The role of nanopores constructed on the micropitted titanium surface in the immune responses of macrophages and the potential mechanisms.

The delayed transition of macrophages (MΦs) from pro-inflammatory M1 to the pro-healing M2 state on implant surfaces is one of the most important reasons for poor osseointegration. This work reports the construction of closely packed nanopores with a small diameter on the micropitted titanium (Ti) surface by two-step acid etching to promote the M1-to-M2 transition of MΦs and pays special attention to the potential mechanisms by which the nanopores decorating the micropits exert immunomodulatory effects. The results show that the structure composed of hybrid nanopores (10-25 nm) and micropits (5-15 µm) can be produced on the Ti surface by a two-step acid etching process. Compared with the unitary micropits, the micropit/nanopore surface could facilitate the switch of MΦs from the pro-inflammatory M1 to the pro-healing M2 phenotype. RNA sequencing reveals that the MAPK, PI3K-AKT and C-type lectin signaling pathways play key roles in the micro/nano-structure-mediated transition. In addition, the micro/nano-structured surface down-regulated CYP1A2 expression, reducing the generation of mitochondrial ROS, in turn restraining the oxidative stress and further attenuating inflammation. This work provides novel insights into the underlying mechanisms of immunomodulation by the nano-structure-decorated micro-structures on the Ti surface, which is significant for designing the surface of orthopedic implants from the perspective of immunomodulation.

Type I collagen decorated nanoporous network on titanium implant surface promotes osseointegration through mediating immunomodulation, angiogenesis, and osteogenesis.

Osseointegration of implants is a complex physiological process that requires temporal and spatial regulation of immune responses, angiogenesis, and osteogenesis. To achieve efficient and long-term osseointegration, type I collagen (COL1) decorated nanoporous network was developed on titanium substrates via alkali treatment, polydopamine coating, and layer-by-layer (LBL) self-assembly. It was noted that the simple physisorbed COL1 could be easily desorbed from the nanostructured surface, however, multilayer COL1 constructed by polydopamine and LBL self-assembly obscured the nanoporous network of the alkali-treated titanium surfaces. Interestingly, the nanostructured surface covalently immobilized with COL1 (T-ADC) could timely convert macrophages (MΦs) from pro-inflammatory M1 to pro-healing M2 phenotype, generating a beneficial osteoimmune microenvironment and promoting angio/osteo-genesis. RNA sequencing revealed that the nanostructure and COL1 could synergistically activate RhoA/ROCK, PI3K-AKT, and classical MAPK signaling pathways in MΦs to sustain the cell cycle, and trigger autocrine feedback-mediated JAK-STAT and FoxO signaling pathways, which in turn motivated autophagy and oxidative stress resistance and attenuated lipopolysaccharide-induced Toll-like receptor signaling pathway and its downstream NF-κB and JNK/p38 MAPK signaling cascades, leading to the inhibition of inflammation and osteoclastic-related gene expression of MΦs. Simultaneously, T-ADC prominently facilitated angiogenesis of endothelial cells and osteogenesis of osteoblasts as well as their cross-talks, further highlighting synergistically positive effects of the nanostructure and COL1 on osseointegration. In vivo experiments revealed that T-ADC could induce abundant new bone mass and ameliorative osseointegration, corroborating the in vitro results. The study elucidated that the COL1 decorated nanoporous network on titanium surfaces could significantly regulate early inflammatory reaction and subsequent angio/osteo-genesis processes, resulting in favorable osseointegration.

Fabrication and characterization of biodegradable Zn scaffold by vacuum heating-press sintering for bone repair.

Bone repair materials with excellent mechanical properties are highly desirable, especially in load-bearing sits. However, the currently used ceramic- and polymer-based ones mainly show poor mechanical properties. Recently, biodegradable metals have attracted extensive attention due to their reliable mechanical strength and degradability. As biodegradable metals, zinc-based materials are promising due to their suitable degradation rate and good biocompatibility. Here, we fabricated biodegradable porous Zn scaffolds with relatively high mechanical properties by vacuum heating-press sintering using NaCl particles as space holders. The microstructure, actual porosity, compressive mechanical properties, in vitro degradation behavior and the vitality of osteoblasts of porous Zn scaffolds were tested and investigated. The results show the porosities of the prepared porous Zn scaffolds are ranging from 11.3 % to 63.3 %, and the pore sizes are similar to the size range of the screened NaCl particles (200-500 µm). Compressive yield strength of 14.2-73.7 MPa and compressive elastic modulus of 1.9-6.7 GPa are shown on porous Zn scaffolds, some of which approach to that of cancellous bone (2-12 MPa and 0.1-5 GPa). Compared to bulk Zn, although the porous structures cause a partial loss of strength, the reliable mechanical properties are still retained. In addition, the porous structures not only greatly increase the degradation rate, but also promote the proliferation of osteoblasts. Based on these results, biodegradable porous Zn scaffolds (porosity in the 40 %-50 %) fabricated by vacuum heating-press sintering method show high application potential for clinical bone repair.

Biocompatible silane adhesion layer on titanium implants improves angiogenesis and osteogenesis.

Silane adhesion layer strategy has been widely used to covalently graft biomolecules to the titanium implant surface, thereby conferring the implant bioactivity to ameliorate osseointegration. However, few researchers pay attention to the effects of silanization parameters on biocompatibility and biofunctionality of the silane adhesion layers. Accordingly, the present study successfully fabricated the silane adhesion layers with different thickness, intactness, and surface morphologies by introducing 3-aminopropyltriethoxysilane on the alkali-treated titanium surface in time-varied processing of silanization. The regulatory effects of the silane adhesion layers on angiogenesis and osteogenesis were assessed in vitro. Results showed that the prolonged silanization processing time increased the thickness and intactness of the silane adhesion layer and significantly improved its biocompatibility. Notably, the silane adhesion layer prepared after 12 h of silanization exhibited a brain-like surface morphology and benefited the adhesion and proliferation of endothelial cells (ECs) and osteoblasts (OBs). Moreover, the layer promoted angiogenesis via stimulating vascular endothelial growth factor (VEGF) secretion and nitric oxide (NO) production of ECs. Simultaneously, it improved osteogenesis by enhancing alkaline phosphatase (ALP) activity, collagen secretion, and extracellular matrix mineralization of OBs. This work systematically investigated the biocompatibility and biofunctionality of the modified silane adhesion layers, thus providing valuable references for their application in covalently grafting biomolecules on the titanium implant surface.

Exosomes derived from magnesium ion-stimulated macrophages inhibit angiogenesis.

Angiogenesis, an essential prerequisite to osteogenesis in bone repair and regeneration, can be mediated by immunoregulation of macrophages. Magnesium and its alloys are promising biodegradable bone implant materials and can affect immunoregulation of macrophages by the degradation products (magnesium ions). Nevertheless, the mechanism of macrophage-derived exosomes stimulated by Mg ions in immunoregulation is still not well understood. Herein, 10-50 mM magnesium ions are shown to inhibit the macrophage viability and proliferation in a dose-dependent manner, but a high concentration results in macrophage apoptosis. The exosomes secreted by macrophages from magnesium ion stimulation inhibit angiogenesis of endothelial cells, as manifested by the suppressed cell viability, proliferation, migration, and tube formation, which arise at least partially from exosome-mediated downregulation of endothelial nitric oxide and the vascular endothelial growth factor. The findings reported in this paper suggest that the bio-functionality of biodegradable magnesium alloys must be considered from the perspective of immunoregulation of macrophage-derived exosomes. Our results also suggest potential cancer therapy by inhibiting tumor-associated angiogenesis.

Microstructure and Corrosion Resistance of Laser-Welded Crossed Nitinol Wires.

Laser welding has been considered to be one of the most promising joining processes for Nitinol medical device manufacturing. Presently, there is still a limited understanding about how laser welding affects the microstructure and the resultant corrosion behaviors. This work aimed to reveal the microstructural factors that influence the corrosion resistance of laser-welded crossed Nitinol joints. The microstructures within various zones of the joints were characterized by using transmission electron microscopy (TEM), and the corrosion behaviors of the joints in 0.9% NaCl and Hank's solutions were studied. The base metal exhibits a single austenite (B2) phase and the highest corrosion resistance. The phase constituent of the fusion zone is the coexistence of the B2 matrix and some precipitates (T2Ni, TiNi3, and Ti3Ni4 particles), resulting in a slight decrease in corrosion resistance. The heat affected zone (HAZ) shows the austenite matrix but with the precipitation of R-phase, which considerably reduces the corrosion potential, making it the weakest zone.