Sanger sequencing as well as confirming their presence as proteins in the byssus. These newly identified proteins greatly expand the palette of mussel holdfast biochemistry and provide new targets for investigation into bioinspired wet adhesion.

Combined effect of mussel-inspired surface modification and topographical cues on the behavior of skeletal myoblasts.

The combined effect of mussel-inspired polydopamine (PDA) surface functionalization and topographical cues on the behavior of skeletal myoblasts is described. On PDA-modified nanofibers, myogenic protein expression and the fusion of myoblasts are increased significantly compared with those on unmodified nanofibers. The multinucleate myotubes on the aligned nanofibers are oriented in a direction parallel to the nanofibers.