New frontiers in salivary extracellular vesicles: transforming diagnostics, monitoring, and therapeutics in oral and systemic diseases.

Salivary extracellular vesicles (EVs) have emerged as key tools for non-invasive diagnostics, playing a crucial role in the early detection and monitoring of diseases. These EVs surpass whole saliva in biomarker detection due to their enhanced stability, which minimizes contamination and enzymatic degradation. The review comprehensively discusses methods for isolating, enriching, quantifying, and characterizing salivary EVs. It highlights their importance as biomarkers in oral diseases like periodontitis and oral cancer, and underscores their potential in monitoring systemic conditions. Furthermore, the review explores the therapeutic possibilities of salivary EVs, particularly in personalized medicine through engineered EVs for targeted drug delivery. The discussion also covers the current challenges and future prospects in the field, emphasizing the potential of salivary EVs in advancing clinical practice and disease management.

The oral microbiome in autoimmune diseases: friend or foe?

The human body is colonized by abundant and diverse microorganisms, collectively known as the microbiome. The oral cavity has more than 700 species of bacteria and consists of unique microbiome niches on mucosal surfaces, on tooth hard tissue, and in saliva. The homeostatic balance between the oral microbiota and the immune system plays an indispensable role in maintaining the well-being and health status of the human host. Growing evidence has demonstrated that oral microbiota dysbiosis is actively involved in regulating the initiation and progression of an array of autoimmune diseases.Oral microbiota dysbiosis is driven by multiple factors, such as host genetic factors, dietary habits, stress, smoking, administration of antibiotics, tissue injury and infection. The dysregulation in the oral microbiome plays a crucial role in triggering and promoting autoimmune diseases via several mechanisms, including microbial translocation, molecular mimicry, autoantigen overproduction, and amplification of autoimmune responses by cytokines. Good oral hygiene behaviors, low carbohydrate diets, healthy lifestyles, usage of prebiotics, probiotics or synbiotics, oral microbiota transplantation and nanomedicine-based therapeutics are promising avenues for maintaining a balanced oral microbiome and treating oral microbiota-mediated autoimmune diseases. Thus, a comprehensive understanding of the relationship between oral microbiota dysbiosis and autoimmune diseases is critical for providing novel insights into the development of oral microbiota-based therapeutic approaches for combating these refractory diseases.

Analysis of the Development Trend in Forensic Odontology Based on CiteSpace.

OBJECTIVES: To realize the dynamic visualization of forensic odontology based on the bibliometrics methods, and capture the research hotspots and identify the future development trend. METHODS: Literature articles published from January 1995 to December 2020 were searched according to specific subject words in the core data set of Web of Science. The visualization analysis of publishing country, institution, discipline, author, co-cited journal and keywords was performed by CiteSpace 5.7.R5W software. RESULTS: The annual analysis of publications showed an upward trend of forensic odontology research literature year by year, with the number of annual publications more than 110 in the last five years. Developed countries were the main source of contributions and the average centrality was greater than 0.2. The research of forensic odontology involved multiple disciplines, including stomatology, biology, computer science and medical imaging, with a distinct interdisciplinary feature. A total of 115 nodes were obtained by keyword cluster analysis. The principal line of forensic odontology mainly included individual identification and age estimation and the emergence of hotspots was closely related to new technologies. Population-based odontology investigation, improvement of traditional dental age estimation method and dental age estimation based on new technology were popular research in forensic odontology. CONCLUSIONS: Developing countries urgently need to increase the focus on related research. It may be an important direction for the development of forensic odontology to establish and enrich the regional dental database, develop new odontology identification technology combined with frontier and high-end technology, and develop the identification program based on advanced information technology.

Bone defects are repaired by enhanced osteogenic activity of the induced membrane: a case report and literature review.

BACKGROUND: The induced membrane technique (IMT) is an effective strategy to repair bone defects and involves a two-stage set of surgical procedures. Although the IM has osteogenic activity, bone grafting is necessary in standard IMT. Bone defects repaired completely by osteogenic activity of the IM alone without bone grafts are rare. CASE PRESENTATION: We present a case of infected fractures and bone defects of the ulna and radius treated with IMT. After the first stage using polymethylmethacrylate (PMMA) beads, X-rays showed that new callus developed after 2 to 4 months, and the defects were repaired completely by 5 months. We also present a literature review on spontaneous osteogenesis of the IM in patients. CONCLUSIONS: We present a case of infected ulnar and radial bone defects that healed by 5 months after the first stage of the IMT using a PMMA spacer. This finding suggests that local associated inflammatory reactions and bone tissue might enhance the osteogenic activity of the IM, causing spontaneous healing of bone defects. This appears to be the first such case reported in the literature.

Porphyromonas gingivalis induces depression via downregulating p75NTR-mediated BDNF maturation in astrocytes.

Many cross-sectional epidemiological studies have shown the incidence of periodontitis is positive correlated with that of depression. However, their causal relationship and underlying mechanism are largely unknown. Porphyromonas gingivalis (Pg) is the main pathogen for periodontitis. Employing female mice treated with Pg every other day for 4 weeks, we found that Pg-mice showed obvious depression-like behavior, an increased number of activated astrocytes and decreased levels of mature brain derived neurotrophic factor (BDNF) and astrocytic p75NTR in the hippocampus. Both hippocampal injection of BDNF and overexpression of p75NTR in astrocytes alleviated Pg-induced depression-like behavior in mice. Moreover, Pg-lipopolysaccharides (LPS) generated similar phenotypes, which were reversed by the TLR-4 inhibitor TAK242. Our results suggest that Pg-LPS decreases the level of astrocytic p75NTR and then downregulates BDNF maturation, leading to depression-like behavior in mice. Our study provides the first evidence that Pg is a modifiable risk factor for depression and uncovers a novel therapeutic target for the treatment of depression.

Visualization and quantitation of fetal movements by real-time three-dimensional ultrasound with live xPlane imaging in the first trimester of pregnancy.

AIM: To prove whether real-time three-dimensional (3D) ultrasound with live xPlane imaging is better in observing fetal movements than standard ultrasound imaging. METHODS: 50 healthy women with singleton pregnancies (22-43 years old) at 11 to 14 weeks of gestation underwent real-time 3D ultrasound examination with live xPlane imaging from July 2014 to February 2015. The incidence and frequency of 10 fetal movement patterns in 10 minutes were evaluated, including general movements (GMs), isolated arm movements, isolated leg movements, hiccup, stretching, breathing, startle, jaw opening, isolated head retroflexion, and isolated head anteflexion. The correlation between gestational age and frequency of each fetal movement pattern was analyzed. RESULTS: GM had the highest incidence (100%), followed by startle (84%) and isolated arm movements (68%). Their median frequency was 5 (IQR 3-6), 5 (IQR 1.75-11.5), and 1 (IQR 0-2), respectively. GM (Z=5.875, P<0.001) and startle (Z=5.302, P<0.001) had significantly higher frequency than isolated arm movements. The other 7 fetal movement patterns had much lower incidence and frequency. The frequency of GM was positively correlated with gestational age (r=0.360, P=0.010). CONCLUSION: Real-time 3D ultrasound with live x Plane imaging was shown to be a feasible tool for observing fetal movements.

Engineering exosomes and biomaterial-assisted exosomes as therapeutic carriers for bone regeneration.

Mesenchymal stem cell-based therapy has become an effective therapeutic approach for bone regeneration. However, there are still limitations in successful clinical translation. Recently, the secretome of mesenchymal stem cells, especially exosome, plays a critical role in promoting bone repair and regeneration. Exosomes are nanosized, lipid bilayer-enclosed structures carrying proteins, lipids, RNAs, metabolites, growth factors, and cytokines and have attracted great attention for their potential application in bone regenerative medicine. In addition, preconditioning of parental cells and exosome engineering can enhance the regenerative potential of exosomes for treating bone defects. Moreover, with recent advancements in various biomaterials to enhance the therapeutic functions of exosomes, biomaterial-assisted exosomes have become a promising strategy for bone regeneration. This review discusses different insights regarding the roles of exosomes in bone regeneration and summarizes the applications of engineering exosomes and biomaterial-assisted exosomes as safe and versatile bone regeneration agent delivery platforms. The current hurdles of transitioning exosomes from bench to bedside are also discussed.

KLF9 Promotes Osteogenic Differentiation of Dental Stem Cells by Negatively Regulating Notch1 Mediated Signaling Pathway.

BACKGROUND: Human dental stem cells (DSCs) are excellent sources of cells for treating dental and craniofacial diseases. However, the mechanisms regulating DSC osteogenic differentiation are still unclear. In this study, we aimed to determine the role of Krüppel-like factor 9 (KLF9) in regulating the biological functions of DSCs and explore the underlying molecular mechanisms. METHODS: Bioinformatic analyses, quantitative real-time polymerase chain reaction (qRT‒PCR) and Western blotting were performed to determine the KLF9 level during osteogenic differentiation of DSCs. The effects of KLF9 depletion or overexpression on DSC osteogenic differentiation were then evaluated. The osteogenic potential and associated mineralized nodule-forming activities of DSCs were monitored via Alizarin red S staining and quantitative analyses of osteogenic markers. The regulatory effect of KLF9 on the Notch1 signaling pathway was analyzed by luciferase reporter assays. RESULTS: KLF9 mRNA expression was consistently increased during mesenchymal stem cell osteogenic differentiation in multiple public datasets, and our qRT‒PCR and Western blotting data further validated this finding. In addition, KLF9 depletion promoted proliferation and suppressed osteogenic differentiation of DSCs, while enforced expression of KLF9 promoted the DSC osteogenic potential. Mechanistically, KLF9 negatively regulated the Notch1-mediated signaling pathway by directly binding to the Notch1 promoter. More importantly, Notch1 inhibition/overexpression partially rescued the suppressive/enhancing effects of KLF9 depletion/overexpression on the osteogenic differentiation of DSCs, indicating that Notch1 is a functional downstream target of KLF9. CONCLUSIONS: In summary, our results strongly demonstrate that KLF9 is a crucial transcription factor that controls the osteogenic differentiation of DSCs by negatively regulating the Notch1 signaling pathway.

Monitoring the Cleanliness of Reusable Surgical Instruments in Central Sterile Supply Department by Adenosine Triphosphate Method.

BACKGROUND: Cleaning is very important in reusable surgical instruments (RSI) reprocessing. The adenosine triphosphate (ATP) method is widely used to assess cleanliness, but few studies focus on using this method on RSIs. OBJECTIVE: To assess the effectiveness of the ATP Bioluminescence Assay for monitoring the cleanliness of RSIs. METHODS: The study was conducted in three central sterile supply departments (CSSD) from 2015 to 2020. Surgical scissors, hemostatic forceps, and other non-lumen instruments were sampled after cleaning. The surface of each instrument was swabbed in a uniform manner, including hinged joints and teeth, and the amount of ATP (amol) was calculated. RESULTS: A total of 871 RSIs were tested, the mean ATP lg (amol) was 2.829 ± 0.539 and the qualified rate was 80.57%. Washer-disinfector cleaning was more efficient than manual cleaning, the mean ATP lg (amol) were 2.776 ± 0.513 and 2.948 ± 0.575, respectively; the qualified rates were 84.97% and 70.59%, respectively. With the time of bare instruments expose to environment increased, the qualified rate was decreased (Ptrend = 0.044). CONCLUSION: ATP levels are stable and repeatable for continuous monitoring of the cleanliness of RSIs. It is a rapid and viable method for assessing the cleanliness of RSIs. Washer-disinfector cleaning is recommended and cleaned instruments should be placed in sterile packaging in good time. HIGHLIGHTS: The ATP method is viable for assessing the cleanliness of RSI in CSSDs.

One-Step Preparation of a Core-Spun Cu/P(VDF-TrFE) Nanofibrous Yarn for Wearable Smart Textile to Monitor Human Movement.

At present, wearable electronic sensors are widely investigated and applied for human life usage especially for the flexible piezoelectric sensor based on piezoelectric fibers. However, most of these fiber-based piezoelectric sensors are thin films, which might had poor air permeability, or do not adapt to complex body movements. In this study, a piezoelectric sensing fabric was proposed based on core-spun Cu/P(VDF-TrFE) nanofibrous yarns. These yarns were fabricated by P(VDF-TrFE) as a piezoelectric material and Cu wire as an inner electrode layer through a one-step conjugate electrospinning process. The Cu/P(VDF-TrFE) fabrics showed good flexibility, breathability, mechanical stability, and sensing capability after continuous running for 60 min or after washing. A 4 cm × 4 cm fabric could generate a current of 38 nA and voltage of 2.7 V under 15 N pressure. Once the fabric was fixed onto the clothes, human motion could be monitored by collecting its generated current, and the signal could be wirelessly transmitted onto a smartphone. Therefore, this study may provide a simple and promising approach to design a smart textile for human motion monitoring.

A novel elastic liposome for skin delivery of papain and its application on hypertrophic scar.

This study aims to investigate the therapeutic effects of papain elastic liposomes (PEL) on hypertrophic scar through topical application. PEL were prepared using the reverse-phase evaporation method and optimized by response surface methodology. The transdermal absorption of optimized PEL was tested by vertical Franz diffusion cells in vitro. The effects of PEL were investigated in rabbit model of hypertrophic scar in vivo, histological analysis and scar-related proteins were detected to reveal potential scar repair mechanism. The best formulation of PEL had EE (43.8±1.4%), particle size (100.9±2.2nm), PDI (0.037±0.003), zeta potential (-26.3±1.3mV), and DI (21.9±3.1). PEL gave the cumulative amounts and steady state fluxes in the receiver solution of 381.9±32.4µg/cm2, 11.4±1.5µg/cm2/h, and showed drug deposition in skin of 19.1±3.2% after 24h. After topical application, the scar elevation index, microvascular density, and collagen fiber were significantly decreased with regular arrangement. The expressions of TGF-ß1, P-Smad-3, P-NF-κB p65, and P-IKBa in hypertrophic scar were significantly down regulated in contrast with those in model group. PEL were proven as an excellent topical preparation for hypertrophic scar treatment.

[Influence of surface roughness on oral streptococcal adhesion forces to dental filling materials].

OBJECTIVE: This study is to determine the common oral streptococcal adhesion forces by using composite resin and glass ionomer cement (GIC) with different degrees of surface roughness via atomic force microscopy (AFM) analysis. The influence of surface roughness on bacterial adhesion force is also discussed. METHODS: Polishing and grinding were applied to obtain 300, 200, 100, and 10 nm surfaces of light-cured composite resin and GIC samples. Surface topography was assessed by AFM analysis. Initial colonizers (Streptococcus sanguinis and Streptococcus mitis) and cariogenic bacterial strains (Streptococcus mutans and Streptococcus sobrinus) were used to obtain bacteria-modified AFM probes. The force-distance curves were also measured by AFM analysis to determine the adhesion forces of bacteria on the surfaces of the composite resin and GIC. Material surface roughness was analyzed using ANOVA, and adhesion forces were subjected to nonparametric analysis (Kruskal-Wallis test). Comparison among groups was performed by Dunn's test. Material surface roughness and bacterial adhesion forces were subjected to correlation analysis. RESULTS: Bacterial adhesion forces increased with increasing material roughness. The adhesion forces of the four bacterial species reached the maximum on the material surface of 300 nm. The adhesion force of Streptococcus mutans increased from 0.578 nN to 2.876 nN on GIC surfaces with 10 and 300 nm roughness. The adhesion forces of the four species on the surface of the composite resin were stronger than that of GIC. The initial colonizers exhibited stronger adhesion forces to different materials than the cariogenic strains. Intergroup differences were evident on the 200 and 300 nm material surfaces. CONCLUSIONS: The surface roughness of the material significantly affected the bacterial adhesion forces, and a significant linear correlation existed between both factors. The bacterial adhesion forces of the GIC were lower than that of the composite resin. Furthermore, surface roughness exhibited less influence on the cariogenic strains than that on the initial colonizers.

Highly efficient uptake of ultrafine mesoporous silica nanoparticles with excellent biocompatibility by Liriodendron hybrid suspension cells.

The characteristics of the interactions co-cultures of ultrafine mesoporous silica nanoparticles (MSNs) and the Liriodendron hybrid suspension cells were systematically investigated using laser scanning confocal microscope (LSCM) and scanning electron microscopy (SEM). Using fluorescein isothiocyanate (FITC) labeling, the LSCM observations demonstrated that MSNs (size, 5-15 nm) with attached FITC molecules efficiently penetrated walled plant cells through endocytic pathways, but free FITC could not enter the intact plant cells. The SEM measurements indicated that MSNs readily aggregated on the surface of intact plant cells, and also directly confirmed that MSNs could enter intact plant cells; this was achieved by determining the amount of silicon present. After 24 h of incubation with 1.0 mg mL(-1) of MSNs, the viability of the plant cells was analyzed using fluorescein diacetate staining; the results showed that these cells retained high viability, and no cell death was observed. Interestingly, after the incubation with MSNs, the Liriodendron hybrid suspension cells retained the capability for plant regeneration via somatic embryogenesis. Our results indicate that ultrafine MSNs hold considerable potential as nano-carriers of extracellular molecules, and can be used to investigate in vitro gene-delivery in plant cells.

Screening of permeable compounds in Flos Lonicerae Japonicae with liposome using ultrafiltration and HPLC.

A new method for the screening of membrane-permeable compounds in traditional Chinese medicines (TCMs), using ultrafiltration and HPLC analysis, has been proposed. We hypothesized that exposure of a TCM extract to a liposome membrane, the concentration of membrane-permeable compounds in the solution should decrease. Using this approach, the permeability of compounds in Flos Lonicerae Japonicae (Japanese honeysuckle) was investigated. By comparing chromatograms of samples prepared before and after interaction with a liposome membrane, eight permeable compounds of Flos Lonicerae Japonicae were identified, and all of them were proven to be biologically active. Based on the significance of these results, this method could be a novel approach for identifying potentially bioactive components in other TCMs.