Few-Layered Black Phosphorus: From Fabrication and Customization to Biomedical Applications.

As a new kind of 2D material, black phosphorus has gained increased attention in the past three years. Although few-layered black phosphorus nanosheets (BPs) degrade quickly under ambient conditions to phosphate anions, which greatly hampers their optical and electronic applications, this property also makes BPs highly biocompatible and biodegradable, and is regarded as an advantage for various biomedical applications. This Concept summarizes the state-of-art progresses of BPs, from fabrication and surface modification to biomedical applications. It is expected that BPs with such fascinating properties will encourage more scientists to engage in expanding its biomedical applications by tackling the scientific challenges involved in their development.

Improved Biocompatibility of Black Phosphorus Nanosheets by Chemical Modification.

Black phosphorus nanosheets (BPs) show great potential for various applications including biomedicine, thus their potential side effects and corresponding improvement strategy deserve investigation. Here, in vitro and in vivo biological effects of BPs with and without titanium sulfonate ligand (TiL4 ) modification are investigated. Compared to bare BPs, BPs with TiL4 modification (TiL4 @BPs) can efficiently escape from macrophages uptake, and reduce cytotoxicity and proinflammation. The corresponding mechanisms are also discussed. These findings may not only guide the applications of BPs, but also propose an efficient strategy to further improve the biocompatibility of BPs.

Adsorption force of fibronectin on various surface chemistries and its vital role in osteoblast adhesion.

The amount, type, and conformation of proteins adsorbed on an implanted biomaterial are believed to influence cell adhesion. Nevertheless, only a few research works have been dedicated to the contribution of protein adsorption force. To verify our hypothesis that the adsorption force of protein on biomaterial is another crucial mediator to cell adhesion, fibronectin (FN) adsorbed on self-assembled monolayers (SAMs) with terminal -OH, -CH3, and -NH2 was quantified for FN adsorption force (F(ad)) by utilizing a sphere/plane adsorption model and parallel plate flow chamber. As revealed, F(ad) on SAMs followed a chemistry-dependence of -NH2 > -CH3 ≫ -OH. It is further demonstrated that F(ad) together with FN conformation could regulate the late osteoblast adhesion and the consequent reorganization of the adsorbed FN and fibrillogenesis of the endogenous FN. Our study suggests that protein adsorption force plays a key role in cell adhesion and should be involved for better biomaterial design.

α-Cyanoacrylate Rapid Medical Adhesive (Medical EC Glue) Localization of Pulmonary Nodules Guided by Computed Tomography before Thoracoscopic Surgery.

OBJECTIVE: The study aims to conduct lung cancer screening by low-dose CT to identify the nature of the pulmonary nodule. The purpose of this study was to evaluate the role of preoperative medical EC glue localization of pulmonary nodules of uncertain nature by minimally invasive surgical resection. METHODS: From December 2017 to December 2019, 18 patients (12 women, 6 men; median age: 54 years)with pulmonary nodules were located using medical EC glue under the guidance of preoperative CT and then resected under video thoracoscopy at Air Force Medical Center of PLA. The clinical characteristics were retrospectively collected to evaluate the effectiveness, safety and feasibility of the operation. RESULTS: The mean value of the maximum diameter of pulmonary nodules on CT images before the operation was 10.8 mm. The average depth was 10.3 mm (1.0-39.5 mm). Among 18 nodules, 8 were pure ground glass nodules, 3 were solid nodules, and 7 were partial solid nodules. The diagnosis rate of medical glue localization under the guidance of CT after the operation was 100%. Postoperative pathological diagnosis showed that there were 10 cases of primary lung adenocarcinoma, 1 case of invasive lung adenocarcinoma, 3 cases of adenocarcinoma in situ, 1 case of metastatic adenocarcinoma, and 3 cases of benign nodules. No obvious serious complications were found after localization. CONCLUSION: This study suggests that CT-guided percutaneous medical EC glue localization is a reliable, safe, feasible and practical method for undiagnosed pulmonary nodules and can significantly improve the rate of resection of small pulmonary nodules. Furthermore, it was considered to be more reasonable to remove pulmonary nodules and maximize the preservation of lung function.

Tuning the arrangement of lamellar nanostructures: achieving the dual function of physically killing bacteria and promoting osteogenesis.

Bacteria killing behavior based on physical effects is preferred for biomedical implants because of the negligible associated side effects. However, our current understanding of the antibacterial activity of nanostructures remains limited and, in practice, nanoarchitectures that are created on orthopedics should also promote osteogenesis simultaneously. In this study, tilted and vertical nanolamellar structures are fabricated on semi-crystalline polyether-ether-ketone (PEEK) via argon plasma treatment with or without pre-annealing. The two types of nanolamellae can physically kill the bacteria that come into contact with them, but the antibacterial mechanisms between the two are different. Specifically, the sharp edges of the vertically aligned nanolamellae can penetrate and damage the bacterial membrane, whereas bacteria are stuck on the tilted nanostructures and are stretched, leading to eventual destruction. The tilted nanolamellae are more desirable than the vertically aligned ones from the perspective of peri-implant bone regeneration. Our study not only reveals the role of the arrangement of nanostructures in orthopedic applications but also provides new information about different mechanisms of physical antibacterial activity.

Balancing the biocompatibility and bacterial resistance of polypyrrole by optimized silver incorporation.

Polypyrrole (PPy) which is a conductive polymer with excellent biocompatibility has enormous potential in implantable electronics. However, pristine PPy does not have sufficient bacterial resistance and hence, bacterial infection poses serious threats in vivo. Silver is an excellent antibacterial agent but the optimal concentration is critical because excessive silver is detrimental to human health. In this study, electrochemical polymerization is carried out to fabricate PPy coatings and silver ions (Ag) are introduced by plasma immersion ion implantation (PIII). The optimal Ag ion fluence is determined by monitoring the antibacterial efficiency and cytotoxicity. Our results show that the optimal balance between the antibacterial ability and cytocompatibility can be attained from sample Ti-PPy@Ag-4 implanted with a silver ion fluence of 4 × 1016 ions cm-2. In addition to retaining good cytocompatibility, 92% of the bacteria Staphylococcus aureus (S. aureus) can be eliminated. The intricate balance between antibacterial effects and biocompatibility arises from the levels of intracellular reactive oxygen species (ROS) in S. aureus and MC3T3-E1 osteoblasts on Ti-PPy@Ag-4. The antibacterial capability and biocompatibility are verified by the subcutaneous infection model in rats in vivo. The results reveal a simple strategy to improve the bacterial resistance of polymers such as PPy while not compromising the inherent biosafety of the materials. To the best of our knowledge, this is the first attempt to functionalize PPy by Ag PIII to create the proper balance between the antibacterial capacity and biosafety of biomedical implants.

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.

Dual-initiating and living frustrated Lewis pairs: expeditious synthesis of biobased thermoplastic elastomers.

Biobased poly(γ-methyl-α-methylene-γ-butyrolactone) (PMMBL), an acrylic polymer bearing a cyclic lactone ring, has attracted increasing interest because it not only is biorenewable but also exhibits superior properties to petroleum-based linear analog poly(methyl methacrylate) (PMMA). However, such property enhancement has been limited to resistance to heat and solvent, and mechanically both types of polymers are equally brittle. Here we report the expeditious synthesis of well-defined PMMBL-based ABA tri-block copolymers (tri-BCPs)-enabled by dual-initiating and living frustrated Lewis pairs (FLPs)-which are thermoplastic elastomers showing much superior mechanical properties, especially at high working temperatures (80-130 °C), to those of PMMA-based tri-BCPs. The FLPs consist of a bulky organoaluminum Lewis acid and a series of newly designed bis(imino)phosphine superbases bridged by an alkyl linker, which promote living polymerization of MMBL. Uniquely, such bisphosphine superbases initiate the chain growth from both P-sites concurrently, enabling the accelerated synthesis of tri-BCPs in a one-pot, two-step procedure. The results from mechanistic studies, including the single crystal structure of the dually initiated active species, detailed polymerizations, and kinetic studies confirm the livingness of the polymerization and support the proposed polymerization mechanism featuring the dual initiation and subsequent chain growth from both P-sites of the superbase di-initiator.

A more defective substrate leads to a less defective passive layer: Enhancing the mechanical strength, corrosion resistance and anti-inflammatory response of the low-modulus Ti-45Nb alloy by grain refinement.

Orthopedic and dental implants made of ß-type Ti alloys have low elastic modulus which can better relieve the stress shielding effects after surgical implantation. Nevertheless, clinical application of ß-type Ti alloys is hampered by the insufficient mechanical strength and gradual release of pro-inflammatory metallic ions under physiological conditions. In this study, the ß-type Ti-45Nb alloy is subjected to high-pressure torsion (HPT) processing to refine the grain size. After HPT processing, the tensile strength increases from 370 MPa to 658 MPa due to grain boundary strengthening and at the same time, the favorable elastic modulus is maintained at a low level of 61-72 GPa because the single ß-phase is preserved during grain refinement. More grain boundaries decrease the work function and facilitate the formation of thicker and less defective passive films leading to better corrosion resistance. In addition, more rapid repair of the passive layer mitigates release of metallic ions from the alloy and consequently, the inflammatory response is suppressed. The results reveal a strategy to simultaneously improve the mechanical and biological properties of metallic implant materials for orthopedics and dentistry. STATEMENT OF SIGNIFICANCE: The low modulus Ti-45Nb alloy is promising in addressing the complication of stress shielding induced by biomedical Ti-based materials with too-high elastic modulus. However, its insufficient strength hampers its clinical application, and traditional strengthening via heat treatments will compromise the low elastic modulus. In the current study, we enhanced the ultimate tensile strength of Ti-45Nb from 370 MPa to 658 MPa through grain-refinement strengthening, while the elastic modulus was maintained at a low value (61-72 GPa). Moreover, substrate grain-refinement has been proved to improve the corrosion resistance of Ti-45Nb with reduced inflammatory response both in vitro and in vivo. A relationship between the substrate microstructure and the surface passive layer has been established to explain the beneficial effects of substrate grain-refinement.

Tuning surface topographies on biomaterials to control bacterial infection.

Microbial contamination and subsequent formation of biofilms frequently cause failure of surgical implants and a good understanding of the bacteria-surface interactions is vital to the design and safety of biomaterials. In this review, the physical and chemical factors that are involved in the various stages of implant-associated bacterial infection are described. In particular, topographical modification strategies that have been employed to mitigate bacterial adhesion via topographical mechanisms are summarized and discussed comprehensively. Recent advances have improved our understanding about bacteria-surface interactions and have enabled biomedical engineers and researchers to develop better and more effective antibacterial surfaces. The related interdisciplinary efforts are expected to continue in the quest for next-generation medical devices to attain the ultimate goal of improved clinical outcomes and reduced number of revision surgeries.

Metal-catechol-(amine) networks for surface synergistic catalytic modification: Therapeutic gas generation and biomolecule grafting.

Regarding the high requirement of cardiac and vascular implants in tissue engineering, a novel concept of surface chemistry strategy featuring multiple functions is proposed in this study, which provides glutathione peroxidase (GPx)-like catalytic activity and allows secondary reactions for grafting functional biomolecules. The suggested strategy is the fabrication of a metal-catechol-(amine) network (MCAN) containing copper ions with GPx-like activity, amine-bearing hexamethylenediamine (HD) and wet adhesive catechol dopamine (DA). With a simple one-step molecular/ion co-assembly, the developed copper-DA-HD (CuII-DA/HD) network can be used to catalyze the generation of therapeutic nitric oxide (NO) gas in a durable and dose-controllable manner. The primary amine groups in the CuII-DA/HD network facilitate the secondary immobilization of bivalirudin (BVLD) to further provide an antithrombotic activity as supplement to the functions of NO. The CuII-DA/HD + BVLD coating functionalized on cardiovascular stents successfully improved thromboresistance, anti-restenosis, and promotes re-endothelialization in vivo. With regard to the ease of operation and low cost, the synergetic modification using MCAN strategy is of great potential for developing multifunctional blood-contacting materials/devices.

Enhanced interfacial adhesion and osseointegration of anodic TiO2 nanotube arrays on ultra-fine-grained titanium and underlying mechanisms.

The poor adhesion of anodic TiO2 nanotubes (TNTs) arrays on titanium (Ti) substrates adversely affects applications in many fields especially biomedical engineering. Herein, an efficient strategy is described to improve the adhesion strength of TNTs by performing grain refinement in the underlying Ti substrate via high-pressure torsion processing, as a larger number of grain boundaries can provide more interfacial mechanical anchorage. This process also improves the biocompatibility and osseointegration of TNTs by increasing the surface elastic modulus. The TNTs in length of 0.4 µm have significantly larger adhesion strength than the 2.0 µm long ones because the shorter TNTs experience less interfacial internal stress. However, post-anodization annealing reduces the fluorine concentration in TNTs and adhesion strength due to the formation of interfacial cavities during crystallization. The interfacial structure of TNTs/Ti system and the mechanism of adhesion failures are further investigated and discussed. STATEMENT OF SIGNIFICANCE: Self-assembled TiO2 nanotubes (TNTs) prepared by electrochemical anodization have a distinct morphology and superior properties, which are commonly used in photocatalytic systems, electronic devices, solar cells, sensors, as well as biomedical implants. However, the poor adhesion between the TNTs and Ti substrate has hampered wider applications. Here in this study, we describe an efficient strategy to improve the adhesion strength of TNTs by performing grain refinement in the underlying Ti substrate via high-pressure torsion (HPT) processing. The interfacial structure of TNTs/Ti system and the mechanism of adhesion failure are systematically studied and discussed. Our findings not only develop the knowledge of TNTs/Ti system, but also provide new insights into the design of Ti-based implants for orthopedic applications.

High-Potential surface on zirconia ceramics for bacteriostasis and biocompatibility.

Bacteria easily adhere, colonize, and form biofilm on oral implants subsequently causing periimplantation periarthritis and mechanical loosening. Previous studies show that a high potential surface on polymeric implants can achieve surface bacteriostasis without side effects. In this study, a high surface potential is introduced to zirconia ceramics to mitigate bacterial infection. Carbon and nitrogen plasma immersion ion implantation (C-PIII and N-PIII) are conducted on zirconia ceramic samples sequentially to elevate the surface potential. The surface with a high potential but without ion leaching exhibits excellent antibacterial effects against oral bacteria and little bacterial resistance is triggered. The surface also has high strength and excellent biocompatibility. The nitrogen-containing inorganic structure with high potential can actualize bacteriostasis and biocompatibility on zirconia ceramics simultaneously and this new strategy can enhance the antibacterial ability of oral implants.

Biomimetic osteogenic peptide with mussel adhesion and osteoimmunomodulatory functions to ameliorate interfacial osseointegration under chronic inflammation.

Bone endoprosthesis in patients with systemic chronic inflammation frequently leads to poor osseointegration after implantation mainly due to the increase in pro-inflammatory cytokines that induce bone resorption and impair bone formation. In this work, peptide-coated implants are designed to create a beneficial bone immune microenvironment around prostheses to promote interfacial osteogenesis. By taking advantage of the spontaneous and stable coordination chemistry, Ti-based implants are coated with the mussel-inspired peptide to mitigate lipopolysaccharide (LPS)-induced inflammation by up-regulating the M2 phenotype of macrophages. In addition, the peptide coating increases the bone-implant contact (BIC) by nearly 3 times resulting in suppressed osteoclastogenesis and promoted osteogenesis by inhibiting the nuclear factor kappa-B (NF-κB) signalling pathway. Our findings indicate that biomimetic peptides with osteoimmunomodulatory bioactivity can be incorporated into Ti-based prostheses to facilitate bone regeneration in patients with chronic inflammatory diseases.

Tuning the surface immunomodulatory functions of polyetheretherketone for enhanced osseointegration.

The adverse macrophage-mediated immune response elicited by the surface of polyetheretherketone (PEEK) is responsible for the formation of fibrous encapsulation and resulting inferior osseointegration of PEEK implants in the dental and orthopedic fields. Therefore, endowing the PEEK surface with immunomodulatory ability is an appealing strategy to enhance implant-bone integration. Herein, a reliable and cost-effective method to construct adherent films with tunable nanoporous structures on PEEK is described. The functionalized surface not only suppresses the acute inflammatory response of macrophages, but also provides a favorable milieu for osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs). Whole genome expression analysis reveals that the suppression effect arises from synergistic inhibition of focal adhesion, Toll-like receptor, and NOD-like receptor signaling pathways, as well as the attenuating loop through the JAK-STAT and TNF signaling pathways in macrophages. Further in vivo studies confirm that the functionalized surface induces less fibrous capsule formation and an improved bone regeneration. The nanoporous films fabricated on PEEK harmonize the early macrophage-mediated inflammatory response and subsequent hBMSCs-centered osteogenic functions consequently yielding superior osseointegration.

Biodegradable near-infrared-photoresponsive shape memory implants based on black phosphorus nanofillers.

In this paper, we propose a new shape memory polymer (SMP) composite with excellent near-infrared (NIR)-photoresponsive shape memory performance and biodegradability. The composite is fabricated by using piperazine-based polyurethane (PU) as thermo-responsive SMP incorporated with black-phosphorus (BP) sheets as NIR photothermal nanofillers. Under 808 nm light irradiation, the incorporated BP sheets with concentration of only 0.08 wt% enable rapid temperature increase over the glass temperature of PU and trigger the shape change of the composite with shape recovery rate of ∼100%. The in vitro and in vivo toxicity examinations demonstrate the good biocompatibility of the PU/BP composite, and it degrades naturally into non-toxic carbon dioxide and water from PU and non-toxic phosphate from BP. By implanting PU/BP columns into back subcutis and vagina of mice, they exhibit excellent shape memory activity to change their shape quickly under moderate 808 nm light irradiaiton. Such SMP composite enable the development of intelligent implantable devices, which can be easily controlled by the remote NIR light and degrade gradually after performing the designed functions in the body.

Near-infrared light-triggered drug delivery system based on black phosphorus for in vivo bone regeneration.

A near-infrared (NIR) light-triggered drug delivery platform is produced by incorporating SrCl2 and BP nanosheets (BPs) into poly(lactic-co-glycolic acid) (PLGA) for bone regeneration. The fabricated BP-SrCl2/PLGA microspheres show efficient NIR absorption and photothermal effects due to the BPs. The NIR-triggered release behavior of Sr2+ by flawing the PLGA shells is investigated and the microspheres exhibit excellent cell viability and biodegradability. Implantation of the BP-SrCl2/PLGA microspheres into a rat femoral defect demonstrates good tissue compatibility and excellent bone regeneration capacity under NIR light irradiation. Our study indicates that local release of Sr2+ at optimal time periods controlled by NIR irradiation improves bone regeneration significantly and this NIR-triggered drug delivery system composed of BPs is suitable for therapies requiring precise control at specific time.

Linker-free covalent immobilization of heparin, SDF-1α, and CD47 on PTFE surface for antithrombogenicity, endothelialization and anti-inflammation.

Small-diameter vascular grafts made of biomedical polytetrafluoroethylene (PTFE) suffer from the poor long-term patency rate originating from thrombosis and intimal hyperplasia, which can be ascribed to the insufficient endothelialization and chronic inflammation of the materials. Hence, bio-functionalization of PTFE grafts is highly desirable to circumvent these disadvantages. In this study, a versatile "implantation-incubation" approach in which the biomedical PTFE is initially modified by plasma immersion ion implantation (PIII) is described. After the N2 PIII treatment, the surface of biomedical PTFE is roughened with nanostructures and more importantly, the abundant free radicals generated underneath the surface continuously migrate to the surface and react with environmental molecules. Taking advantage of this mechanism, various biomolecules with different functions can be steadily immobilized on the surface of PTFE by simple solution immersion. As examples, three typical biomolecules, heparin, SDF-1α, and CD47, are covalently grafted onto the PTFE. In addition to retaining the bioactivity, the surface-functionalized PTFE exhibits reduced thrombogenicity, facilitates the recruitment of endothelial progenitor cells, and even alleviates the inflammatory immune responses of monocytes-macrophages and is thus promising to the development of small-diameter prosthetic vascular grafts with good long-term patency.

Enhanced cytocompatibility and reduced genotoxicity of polydimethylsiloxane modified by plasma immersion ion implantation.

Polydimethylsiloxane(PDMS) is a common industrial polymer with advantages such as ease of fabrication, tunable hardness, and other desirable properties, but the basic (-OSi(CH3)2-)n structure in PDMS is inherently hydrophobic thereby hampering application to biomedical engineering. In this study, plasma immersion ion implantation (PIII) is conducted on PDMS to improve the biological properties. PIII forms wrinkled "herringbone" patterns and abundant O-containing functional groups on PDMS to alter the surface hydrophilicity. The biocompatibility of the modified PDMS is assessed with Chinese hamster ovarian cells and compared to that of the untreated PDMS. Our results reveal that the PDMS samples after undergoing PIII have better cytocompatibility and lower genotoxicity. PIII which is a non-line-of-sight technique extends the application of PDMS to the biomedical field.

PLLA nanofibrous paper-based plasmonic substrate with tailored hydrophilicity for focusing SERS detection.

We report a new paper-based surface enhanced Raman scattering (SERS) substrate platform contributed by a poly(l-lactic acid) (PLLA) nanofibrous paper adsorbed with plasmonic nanostructures, which can circumvent many challenges of the existing SERS substrates. This PLLA nanofibrous paper has three-dimensional porous structure, extremely clean surface with good hydrophobicity (contact angle is as high as 133.4°), and negligible background interference under Raman laser excitation. Due to the strong electrostatic interaction between PLLA nanofiber and cetyltrimethylammonium bromide (CTAB) molecules, the CTAB-coated gold nanorods (GNRs) are efficiently immobilized onto the fibers. Such a hydrophobic paper substrate with locally hydrophilic SERS-active area can confine analyte molecules and prevent the random spreading of molecules. The confinement leads to focusing effect and the GNRs-PLLA SERS substrate is found to be highly sensitive (0.1 nM Rhodamine 6G and malachite green) and exhibit excellent reproducibility (∼8% relative standard deviation (RSD)) and long-term stability. Furthermore, it is also cost-efficient, with simple fabrication methodology, and demonstrates high sample collection efficiency. All of these benefits ensure that this GNRs-PLLA substrate is a really perfect choice for a variety of SERS applications.