Low-intensity pulsed ultrasound enhances immunomodulation and facilitates osteogenesis of human periodontal ligament stem cells by inhibiting the NF-κB pathway.

Human periodontal ligament stem cells (hPDLSCs) are vital in cellular regeneration and tissue repair due to their multilineage differentiation potential. Low intensity pulsed ultrasound (LIPUS) has been applied for treating bone and cartilage defects. This study explored the role of LIPUS in the immunomodulation and osteogenesis of hPDLSCs. hPDLSCs were cultured in vitro, and the effect of different intensities of LIPUS (30, 60, and 90 mW/cm2) on hPDLSC viability was measured. hPDLSCs irradiated by LIPUS and stimulated by lipopolysaccharide (LPS) and LIPUS (90 mW/cm2) were co-cultured with peripheral blood mononuclear cells (PBMCs). Levels of immunomodulatory factors in hPDLSCs and inflammatory factors in PBMCs were estimated, along with determination of osteogenesis-related gene expression in LIPUS-irradiated hPDLSCs. The mineralized nodules and alkaline phosphatase (ALP) activity of hPDLSCs and levels of IκBα, p-IκBα, and p65 subunits of NF-κB were determined. hPDLSC viability was increased as LIPUS intensity increased. Immunomodulatory factors were elevated in LIPUS-irradiated hPDLSCs, and inflammatory factors were reduced in PBMCs. Osteogenesis-related genes, mineralized nodules, and ALP activity were promoted in LIPUS-irradiated hPDLSCs. The cytoplasm of hPDLSCs showed increased IκBα and p65 and decreased p-IκBα at increased LIPUS intensity. After LPS and LIPUS treatment, the inhibitory effect of LIPUS irradiation on the NF-κB pathway was partially reversed, and the immunoregulation and osteogenic differentiation of hPDLSCs were decreased. LIPUS irradiation enhanced immunomodulation and osteogenic differentiation abilities of hPDLSCs by inhibiting the NF-κB pathway, and the effect is dose-dependent. This study may offer novel insights relevant to periodontal tissue engineering.

miR-200a-3p represses osteogenesis of human periodontal ligament stem cells by targeting ZEB2 and activating the NF-κB pathway.

OBJECTIVES: Human periodontal ligament stem cells (hPDLSCs) bear multilineage differentiation potential and represent the cytological basis of periodontal tissue regeneration. microRNA (miR) is accepted as a critical regulator of cell differentiation. This study explored the molecular mechanism of miR-200a-3p in osteogenesis of hPDLSCs. MATERIAL AND METHODS: hPDLSCs were cultured and identified in vitro. miR-200a-3p expression during osteogenic differentiation of hPDLSCs was detected. hPDLSCs were transfected with miR-200a-3p mimic or miR-200a-3p inhibitor. Alkaline phosphatase (ALP) activity, calcified nodules and osteogenesis-related genes of hPDLSCs were measured. The binding relationship between miR-200a-3p and ZEB2 was predicted and verified. hPDLSCs were infected with sh-ZEB2, and then the osteogenic capacity was examined. miR-200a-3p inhibitor-transfected hPDLSCs were infected with sh-ZEB2. The key proteins of the NF-κB pathway were measured. RESULTS: miR-200a-3p expression was downregulated during osteogenic differentiation of hPDLSCs. Upregulation of miR-200a-3p reduced ALP activity, calcified nodules and osteogenesis-related genes of hPDLSCs, while downregulation of miR-200a-3p facilitated the osteogenesis of hPDLSCs. miR-200a-3p targeted ZEB2. ZEB2 silencing repressed osteogenesis of hPDLSCs. ZEB2 silencing attenuated the promoting effect of miR-200a-3p inhibitor on osteogenesis of hPDLSCs. miR-200a-3p activated the NF-κB pathway by targeting ZEB2. CONCLUSION: miR-200a-3p repressed osteogenesis of hPDLSCs by targeting ZEB2 and activating the NF-κB pathway. This study may offer insights for periodontal tissue regeneration engineering.

Enhancing osseointegration of titanium implants through large-grit sandblasting combined with micro-arc oxidation surface modification.

PURPOSE: The demand for titanium dental implants has risen sharply. However, the clinical success rate of implant surgery needs to be improved. In this paper, we report a novel surface modification strategy, large-grit sandblasting combined with micro-arc oxidation (SL-MAO), aiming to promote peri-implant bone formation and osseointegration of titanium implants. MATERIALS AND METHODS: Modified titanium samples were prepared by large-grit sandblasting and acid etching (SLA), micro-arc oxidation (MAO), and SL-MAO. The resulting topographical changes and chemical composition of the samples were examined by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS), respectively, and the biocompatibility and bioactivity were analyzed by bone-marrow mesenchymal stem cells (BMMSC) adhesion tests. Modified titanium implants were also inserted into the femurs of beagle dogs, and their competence of osseointegration was appraised by quantitative histomorphometry and micro-computed-tomography (micro-CT) analyses. RESULTS: Compared to SLA and MAO techniques, SL-MAO surface modification further enhanced titanium surfaces by creating a topographic morphology characterized by both micron-sized craters and sub-micron-scale pits, and resulted in superior chemical composition, which promoted cell adhesion, proliferation, and osteogenic differentiation. SL-MAO-modified titanium implants osseointegrated more efficiently than SLA or MAO controls, with significantly higher bone-area (BA) ratio and bone-implant contact (BIC) in the peri-implant region. CONCLUSIONS: The SL-MAO surface modification technique optimized the surface properties of titanium implants and enhanced peri-implant bone formation and osseointegration.

Aloe-emodin induces apoptosis in human oral squamous cell carcinoma SCC15 cells.

BACKGROUND: Oral and pharyngeal cancer is the most common malignant human cancers. Chemotherapy is an effective approach for anti-oral cancer therapy, while the drug tolerance and resistance remain a problem for oral cancer patients. Aloe-emodin, rhein and physcion are classified as anthraquinones, which are the main pharmacodynamic ingredients of Rheum undulatum L.. This study was undertaken to investigate whether aloe-emodin, rhein and physcion show inhibiting growth and inducing apoptosis in oral squamous cell carcinoma SCC15 cells. We found that aloe-emodin show inhibiting growth and inducing apoptosis in oral squamous cell carcinoma SCC15 cells, we also investigated the underlying mechanisms of apoptosis induced by aloe-emodin. METHODS: Thiazolyl blue tetrazolium bromide (MTT) test was used to detect cell proliferation. Cell apoptosis was detected by flow cytometry. We also used western blot analysis to detect the potential mechanisms of apoptosis. RESULTS: Aloe-emodin, rhein and physcion inhibit the proliferation of SCC15 cells and the order of inhibition level are aloe-emodin > Rhein > Physcion, the half maximal inhibitory concentrations (IC50) value of aloe-emodin was 60.90 µM at 48 h of treatment. Aloe-emodin treatment resulted in a time- and dose-dependent decrease in cell viability and increased the apoptotic cell ratio. The results of western blotting showed the expression levels of caspase-9 and caspase-3 proteins increased following aloe-emodin treatment. CONCLUSIONS: Our results revealed that aloe-emodin treatment could inhibit cell viability of SCC15 cells and the potential mechanism of inhibition might be through the induction of apoptosis by regulation of the expression levels of caspase-9 and caspase-3. This indicates that aloe-emodin may be a good agent for anti-oral cancer drug exploring.

Four-Dimensional Superimposition Techniques to Compose Dental Dynamic Virtual Patients: A Systematic Review.

Four-dimensional virtual patient is a simulation model integrating multiple dynamic data. This study aimed to review the techniques in virtual four-dimensional dental patients. Searches up to November 2022 were performed using the PubMed, Web of Science, and Cochrane Library databases. The studies included were based on the superimposition of two or more digital information types involving at least one dynamic technique. Methodological assessment of the risk of bias was performed according to the Joanna Briggs Institute Critical Appraisal Checklist. Methods, programs, information, registration techniques, applications, outcomes, and limitations of the virtual patients were analyzed. Twenty-seven full texts were reviewed, including 17 case reports, 10 non-randomized controlled experimental studies, 75 patients, and 3 phantoms. Few studies showed a low risk of bias. Dynamic data included real-time jaw motion, simulated jaw position, and dynamic facial information. Three to five types of information were integrated to create virtual patients based on diverse superimposition methods. Thirteen studies showed acceptable dynamic techniques/models/registration accuracy, whereas 14 studies only introduced the feasibility. The superimposition of stomatognathic data from different information collection devices is feasible for creating dynamic virtual patients. Further studies should focus on analyzing the accuracy of four-dimensional virtual patients and developing a comprehensive system.

Periodontal regeneration with stem cells-seeded collagen-hydroxyapatite scaffold.

Re-establishing compromised periodontium to its original structure, properties and function is demanding, but also challenging, for successful orthodontic treatment. In this study, the periodontal regeneration capability of collagen-hydroxyapatite scaffolds, seeded with bone marrow stem cells, was investigated in a canine labial alveolar bone defect model. Bone marrow stem cells were isolated, expanded and characterized. Porous collagen-hydroxyapatite scaffold and cross-linked collagen-hydroxyapatite scaffold were prepared. Attachment, migration, proliferation and morphology of bone marrow stem cells, co-cultured with porous collagen-hydroxyapatite or cross-linked collagen-hydroxyapatite, were evaluated in vitro. The periodontal regeneration capability of collagen-hydroxyapatite scaffold with or without bone marrow stem cells was tested in six beagle dogs, with each dog carrying one sham-operated site as healthy control, and three labial alveolar bone defects untreated to allow natural healing, treated with bone marrow stem cells - collagen-hydroxyapatite scaffold implant or collagen-hydroxyapatite scaffold implant, respectively. Animals were euthanized at 3 and 6 months (3 animals per group) after implantation and the resected maxillary and mandibular segments were examined using micro-computed tomography scan, H&E staining, Masson's staining and histometric evaluation. Bone marrow stem cells were successfully isolated and demonstrated self-renewal and multi-potency in vitro. The porous collagen-hydroxyapatite and cross-linked collagen-hydroxyapatite had average pore sizes of 415 ± 20 µm and 203 ± 18 µm and porosity of 69 ± 0.5% and 50 ± 0.2%, respectively. The attachment, proliferation and migration of bone marrow stem cells were satisfactory on both porous collagen-hydroxyapatite and cross-linked collagen-hydroxyapatite scaffolds. Implantation of bone marrow stem cells - collagen-hydroxyapatite or collagen-hydroxyapatite scaffold in beagle dogs with experimental periodontal defects resulted in significantly enhanced periodontal regeneration characterized by formation of new bone, periodontal ligament and cementum, compared with the untreated defects, as evidenced by histological and micro-computed tomography examinations. The prepared collagen-hydroxyapatite scaffolds possess favorable bio-compatibility. The bone marrow stem cells - collagen-hydroxyapatite and collagen-hydroxyapatite scaffold - induced periodontal regeneration, with no aberrant events complicating the regenerative process. Further research is necessary to improve the bone marrow stem cells behavior in collagen-hydroxyapatite scaffolds after implantation.

[Periodontal therapy for rheumatoid arthritis: a systematic review].

OBJECTIVE: To assess the effect and safety of periodontal therapy in relieving the symptoms and clinical signs of rheumatoid arthritis (RA). METHODS: The electronic search was conducted in Medline (OVID, 1950-2010 Sep), EMBASE (1984-2010 Sep), CENTRAL (2010, Issue 3), CBM (1978-2010 Sep) and the Chinese journals on stomatology were hand-searched. Clinical randomized controlled trials as well as clinical controlled trials were selected regarding the targeted issue. Two investigators evaluated the reporting quality and risk of bias of those included trials in accordance with CONSORT statement and Cochrane risk of bias assessment tools, and collected data of included studies in duplicate. Revman 5.0.23 was applied for Meta-analysis. RESULTS: Four trials met the inclusion criteria and a total of 150 patients were enrolled in the trials, one had low risk of bias and others had moderate risk of bias. Meta-analysis showed that pure periodontal therapy could not decrease disease activity score in 28 joints (DAS28) (P=0.06), and there was no statistically significant difference between periodontal therapy with anti-tumor necrosis factor-alpha (TNF-alpha) medication and pure anti-TNF-alpha medication (P=0.24). But the subgroup analysis showed that a significantly decreased DAS28 was achieved by periodontal therapy (P=0.03), and the interventions provided a remarkable effect on alleviating clinical signs and erythrocyte sedimentation rate of RA (P<0.05). Results of the symptoms relief differed from the studies. No adverse events were reported. CONCLUSION: The evidence available currently indicates that periodontal therapy may play a positive role in remitting the clinical signs and periodontal status of RA except the relief of the symptoms.