Surface modification of titanium with antibacterial porous N-halamine coating to prevent peri-implant infection.

Peri-implant infection remains one of the greatest threats to orthopedics. The construction of bone implants with good antibacterial and osteogenic properties is beneficial for reducing the risk of implant-related infections and healing bone defects. In this study, N-halamine coating (namely N-Cl) was grafted onto alkali-heat treated titanium (Ti) using polydopamine to endow Ti-based orthopedic implants with strong bactericidal activity. Surface characterization revealed that the N-Cl coating has porous structure loaded with active chlorine (Cl+). The N-Cl coating also provided micro/nano-structured Ti surfaces with excellent antibacterial ability via transformation between N-H and N-Cl, and approximately 100% disinfection was achieved. Furthermore, the as-prepared N-Cl coating exhibited good biocompatibility and osteogenesis abilityin vitro. These results indicate that applying N-Cl coatings on Ti could prevent and treat peri-implant infections.

A Novel Nomogram for Predicting Morbidity Risk in Patients with Secondary Malignant Neoplasm of Bone and Bone Marrow: An Analysis Based on the Large MIMIC-III Clinical Database.

Objective: Bone and bone marrow are the third most frequent sites of metastases from many cancers and are associated with low survival and high morbidity rates. Currently, there are no effective bedside tools to predict the morbidity risk of these patients in general intensive care units (ICUs). The main objective of this study was to establish and validate a nomogram to predict the morbidity risk of patients with bone and bone marrow metastases. Methods: Data on patients with bone and bone marrow metastases were extracted from the Medical Information Mart for Intensive Care III (MIMIC-III) database. The patients were divided into training and validation cohorts. The data were analyzed using univariate and multivariate Cox regression methods. Factors significantly and independently prognostic of survival were used to construct a nomogram predicting 30-day morbidity. The nomogram was validated by various methods, including Harrell's concordance index (C-index), area under the receiver operating characteristic curve (AUC), calibration curve, integrated discrimination improvement (IDI), net reclassification index (NRI), and decision curve analysis (DCA). Results: The study included 610 patients in the training cohort and 262 in the validation cohort. Multivariate Cox regression analysis showed that temperature, SpO2, Sequential Organ Failure Assessment (SOFA) score, Oxford Acute Severity of Illness Score (OASIS), comorbidities with coagulopathy, white blood cell count, heart rate, and respiratory rate were independent predictors of patient survival. The resulting nomogram had good discriminative ability, as shown by high AUCs, and was well calibrated, as demonstrated by calibration curves. Improvements in NRI and IDI values suggested that the nomogram was superior to the SOFA scoring system. DCA curves revealed that the nomogram showed good value in clinical applications. Conclusion: This prognostic nomogram, based on demographic and laboratory parameters, was predictive of the 30-day morbidity rate in patients with secondary malignant neoplasms of the bone and bone marrow, suggesting its applicability in clinical practice.

[Effect of different functional groups on self-assembled monolayers on the biological characteristics of skeletal muscle cells in vitro].

OBJECTIVE: To explore the effect of different functional groups on self-assembled monolayers on the biological characteristics of rabbit skeletal muscle cells in vitro. METHODS: Rabbit skeletal muscle cells were cultured on self-assembled monolayers of gold on which different terminal chemical groups including methyl groups (-CH(3)), amino(-NH(2)), hydroxyl(-OH) and carboxyl (-COOH ) were anchored with self-assembled methods. Contact angle measurements and atomic force microscopy were employed to confirm the similar density of different functional groups occupation. Fluorescence microscopy, MTT assay, flow cytometry, and scanning electron microscopy (SEM) were used to analyze the morphological and biological alterations of the cells. RESULTS: SEM results revealed that the chemical groups on the surface of the monolayer modulated the structure of skeletal muscle cells and the cell morphology. Skeletal muscle cells cultured on the monolayer with -CH3 exhibited the smallest contact area with a spherical morphology, while the cells on the monolayers with -NH(2), -OH and -COOH showed much larger contact area and flatter morphology. The functional groups -NH(2) and -COOH obviously promoted cell adhesion and proliferation, while -CH(3) group produced significantly greater toxicity than -NH(2), -OH and -COOH groups to inhibit the cell growth and adhesion and promote cell death. Cell attachment and growth was enhanced, in the order the magnitude of the effect, by -NH(2)>-COOH>-OH>-CH(3), and the toxicity decreased in the order of -NH(2)>-COOH>-OH>-CH(3). CONCLUSION: The terminal chemical groups can obviously affect the phenotype of skeletal muscle cells in vitro, and this finding provides a theoretical basis for surface design of biomaterials.

Controlled oxidative nanopatterning of microrough titanium surfaces for improving osteogenic activity.

To further enhance the biological properties of acid-etched microrough titanium surfaces, titania nanotextured thin films were produced by simple chemical oxidation, without significantly altering the existing topographical and roughness features. The nanotextured layers on titanium surfaces can be controllably varied by tuning the oxidation duration time. The oxidation treatment significantly reduced water contact angles and increased the surface energy compared to the surfaces prior to oxidation. The murine bone marrow stromal cells (BMSCs) were used to evaluate the bioactivity. In comparison, oxidative nanopatterning of microrough titanium surfaces led to improved attachment and proliferation of BMSCs. The rate of osteoblastic differentiation was also represented by the increased levels of alkaline phosphatase activity and mineral deposition. These data indicated that oxidative nanopatterning enhanced the biological properties of the microrough titanium surfaces by modulating their surface chemistry and nanotopography. Based on the proven mechanical interlocking ability of microtopographies, enhancement of multiple osteoblast functions attained by this oxidative nanopatterning is expected to lead to better implant osseointegration in vivo.

[Biocompatibility of silicon containing micro-arc oxidation coated magnesium alloy ZK60 with osteoblasts cultured in vitro].

OBJECTIVE: To research in vitro biocompatibility of silicon containing micro-arc oxidation (MAO) coated magnesium alloy ZK60 with osteoblasts. METHODS: The surface microstructure of silicon containing MAO coated magnesium alloy ZK60 was observed by a scanning electron microscopy (SEM), and chemical composition of the coating surface was determined by energy dispersive spectrum analysis. The experiments were divided into 4 groups: silicon containing MAO coated magnesium alloy ZK60 group (group A), uncoated magnesium alloy ZK60 group (group B), titanium alloy group (group C), and negative control group (group D). Extracts were prepared respectively with the surface area to extraction medium ratio (1.25 cm(2)/mL) according to ISO 10993-12 standard in groups A, B, and C, and were used to culture osteoblasts MC3T3-E1. The a-MEM medium supplemented with 10% fetal bovine serum was used as negative control in group D. The cell morphology was observed by inverted phase contrast microscopy. MTT assay was used to determine the cell viability. The activity of alkaline phosphatase (ALP) was detected. Cell attachment morphology on the surface of different samples was observed by SEM. The capability of protein adsorption of the coating surface was assayed, then DAPI and calcein-AM/ethidium homodimer 1 (calcein-AM/EthD-1) staining were carried out to observe cell adhesion and growth status. RESULTS: The surface characterization showed a rough and porous layer with major composition of Mg, O, and Si on the surface of silicon containing MAO coated magnesium alloy ZK60 by SEM. After cultured with the extract, cells grew well and presented good shape in all groups by inverted phase contrast microscopy, group A was even better than the other groups. At 5 days, MTT assay showed that group A presented a higher cell proliferation than the other groups (P < 0.05). Osteoblasts in groups A and C presented a better cell extension than group B under SEM, and group A exhibited better cell adhesion and affinity. Protein adsorption in group A [ (152.7 +/- 6.3) microg/mL] was significantly higher than that of group B [(96.3 +/-3.9) microg/mL] and group C [ (96.1 +/-8.7) microg/mL] (P < 0.05). At each time point, the adherent cells on the sample surface of group A were significantly more than those of groups B and C (P < 0.05). The calcein-AM/EthD-1 staining showed that groups A and C presented better cell adhesion and growth status than group B. The ALP activities in groups A and B were 15.55 +/-0.29 and 13.75 +/-0.44 respectively, which were significantly higher than those in group C (10.43 +/-0.79) and group D (10.73 +/-0.47) (P < 0.05), and group A was significantly higher than group B (P < 0.05). CONCLUSION: The silicon containing MAO coated magnesium alloy ZK60 has obvious promoting effects on the proliferation, adhesion, and differentiation of osteoblasts, showing a good biocompatibility, so it might be an ideal surface modification method on magnesium alloys.

MiR-133b is down-regulated in human osteosarcoma and inhibits osteosarcoma cells proliferation, migration and invasion, and promotes apoptosis.

MicroRNAs (miRNAs) decrease the expression of specific target oncogenes or tumor suppressor genes and thereby play crucial roles in tumorigenesis and tumor growth. To date, the potential miRNAs regulating osteosarcoma growth and progression are not fully identified yet. In this study, the miRNA microarray assay and hierarchical clustering analysis were performed in human osteosarcoma samples. In comparison with normal human skeletal muscle, 43 miRNAs were significantly differentially expressed in human osteosarcomas (fold change ≥2 and p≤0.05). Among these miRNAs, miR-133a and miR-133b expression was decreased by 135 folds and 47 folds respectively and the decreased expression was confirmed in both frozen and paraffin-embedded osteosarcoma samples. The miR-133b precursor expression vector was then transfected into osteosarcoma cell lines U2-OS and MG-63, and the stable transfectants were selected by puromycin. We found that stable over-expression of miR-133b in osteosarcoma cell lines U2-OS and MG-63 inhibited cell proliferation, invasion and migration, and induced apoptosis. Further, over-expression of miR-133b decreased the expression of predicted target genes BCL2L2, MCL-1, IGF1R and MET, as well as the expression of phospho-Akt and FAK. This study provides a new insight into miRNAs dysregulation in osteosarcoma, and indicates that miR-133b may play as a tumor suppressor gene in osteosarcoma.

[Prevention and control strategies of common post-operative complications of microwave ablation in situ in treatment of bone tumors].

OBJECTIVE: To analyze the common post-operative complications of microwave ablation in situ in the treatment of bone tumors and the prevention and control strategies. METHODS: Between March 2009 and July 2012, 73 cases of bone tumors were treated with microwave ablation in situ, of which 54 cases met the inclusion criteria. There were 31 males and 23 females with a median age of 27 years (range, 9-74 years), including 37 malignant tumors and 17 benign tumors. In 49 primary bone tumors, 17 cases were in stage 3, 13 in stage IIA, and 19 in stage IIB according to Enneking grading system. The postoperative complications, managements, and outcomes in patients were analyzed. RESULTS: After operation, 54 patients were followed up 12-40 months (mean, 24 months). Seven kinds of complications occurred in 21 patients (38.9%) with 3 cases suffered from more than one, including pathologic fracture in 4 cases (7.4%), deep infection in 2 cases (3.7%), nerve injury in 7 cases (13.0%), deep vein thrombosis of the lower extremity in 1 case (1.9%), medial collateral ligament heat injury of the knee in 1 case (1.9%), hematoma in 2 cases (3.7%), and fat liquefaction of incision and flap necrosis in 8 cases (14.8%). CONCLUSION: Pathologic fracture is the primary complication which results in a second surgery. Deep infection is the main complication that often leads to failure of the limb salvage. Nerve injury and poor wound healing are the most common complications. Good control of microwave temperature is the key to successful operation, and the related preventive strategies could reduce complications.