Exosomal circ_0000722 derived from periodontal ligament stem cells undergoing osteogenic differentiation promotes osteoclastogenesis.

Periodontal ligament stem cells (PDLSCs), which are considered promising stem cells for regeneration of periodontal bony tissue, can also manipulate alveolar bone remodeling by exosomes. In this study, we investigated interactions between PDLSCs under osteogenic differentiation and osteoclast precursors. The results showed that conditioned medium from PDLSCs under 5d osteogenic induction promoted osteoclastogenesis of RAW264.7 cells. The exosomes extracted from those conditioned media showed similar effects on osteoclastogenesis. Furthermore, exosomes from PDLSCs under 5d of osteogenic induction showed significantly high expression of circ_0000722, compared with exosomes from PDLSCs before osteogenic induction. Downregulation of circ_0000722 significantly attenuated the effect of PDLSC-derived exosomes on the osteoclastogenesis of RAW264.7 cells. Our findings suggested that exosomal circ_0000722 derived from periodontal ligament stem cells undergoing osteogenic differentiation might promote osteoclastogenesis by upregulating TRAF6 expression and activating downstream NF-κB and AKT signaling pathways.

Inhaled RNA drugs to treat lung diseases: Disease-related cells and nano-bio interactions.

In recent years, RNA-based therapies have gained much attention as biomedicines due to their remarkable therapeutic effects with high specificity and potency. Lung diseases offer a variety of currently undruggable but attractive targets that could potentially be treated with RNA drugs. Inhaled RNA drugs for the treatment of lung diseases, including asthma, chronic obstructive pulmonary disease, cystic fibrosis, and acute respiratory distress syndrome, have attracted more and more attention. A variety of novel nanoformulations have been designed and attempted for the delivery of RNA drugs to the lung via inhalation. However, the delivery of RNA drugs via inhalation poses several challenges. It includes protection of the stability of RNA molecules, overcoming biological barriers such as mucus and cell membrane to the delivery of RNA molecules to the targeted cytoplasm, escaping endosomal entrapment, and circumventing unwanted immune response etc. To address these challenges, ongoing researches focus on developing innovative nanoparticles to enhance the stability of RNA molecules, improve cellular targeting, enhance cellular uptake and endosomal escape to achieve precise delivery of RNA drugs to the intended lung cells while avoiding unwanted nano-bio interactions and off-target effects. The present review first addresses the pathologic hallmarks of different lung diseases, disease-related cell types in the lung, and promising therapeutic targets in these lung cells. Subsequently we highlight the importance of the nano-bio interactions in the lung that need to be addressed to realize disease-related cell-specific delivery of inhaled RNA drugs. This is followed by a review on the physical and chemical characteristics of inhaled nanoformulations that influence the nano-bio interactions with a focus on surface functionalization. Finally, the challenges in the development of inhaled nanomedicines and some key aspects that need to be considered in the development of future inhaled RNA drugs are discussed.

miR-148a-3p regulates proliferation and apoptosis of idiopathic gingival fibroma by targeting NPTX1.

OBJECTIVE: Idiopathic gingival fibromatosis (IGF) is a rare heterogeneous disease that results in the progressive and diffuse hyperplasia of gingival tissues. MicroRNAs are implicated in the development and progression of various tumors. The present study aimed to explore the potential roles and mechanisms of miR-148a-3p in IGF. METHODS: Gingival fibroblasts (GFs) were transfected with miR-148a-3p mimics, miR-148a-3p inhibitors, or siNPTX1, and then, the proliferation and apoptosis of GFs and the expression of related genes were evaluated using Cell Counting Kit-8 assays, 5-ethynyl-2'-deoxyuridine assays, flow cytometry, reverse transcription-quantitative polymerase chain reaction, and western blot analysis, respectively. RESULTS: miR-148a-3p was highly expressed in GFs of IGF (IGF-GFs) as compared with normal GFs (N-GFs). Overexpression of miR-148a-3p promoted the proliferation and inhibited the apoptosis of N-GFs, whereas downregulation of miR-148a-3p had the opposite effect in IGF-GFs. Knockdown of NPTX1 reversed miR-148a-3p-mediated effects in IGF-GFs. Dual-luciferase reporter assay confirmed that NPTX1 is a direct target of miR-148a-3p. CONCLUSION: These findings identify that miR-148a-3p could regulate cell proliferation and apoptosis by targeting NPTX1, providing new insights for the further study of the molecular mechanism and treatment of IGF.

Capsella bursa-pastoris (L.) Medic. extract alleviate cataract development by regulating the mitochondrial apoptotic pathway of the lens epithelial cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Capsella bursa-pastoris (L.) Medic. (CBP) is a cruciferous plant valuable in reducing fever, improving eyesight and calming the liver. This herb was recorded in the Compendium of Materia Medica for cataract treatment. AIM OF THE STUDY: To determine the effects and mechanism of CBP on cataract prevention and treatment using a selenite cataract model. MATERIALS AND METHODS: The main compounds in CBP extract were analyzed by UPLC, 1H-NMR and 13C-NMR spectroscopic techniques. Flavonoids formed a significant proportion of its compounds, thus necessitating an evaluation of their inhibitory effects on the development of cataract using a selenite cataract model. The protective effects of CBP flavonoids (CBPF) against oxidative damage and the modulation of mitochondrial apoptotic pathway were subsequently verified on H2O2-treated SRA01/04 lens epithelial cells. RESULTS: CBPF significantly alleviated the development of cataract by decreasing the MDA level and increasing the GSH-Px and SOD levels in the lens. It also inhibited H2O2-induced apoptosis in SRA01/04 cells, increased the expression of Bcl-2 protein and decreased the expressions of Caspase-3 and Bax proteins. CONCLUSION: CBPF exerts a significant preventive effect on cataract development by regulating the mitochondrial apoptotic pathway of the lens epithelial cells. It is thus a potent traditional Chinese medicine (TCM) whose application should be further developed for the clinical treatment of cataract.

Alteration of circRNA and lncRNA expression profile in exosomes derived from periodontal ligament stem cells undergoing osteogenic differentiation.

OBJECTIVE: This study investigated circRNA and lncRNA expression profile in exosomes derived from periodontal ligament stem cell (PDLSC) before and after its osteogenic differentiation. DESIGN: Exosomes derived from PDLSCs before (EX0) and after osteogenic induction for 5 (EX5) and 7 (EX7) days were harvested and exosomal circRNAs and lncRNAs were analyzed by RNA sequencing. Certain RNAs showing significantly altered expression were selected for qRT-PCR verification. The circRNA-miRNA-mRNA network and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed. RESULTS: All groups of exosomes showed typical characteristics under nanoparticle tracking analysis, flow cytometry assay and transmission electron microscopy. 69-557 circRNAs and 2907-11581 lncRNAs were found in EX0, EX5 and EX7, which were broadly distributed across the 24 pairs of human chromosomes. Compared with EX0, 3 circRNAs and 2 lncRNAs were up-regulated and 39 circRNAs and 5 lncRNAs down-regulated consistently through out of EX5 and EX7, p < 0.05. qRT-PCR confirmed certain those consistently expressed RNAs, such as circ lysophosphatidic acid receptor 1 (LPAR1). KEGG analysis showed that those consistent expressed RNAs closely related to TGF-beta pathway, MAPK pathway, mTOR pathway and FoxO signaling pathways regulating pluripotency of stem cells. CONCLUSIONS: Exosomal circRNAs and lncRNAs had significant expression changes during the early phase of osteogenic differentiation of PDLSCs. Further study would be taken for understanding the roles of exosomal circRNAs and lncRNAs playing in osteogenic differentiation of PDLSCs.

Exosomes derived from idiopathic gingival fibroma fibroblasts regulate gingival fibroblast proliferation and apoptosis.

OBJECTIVE: Exosomes have been proved to play an essential role in intercellular information transmission. However, few researches focused on exosomes derived from gingival fibroblasts (GFs) of IGF. The aim of this study is to investigate the effect of exosomes derived from GFs of IGF (IGF-GFs) on the proliferation and apoptosis of normal gingival fibroblasts (N-GFs). METHODS: Gingival fibroblasts were cultured and identified using immunocytochemistry. Exosomes were isolated with exosomes extraction kit and characterized by transmission electron microscopy (TEM) and flow cytometry. PKH67 labeling was further used to trace the intracellular distribution of the exosomes. And MTS assay was used to test the effective concentration and time course of IGF-GFs-derived exosomes. Furthermore, the expression of PCNA, Ki67, Bcl-2, and Bax in N-GFs was analyzed by qRT-PCR and Western blot. RESULTS: Exosomes were isolated from IGF-GFs; the identification of exosomes and gingival fibroblasts was successfully finished. Moreover, we found that N-GFs co-cultured with exosomes showed a great increase in PCNA and Bcl-2 levels, and a moderate increase in Ki67 levels. By contrast, the levels of Bax were significantly reduced. CONCLUSION: These results suggest that exosomes derived from idiopathic gingival fibroma fibroblasts are involved in the regulation of gingival fibroblast proliferation and apoptosis.

Exosomal circLPAR1 Promoted Osteogenic Differentiation of Homotypic Dental Pulp Stem Cells by Competitively Binding to hsa-miR-31.

Human dental pulp stem cells (DPSCs) hold great promise in bone regeneration. However, the exact mechanism of osteogenic differentiation of DPSCs remains unknown, especially the role of exosomes played in. The DPSCs were cultured and received osteogenic induction; then, exosomes from osteogenic-induced DPSCs (OI-DPSC-Ex) at different time intervals were isolated and sequenced for circular RNA (circRNA) expression profiles. Gradually, increased circular lysophosphatidic acid receptor 1 (circLPAR1) expression was found in the OI-DPSC-Ex coincidentally with the degree of osteogenic differentiation. Meanwhile, results from osteogenic differentiation examinations showed that the OI-DPSC-Ex had osteogenic effect on the recipient homotypic DPSCs. To investigate the mechanism of exosomal circLPAR1 on osteogenic differentiation, we verified that circLPAR1 could competently bind to hsa-miR-31, by eliminating the inhibitory effect of hsa-miR-31 on osteogenesis, therefore promoting osteogenic differentiation of the recipient homotypic DPSCs. Our study showed that exosomal circRNA played an important role in osteogenic differentiation of DPSCs and provided a novel way of utilization of exosomes for the treatment of bone deficiencies.

Yunnan Baiyao diminishes lipopolysaccharide-induced inflammation in osteoclasts.

Yunnan Baiyao (YNBY) has been refined for hundreds of years and has become a treasure of proprietary Chinese medicine that has significant curative effects in the field of hemostasis, blood circulation, and callus. In past years, YNBY has been demonstrated to play an anti-inflammatory role in bone-related diseases, such as rheumatoid arthritis and osteoporosis. However, the osteoclasts are multinucleated giant cells that resorb bone and participate in the occurrence, development, and progression of these bone-related diseases. Previous studies have reported that the inflammatory function is closely associated with arachidonic acid (AA) metabolism, as well as some inflammatory-related pathways, including the nuclear factor кB (NF-кB), mitogen-activated protein kinase (MAPK), and Wnt5a pathways. Therefore, we speculated that the anti-inflammatory effect of YNBY might be associated with the NF-кB, MAPK, and Wnt5a pathways. In order to further excavate the anti-inflammatory roles of YNBY, lipopolysaccharide (LPS) with an optimal concentration of 1,000 pg/ml was used to induce inflammation in osteoclasts. Our results showed that YNBY with a time- and dose-dependent method decreased the concentration of pro-inflammatory cytokines and the expression levels of cyclooxygenase-1 (COX-1), COX-2, 5-lipoxygenase, and prostaglandin E2. Moreover, it was found that COX-2 was the target gene regulated by YNBY. Finally, using NF-кB and MAPK pathway inhibitors or miRNA101b (involved in the Wnt5a pathway) in tandem with YNBY and the results exhibited that these groups caused a reduction in COX-1 and COX-2 expression, indicating that the anti-inflammatory function of YNBY might directly affect the NF-кB, MAPK, and Wnt5a pathways. PRACTICAL APPLICATIONS: Yunnan Baiyao (YNBY) is mainly extracted from precious Chinese medicines such as Panax notoginseng, borneol, musk, and yam and has a wide range of clinical applications. It is not only used to treat various types of traumatic injuries, but also used for upper gastrointestinal bleeding and wound ulcers, neonatal umbilitis, recurrent oral ulcers, esophagitis, bacterial dysentery, and so on. Although the detailed mechanism of action is not clear at present, it is believed that this is related to its anti-inflammatory, hemostatic, and immune-enhancing effects. Many bone-related diseases, such as rheumatoid arthritis and osteoporosis, are regarded to be intimately related to the inflammatory reaction. Thus, this study aimed to explore the underlying mechanisms of YNBY at anti-inflammatory roles. And our results suggested that YNBY directly affected the inflammatory cytokines and AA metabolic products which referred to the NF-кB, MAPK, and Wnt5a pathways, as well as AA metabolism, respectively. Hence, the practical applications of YNBY are the anti-inflammatory effects used to treat for bone-related diseases.

Cone-beam CT assessment of implant-related anatomy landmarks of the anterior mandible in a Chinese population.

PURPOSE: To investigate the prevalence, location, and morphology of the mandibular lingual foramen (MLF), mandibular incisive canal (MIC), and anterior loop of the inferior alveolar canal (ALC) in a Chinese population using cone-beam CT (CBCT). MATERIALS AND METHODS: From 2014 to 2016, CBCT images from patients with various scanning purposes were obtained from the database of the Affiliated Stomatology Hospital of Kunming Medical University, China. Imaging analyses of the MLF, MIC, and ALC were performed via the NNT viewer software. The prevalence, location, length, classification of MLF, and its distances to the alveolar crest and the lower border of mandible were investigated, and the prevalence and length of MIC and the prevalence of ALC were also studied. RESULTS: This study examined 1008 subjects, 521 (51.7%) males, and 487 (48.3%) females. 916 (90.9%) subjects showed the medial lingual foramina (LF), a single medial LF with the supraspinous-type predominating. Lateral LF were observed in 547 (54.3%) subjects mostly located in the premolar areas. 876 (86.9%) subjects had the MIC on the left side, whereas 877 (87.0%) had the MIC on the right side. The ALC was present in 147 (14.6%) subjects. CONCLUSIONS: This study showed a high prevalence of LF and MIC in the Southwest Chinese population. Therefore, caution should be taken during the implant treatment at the anterior mandible region.

[Applications of minimally invasive bone splitting technique in cases with missing maxillary anterior teeth and obvious labial alveolar bone depression for dental implantation].

PURPOSE: The purpose of this study was to introduce a minimally invasive bone splitting technique which is suitable for cases with missing anterior teeth and obvious depression of alveolar bone in labial side, and to evaluate its clinical results. METHODS: Minimally invasive bone splitting technique was used in 8 healthy adults with bone defects in the aesthetic zone. The labial alveolar bone incisions were confined around the bone defects which were smaller than traditional incisions. The other procedures were the same as conventional bone splitting technique. Cone-beam CT (CBCT) for missing anterior teeth was taken before surgery, after the surgery and 6 months after surgery and alveolar bone height and width were recorded with landmark identification designed by ourselves in this study. The data were analyzed with SPSS 21.0 software package for paired t test. RESULTS: Paired t test indicated that after surgery and six months after surgery, the labial bone defect was significant improved (P<0.05), but the height of the alveolar ridge bone didn't increase significantly (P>0.05) while the width of the alveolar ridge bone significant improved (P>0.05). CONCLUSIONS: This minimally invasive technique can achieve good clinical results for not only intact labial alveolar ridge bones but also good bone grafts, which is beneficial to implantation and prosthetic aesthetics. The long-term outcome needs to be observed.

[Clinical observation of assisted occlusion therapy for repositioning temporomandibular joint discs].

PURPOSE: To investigate the occurrence of postoperative malocclusion of patients with temporomandibular joint disc repositioning and the necessity of postoperative orthodontic treatment. METHODS: One hundred and eight patients who received temporomandibular joint disc repositioning from 2010.10 to 2015.10 were selected in this study. The patients' occlusion was recorded preoperatively and postoperatively. All patients received functional appliance or orthodontic treatment after surgery. The occlusion and the relative position of the articular disc and condyle were evaluated at regular follow-up. RESULTS: Postoperative malocclusion occurred in all patients. The use of functional appliance for 3-6 months may decrease the proportion of malocclusion. After orthodontic treatment, all patients had a complete recovery of malocclusion and remained good articular disc and condyle relationship during long term follow-up. CONCLUSIONS: Postoperatively malocclusion may occur after temporomandibular joint disc repositioning, and the use of functional appliance and orthodontic treatment are strongly recommended to retain good articular disc and condyle relationship.