Expression patterns of mechanosensitive ion channel PIEZOs in irreversible pulpitis.

BACKGROUND: Mechanosensitive ion channel PIEZOs have been widely reported to involve inflammation and pain. This study aimed to clarify expression patterns of PIEZOs and their potential relations to irreversible pulpitis. MATERIALS AND METHODS: Normal pulp tissues (n = 29) from patients with impacted third molars and inflamed pulp tissues (n = 23) from patients with irreversible pulpitis were collected. Pain levels were assessed using a numerical rating scale. PIEZO expressions were measured using real-time PCR and then confirmed using GEO datasets GSE77459, immunoblot, and immunohistochemistry staining. Correlations of PIEZO mRNA expression with inflammatory markers, pain markers, or clinical pain levels were evaluated using Spearman's correlation analysis. Univariate analysis was conducted to analyze PIEZO expressions based on pain description and clinical examinations of cold test, percussion, palpation, and bite test. RESULTS: Compared with normal pulp tissues, mRNA expression levels of PIEZO1 were significantly increased in inflamed pulp tissues, while PIEZO2 was significantly decreased, which was further confirmed in GSE77459 and on a protein and histological level. The positive correlation of the mRNA expression levels between PIEZO1 and inflammatory markers, as well as between PIEZO2 and pain markers, was verified. PIEZO2 expression was also positively correlated with pain levels. Besides, irreversible pulpitis patients who reported continuous pain and who detected a positive response to cold stimulus exhibited a higher expression level of PIEZO2 in the inflamed pulp tissues. By contrast, patients reporting pain duration of more than one week showed a higher expression level of PIEZO1. CONCLUSIONS: This study demonstrated the upregulation of PIEZO1 and the downregulation of PIEZO2 in irreversible pulpitis and revealed the potential relation of PIEZO1 and PIEZO2 to inflammation and pain. These findings suggested that PIEZOs might play critical roles in the progression of irreversible pulpitis and paved the way for further investigations aimed at novel therapies of irreversible pulpitis by targeting PIEZOs.

Effects of immediate loading directionality on the mechanical sensing protein PIEZO1 expression and early-stage healing process of peri-implant bone.

BACKGROUND: The reduced treatment time of dental implants with immediate loading protocol is an appealing solution for dentists and patients. However, there remains a significant risk of early peri-implant bone response following the placement of immediately loaded implants, and limited information is available regarding loading directions and the associated in vivo characteristics of peri-implant bone during the early stages. This study aimed to investigate the effects of immediate loading directionality on the expression of mechanical sensing protein PIEZO1 and the healing process of peri-implant bone in the early stage. METHODS: Thirty-two implants were inserted into the goat iliac crest models with 10 N static lateral immediate loading applied, followed by histological, histomorphological, immunohistochemical, X-ray microscopy and energy dispersive X-ray spectroscopy evaluations conducted after 10 days. RESULTS: From evaluations at the cellular, tissue, and organ levels, it was observed that the expression of mechanical sensing protein PIEZO1 in peri-implant bone was significantly higher in the compressive side compared to the tensile side. This finding coincided with trends observed in interfacial bone extracellular matrix (ECM) contact percentage, bone mass, and new bone formation. CONCLUSIONS: This study provides a novel insight into the immediate loading directionality as a potential influence factor for dental implant treatments by demonstrating differential effects on the mechanical sensing protein PIEZO1 expression and related early-stage healing processes of peri-implant bone. Immediate loading directions serve as potential therapeutic influence factors for peri-implant bone during its early healing stage.

Role of αENaC in root resorption of adjacent teeth due to entirely impacted mandibular third molars.

BACKGROUND: Entirely impacted mandibular third molar (EIM3M) concerns the pathological external root resorption (ERR) of the adjacent mandibular second molar (M2M) and formation of granulation tissue between two molars. The study aimed to clarify the effect of αENaC, a mechano-sensitive molecule, to explore the mechanical mechanism in this scenario. METHODS: The force EIM3M exerted on M2M was proved by finite element analysis. αENaC expressions were tested by real-time polymerase chain reaction (PCR), immunoblotting and immunofluorescence. Inflammatory and epithelial-mesenchymal transition (EMT)-related molecules expressions were also detected by real-time PCR. The correlation was analyzed by Spearman's correlation analysis, and receiver-operator characteristic (ROC) curve was further exhibited. RESULTS: The force was concentrated in the ERR area. αENaC was upregulated, positively correlated with ERR degree and localized to the fibroblasts in ERR granulation tissues. Moreover, αENaC was respectively and positively associated with elevated TNF-α and N-cadherin in ERR granulation tissues. More importantly, ROC analysis verified αENaC as a novel indication of the incidence of this disease. CONCLUSIONS: Our finding revealed the force from EIM3M causing ERR of M2M, and elucidated the expression and localization of αENaC and its positive correlation with inflammation, EMT and disease severity, suggesting a novel indication in this disease.

The crucial regulatory role of type I interferon in inflammatory diseases.

Type I interferon (IFN-I) plays crucial roles in the regulation of inflammation and it is associated with various inflammatory diseases including systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and periodontitis, impacting people's health and quality of life. It is well-established that IFN-Is affect immune responses and inflammatory factors by regulating some signaling. However, currently, there is no comprehensive overview of the crucial regulatory role of IFN-I in distinctive pathways as well as associated inflammatory diseases. This review aims to provide a narrative of the involvement of IFN-I in different signaling pathways, mainly mediating the related key factors with specific targets in the pathways and signaling cascades to influence the progression of inflammatory diseases. As such, we suggested that IFN-Is induce inflammatory regulation through the stimulation of certain factors in signaling pathways, which displays possible efficient treatment methods and provides a reference for the precise control of inflammatory diseases.

Proteomic and single-cell analysis shed new light on the anti-inflammatory role of interferonß in chronic periodontitis.

Periodontitis, a condition that results in periodontal attachment loss and alveolar bone resorption, contributes to the global burden of oral disease. The underlying mechanism of periodontitis involves the dysbiosis and dyshomeostasis between host and oral microbes, among which the macrophage is one of the major innate immune cell players, producing interferon ß (IFNß) in response to bacterial infection. The objective of this research was to examine the interaction of macrophages with periodontitis and the role and mechanism of IFNß on macrophages. IFNß has been shown to have the potential to induce the differentiation of M1 to M2 macrophages, which are stimulated by low levels of lipopolysaccharide (LPS). Additionally, IFNß has been demonstrated to promote the production of ISG15 by macrophages, which leads to the inhibition of the innate immune response. Moreover, our investigation revealed that IFNß has the potential to augment the secretion of ISG15 and its downstream cytokine, IL10, in LPS-stimulated macrophages. Single-cell analysis was conducted on the gingival tissues of patients with periodontitis, which revealed a higher proportion of macrophages in the periodontitis-diseased tissue and increased expression of IFNß, ISG15, and IL10. Gene Set Enrichment Analysis indicated that bacterial infection was associated with upregulation of IFNß, ISG15, and IL10. Notably, only IL10 has been linked to immunosuppression, indicating that the IFNß-ISG15-IL10 axis might promote an anti-inflammatory response in periodontitis through IL10 expression. It is also found that macrophage phenotype transitions in periodontitis involve the release of higher levels of IFNß, ISG15, and IL10 by the anti-inflammatory M2 macrophage phenotype compared to the pro-inflammatory M1 phenotype and myeloid-derived suppressor cells (MDSCs). This implies that the IFNß-induced production of IL10 might be linked to the M2 macrophage phenotype. Furthermore, cell communication analysis demonstrated that IL10 can promote fibroblast proliferation in periodontal tissues via STAT3 signaling.

Profiling of bile acids and activated receptor S1PR2 in gingival tissues of periodontitis patients.

BACKGROUND: Bile acids, as a group of cholesterol metabolites, play important roles in inflammation and bone metabolism. However, the possible link between bile acids and periodontitis is still unclear. This study aimed to clarify the alterations of the bile acid profile and corresponding receptor expression levels in periodontitis patients, and evaluate their association with periodontitis severity. METHODS: The concentrations of 15 bile acids in gingival tissues from 16 periodontitis patients and 16 healthy individuals were tested by metabolomics. Sphingosine-1-phosphate receptor 2 (S1PR2) expression was determined by real-time PCR and immunohistochemistry, which was also validated in two datasets, GSE16134 and GSE10334. The correlation between bile acids, S1PR2, and clinical parameters was analyzed by Spearman's correlation analysis, and receiver-operator characteristic (ROC) curves were examined to access the ability of bile acids and S1PR2 for defining local periodontitis status. RESULTS: In the periodontitis group, concentrations of total bile acids were elevated by increases of all bile acid forms, and five conjugated bile acids were significantly increased. Meanwhile, the expression of their receptor, S1PR2, was also upregulated in the periodontitis group. Positive correlations were further observed between glycocholic acid (GCA), taurochenodeoxycholic acid (TCDCA), taurocholic acid (TCA), S1PR2, and periodontal clinical parameters. ROC analysis also showed combinations of two bile acids (GCA and TCDCA) with S1PR2 as novel signatures for indicating local periodontitis status. CONCLUSION: Our findings demonstrated the alterations of the bile acid profile and receptor S1PR2 expression in periodontitis patients, and provided evidence of association between bile acids and periodontitis status.

Upregulated Vanins and their potential contribution to periodontitis.

BACKGROUND: Although Vanins are closely related to neutrophil regulation and response to oxidative stress, and play essential roles in inflammatory diseases with clinical significance, their contribution to periodontitis remains to be determined. This research was designed to assess the expression of Vanins in human gingiva, and to define the relationship between Vanins and periodontitis. METHODS: Forty-eight patients with periodontitis and forty-two periodontal healthy individuals were enrolled for gingival tissue sample collection. Expression levels of VNN1, VNN2 and VNN3 were evaluated by RT-qPCR and validated in datasets GSE10334 and GSE16134. Western blot and immunohistochemistry identified specific proteins within gingiva. The histopathological changes in gingival sections were investigated using HE staining. Correlations between Vanins and clinical parameters, PD and CAL; between Vanins and inflammation, IL1B; and between Vanins and MPO in periodontitis were investigated by Spearman's correlation analysis respectively. Associations between VNN2 and indicators of neutrophil adherence and migration were further validated in two datasets. RESULTS: Vanins were at higher concentrations in diseased gingival tissues in both RT-qPCR and dataset analysis (p < 0.01). Assessment using western blot and immunohistochemistry presented significant upregulations of VNN1 and VNN2 in periodontitis (p < 0.05). The higher expression levels of Vanins, the larger the observed periodontal parameters PD and CAL (p < 0.05), and IL1B (p < 0.001). Moreover, positive correlations existed between VNN2 and MPO, and between VNN2 and neutrophil-related indicators. CONCLUSION: Our study demonstrated upregulation of Vanins in periodontitis and the potential contribution of VNN2 to periodontitis through neutrophils-related pathological processes.

Mechanobiologically optimized Ti-35Nb-2Ta-3Zr improves load transduction and enhances bone remodeling in tilted dental implant therapy.

The tilted implant with immediate function is increasingly used in clinical dental therapy for edentulous and partially edentulous patients with excessive bone resorption and the anatomic limitations in the alveolar ridge. However, peri-implant cervical bone loss can be caused by the stress shielding effect. Herein, inspired by the concept of "materiobiology", the mechanical characteristics of materials were considered along with bone biology for tilted implant design. In this study, a novel Ti-35Nb-2Ta-3Zr alloy (TNTZ) implant with low elastic modulus, high strength and favorable biocompatibility was developed. Then the human alveolar bone environment was mimicked in goat and finite element (FE) models to investigate the mechanical property and the related peri-implant bone remodeling of TNTZ compared to commonly used Ti-6Al-4V (TC4) in tilted implantation under loading condition. Next, a layer-by-layer quantitative correlation of the FE and X-ray Microscopy (XRM) analysis suggested that the TNTZ implant present better mechanobiological characteristics including improved load transduction and increased bone area in the tilted implantation model compared to TC4 implant, especially in the upper 1/3 region of peri-implant bone that is "lower stress". Finally, combining the static and dynamic parameters of bone, it was further verified that TNTZ enhanced bone remodeling in "lower stress" upper 1/3 region. This study demonstrates that TNTZ is a mechanobiological optimized tilted implant material that enhances load transduction and bone remodeling.

Identification of potentially functional circRNAs and prediction of circRNA-miRNA-mRNA regulatory network in periodontitis: Bridging the gap between bioinformatics and clinical needs.

BACKGROUND AND OBJECTIVE: Periodontitis is a multifactorial chronic inflammatory disease that can lead to the irreversible destruction of dental support tissues. As an epigenetic factor, the expression of circRNA is tissue-dependent and disease-dependent. This study aimed to identify novel periodontitis-associated circRNAs and predict relevant circRNA-periodontitis regulatory network by using recently developed bioinformatic tools and integrating sequencing profiling with clinical information for getting a better and more thorough image of periodontitis pathogenesis, from gene to clinic. MATERIAL AND METHODS: High-throughput sequencing and RT-qPCR were conducted to identify differentially expressed circRNAs in gingival tissues from periodontitis patients. The relationship between upregulated circRNAs expression and probing depth (PD) was performed using Spearman's correlation analysis. Bioinformatic analyses including GO analysis, circRNA-disease association prediction, and circRNA-miRNA-mRNA network prediction were performed to clarify potential regulatory functions of identified circRNAs in periodontitis. A receiver-operating characteristic (ROC) curve was established to assess the diagnostic significance of identified circRNAs. RESULTS: High-throughput sequencing identified 70 differentially expressed circRNAs (68 upregulated and 2 downregulated circRNAs) in human periodontitis (fold change >2.0 and p < .05). The top five upregulated circRNAs were validated by RT-qPCR that had strong associations with multiple human diseases, including periodontitis. The upregulation of circRNAs were positively correlated with PD (R = .40-.69, p < .05, moderate). A circRNA-miRNA-mRNA network with the top five upregulated circRNAs, differentially expressed mRNAs, and overlapped predicted miRNAs indicated potential roles of circRNAs in immune response, cell apoptosis, migration, adhesion, and reaction to oxidative stress. The ROC curve showed that circRNAs had potential value in periodontitis diagnosis (AUC = 0.7321-0.8667, p < .05). CONCLUSION: CircRNA-disease associations were predicted by online bioinformatic tools. Positive correlation between upregulated circRNAs, circPTP4A2, chr22:23101560-23135351+, circARHGEF28, circBARD1 and circRASA2, and PD suggested function of circRNAs in periodontitis. Network prediction further focused on downstream targets regulated by circRNAs during periodontitis pathogenesis.

miR-99a regulates CD4+ T cell differentiation and attenuates experimental autoimmune encephalomyelitis by mTOR-mediated glycolysis.

Multiple microRNAs exhibit diverse functions to regulate inflammatory and autoimmune diseases. MicroRNA-99a (miR-99a) has been shown to be involved in adipose tissue inflammation and to be downregulated in the inflammatory lesions of autoimmune diseases rheumatoid arthritis and systemic lupus erythematosus. In this study, we found that miR-99a was downregulated in CD4+ T cells from experimental autoimmune encephalomyelitis (EAE) mice, an animal model of multiple sclerosis. Overexpression of miR-99a alleviated EAE development by promoting regulator T cells and inhibiting T helper type 1 (Th1) cell differentiation. Bioinformatics and functional analyses further revealed that the anti-inflammatory effects of miR-99a was attributable to its role in negatively regulating glycolysis reprogramming of CD4+ T cells by targeting the mTOR pathway. Additionally, miR-99a expression was induced by transforming growth factor ß (TGF-ß) to regulate CD4+ T cell glycolysis and differentiation. Taken together, our results characterize a pivotal role of miR-99a in regulating CD4+ T cell differentiation and glycolysis reprogramming during EAE development, which may indicate that miR-99a is a promising therapeutic target for the amelioration of multiple sclerosis and possibly other autoimmune diseases.

Associations of dietary and drinking water habits with number of natural teeth: a longitudinal study in the Chinese elderly population.

BACKGROUND: The relationship between dietary and drinking water habits and oral health are still unclear. We aimed at evaluating the association of dietary and drinking water habits with number of teeth in the elderly adults. METHODS: We conducted a longitudinal study based on the Chinese Longitudinal Healthy Longevity Survey from 1998 to 2018. The data of dietary and drinking water habits at baseline were collected using a questionnaire. The number of teeth at baseline and follow-up was collected for each subject. We used the linear mixed-effect model to analyze the associations of dietary habits and drinking water sources with tooth number. RESULTS: Among 19,896 participants at baseline, the mean age of the participants was 83.87 years, with the average number of natural teeth of 9.37, 8.26, 8.38, 8.68, 4.05, 1.92, 1.12, 2.20 for the first to eighth waves of survey. Compared with subjects drinking tap water, 1.036 (95 % CI: -1.206, -0.865), 0.880 (95 % CI: -1.122, -0.637) and 1.331 (95 % CI: -1.715, -0.947) fewer natural teeth were reported for those drinking well, surface water and spring at baseline survey. Compared with participants with rice intake as the staple food, those with wheat intake (ß = -0.684; 95 % CI: -0.865, -0.503) tended to have fewer natural teeth. Compared with participants with fresh fruit intake almost every day, those with quite often intake of fresh fruit tended to have fewer teeth with a significant dose-response trend (Ptrend <0.001). Similar decreased trend for number of teeth was also indicated for increased frequency of vegetable intake (Ptrend <0.001). Fewer number of teeth was found for subjects with less frequency of meat and fish intakes. CONCLUSIONS: The study suggested that drinking well, surface water, and spring, intakes of wheat as staple food, as well as less frequency of fresh fruit, vegetable, meat and fish intakes were associated with significantly fewer number of teeth in the Chinese elderly population.

Metal Material, Properties and Design Methods of Porous Biomedical Scaffolds for Additive Manufacturing: A Review.

Design an implant similar to the human bone is one of the critical problems in bone tissue engineering. Metal porous scaffolds have good prospects in bone tissue replacement due to their matching elastic modulus, better strength, and biocompatibility. However, traditional processing methods are challenging to fabricate scaffolds with a porous structure, limiting the development of porous scaffolds. With the advancement of additive manufacturing (AM) and computer-aided technologies, the development of porous metal scaffolds also ushers in unprecedented opportunities. In recent years, many new metal materials and innovative design methods are used to fabricate porous scaffolds with excellent mechanical properties and biocompatibility. This article reviews the research progress of porous metal scaffolds, and introduces the AM technologies used in porous metal scaffolds. Then the applications of different metal materials in bone scaffolds are summarized, and the advantages and limitations of various scaffold design methods are discussed. Finally, we look forward to the development prospects of AM in porous metal scaffolds.

Expression profile of lipoxygenases in gingival tissues of human periodontitis.

OBJECTIVES: This study aimed to clarify the expression profile and significance of lipoxygenases in periodontitis. MATERIALS AND METHODS: The mRNA levels of lipoxygenases in gingival tissues from 14 patients with periodontitis and 14 healthy individuals were determined by real-time PCR, and validated in datasets, GSE16134 and GSE10334, and by Western blotting. Correlation of differentially expressed lipoxygenases with clinical parameters and expression of tumor necrosis factor-α (TNF-α), interleukin-1ß, matrix metalloproteinase (MMP)-8, MMP-9, and receptor activator of nuclear factor-κB ligand (RANKL) was investigated in patients with periodontitis by Spearman's correlation analysis. RESULTS: The expression of ALOX5 (2.1-fold, p < .05), ALOX12B (2.9-fold, p < .001), and ALOX15B (9.4-fold, p < .001) was upregulated in gingival tissues from patients with periodontitis, which was validated by dataset analysis and Western blotting. Positive correlations were observed between ALOX5 and probing depth, and ALOX15B and probing depth and clinical attachment loss. Furthermore, ALOX5 expression was positively correlated with TNF-α, MMP-8, MMP-9, and RANKL expression, and ALOX15B was positively correlated with MMP-8 and RANKL. CONCLUSIONS: Our findings indicated the upregulation of ALOX5 and ALOX15B in periodontitis and suggested that ALOX5 and ALOX15B may be involved in periodontitis pathogenesis, including inflammation, connective tissue destruction, and abnormal bone metabolism.

Eleven-year experience implementing a dental undergraduate research programme in a prestigious dental school in China: Lessons learned and future prospects.

OBJECTIVE: To summarise the lessons learned from implementing a dental undergraduate research programme over the past eleven years and identify key elements to guarantee the success of dental undergraduate research in the future. MATERIAL AND METHODS: An overview of 80 research projects from 2007 to 2017 was provided, consisting of 239 participating undergraduates and 47 faculty advisors. Students' academic performance was compared between participating and non-participating undergraduates. An anonymous questionnaire was designed and distributed to participants, with VAS satisfaction assessment, Likert-scale items and open-ended questions. Questions focused on overall satisfaction, motivation, benefits to student career development and essential elements for the dental undergraduate research programme in the future. RESULTS: The undergraduate participants had significantly better scores of GPAs (3.41 vs 3.21; P < 0.0001), obtained more awards on the honour rolls (0.53 vs 0.30; P = 0.0171) and published more peer-reviewed articles (1.62 vs 1.31; P = 0.0253) than non-participants. Seventy-two undergraduates (75.79%) and thirty-eight advisors (80.85%) responded to the questionnaire. The overall satisfaction was scored as 78.18/100 and 72.36/100 amongst advisors and students, respectively. Interest was considered the best motivation for participation by students (3.81/5.00) and advisors (4.00/5.00). The research programme was beneficial to students' overall career development. Essential roles played by the dental school, faculty and undergraduates were suggested to improve the research environment. CONCLUSION: The dental undergraduate research programme has positive effects on students' academic performance. The support of the dental school, the engagement of qualified faculty, the interest and autonomy of students and camaraderie amongst these three key elements are essential for the research programme in the future.

Fabrication of PCL Scaffolds by Supercritical CO2 Foaming Based on the Combined Effects of Rheological and Crystallization Properties.

Polycaprolactone (PCL) scaffolds have recently been developed via efficient and green supercritical carbon dioxide (scCO2) melt-state foaming. However, previously reported gas-foamed scaffolds sometimes showed insufficient interconnectivity or pore size for tissue engineering. In this study, we have correlated the thermal and rheological properties of PCL scaffolds with their porous morphology by studying four foamed samples with varied molecular weight (MW), and particularly aimed to clarify the required properties for the fabrication of scaffolds with favorable interconnected macropores. DSC and rheological tests indicate that samples show a delayed crystallization and enhanced complex viscosity with the increasing of MW. After foaming, scaffolds (27 kDa in weight-average molecular weight) show a favorable morphology (pore size = 70-180 µm, porosity = 90% and interconnectivity = 96%), where the lowest melt strength favors the generation of interconnected macropore, and the most rapid crystallization provides proper foamability. The scaffolds (27 kDa) also possess the highest Young's modulus. More importantly, owing to the sufficient room and favorable material transportation provided by highly interconnected macropores, cells onto the optimized scaffolds (27 kDa) perform vigorous proliferation and superior adhesion and ingrowth, indicating its potential for regeneration applications. Furthermore, our findings provide new insights into the morphological control of porous scaffolds fabricated by scCO2 foaming, and are highly relevant to a broader community that is focusing on polymer foaming.

Supercritical CO2 foamed composite scaffolds incorporating bioactive lipids promote vascularized bone regeneration via Hif-1α upregulation and enhanced type H vessel formation.

Bone tissue engineering has substantial potential for the treatment of massive bone defects; however, efficient vascularization coupled with bone regeneration still remains a challenge in this field. In the current study, supercritical carbon dioxide (scCO2) foaming technique was adopted to fabricate mesoporous bioactive glasses (MBGs) particle-poly (lactic-co-glycolic acid) (PLGA) composite scaffolds with appropriate mechanical and degradation properties as well as in vitro bioactivity. The MBG-PLGA scaffolds incorporating the bioactive lipid FTY720 (designated as FTY/MBG-PLGA) exhibited simultaneously sustained release of the bioactive lipid and ions. In addition to providing a favorable microenvironment for cellular adhesion and proliferation, FTY/MBG-PLGA scaffolds significantly facilitated the in vitro osteogenic differentiation of rBMSCs and also markedly stimulated the upregulation of Hif-1α expression via the activation of the Erk1/2 pathway, which mediated the osteogenic and pro-angiogenic effects on rBMSCs. Furthermore, FTY/MBG-PLGA extracts induced superior in vitro angiogenic performance of HUVECs. In vivo evaluation of critical-sized rat calvarial bone defects indicated that FTY/MBG-PLGA scaffolds potently promoted vascularized bone regeneration. Notably, the significantly enhanced formation of type H vessels (CD31hiEmcnhi neo-vessels) was observed in newly formed bone tissue in FTY/MBG-PLGA group, strongly suggesting that FTY720 and therapeutic ions released from the scaffolds synergistically induced more type H vessel formation, which indicated the coupling of angiogenesis and osteogenesis to achieve efficiently vascularized bone regeneration. Overall, the results indicated that the foamed porous MBG-PLGA scaffolds incorporating bioactive lipids achieved desirable vascularization-coupled bone formation and could be a promising strategy for bone regenerative medicine. STATEMENT OF SIGNIFICANCE: Efficacious coupling of vascularizationandbone formation is critical for the restoration of large bone defects. Anoveltechnique was used to fabricate composite scaffolds incorporating bioactive lipids which possessedsynergistic cues of bioactive lipids and therapeutic ions to potently promotebone regenerationas well as vascularization. The underlying molecular mechanism for the osteogenic and pro-angiogenic effects of the compositescaffolds was unveiled. Interestingly, the scaffolds were furtherfoundto enhance the formation oftype H capillarieswithin the bone healing microenvironment to couple angiogenesis to osteogenesis to achieve satisfyingvascularizedbone regeneration.These findings provide a novel strategy to develop efficiently vascularized engineering constructs to treat massive bone defects.

Exosomal miR-196a derived from cancer-associated fibroblasts confers cisplatin resistance in head and neck cancer through targeting CDKN1B and ING5.

BACKGROUND: Cisplatin resistance is a major challenge for advanced head and neck cancer (HNC). Understanding the underlying mechanisms and developing effective strategies against cisplatin resistance are highly desired in the clinic. However, how tumor stroma modulates HNC growth and chemoresistance is unclear. RESULTS: We show that cancer-associated fibroblasts (CAFs) are intrinsically resistant to cisplatin and have an active role in regulating HNC cell survival and proliferation by delivering functional miR-196a from CAFs to tumor cells via exosomes. Exosomal miR-196a then binds novel targets, CDKN1B and ING5, to endow HNC cells with cisplatin resistance. Exosome or exosomal miR-196a depletion from CAFs functionally restored HNC cisplatin sensitivity. Importantly, we found that miR-196a packaging into CAF-derived exosomes might be mediated by heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1). Moreover, we also found that high levels of plasma exosomal miR-196a are clinically correlated with poor overall survival and chemoresistance. CONCLUSIONS: The present study finds that CAF-derived exosomal miR-196a confers cisplatin resistance in HNC by targeting CDKN1B and ING5, indicating miR-196a may serve as a promising predictor of and potential therapeutic target for cisplatin resistance in HNC.

Evaluation of highly carbonated hydroxyapatite bioceramic implant coatings with hierarchical micro-/nanorod topography optimized for osseointegration.

BACKGROUND: Optimal osseointegration has been recognized as a pivotal factor in determining the long-term success of biomedical implants. MATERIALS AND METHODS: In the current study, highly carbonated hydroxyapatite (CHA) with carbonate contents of 8, 12 and 16 wt% and pure hydroxyapatite (HA) were fabricated via a novel hydrothermal method and deposited on the titanium substrates to generate corresponding CHA bioceramic coatings (designated as C8, C12 and C16, respectively) and HA bioceramic coatings (designated as C0). RESULTS: C8, C12 and C16 were endowed with nanoscale, hierarchical hybrid micro-/nanoscale and microscale surface topographies with rod-like superstructures, respectively. Compared with C0, the micro-/nanotextured CHA bioceramic coatings (C8, C12 and C16) possessed excellent surface bioactivity and biocompatibility, as well as better wettability, which mediated improved protein adsorption, giving rise to simultaneous enhancement of a biological cascade of events of rat bone-marrow-derived mesenchymal stem cells including cell adhesion, proliferation, osteogenic differentiation and, notably, the production of the pro-angiogenic growth factor, vascular endothelial growth factor-A. In particular, C12 with biomimetic hierarchical hybrid micro-/nanorod topography exhibited superior fractal property and predominant performance of protein adsorption, cell adhesion, proliferation and osteogenesis concomitant with angiogenesis. CONCLUSION: All these results suggest that the 12 wt% CHA bioceramic coating with synergistic modification of surface chemistry and topography has great prospect for future use as implant coating to achieve optimum osseointegration for orthopedic and dental applications.

Microstructures, mechanical, and biological properties of a novel Ti-6V-4V/zinc surface nanocomposite prepared by friction stir processing.

BACKGROUND: The interaction between the material and the organism affects the survival rate of the orthopedic or dental implant in vivo. Friction stir processing (FSP) is considered a new solid-state processing technology for surface modification. PURPOSE: This study aims to strengthen the surface mechanical properties and promote the osteogenic capacity of the biomaterial by constructing a Ti-6Al-4V (TC4)/zinc (Zn) surface nanocomposites through FSP. METHODS: FSP was used to modify the surface of TC4. The microstructures and mechanical properties were analyzed by scanning electron microscopy, transmission electron microscopy, nanoindentation and Vickers hardness. The biological properties of the modified surface were evaluated by the in vitro and in vivo study. RESULTS: The results showed that nanocrystalline and numerous ß regions, grain boundary α phase, coarser acicular α phase and finer acicular martensite α' appeared because of the severe plastic deformation caused by FSP, resulting in a decreased elastic modulus and an increased surface hardness. With the addition of Zn particles and the enhancement of hydrophilicity, the biocompatibility was greatly improved in terms of cell adhesion and proliferation. The in vitro osteogenic differentiation of rat bone marrow stromal cells and rapid in vivo osseointegration were enhanced on the novel TC4/Zn metal matrix nanocomposite surface. CONCLUSION: These findings suggest that this novel TC4/Zn surface nanocomposite achieved by FSP has significantly improved mechanical properties and biocompatibility, in addition to promoting osseointegration and thus has potential for dental and orthopedic applications.