Design and biological evaluation of dual tubulin/HDAC inhibitors based on millepachine for treatment of prostate cancer.

In this work, a novel of dual tubulin/HDAC inhibitors were designed and synthesized based on the structure of natural product millepachine, which has been identified as a tubulin polymerization inhibitor. Biological evaluation revealed that compound 9n exhibited an impressive potency against PC-3 cells with the IC50 value of 16 nM and effectively inhibited both microtubule polymerization and HDAC activity. Furthermore, compound 9n not only induced cell cycle arrest at G2/M phase, but also induced PC- 3 cells apoptosis. Further study revealed that the induction of cell apoptosis by 9n was accompanied by a decrease in mitochondrial membrane potential and an elevation in reactive oxygen species levels in PC-3 cells. Additionally, 9n exhibited inhibitory effects on tumor cell migration and angiogenesis. In PC-3 xenograft model, 9n achieved a remarkable tumor inhibition rate of 90.07%@20 mg/kg, significantly surpassing to that of CA-4 (55.62%@20 mg/kg). Meanwhile, 9n exhibited the favorable drug metabolism characteristics in vivo. All the results indicate that 9n is a promising dual tubulin/HDAC inhibitor for chemotherapy of prostate cancer, deserving the further investigation.

Design, synthesis and biological evaluation of CDC20 inhibitors for treatment of triple-negative breast cancer.

The involvement of CDC20 in promoting tumor growth in different types of human cancers and it disturbs the process of cell division and impedes tumor proliferation. In this work, a novel of Apcin derivatives targeting CDC20 were designed and synthesized to evaluate for their biological activities. The inhibitory effect on the proliferation of four human tumor cell lines (MCF-7, MDA-MB-231, MDA-MB-468 and A549) was observed. Among them, compound E1 exhibited the strongest inhibitory effect on the proliferation of MDA-MB-231 cells with an IC50 value of 1.43 µM, which was significantly superior to that of Apcin. Further biological studies demonstrated that compound E1 inhibited cancer cell migration and colony formation. Furthermore, compound E1 specifically targeted CDC20 and exhibited a higher binding affinity to CDC20 compared to that of Apcin, thereby inducing cell cycle arrest in the G2/M phase of cancer cells. Moreover, it has been observed that compound E1 induces autophagy in cancer cells. In 4T1 Xenograft Models compound E1 exhibited the potential antitumor activity without obvious toxicity. These findings suggest that E1 could be regarded as a CDC20 inhibitor deserved further investigation.

Discovery of Novel Isoquinoline Analogues as Dual Tubulin Polymerization/V-ATPase Inhibitors with Immunogenic Cell Death Induction.

Cancer immunotherapy has revolutionized clinical advances in a variety of cancers. Due to the low immunogenicity of the tumor, only a few patients can benefit from it. Specific microtubule inhibitors can effectively induce immunogenic cell death and improve immunogenicity of the tumor. A series of isoquinoline derivatives based on the natural products podophyllotoxin and diphyllin were designed and synthesized. Among them, F10 showed robust antiproliferation activity against four human cancer cell lines, and it was verified that F10 exerted antiproliferative activity by inhibiting tubulin and V-ATPase. Further studies indicated that F10 is able to induce immunogenic cell death in addition to apoptosis. Meanwhile, F10 inhibited tumor growth in an RM-1 homograft model with enhanced T lymphocyte infiltration. These results suggest that F10 may be a promising lead compound for the development of a new generation of microtubule drugs.

Discovery of Novel Potent Covalent Glutathione Peroxidase 4 Inhibitors as Highly Selective Ferroptosis Inducers for the Treatment of Triple-Negative Breast Cancer.

Glutathione peroxidase 4 (GPX4) is a promising target to induce ferroptosis for the treatment of triple-negative breast cancer (TNBC). We designed and synthesized a novel series of covalent GPX4 inhibitors based on RSL3 and ML162 by structural integration and simplification strategies. Among them, compound C18 revealed a remarkable inhibitory activity against TNBC cells and significantly inhibited the activity of GPX4 compared to RSL3 and ML162. Moreover, it was identified that C18 could notably induce ferroptosis with high selectivity by increasing the accumulation of lipid peroxides (LPOs) in cells. Further study demonstrated that C18 covalently bound to the Sec46 of GPX4. Surprisingly, C18 exhibited an outstanding potency of tumor growth inhibition in the MDA-MB-231 xenograft model with a TGI value of 81.0%@20 mg/kg without obvious toxicity. Overall, C18 could be a promising GPX4 covalent inhibitor to induce ferroptosis for the treatment of TNBC.

Discovery of Novel N-Heterocyclic-Fused Deoxypodophyllotoxin Analogues as Tubulin Polymerization Inhibitors Targeting the Colchicine-Binding Site for Cancer Treatment.

Natural products are a major source of anticancer agents and play critical roles in anticancer drug development. Inspired by the complexity-to-diversity strategy, novel deoxypodophyllotoxin (DPT) analogues were designed and synthesized. Among them, compound C3 exhibited the potent antiproliferative activity against four human cancer cell lines with IC50 values in the low nanomolar range. Additionally, it showed marked activity against paclitaxel-resistant MCF-7 cells and A549 cells. Moreover, compound C3 can inhibit tubulin polymerization by targeting the colchicine-binding site of tubulin. Further study revealed that compound C3 could arrest cancer cells in the G2/M phase and disrupt the angiogenesis in human umbilical vein endothelial cells. Meanwhile, C3 remarkably inhibited cancer cell motility and migration, as well as considerably inhibited tumor growth in MCF-7 and MCF-7/TxR xenograft model without obvious toxicity. Collectively, these results indicated that compound C3 may be a promising tubulin polymerization inhibitor development for cancer treatment.

Microstructure and Isothermal Oxidation of Ir-Rh Spark Plug Electrodes.

High-temperature oxidation tests were performed on pure iridium, rhodium, and the iridium alloys, IrRh10, IrRh25, and IrRh40, at 1100 °C in a stable air environment for 60 h. The results of the oxidation were analyzed by X-ray photoelectron spectroscopy (XPS). Microstructural changes of the Ir-Rh alloys were characterized by scanning electron microscopy (SEM). XPS analysis results show that the main oxide of the Ir-Rh alloy in a 1100 °C environment was Rh2O3, and SEM analysis shows that the surfaces of the Ir-Rh alloys after oxidation formed both linear and ellipse-shaped corrosion pits, and had the same direction with the wire-drawing process. The oxidation behavior of Ir-Rh alloys, including the mass change, the reason for the mass loss, and the role of Rh in improving oxidation resistance performance, are discussed.

In Situ, Light-Guided Axon Growth on Biomaterials via Photoactivatable Laminin Peptidomimetic IK(HANBP)VAV.

The ability to guide the growth of neurites is relevant for reconstructing neural networks and for nerve tissue regeneration. Here, a biofunctional hydrogel that allows light-based directional control of axon growth in situ is presented. The gel is covalently modified with a photoactivatable derivative of the short laminin peptidomimetic IKVAV. This adhesive peptide contains the photoremovable group 2-(4'-amino-4-nitro-[1,1'-biphenyl]-3-yl)propan-1-ol (HANBP) on the Lys rest that inhibits its activity. The modified peptide is highly soluble in water and can be simply conjugated to -COOH containing hydrogels via its terminal -NH2 group. Light exposure allows presentation of the IKVAV adhesive motif on a soft hydrogel at desired concentration and at defined position and time point. The photoactivated gel supports neurite outgrowth in embryonic neural progenitor cells culture and allows site-selective guidance of neurites extension. In situ exposure of cell cultures using a scanning laser allows outgrowth of neurites in desired pathways.

Toward Light-Regulated Living Biomaterials.

Living materials are an emergent material class, infused with the productive, adaptive, and regenerative properties of living organisms. Property regulation in living materials requires encoding responsive units in the living components to allow external manipulation of their function. Here, an optoregulated Escherichia coli (E. coli)-based living biomaterial that can be externally addressed using light to interact with mammalian cells is demonstrated. This is achieved by using a photoactivatable inducer of gene expression and bacterial surface display technology to present an integrin-specific miniprotein on the outer membrane of an endotoxin-free E. coli strain. Hydrogel surfaces functionalized with the bacteria can expose cell adhesive molecules upon in situ light-activation, and trigger cell adhesion. Surface immobilized bacteria are able to deliver a fluorescent protein to the mammalian cells with which they are interacting, indicating the potential of such a bacterial material to deliver molecules to cells in a targeted manner.

Photoactivatable Adhesive Ligands for Light-Guided Neuronal Growth.

Neuro-regeneration after trauma requires growth and reconnection of neurons to reestablish information flow in particular directions across the damaged tissue. To support this process, biomaterials for nerve tissue regeneration need to provide spatial information to adhesion receptors on the cell membrane and to provide directionality to growing neurites. Here, photoactivatable adhesive peptides based on the CASIKVAVSADR laminin peptidomimetic are presented and applied to spatiotemporal control of neuronal growth to biomaterials in vitro. The introduction of a photoremovable group [6-nitroveratryl (NVOC), 3-(4,5-dimethoxy-2-nitrophenyl)butan-2-yl (DMNPB), or 2,2'-((3'-(1-hydroxypropan-2-yl)-4'-nitro-[1,1'-biphenyl]-4-yl)azanediyl)bis(ethan-1-ol) (HANBP)] at the amino terminal group of the K residue temporally inhibited the activity of the peptide. The bioactivity was regained through controlled light exposure. When used in neuronal culture substrates, the peptides allowed light-based control of the attachment and differentiation of neuronal cells. Site-selective irradiation activated adhesion and differentiation cues and guided seeded neurons to grow in predefined patterns. This is the first demonstration of ligand-based light-controlled interaction between neuronal cells and biomaterials.

Microenvironments to study migration and somal translocation in cortical neurons.

Migrating post-mitotic neurons of the developing cerebral cortex undergo terminal somal translocation (ST) when they reach their final destination in the cortical plate. This process is crucial for proper cortical layering and its perturbation can lead to brain dysfunction. Here we present a reductionist biomaterials platform that faithfully supports and controls the distinct phases of terminal ST in vitro. We developed microenvironments with different adhesive molecules to support neuronal attachment, neurite extension, and migration in distinct manners. Efficient ST occurred when the leading process of migratory neurons crossed from low-to high-adhesive areas on a substrate, promoting spreading of the leading growth cone. Our results indicate that elementary adhesive cell-substrate interactions strongly influence migratory behavior and the final positioning of neurons during their developmental journey. This in vitro model allows advanced experimentation to reveal the microenvironmental requirements underlying cortical layer development and disorders.

Mechanically Reinforced Catechol-Containing Hydrogels with Improved Tissue Gluing Performance.

In situ forming hydrogels with catechol groups as tissue reactive functionalities are interesting bioinspired materials for tissue adhesion. Poly(ethylene glycol) (PEG)⁻catechol tissue glues have been intensively investigated for this purpose. Different cross-linking mechanisms (oxidative or metal complexation) and cross-linking conditions (pH, oxidant concentration, etc.) have been studied in order to optimize the curing kinetics and final cross-linking degree of the system. However, reported systems still show limited mechanical stability, as expected from a PEG network, and this fact limits their potential application to load bearing tissues. Here, we describe mechanically reinforced PEG⁻catechol adhesives showing excellent and tunable cohesive properties and adhesive performance to tissue in the presence of blood. We used collagen/PEG mixtures, eventually filled with hydroxyapatite nanoparticles. The composite hydrogels show far better mechanical performance than the individual components. It is noteworthy that the adhesion strength measured on skin covered with blood was >40 kPa, largely surpassing (>6 fold) the performance of cyanoacrylate, fibrin, and PEG⁻catechol systems. Moreover, the mechanical and interfacial properties could be easily tuned by slight changes in the composition of the glue to adapt them to the particular properties of the tissue. The reported adhesive compositions can tune and improve cohesive and adhesive properties of PEG⁻catechol-based tissue glues for load-bearing surgery applications.

[Application of modified lateral window for maxillary sinus floor augmentation].

Objective: To evaluate the clinical efficacy of modified lateral window for maxillary sinus floor augmentation (MSFA). Methods: Fifty-five patients who visited the Stomatology Hospital Affiliated to Zhejiang University School of Medicine between June 2012 and October 2014 were enrolled in the study. Patients underwent MSFA with Bio-Oss grafts based on modified access window. During the operation the vertical height of the bony window was reduced from 6-8 mm of conventional oval window to 4-5 mm of slot-shaped window. The sinus membrane was detached completely via the lateral access and large particle Bio-Oss graft was placed in the sub-mucosal space. The implant survival, graft height, graft volume and resorption rates were measured. Intra-op and post-op complications were recorded. Results: There were 86 implants inserted. The 2-4 year cumulative survival rates were 97.67% by implant-based analysis and 96.36% by patient-based analysis. The residual bone height was (4.7±2.6) mm and bone width was (8.4±2.7) mm. The bone height of implantation site immediately after operation was (16.1±2.5) mm and it was (16.2±2.2) mm at restoration. The bone heights at 1 and 2 years after operation were (14.9±2.5) mm and (13.6±2.6) mm, respectively. The graft height was (10.6±2.8) mm and the graft volume was (1569±745) mm3 immediately after operation. The resorption rate of graft height 6 months after operation was 3.79% and that of graft volume was 7.87%. The 1-year accumulative resorption rate of graft height was 6.63% and that of graft volume was 10.89%. The 2-year accumulative resorption rate of graft height was 7.58% and that of graft volume was 15.26%. Small membrane perforation during MSFA was observed in 5 cases and all were successfully repaired by a collagen Bio-Gide membrane. Conclusion: The modified lateral technique obtains high implant survival rate, excellent graft stability and low complication rate at 2-4 year clinical follow-up, indicating that it is a safe, predictable and minimally invasive surgical method for severe atrophic maxillary posterior dentition.

Biological and immunotoxicity evaluation of antimicrobial peptide-loaded coatings using a layer-by-layer process on titanium.

The prevention and control of peri-implantitis is a challenge in dental implant surgery. Dental implants with sustained antimicrobial coating are an ideal way of preventing peri-implantitis. This study reports development of a non- immunotoxicity multilayered coating on a titanium surface that had sustained antimicrobial activity and limited early biofilm formation. In this study, the broad spectrum AMP, Tet213, was linked to collagen IV through sulfo-SMPB and has been renamed as AMPCol. The multilayer AMPCol coatings were assembled on smooth titanium surfaces using a LBL technique. Using XPS, AFM, contact angle analysis, and QCM, layer-by-layer accumulation of coating thickness was measured and increased surface wetting compared to controls was confirmed. Non-cytotoxicity to HaCaT and low erythrocyte hemolysis by the AMPCol coatings was observed. In vivo immunotoxicity assays showed IP administration of AMPCol did not effect serum immunoglobulin levels. This coating with controlled release of AMP decreased the growth of both a Gram-positive aerobe (Staphylococcus aureus) and a Gram-negative anaerobe (Porphyromonas gingivalis) up to one month. Early S. aureus biofilm formation was inhibited by the coating. The excellent long-term sustained antimicrobial activity of this multilayer coating is a potential method for preventing peri-implantitis through coated on the neck of implants before surgery.

Characterization and In Vivo Evaluation of Trace Element-Loaded Implant Surfaces in Ovariectomized Rats.

PURPOSE: The purpose of this work was to investigate the effect of a hydroxyapatite (HA) coating doped with zinc (Zn), magnesium (Mg), or strontium (Sr) on implant osseointegration in ovariectomized rats. MATERIALS AND METHODS: The HA coating doped with trace elements was produced by electrochemical deposition. Surface properties were tested by field emission scanning electron microscopy, x-ray diffraction, x-ray photoelectron spectroscopy, and measurement of ionic release. A total of 36 ovariectomized rats were randomly divided into four groups: the HA coating group, the Zn-HA coating group, the Mg-HA coating group, and the Sr-HA coating group. The corresponding implants were then inserted into the tibiae. Histomorphometric analysis was performed after 4, 8, and 12 weeks. RESULTS: HA coatings doped with 2.5% Zn, Mg, or Sr by electrochemical deposition showed almost identical surface morphologies. Bone-implant contact (BIC) in the Zn-HA group was significantly greater than that in the HA group for all time periods. BIC values in the Sr-HA group significantly exceeded those in the HA group at 8 and 12 weeks after implantation. Bone ingrowth area values of both the Zn-HA and Sr-HA groups were statistically significantly different from those in the HA group after 8 and 12 weeks. CONCLUSION: The Zn-HA and Sr-HA coatings have the potential to improve implant osseointegration in osteoporosis.

Influence of simvastatin-loaded implants on osseointegration in an ovariectomized animal model.

The success of bone implants in the presence of osteoporosis is limited by lack of osseointegration between the implant and the natural bone. This study applied an electrochemical process to deposit simvastatin-nanohydroxyapatite (HA) coatings on porous implant surfaces and investigated the effects of these simvastatin-HA coatings on implant surfaces in an animal model of osteoporosis. In this study, simvastatin-HA coated implants were inserted into the tibia of osteoporotic rats. After 2, 4, and 12 weeks, tissue was retrieved for histomorphometric evaluation. The results indicated that the simvastatin-HA coatings increased bone-implant contact and new bone formation around implant surfaces. In conclusion, implants loaded with simvastatin by an electrochemical process improved implant osseointegration in osteoporotic rats. Furthermore, the increased concentration of simvastatin could affect the osseointegration, but the dose-effects also need further investigation.

Topography-dependent antibacterial, osteogenic and anti-aging properties of pure titanium.

After nearly half a century of development under the guidance of the osseointegration theory, the major dilemmas for current implant dentistry are the implant associated infection and insufficient osseointegration. Moreover, biological aging of titanium (Ti) implants also brings great uncertainty to clinical results. In the present study, a novel nano-micro-hierarchical topography pattern is created by sandblasting and dual acid-etching on a Ti surface. The physico-chemical properties of the surfaces were characterized by scanning electron microscopy, contact angle measurement, X-ray photoelectron spectroscopy and X-ray diffraction. The effects of the hierarchical surfaces on osteoprogenitor cell growth and bacterial activities were separately evaluated. The optimized nano-micro-hierarchical Ti surface exhibits surprisingly topography-dependent antibacterial capacity via inhibiting bacterial adhesion of several species in the early stage and better osteogenesis ability than the microscaled surface. Aging studies demonstrate that, compared with the surface with a microscale structure, the nano-micro-hierarchical Ti surface has greater anti-aging ability manifested as being more capable to retain hydrophilicity and bioactivity during aging. Furthermore, the present study reveals that the biological aging of the Ti implant is attributed to two decisive factors during the aging period: the progressively thickened amorphous TiO2 layer by autoxidation and the unavoidable accumulation of hydrocarbons on the Ti implant surface.

The construction of hierarchical structure on Ti substrate with superior osteogenic activity and intrinsic antibacterial capability.

The deficient osseointegration and implant-associated infections are pivotal issues for the long-term clinical success of endosteal Ti implants, while development of functional surfaces that can simultaneously overcome these problems remains highly challenging. This study aimed to fabricate sophisticated Ti implant surface with both osteogenic inducing activity and inherent antibacterial ability simply via tailoring surface topographical features. Micro/submciro/nano-scale structure was constructed on Ti by three cumulative subtractive methods, including sequentially conducted sandblasting as well as primary and secondary acid etching treatment. Topographical features of this hierarchical structure can be well tuned by the time of the secondary acid treatment. Ti substrate with mere micro/submicro-scale structure (MS0-Ti) served as a control to examine the influence of hierarchical structures on surface properties and biological activities. Surface analysis indicated that all hierarchically structured surfaces possessed exactly the same surface chemistry as that of MS0-Ti, and all of them showed super-amphiphilicity, high surface free energy, and high protein adsorption capability. Biological evaluations revealed surprisingly antibacterial ability and excellent osteogenic activity for samples with optimized hierarchical structure (MS30-Ti) when compared with MS0-Ti. Consequently, for the first time, a hierarchically structured Ti surface with topography-induced inherent antibacterial capability and excellent osteogenic activity was constructed.

Multifocal Langerhans cell histiocytosis in an adult with a pathological fracture of the mandible and spontaneous malunion: A case report.

Langerhans cell histiocytosis (LCH) is rare in the adult population and even rarer with jaw involvement. The current study presents the case of a 39-year-old male who complained of recurrent pain, swelling of the gingiva and an occasional pus-like discharge in the right mandible for one year. The patient was previously prescribed antibiotics, but this did not resolve the problem. An initial panoramic radiograph showed an osteolytic lesion and bone fracture in the right mandible. Eight months later, a new radiograph showed the spontaneous malunion of the fractured mandible. The patient was eventually diagnosed with Langerhans cell histiocytosis by histopathology and immunohistochemistry. Further lesions were found in the ribs and ilium by nuclear bone scanning. The patient was subsequently treated with systemic chemotherapy, and the lesions are currently effectively being controlled. This study is the first to show that spontaneous intralesional bone regeneration may lead to reunification of the mandible fracture caused by LCH in an adult.

Enucleation combined with peripheral ostectomy: its role in the management of large cystic ameloblastomas of the mandible.

AIM: Aggressive resection of a large mandibular cystic ameloblastoma may cause severe deformity and dysfunction, while simple enucleation may carry a high risk of recurrence. The purpose of this study was to evaluate, in terms of both recurrence and the preservation of contour and function, the effectiveness of enucleation combined with peripheral ostectomy (Enu/PO) in managing large mandibular cystic ameloblastomas. MATERIALS AND METHODS: Fourteen patients who had large mandibular cystic ameloblastoma (11 unicystic, 3 multicystic) and had been treated with the Enu/PO were reviewed. RESULTS: The follow-up period was 19-117 months. Using Enu/PO, mandibular continuity was preserved in all patients, and only one patient complained of permanent lip numbness. With bone regeneration, the thinned bone plate significantly thickened and the residual cavity shrank. No pathological fracture occurred. There was recurrence in 3 patients. Two recurrences were treated with radical surgery, and the other with Enu/PO. No new recurrence was found thereafter. All patients were satisfied with their facial appearance. CONCLUSIONS: The Enu/PO technique is excellent in preserving appearance and functions and with a low risk of recurrence, is a reliable option for the management of large mandibular cystic ameloblastomas.

In vitro and in vivo evaluation of the osteogenic ability of implant surfaces with a local delivery of simvastatin.

PURPOSE: This study established local delivery with a calcium phosphate (CaP) coating and investigated effects of delivery on implant osseointegration by in vitro and in vivo experiments. MATERIAL AND METHODS: Simvastatin was prepared onto titanium surfaces with varying concentration (10⁻7, 10⁻6, 10⁻5, and 10⁻4 mol/L). Surface characteristics were performed by field-emission scanning electron microscope (FSEM), x-ray diffractometer (XRD), and fourier transform infrared spectroscopy (FTIR). Alkaline phosphatase activity (ALP) and osteocalcin release were used to measure osteoblastic activities. Ovariectomized rats randomly received control and test implants in both tibiae. After 4 and 12 weeks of implantation, the tibiae were retrieved and prepared for histomorphometric evaluation. RESULTS: FSEM observation showed that the size of flakes decreased with an increase of simvastatin concentrations. XRD and FTIR examinations demonstrated that all coatings were composed of octacalcium phosphate (OCP). Simvastatin-loaded titanium surface had an increased effect on ALP activities at different concentrations on day 4 and day 7, and only the 10⁻6 mol/L group showed significant differences on day 14 (P<.05). The 10⁻6 mol/L group showed significant expression of osteocalcin (P<.05). Test implants (10⁻6 mol/L) showed a significantly greater bone area and bone-implant contact compared to control implants during the observation periods (P<.05). CONCLUSIONS: It was concluded that the local delivery of simvastatin was established onto implant surfaces using the biomimetic CaP coating and could improve osteoblast function and implant osseointegration in ovariectomized rats.