On-demand engineering of cell membrane receptors to nongenetically intervene in cellular behaviors is still a challenge. Herein, a membraneless enzyme biofuel cell-based self-powered biosensor (EBFC-SPB) was developed for autonomously and precisely releasing Zn2+ to initiate DNAzyme-based reprogramming of cell membrane receptors, which further mediates signal transduction to regulate cellular behaviors. The critical component of EBFC-SPB is a hydrogel film on a biocathode which is prepared using a Fe3+-cross-linked alginate hydrogel film loaded with Zn2+ ions. In the working mode in the presence of glucose/O2, the hydrogel is decomposed due to the reduction of Fe3+ to Fe2+, accompanied by rapid release of Zn2+ to specifically activate a Zn2+-responsive DNAzyme nanodevice on the cell surface, leading to the dimerization of homologous or nonhomologous receptors to promote or inhibit cell proliferation and migration. This EBFC-SPB platform provides a powerful \"sensing-actuating-treating\" tool for chemically regulating cellular behaviors, which holds great promise in precision biomedicine.

Oligomerization of cellular membrane receptors plays crucial roles in activating intracellular downstream signaling cascades for controlling cellular behaviors in physiological and pathological processes. However, the reversible and controllable regulation of receptors in a user-defined manner remains challenging. Herein, we developed a versatile DNA nanorobot (nR) with installed aptamers and hairpin structures to reversibly and controllably regulate cell migration. This was achieved by dimerization and de-dimerization of mesenchymal-epithelial transition (Met) receptors through DNA strand displacement reactions. The functionalized DNA nR not only plays similar roles as hepatocyte growth factor (HGF) in inducing cell migration but also allows a downgrade to the original state of cell migration. The advanced DNA nanomachines can be flexibly designed to target other receptors for manipulating cellular behaviors and thus represent a powerful tool for the future of biological and medical engineering.

Fairly high concentrations of magnesium and lithium are conducive to improving the osteogenic and angiogenic capacities. In the current study, lithium-containing magnesium phosphate-based ceramics (AMP/LMPGs) were prepared from amorphous magnesium phosphate (AMP) at a low sintering temperature (650 °C), and the lithium/magnesium-containing phosphate glasses (LMPGs) were utilized as sintering additives. During the sintering procedure of AMP/LMPGs, the AMP reacted with LMPGs, producing new compounds. The AMP/LMPGs displayed nano-size grains and plentiful micropores. The addition of LMPGs noticeably increased the porosity as well as compressive strength of the AMP/LMPGs ceramics. The AMP/LMPGs sustainedly released Mg, P and Li ions, forming Mg-rich ionic microenvironment, which ameliorated cellular proliferation, osteogenic differentiation and proangiogenic capacities. The AMP/LMPGs ceramics with considerably high compressive strength, osteostimulation and proangiogenic effects were expected to efficiently regenerate the bone defects.

Liquid-liquid phase separation is a multicomponent system separated into phases with different compositions and structures. It has been identified and explored in organisms after being introduced from the thermodynamic field. Condensate, the product of phase separation, exists in different scales of cellular structures, such as nucleolus, stress granules, and other organelles in nuclei or cytoplasm. And also play critical roles in different cellular behaviors. Here, we review the concept, thermodynamical and biochemical principles of phase separation. We summarized the main functions including the adjustment of biochemical reaction rates, the regulation of macromolecule folding state, subcellular structural support, the mediation of subcellular location, and intimately linked to different kinds of diseases, such as cancer and neurodegeneration. Advanced detection methods to investigate phase separation are collected and analyzed. We conclude with the discussion of anxiety of phase separation, and thought about how progress can be made to develop precise detection methods and disclose the potential application of condensates.

Porous Nb-25Ta-25Ti alloys (60% porosity and 100-600 μm pore size) for bone implant applications were manufactured combining impregnation and sintering methods. Surfaces with porous micro-nanostructured networks on Nb-Ta-Ti alloys were successfully modified by various surface pre-treatments (acid etching, alkali-heat treatment and annealing treatment). Surface characteristics and Ca-P layer deposition behaviors of the multilevel structured porous Nb-Ta-Ti alloys were investigated by conducting various tests, including x-ray diffraction, scanning electron microscopy, energy-dispersive x-ray, atomic force microscopy and optical contact angle measurement. In particular, bulk Nb-Ta-Ti alloys were also used as mutual control. The results demonstrated that the porous alloy exhibited a unique multilevel porous structure with macro-networks and micro-pits after pre-treatments. The surface passive TiO2/Nb2O5/Ta2O5layers on Nb-Ta-Ti alloys were partially dissolved by the corrosive attack of hydroxyl ions during alkali heat treatment. In addition, subsequent annealing treatment increased the density of the gel layers formed during alkali heat treatment. After immersion in SBF for 14 d, a continuous relatively uniform apatite layer was formed on the multilevel structured surfaces. Moreover, the mechanism of surface mineralization can be construed as electrostatic interactions between substrates and ions. Furthermore,in vitrocell culture showed that Nb-Ta-Ti alloys had a good biocompatibility and the multilevel porous structure could enhance the cellular behaviors including: cell adhesion and spreading.

In the process of tumorigenesis and development, cancer cells must integrate and respond to complex and dynamic signals in the tumor microenvironment. Nevertheless, simulating the original biomechanical and biochemical microenvironment of different cells in vitro is a fundamental challenge in studying synergistic effects. To address this issue, we have proposed a biomimetic multi-factor stimulation platform, which conveniently creates a two-dimensional matrix environment with controllable stiffness as biomechanical factor and can smoothly introduce biochemical cue. Our results indicated that the extracellular matrix (ECM) stiffness could enhance the cell stretching, which further lead to the amplification of cell-matrix adhesion and proliferation. And there existed obvious differences of endocytosis efficiency response of cells on matrix stiffness. Nanoparticles (NPs) with the same size and shape, but differs in electrical charges showed more uptake on harder matrix. Besides, the inorganic polyphosphates (polyP), which acts as energy storage and producer in the in-extracellular space, was proven to synergistically promote aforementioned cell biomechanical behaviors by increasing ATP metabolism for the first time. These results explored the impact of microenvironmental performance on the glioma mechanoresponses, and we believe this biomimetic multifactor stimulation method would exhibit a profound impact on researches of in vitro biomimetic cell culture and NPs-like drug phagocytosis.

The precise regulation of fundamental cellular behaviors, such as cell adhesion, proliferation, migration, differentiation, and cell-cell interactions, is essential in multicellular organisms. The regulation of cellular behaviors largely relies on the receptor-mediated sensing and transduction of the extracellular ligands. The precise manipulation of ligand-induced receptor activation via molecular engineering approaches will offer fundamental insight into physiological processes, facilitating the rational design of cell-based therapeutics to treat diseases. DNA is a highly attractive biomolecule as it works through either genetically encoding the functional protein or rationally fabricating functional nanomaterials. Recently accumulated research in functional nucleic acids and DNA nanotechnology makes it possible to construct dynamic and complex nanodevices to modulate receptor-mediated signaling and cellular behaviors. This review summarizes the recent advances in state-of-art of DNA technologies in engineering the interaction between ligand and cell-surface receptor, mainly highlighting the design principles and the emerging applications of DNA-based nongenetic engineering strategies for receptor-mediated cellular behaviors. We expect that significant advancements in DNA-based engineering strategies will enable the emergence of applications in cell-based therapy in regenerative medicine and cancer immunotherapy.

Electrospun fiber scaffolds, due to their mimicry of bone extracellular matrix (ECM), have become an important biomaterial widely applied in bone tissue engineering in recent years. While topographic cues of electrospun membranes such as alignment and diameter played vital roles in determining cellular behaviors. Yet few researches about the effects of these two significant parameters on osteogenesis have been reported. Thus, the present work explored the influence of aligned and random poly (L-lactic acid) (PLLA) fiber matrices with diameters of nanoscale (0.6 μm) and microscale (1.2 μm), respectively, on cellular responses of bone marrow mesenchymal stem cells (BMSCs), such as cell adhesion, migration, proliferation and osteogenesis. Our results revealed that aligned nanofibers (AN) could affect cell morphology and promote the migration of BMSCs after 24 h of cell culturing. Besides, AN group was observed to possess excellent biocompatibility and have significantly improved cell growth comparing with random nanofibers. More importantly, in vitro osteogenesis researches including ALP and Alizarin Red S staining, qRT-PCR and immunofluorescence staining demonstrated that BMSCs culturing on AN group exhibited higher osteogenic induction proficiency than that on aligned microfibers (AM) and random fiber substrates (RN and RM). Accordingly, aligned nanofiber scaffolds have greater application potential in bone tissue engineering.

The grain structure and surface morphology of bio-implants act as a pivotal part in altering cell behavior. Titanium alloy bone screws, as common implants, are prone to screws loosening and complications threat in the physiological environment due to their inferior anti-wear and surface inertia. Manufacturing bone screws with high wear resistance and ideal biocompatibility has always been a challenge. In this study, a series of overlapping morphologies inspired by the hierarchical structure of fish scales and micro bulges of shrimp were structured on Ti-6Al-4V implant by laser texturing. The results indicate that the textured patterns could improve cell attachment, proliferation, and osteogenic differentiation. The short-term response of human bone marrow-derived mesenchymal stem cells (hBMSCs) on the textured surface are more sensitive to the microstructure than the surface roughness, wettability, grain size and surface chemical elements of the textured surfaces. More importantly, the friction-increasing and friction-reducing type overlapping structures exhibit excellent friction stability at different stages of modified simulated body fluid (m-SBF) soaking. The overlapping structure (Micro-smooth stacked ring: MSSR) is more beneficial to promote the formation of apatite. Deposited spherical-like apatite particles can act as a \"lubricant\" on the MSSR surface during the friction process to alleviate the adhesion wear of the surface. Meanwhile, apatite particles participate in the formation of friction film, which plays an effective role in reducing friction and antiwear in corrosion solution (m-SBF) for a long time. These features show that the combination of soaking treatment in m-SBF solution with laser-textured MSSR structure is expected to be an efficient and environmentally friendly strategy to prolong the service life of bone screws and reducing the complications of mildly osteoporotic implants.

BACKGROUND: CHIP is an E3 ubiquitin ligase that plays contrast roles in diverse human malignancies, depending on its targets. To date, the mechanisms underlying the function of CHIP in gastric cancer remains unclear. Here, we aim to further clarify the effects of CHIP on the development and progression of gastric cancer and explore its potential target.
METHODS: Stably transfected CHIP-shRNA and TRAF2-shRNA AGS gastric cancer cell lines were established using Lipofectamine 2000. Cell growth was measured by an xCelligence real-time monitoring system and colony formation assay. Cell proliferation was detected using CCK-8, Ki-67, or CFSE assays. Apoptosis was detected by TUNEL assay or Annexin V/PI-staining followed by flow cytometric analysis. Cell cycle distribution was detected by PI-staining followed by flow cytometric analysis. Cell migration and invasion abilities were measured by a real-time xCelligence system, Transwell insert, and scratch assays. The expression of cell cycle-related proteins, apoptosis-related proteins, AKT, ERK, NF-κB signaling subunits, MMP2, MMP9, and Integrin β-1 were detected by Western blotting analysis. NF-κB DNA-binding capability was quantified using an ELISA-based NF-κB activity assay. Gastric cancer tissue microarray was analyzed to investigate the expression of both CHIP and TRAF2, and their clinical significance.
RESULTS: The CHIP-silencing in the AGS cells was oncogenic evidenced by the appearance of capable of anchorage-independent growth. The CHIP-silencing significantly enhanced the AGS cell proliferation capability likely due to the induced phosphorylation of ERK. The CHIP-silencing significantly inhibited apoptosis due to increased expression of Bcl-2. The CHIP-silencing promoted the AGS cell migration and invasion abilities, likely by regulating the expression of Integrin β-1. TRAF2 expression was markedly decreased in the CHIP-overexpressing cells at protein level, but not at mRNA level. The TRAF2-silencing markedly inhibited the proliferation ability of the AGS cells, the defected cell proliferation and enhanced apoptosis were involved in. The TRAF2-silencing also attenuated the cell migration and invasion capacities of the AGS cells. Furthermore, the expression of CHIP was downregulated while the expression of TRAF2 was upregulated in gastric cancer tissues. TRAF2 expression is independent prognostic factors of gastric cancer. The expression of CHIP and TRAF2 was negatively correlated in the gastric cancer tissue. Lower CHIP or higher TRAF2 was significantly linked to shorter overall survival in gastric cancer patients.
CONCLUSIONS: TRAF2 influenced diverse aspects of cellular behavior of gastric cancer cells, including cell growth, migration, and invasion, which was in contrast to the functions of CHIP. TRAF2 could be considered as an independent prognostic factor in gastric cancer patients. It is possible that TRAF2 was a substrate of CHIP and CHIP regulated the TRAF2/NF-κB axis, which modulated diverse cellular behaviors in the AGS gastric cancer cells.