Black phosphorus-incorporated novel Ti-12Mo-10Zr implant for multimodal treatment of osteosarcoma.

The repair and reconstruction of large bone defects after bone tumor resection is still a great clinical challenge. At present, orthopedic implant reconstruction is the mainstream treatment for repairing bone defects. However, according to clinical feedback, local tumor recurrence and nonunion of bone graft are common reasons leading to the failure of bone defect repair and reconstruction after bone tumor resection, which seriously threaten the physical and mental health of patients. On this basis, here the self-developed low modulus Ti-12Mo-10Zr alloy (TMZ) was chosen as substrate material. To improve its biological activity and osteointegration, calcium, oxygen, and phosphorus co-doped microporous coating was prepared on TMZ alloy by microarc oxidation (MAO). Then, black phosphorus (BP) nanosheets were incorporated onto MAO treated TMZ alloy to obtain multifunctional composites. The obtained BP-MAO-TMZ implant exhibited excellent photothermal effects and effective ablation of osteosarcoma cancer cells under the irradiation of 808 nm near infrared laser, while no photothermal or therapeutic effects were observed for TMZ alloy. Meanwhile, the structure/component bionic coating obtained after MAO treatment as well as the P-driven in situ biomineralization performance after incorporation of BP nanosheets endowed BP-MAO-TMZ implant with synergistic promoting effect on MC3T3-E1 osteoblasts' activity, proliferation and differentiation ability. This study is expected to provide effective clinical solutions for problems of difficult bone regeneration and tumor recurrence after tumor resection in patients with bone tumors and to solve a series of medical problems such as poor prognosis and poor postoperative quality of patients life with malignant bone tumors.

Surface modification of new innocuous Ti-Mo-Zr based alloys for biomedical applications.

To address the clinical challenges of modulus mismatch, lack of initial osteointegration and contain toxic elements towards traditional titanium and its alloys with surrounding bone tissue, a new ß-type titanium alloy (Ti-12Mo-10Zr) designed by our group will be chosen as dental implant in this proposal due to its excellent properties, e.g. low young's modulus (~ 50.8 GPa) and excellent compressive yield strength (~ 430.89 MPa). A modified hydrothermal and pressure method will be deployed to create tailored micro/nano topography and chemistry (phosphorus) on implant surface with the aim of promoting osteointegration. The formation process and mechanism of micro/nano-scaled hierarchical hybrid coating containing phosphorous will be revealed from the perspective of energetics and crystallography to realize co-design of multiple structure and chemical on Ti-12Mo-10Zr surface. The in vitro cytological performance of this hierarchical hybrid coating containing phosphorous will also be evaluated by co-culturing with rat bone marrow stromal cells This proposal will not only provide guidance and experimental database for next generation potential implant named Ti-12Mo-10Zr, but also display new insights to improve long-lasting stability for dental implant which demonstrate tremendous scientific significance.

3D Printed Multifunctional Ti6Al4V-Based Hybrid Scaffold for the Management of Osteosarcoma.

Osteosarcoma is a challenging bone disease which is commonly associated with critically sized bone defects and cancer recurrence. Here, we designed and developed a multifunctional, hierarchical structured bone scaffold which can meet the demanding requirements for osteosarcoma management. The 3D printed Ti6Al4V scaffold with hydrothermally induced TiO2/TiP coating can offer a unique photothermal conversion property for in vitro bone cancer ablation. The scaffold is also infused with drug-laden gelatin/hydroxyapatite nanocomposite, which provides the ideal porous structure for cell adhesion/bone ingrowth and promotes bone regeneration. The scaffold has been thoroughly characterized by SEM/EDX, TEM, XPS, XRD, TGA, and UV-vis, and its in vitro bone cancer ablation efficiency has been validated using MG-63 cells. The hybrid scaffold showed excellent biocompatibility, and its osteointegration function has been demonstrated using an animal model.

Bioinspired Fabrication of Calcium-Doped TiP Coating with Nanofibrous Microstructure to Accelerate Osseointegration.

Bioinspired by the morphology of osteoclast-resorbed bone surfaces, we prepared a calcium-doped titanium phosphate (Ca-TiP) coating, which consists of a nanofibrous network, on titanium (Ti) substrate via a simple two-step hydrothermal method, trying to mimic natural bone compositionally and microstructurally. The in vitro studies show that the Ca-TiP coating with synergistic features of nanofibrous biomimetic topography and surface chemistry could elicit intensively osteogenic behavior and responses including enhanced cell adhesion, spreading, and proliferation as well as alkaline phosphatase (ALP) activity and up-regulated expression of bone-related genes, which inevitably benefit the formation of new bone and the quality of osseointegration. When the two control groups are compared in vivo, the significantly improved new bone formation in the early stage and the much stronger interfacial bonding with the surrounding bone for Ca-TiP coating suggest that Ca-TiP coating modified Ti implants hold great potential for orthopedic and dental applications.

Ultrasonic Nakagami imaging for automatically positioning and identifying the treated lesion induced by histotripsy.

Histotripsy has been proposed as a non-invasive surgical procedure for clinical use that liquefies the tissue into acellular debris by utilizing the mechanical mechanism of bubbles. Accurate and reliable imaging guidance is essential for successful clinical histotripsy implementation. Nakagami imaging is a promising method to evaluate the microstructural change induced by high intensity focused ultrasound. However, practically, it is difficult for the Nakagami imaging to distinguish the treated lesion induced by histotripsy from the surrounding normal biological tissues. In this study, we introduce the use of noise-assisted correlation algorithm (NCA) in Nakagami images as a solution to suppress the background normal tissue and identify the treated lesion induced by histotripsy. Experiments are conducted on fresh porcine liver ex vivo by cavitation-cloud histotripsy. Results show that the contrast-to-noise ratio between the treated lesion and surrounding tissue corresponding to the Nakagami image after NCA and original Nakagami image is 3.434 and 0.505, respectively. The optimal artificial noise level is 1-fold of the background normal tissue amplitude, and the corresponding optimal threshold of correlation coefficient should be between 0.6 and 0.8 in the application of NCA. Therefore, the use of NCA in Nakagami image can suppress the background normal tissues without affecting the information of treated lesion for an appropriate artificial noise level and threshold used in the NCA. Moreover, the Nakagami images after the application of the NCA can also be used for automatically distinguishing and measuring the tissue fractionation accurately using binarization. The proposed Nakagami images overlaid on the B-mode images can provide a promising method for positioning and visualizing the treated lesion to achieve precise histotripsy treatment.

Comparative Study on 3D Printed Ti6Al4V Scaffolds with Surface Modifications Using Hydrothermal Treatment and Microarc Oxidation to Enhance Osteogenic Activity.

Titanium (Ti) and its alloys have been widely used in clinics as preferred materials for bone tissue repair and replacement. However, the lack of biological activity of Ti limits its clinical applications. Surface modification of Ti with bioactive elements has always been a research hotspot. In this study, to promote the osseointegration of Ti6Al4V (Ti64) implants, calcium (Ca), oxygen (O), and phosphorus (P) codoped multifunctional micro-nanohybrid coatings were prepared on a three-dimensional (3D) printed porous Ti64 surface by microarc oxidation (MAO) and a hydrothermal method (HT). The surface morphologies, chemical compositions, and surface/cell interactions of the obtained coatings were studied. In vitro experiments indicated that all hybrid coating-modified Ti64 implants could enhance protein adsorption and MC3T3 osteoblasts' activity, adhesion, and differentiation ability. In vivo experiments showed that the hybrid coating promoted early osseointegration. By comparison, microarc oxidation-treated Ti64 (M-Ti) has the best biological activity and the strongest ability of osseointegration. It provides important theoretical significance and potential application prospects for improving the biological activity of Ti implants.

The custom making of hierarchical micro/nanoscaled titanium phosphate coatings and their formation mechanism analysis.

In this study, a series of hierarchical micro/nanoscaled titanium phosphate (TiP) coatings possessing various surface morphologies were successfully fabricated on titanium (Ti) discs. The hydrothermal reactions of Ti discs in hydrogen peroxide (H2O2) and phosphoric acid (H3PO4) mixed solution yield diverse topographies such as hemispheric clump, cylindrical rod, spherical walnut, micro/nano grass, micro/nano sheet, and fibrous network. And their crystal structures were mainly composed of Ti(HPO4)2·0.5H2O, (TiO)2P2O7, H2TiP2O8, Ti(HPO4)2 and TiO2. The morphology and crystal shape of the TiP coatings depend strongly on the mass ratio of H2O2/H3PO4, reaction temperature and water content. Besides, the formation mechanism of TiP coatings with diverse morphologies was explored from the perspective of energetics and crystallography. The mechanism exploration paved the way for custom-making TiP coatings with desirable micro/nanoscaled morphologies to meet specific application purposes. The in vitro cytological performances of TiP coatings were also evaluated by co-culturing with rat bone marrow stromal cells (BMSCs), demonstrating a positive prospect for their use in bone tissue engineering.

RGD/CTX-conjugated multifunctional Eu-Gd2O3 NRs for targeting detection and inhibition of early tumor.

Glioblastoma is currently the most common and lethal brain tumor, so accurate detection and effective therapy at the early glioblastoma stage is crucial. Herein, multifunctional Eu-Gd2O3 nanorods (NRs) with good paramagnetic and luminescence properties were fabricated through a hydrothermal method and a subsequent calcination technique, and exhibited good T1-weighted magnetic resonance (MR) imaging (r1 = 5.13 Gd mM-1 s-1) and cell-luminescence imaging properties. Then, Eu-Gd2O3 NRs were coated with difunctionalized polyethylene glycol (Mal-PEG-NHS), and subsequently conjugated through thiolation with arginine-glycine-aspartic (RGD) and chlorotoxin (CTX), respectively. The results of the cell test indicated that RGD/CTX-conjugated Eu-Gd2O3 NRs (RGD-NRs-CTX) could specifically target and adhere on U251 cells, leading to cellular apoptosis. The in vivo investigation revealed that RGD-NRs-CTX possessed no tissue/organ toxicity and a long blood circulation time. The results of in vivo MR imaging showed a significant preferential targeting and accumulation of RGD-NRs-CTX onto the early tumor, and in vivo luminescence imaging displayed a good infiltration capacity in tumor regions. Based on a superimposed targeting property of CTX and RGD, the results of everyday tail-vein injection suggested that RGD-NRs-CTX could effectively inhibit early tumor growth, without any damage to normal tissues/organs. Therefore, these results demonstrate that RGD-NRs-CTX are promising candidates for simultaneous targeting detection and therapy for early tumor.

Fabrication of RGD-conjugated Gd(OH)3:Eu nanorods with enhancement of magnetic resonance, luminescence imaging and in vivo tumor targeting.

The development of multimodal probes with magnetic resonance imaging (MRI) and intraoperative fluorescence imaging is the most challenging task in the field of tumor diagnosis. Herein, a simple one-pot hydrothermal method is used to prepare Eu-doped Gd(OH)3 nanorods (Gd(OH)3:Eu NRs) with good fluorescence and the longitudinal relaxivity r1 value of 4.78 (Gd mM s-1). After dual-functionalized maleimide-polyethylene glycol-succinimide (Mal-PEG-NHS) macromolecules are coated on the surface of Gd(OH)3:Eu NRs (PEG-NRs), the results of a lower degradation ratio in newborn calf serum (NCS), reactive oxygen species (ROS) generation in L929 cells and the hemolytic rate of PEG-NRs show their good cyto-compatibility and longer blood circulation time. Moreover, the actively tumor-targeting properties are endowed to NRs through the conjugation of cyclic arginine-glycine-aspartic acid (cRGD) (denoted RGD-NRs). The bio-distributions of RGD-NRs in tumor-bearing nude mice via tail-vein injection indicate that RGD-NRs are specifically taken-up by gliomas. The tests of in vivo T1-weighted MR imaging via tail-vein injection confirm that RGD-NRs possess a higher positive signal-enhancement ability in gliomas. Besides, the better luminescence imaging of living cells under a fluorescence microscope and the clear in vivo fluorescence imaging further confirm the targeting properties and better in vivo optical imaging behavior of RGD-NRs.