Long Noncoding RNA X-Inactive Specific Transcript (XIST) Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells by Sponging MicroRNA-214-3p.

BACKGROUND Osteogenic differentiation of periodontal ligament stem cells (PDLSCs) is associated with periodontitis. It has been reported that long noncoding RNA X-inactive specific transcript (lncRNA XIST) is upregulated and microRNA-214-3p (miR-214-3p) is downregulated in PDLSCs after osteogenic induction. However, whether XIST is involved in osteogenic differentiation of PDLSCs via miR-214-3p has not been reported. MATERIAL AND METHODS The protein expressions of osteogenic markers alkaline phosphatase (ALP), osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2) were examined by Western blot. The levels of miR-214-3p and XIST were determined by qRT-PCR. The relationship between miR-214-3p and XIST was evaluated by luciferase reporter, RNA immunoprecipitation, and RNA pulldown assays. RESULTS We found that XIST was increased and miR-214-3p was decreased in PDLSCs after osteogenic stimulation. Silencing of XIST decreased the protein expressions of ALP, OCN, and RUNX2, and also decreased ALP activity. Higher miR-214-3p levels also inhibited osteogenic differentiation of PDLSCs. XIST interacted with miR-214-3p and depletion of miR-214-3p mitigated XIST absence-mediated suppression of osteogenic differentiation. CONCLUSIONS XIST participates in osteogenic differentiation of PDLSCs by sponging miR-214-3p.

Intrinsic Surface Effects of Tantalum and Titanium on Integrin α5ß1/ ERK1/2 Pathway-Mediated Osteogenic Differentiation in Rat Bone Mesenchymal Stromal Cells.

BACKGROUND/AIMS: Accumulating evidence demonstrates the superior osteoinductivity of tantalum (Ta) to that of titanium (Ti); however, the mechanisms underlying these differences are unclear. Thus, the objective of the present study was to examine the effects of Ta and Ti surfaces on osteogenesis using rat bone mesenchymal stromal cells (rBMSCs) as a model. METHODS: Ta and Ti substrates were polished to a mirror finish to minimize the influences of structural factors, and the intrinsic surface effects of the two materials on the integrin α5ß1/mitogen-activated protein kinases 3 and 1 (ERK1/2) cascade-mediated osteogenesis of rBMSCs were evaluated. Alkaline phosphatase (ALP) activity, Alizarin Red staining, real-time polymerase chain reaction, and western blot assays of critical osteogenic markers were conducted to evaluate the effects of the two substrates on cell osteogenesis. Moreover, the role of the integrin α5ß1/ERK1/2 pathway on the osteoinductive performance of Ta and Ti was assessed by up- and down-regulation of integrin α5 and ß1 with RNA interference, as well as through ERK1/2 inhibition with U0126. RESULTS: Osteogenesis of rBMSCs seeded on the Ta surface was superior to that of cells seeded on the Ti surface in terms of ALP activity, extracellular matrix calcification, and the expression of integrin α5, integrin ß1, ERK1/2, Runt-related transcription factor 2, osteocalcin, collagen type I, and ALP at both the mRNA and protein levels. Moreover, down-regulation of integrin α5 or integrin ß1, or ERK1/2 inhibition severely impaired the osteoblastic differentiation on the Ta surface. By contrast, over-expression of integrin α5 or integrin ß1 improved osteogenesis on the Ti substrates, while subsequent ERK1/2 inhibition abrogated this effect. CONCLUSION: The integrin α5ß1/ERK1/2 pathway plays a crucial role in regulating rBMSCs osteogenic differentiation; thus, the greater ability of a Ta surface to trigger integrin α5ß1/ERK1/2 signaling may explain its better osteoinductivity. The different effects of Ta and Ti surfaces on rBMSC osteogenesis are considered to be related to the conductive behaviors between integrin α5ß1 and the oxides spontaneously formed on the two metals. These results should facilitate the development of engineering strategies with Ta and Ti surfaces for improved osteogenesis in endosteal implants.

A Novel Lipopeptide-Functionalized Metal-Organic Framework for Periodontitis Therapy through the Htra1/FAK/YAP Pathway.

Periodontitis is a chronic inflammatory disease characterized by plaque accumulation, resulting in immune microenvironment disorders and resorption of alveolar bone. To promote bone healing under inflammatory environments, a functional biomaterial based on disease pathophysiology is designed. A novel fatty acid C10-modified polypeptide, C10-KR8, is discovered to have excellent abilities in modulating macrophage repolarization and promoting bone regeneration in periodontitis. To build a multifunctional material localized drug delivery system, C10-KR8@ZIF-8 (C10-KR8-loaded zeolitic imidazolate framework-8) nanoparticles are constructed to sustainedly release the C10-KR8 peptide and Zn elements. By synergistic effects of providing a dynamic immuno-modulatory environment and promoting osteogenesis under pathological conditions, the obtained pH-responsive nanoparticles display excellent bone regeneration capability. Furthermore, coimmunoprecipitation/liquid chromatography-tandem mass spectrometry analysis and proteomics analysis revealed that the C10-KR8 peptide directly interacts with the high-temperature requirement protein A1 (Htra1), and C10-KR8@ZIF-8 nanoparticles promote the osteogenic differentiation of bone mesenchymal stem cells by activating the focal adhesion kinase (FAK)/phosphatidylinositide 3-kinase (PI3K)/AKT pathway and enhancing the nuclear localization of Yes-associated protein (YAP). Taken together, this study demonstrates C10-KR8 peptide regulate osteoimmunology and bone regeneration by Htra1/FAK/YAP pathway and that ZIF-8-based peptide loading platform is a promising strategy for periodontitis.

Multifunctional nanoparticles for enhanced sonodynamic-chemodynamic immunotherapy with glutathione depletion.

Aim: This study aimed to develop a sonodynamic-chemodynamic nanoparticle functioning on glutathione depletion in tumor immunotherapy. Materials & methods: The liposome-encapsulated 2,2-azobis[2-(2-imidazolin-2-yl) propane] dihydrochloride (AIPH) and copper-cysteine nanoparticles, AIPH/Cu-Cys@Lipo, were synthesized with a one-pot method. 4T1 cells were injected into female BALB/c mice for modeling. Results: AIPH/Cu-Cys@Lipo was well synthesized. It generated alkyl radicals upon ultrasound stimulation. AIPH/Cu-Cys@Lipo promoted the generation of -OH via a Fenton-like reaction. Both in vitro and in vivo experiments verified that AIPH/Cu-Cys@Lipo significantly inhibited tumor development by decreasing mitochondrial membrane potential, activating CD4+ and CD8+ T cells and promoting the expression of IL-2 and TNF-α. Conclusion: AIPH/Cu-Cys@Lipo provides high-quality strategies for safe and effective tumor immunotherapy.

The Inhibition of MicroRNA-139-5p Promoted Osteoporosis of Bone Marrow-Derived Mesenchymal Stem Cells by Targeting Wnt/Beta-Catenin Signaling Pathway by NOTCH1.

We investigated the therapeutic effects of microRNA-139-5p in relation to osteoporosis of bone marrow-derived mesenchymal stem cell (BMSCs) and its underlying mechanisms. In this study we used a dexamethasone-induced in vivo model of osteoporosis and BMSCs were used for the in vitro model. Real-time quantitative polymerase chain reaction (RT-PCR) and gene chip were used to analyze the expression of microRNA-139-5p. In an osteoporosis rat model, the expression of microRNA-139-5p was increased, compared with normal group. Downregulation of microRNA-139-5p promotes cell proliferation and osteogenic differentiation in BMSCs. Especially, up-regulation of microRNA-139-5p reduced cell proliferation and osteogenic differentiation in BMSCs. Overexpression of miR-139-5p induced Wnt/ß-catenin and down-regulated NOTCH1 signaling in BMSCs. Down-regulation of miR-139-5p suppressed Wnt/ß-catenin and induced NOTCH1 signaling in BMSCs. The inhibition of NOTCH1 reduced the effects of anti-miR-139-5p on cell proliferation and osteogenic differentiation in BMSCs. Activation of Wnt/ß-catenin also inhibited the effects of anti-miR-139-5p on cell proliferation and osteogenic differentiation in BMSCs. Taken together, our results suggested that the inhibition of microRNA-139-5p promotes osteogenic differentiation of BMSCs via targeting Wnt/ß-catenin signaling pathway by NOTCH1.

Enhanced antibacterial properties of biocompatible titanium via electrochemically deposited Ag/TiO2 nanotubes and chitosan-gelatin-Ag-ZnO complex coating.

A novel double-layered antibacterial coating was fabricated on pure titanium (Ti) via a simple three-step electrodeposition process. Scanning electronic microscopy (SEM) images show that the coating was constructed with the inner layer of TiO2 nanotubes doped with silver nanoparticles (TNTs/Ag) and the outer layer of chitosan-gelatin mixture with zinc oxide and silver nanoparticles (CS-Gel-Ag-ZnO). In comparison, we also investigated the composition, structure and antibacterial properties of pure Ti coated with TNTs, TNTs/Ag or TNTs/Ag + CS-Gel-Ag-ZnO, respectively. The TNTs was about 100 nm wide and 240 nm to 370 nm tall, and most Ag nanoparticles (Ag NPs) with diameter smaller than 20 nm were successfully deposited inside the tubes. The CS-Gel-Ag-ZnO layer was continuous and uniform. Antibacterial activity against planktonic and adherent bacteria were both investigated. Agar diffusion test against Staphylococcus aureus (S. aureus) shows improved antibacterial capacity of the TNTs/Ag + CS-Gel-Ag-ZnO coating, with a clear zone of inhibition (ZOI) up to 14.5 mm wide. Dead adherent bacteria were found on the surface by SEM. The antibacterial rate against planktonic S. aureus was as high as 99.2% over the 24 h incubation period.

Postischemic Housing Environment on Cerebral Metabolism and Neuron Apoptosis after Focal Cerebral Ischemia in Rats.

The purpose of this study was to evaluate the roles of different housing environments in neurological function, cerebral metabolism, cerebral infarction and neuron apoptosis after focal cerebral ischemia. Twenty-eight Sprague-Dawley rats were divided into control group (CG) and cerebral ischemia group, and the latter was further divided into subgroups of different housing conditions: standard environment (SE) subgroup, individual living environment (IE) subgroup, and enriched environment (EE) subgroup. Focal cerebral ischemia was induced by the middle cerebral artery occlusion (MCAO). Beam walking test was used to quantify the changes of overall motor function. Cerebral infarction and cerebral metabolism were studied by in vivo magnetic resonance imaging and 1H-magnetic resonance spectra, respectively. Neuron necrosis and apoptosis were detected by hematoxylin-eosin and TUNEL staining methods, respectively. The results showed that performance on the beam-walk test was improved in EE subgroup when compared to SE subgroup and IE subgroup. Cerebral infarct volume in IE subgroup was significantly larger than that in SE subgroup (P<0.05) and EE subgroup (P<0.05) on day 14 after MCAO. NAA/Cr and Cho/Cr ratios were lower in MCAO groups under different housing conditions as compared to those in CG (P<0.05). NAA/Cr ratio was lower in IE subgroup (P<0.05) and higher in EE subgroup (P<0.05) than that in SE subgroup. NAA/ Cr ratio in EE was significantly higher than that in IE subgroup (P<0.05). Cho/Cr ratio was decreased in MCAO groups as compared to that in CG (P<0.05). A significant decrease in normal neurons in cerebral cortex was observed in MCAO groups as compared to CG (P<0.05). The amount of normal neurons was less in IE subgroup (P<0.05), and more in EE subgroup (P<0.05) than that in SE subgroup after MCAO. The amount of normal neurons in EE subgroup was significantly more than that in IE subgroup after MCAO (P<0.05). The ratio of TUNEL-positive neurons in EE was significantly lower than that in SE subgroup (P<0.05) and IE subgroup (P<0.05). Correlation analysis showed that the beam walking test was negatively correlated with NAA/Cr ratio (P<0.05). Cerebral infarct volume was negatively correlated with both NAA/Cr ratio (P<0.01) and Cho/Cr ratio (P<0.01). The amount of normal cortical neurons was positively correlated with both NAA/Cr ratio (P<0.01) and Cho/Cr ratio (P<0.05). The TUNEL-positive neurons showed a negative correlation with both NAA/Cr ratio (P<0.01) and Cho/Cr ratio (P<0.01). This study goes further to show that EE may improve neurological functional deficit and cerebral metabolism, decrease cerebral infarct volume, neuron necrosis and apoptosis, while IE may aggravate brain damage after MCAO.

[Research on periodontal changes on the compression side during rapid tooth movement into newly distracted bone].

PURPOSE: To investigate periodontal remodeling mechanism on the compression side during early tooth movement into newly distracted bone. METHODS: Ten male Beagle dogs were selected. Distraction osteogenesis was performed on randomized side as experimental group, while the fourth premolars were extracted on the other side as control group. Then the third premolars were distalized with 30g orthodontic force instantly after the cessation of distraction or extraction. The distance of the tooth movement was measured with a sliding caliper every week. Each distance was measured 3 times and the mean value was recorded. The measurement data were analysed with paired t test by SPSS 18.0 software package. Beagle dogs were killed in the first, second, fourth week after tooth movement. Slices were obtained for HE staining and TRAP staining to observe the periodontal tissue on the compression side. RESULTS: The average moving velocity of the teeth in the distracted bone was (1.055±0.054)mm per week, which was significantly faster than that in the control group(P<0.05).There was no apparent lag period in the experimental group. In addition, there was no hyalinization observed on the compression side of the periodontal tissue in the experimental group, while the amount and area of distribution of the TRAP-positive cells on the compression side was significantly larger and strongly expressed. CONCLUSIONS: The moving velocity of the teeth in the newly distracted bone was significantly faster, and no apparent lag period, which may be related to no hyalinization and more early-appeared, vigorous and wide-distributed osteoclasts on the compression side of the periodontal tissue.