Validation of the "obturator functioning scale" for Chinese-speaking patients with obturator prostheses after cancer-related maxillectomy.

Objective: The Obturator Functioning Scale (OFS) is a scale without formal measures of validity in any language. This study aimed to translate and adapt the OFS from English to Chinese and check its reliability and validity in Chinese-speaking patients with obturator prostheses after cancer-related maxillectomy. Methods: The 15-item Chinese preversion of the OFS was completed by 133 patients in three tertiary stomatological hospitals. Of these, 41 completed it again one week after the first measurement. The patients also completed the Chinese version of the University of Washington quality of life scale (UW-QOL, Version 4). Results: Item 12 ("upper lip feels numb") was deleted to achieve a better statistical fit. The 14-item Chinese version of the OFS (OFS-Ch) demonstrated high internal consistency (Cronbach's alpha = 0.908). The test-retest reliability coefficients for most items exceeded 0.90, indicating substantial reproducibility. Confirmatory factor analysis found that the scale consisted of three correlated factors: 1) eating (four items), 2) speech (five items), and 3) other problems (five items). This explained 70.2 % of the total variance using exploratory factor analysis. The scale was significantly convergent and discriminant and could validly discriminate between patients with Brown I and IId maxillary defects. Conclusions: Our results showed that the OFS-Ch scale is a valid tool for evaluating oral dysfunction and satisfaction with appearance for patients with the obturator prosthesis and identifying those at risk of poor obturator function in clinical settings.

Sesquiterpenes from Chloranthus holostegius with anti-inflammatory activities.

The phytoconstituents of the whole plants of Chloranthus holostegius were investigated. As a result, thirteen undescribed sesquiterpenes (chloranholosins A-M, 1-13), including ten acorane-type sesquiterpenes (1-10), one germacrene-type sesquiterpene (11), and two lindenane-type sesquiterpenes (12-13), together with fifteen known sesquiterpenes were isolated. Their structures and absolute configurations were elucidated by a comprehensive method including the spectroscopic data, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction. Chloranholosin L (12) was elucidated as a rare lindenane-type sesquiterpene featuring 14α-Me and 5-OH moieties. And chloranholosin M (13) was the first lindenane-type sesquiterpene possessing ß-cyclopropane, 14α-Me, and 5ß-H configuration from the family Chloranthaceae. Furthermore, twelve new isolates and some known sesquiterpenes were evaluated for their inhibitory activity against LPS-induced nitric oxide (NO) production in RAW 264.7 macrophage cells. Among them, compounds 12, 16, and 23 showed comparable inhibitory activity to that of the positive control, with IC50 values of 47.9, 41.5, and 48.3 µM, respectively.

Cinobufotalin prevents bone loss induced by ovariectomy in mice through the BMPs/SMAD and Wnt/ß-catenin signaling pathways.

BACKGROUND: Osteoporosis is a chronic bone disease characterized by bone loss and decreased bone strength. However, current anti-resorptive drugs carry a risk of various complications. The deep learning-based efficacy prediction system (DLEPS) is a forecasting tool that can effectively compete in drug screening and prediction based on gene expression changes. This study aimed to explore the protective effect and potential mechanisms of cinobufotalin (CB), a traditional Chinese medicine (TCM), on bone loss. METHODS: DLEPS was employed for screening anti-osteoporotic agents according to gene profile changes in primary osteoporosis. Micro-CT, histological and morphological analysis were applied for the bone protective detection of CB, and the osteogenic differentiation/function in human bone marrow mesenchymal stem cells (hBMMSCs) were also investigated. The underlying mechanism was verified using qRT-PCR, Western blot (WB), immunofluorescence (IF), etc. RESULTS: A safe concentration (0.25 mg/kg in vivo, 0.05 µM in vitro) of CB could effectively preserve bone mass in estrogen deficiency-induced bone loss and promote osteogenic differentiation/function of hBMMSCs. Both BMPs/SMAD and Wnt/ß-catenin signaling pathways participated in CB-induced osteogenic differentiation, further regulating the expression of osteogenesis-associated factors, and ultimately promoting osteogenesis. CONCLUSION: Our study demonstrated that CB could significantly reverse estrogen deficiency-induced bone loss, further promoting osteogenic differentiation/function of hBMMSCs, with BMPs/SMAD and Wnt/ß-catenin signaling pathways involved.

Synthesis and Biological Activity Evaluation of 2-Cyanopyrrole Derivatives as Potential Tyrosinase Inhibitors.

In order to find potential inhibitors of tyrosinase, two series of pyrrole derivatives A (1-17) and B (1-8) were synthesized and screened for their inhibitory activities on tyrosinase. Most of the 2-cyanopyrrole derivatives exhibited effective inhibitory activities. In particular, A12 exhibited the strongest inhibitory activities, with the IC50 values of 0.97 µM, which is ∼30 times stronger than the reference inhibitor kojic acid (IC50: 28.72 µM). The inhibitory mechanism analysis results revealed that A12 was a reversible and mixed-type inhibitor. Molecular docking experiments clarified the interaction between A12 with tyrosinase. Furthermore, A12 (100 µM) presented effective inhibitory effect on tyrosinase in B16 melanoma cells with inhibition of 33.48%, which was equivalent to that of Kojic acid (39.81%). Accordingly, compound A12 may serve as the lead structure for the further design of potent tyrosinase inhibitors. Molecular docking studies confirmed the interaction between the compound and tyrosinase.

Quality of life in patients with cancer-related Brown IIb maxillary defect: A comparison between conventional obturation rehabilitation and submental flap reconstruction.

OBJECTIVES: The purpose of this retrospective study was to compare the differences in quality of life (QOL) outcomes between the conventional obturator prostheses (COP) and the pedicled submental artery island flap (SAIF) in the reconstruction of Brown IIb maxillary defects. MATERIALS AND METHODS: The QOL of 116 eligible patients who had a lapse ≥ 12 months after the cancer-related maxilla ablation was evaluated by the University of Washington quality of life scale (UW-QOL), Performance Status Scale for Head and Neck (PSS-HN), and Obturator Functioning Scale (OFS). RESULTS: Patients in the SAIF group reported statistically and clinically significant higher overall QOL scores but lower chewing scores in the UW-QOL scale when compared with those in the COP group (P < 0.05). Clinically significantly higher scores were also observed in the recreation and anxiety domains in the UW-QOL scale for the SAIF group, but there was no statistical significances. The COP group reported more complaints about the nasal leakage when swallowing and the shape of the upper lip, and had a stronger willingness to avoid family or social events in the OFS (P < 0.05). CONCLUSIONS: For patients with Brown IIb defects, SAIF reconstruction can achieve reduced nasal leakage when swallowing, improved upper-lip contour, increased social activity, and superior overall QOL than COP. The inferior chewing function in the SAIF group indicated the need for dental rehabilitation with a conventional denture or osseointegrated implants.

Luteolin relieves lung cancer-induced bone pain by inhibiting NLRP3 inflammasomes and glial activation in the spinal dorsal horn in mice.

BACKGROUND: Bone cancer pain (BCP) is one of the most severe complications in cancer patients. However, the pharmacological therapeutic approaches are limited. Luteolin, a major component of flavones, is widely distributed in plants and plays a critical role in the antinociceptive effects, but whether luteolin could alleviate cancer pain and its underlying mechanisms are not known. HYPOTHESIS/PURPOSE: This study investigated the molecular mechanisms by which luteolin reduced BCP. METHODS: Behavioral, pharmacological, immunohistochemical, and biochemical approaches were used to investigate the effect of luteolin on BCP. RESULTS: Luteolin treatment ameliorated Lewis lung cancer (LLC)-induced bone pain in mice in a dose-dependent manner. Luteolin treatment could inhibit the activation of neurons, glial cells, and NOD-like receptor protein 3 (NLRP3) inflammasomes in the dorsal spinal cord in the BCP mouse model. Furthermore, phosphorylated p-38 mitogen-activated protein kinase (MAPK) in the spinal dorsal horn (SDH) was suppressed by luteolin treatment that could influence the analgesic and glial inhibition effects of luteolin. CONCLUSION: Our results demonstrated that luteolin inhibited neuroinflammation by obstructing glial cell and NLRP3 inflammasome activation via modulating p38 MAPK activity in SDH, ultimately improving LLC-induced BCP.

The Creation of an Experimental Data Set Containing Coronal Section Images of a Human Head.

OBJECTIVES: The aim of the research is to create an experimental data set of coronal section images of a human head. METHODS: The head of a 49-year-old male cadaver was scanned by computed tomography (CT), then perfused with a green filling material via the bilateral common carotid artery, before being frozen and embedded. The head was sectioned along the coronal plane by a computer-controlled 5520 engraving and milling machine, capable of either 0.03-mm or 0.06-mm interspacing. All images were captured with a Canon 5D-Mk III digital camera. RESULTS: A total of 3854 section images were obtained, each with a resolution of 5760 × 3840 pixels. The number of section images at 0.03- and 0.06-mm interspacing were 1437 and 2417, respectively. All the images were stored in JPG and RAW formats. The image size of each RAW format was about 24.5 MB, whereas for JPG format, the equivalent size was about 5.9 MB. All the RAW and JPG images together occupied 117.35 GB of disk space. CONCLUSIONS: The interspacing of this data set section was thinner than those of any comparable studies, and the image resolution was higher, too. This data set was also the first to take coronal sections of the human head. The data set contains image information from the smallest structures within the human head and can satisfy the needs of future developments and applications, such as the virtual operation training systems for otolaryngology, ophthalmology, stomatology, and neurosurgery, and help develop medical teaching software and maps.

Regular Aerobic Exercise Attenuates Pain and Anxiety in Mice by Restoring Serotonin-Modulated Synaptic Plasticity in the Anterior Cingulate Cortex.

PURPOSE: Clinical studies found that regular aerobic exercise has analgesic and antianxiety effects; however, the underlying neural mechanisms remain unclear. Multiple studies have suggested that regular aerobic exercise may exert brain-protective effects by promoting the release of serotonin, which may be a pain modulator. Anterior cingulate cortex (ACC) is a key brain area for pain information processing, receiving dense serotonergic innervation. As a result, we hypothesized that exercise may increase the release of serotonin in the ACC, thus improving pain and anxiety behaviors. METHODS: Integrative methods were used, including behavioral, electrophysiological, pharmacological, biochemical, and genetic approaches, to explore the effects of regular aerobic exercise and the underlying neural mechanisms. RESULTS: Regular aerobic exercise in the form of voluntary wheel running for 30 min daily for 15 d showed significant effectiveness in relieving pain and concomitant anxiety in complete Freund's adjuvant-induced chronic inflammation pain models. c-Fos staining and multielectrode array recordings revealed alterations in neuronal activities and synaptic plasticity in the ACC. Moreover, systemic pharmacological treatment with 4-chloro-dl-phenylalanine (PCPA) to deplete endogenous serotonin and local delivery of serotonin to the ACC revealed that exercise-related serotonin release in the ACC bidirectionally modulates pain sensitization and anxiety behaviors by modulating synaptic plasticity in the ACC. Furthermore, we found that 5-HT1A and 5-HT7 receptors mediated the serotonin modulation effects under conditions of regular aerobic exercise through local infusion of a selective antagonist and shRNA in the ACC. CONCLUSIONS: Our results reveal that regular aerobic exercise can increase serotonin release and modulate synaptic plasticity in the ACC, ultimately improving pain and concomitant anxiety behaviors through the functions of the 5-HT1A and 5-HT7 receptors.

Clinical Evaluation of 532-nm Green Laser on Dentin Hypersensitivity: A Randomized Double-Blind Clinical Trial.

Objective: The aim of this study was to evaluate the therapies of low-level green laser and chemical desensitizer in the treatment of dentin hypersensitivity (DH). Methods: Forty-eight patients with 96 sensitive teeth were invited to participate in this clinical trial and were randomly divided into three groups. One group was treated with low-level green laser, the second group was treated with desensitizer [sodium fluoride (NaF)], and the third group acted as the placebo group and was treated with distilled water and placebo laser. The wavelength of green laser was 532 nm and the irradiance was 15 J/cm2 per treatment site. Hypersensitivity was assessed by visual analogue scale (VAS) according to cold test and probing at baseline. Immediately, 2 weeks, and 3 months after the application of green laser, NaF, and placebo, the participants' sensitivity level was accessed by new VAS analysis. Results: Forty-five patients with 90 teeth (n = 15 patients/group; 30 teeth/group) were followed up for 2 weeks and 3 months after treatment. There were significant differences in VAS scores between the placebo group and intervention group (green laser group and NaF group; analysis of variance, p < 0.05) at all three time points. The mean pain scores in DH reduced significantly immediately after treatment in the green laser group and NaF group when stimulated by cold and probing, whereas no significant difference was observed with these two therapies after 2 weeks (p > 0.05). After 3 months, mean VAS scores of the NaF group were higher than those of the green laser group (p < 0.01). Conclusions: Therefore, the green laser displayed similar effectiveness as NaF in treatment of DH and could be a promising new therapy to reduce DH.

Clinical application and research progress of digital complete denture.

Digital technologies use high-precision three-dimensional scanning, intelligence-aided design software, and multi-axis numerical control milling or 3D printing, which can produce restorations with reliable precision and suitable function. However, the development of digital technologies in the field of complete denture restoration has been slow due to the complexity of prosthesis. This review article introduces the current research status and clinical applications of digital complete dentures in prosthodontic clinics and dental laboratories to provide beneficial references to prosthodontists and dental technicians.

TRIB3 Promotes Osteogenic Differentiation of Human Adipose-derived Mesenchymal Stem Cells Levelled by Post-transcriptional Regulation of miR-24-3p.

OBJECTIVE: To explore the effect of TRIB3 on the osteogenic differentiation of human adipose-derived mesenchymal stem cells (hASCs) and reveal the potential role of TRIB3 in bone regeneration. METHODS: TRIB3-knockdown and TRIB3-overexpression hASCs were used to explore the effect of TRIB3 on osteogenic differentiation by alkaline phosphatase (ALP) staining, alizarin red S (ARS) staining, quantitative real-time polymerase chain reaction (qRT-PCR) and heterotopic bone formation. The regulation of miR-24-3p on TRIB3 was detected by qRT-PCR and western blot. Ribonucleic acid (RNA) sequencing was performed to investigate the downstream regulatory network of TRIB3. RESULTS: TRIB3 promoted the osteogenic differentiation of hASCs both in vitro and in vivo. This process was regulated epigenetically by the post-transcriptional regulation of miR-24-3p, which could bind directly to the three prime untranslated region (3'UTR) of TRIB3 and inhibit TRIB3 expression. The downstream regulatory network of TRIB3-mediated osteogenic differentiation was related to calcium ion binding and cell metabolism, extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) and nuclear factor-κB (NF-κB) signalling pathways. CONCLUSION: TRIB3 is a promising therapeutic target for hASC-based bone tissue engineering and the epigenetic regulation of TRIB3 through miR-24-3p permits regulatory controllability, thus promoting osteogenesis through an important metabolic target while obtaining a safe and controllable effect via post-transcriptional epigenetic regulation.

Musculoskeletal Pain during the Menopausal Transition: A Systematic Review and Meta-Analysis.

Musculoskeletal pain (MSP) is one of the most severe complaints in women undergoing menopause. The prevalence of MSP varied when taking the menopausal state and age factor into consideration. This study investigated the prevalence of MSP in perimenopausal women and its association with menopausal state. The MEDLINE, Embase, Web of Science, and PubMed databases were searched from inception to July 2020, and 16 studies were retrieved for the current meta-analysis. The primary outcome measure was the MSP Odds Ratio (OR). The estimated overall prevalence of MSP among perimenopausal women was 71% (4144 out of 5836, 95% confidence interval (CI): 64%-78%). Perimenopausal women demonstrated a higher risk for MSP than premenopausal ones (OR: 1.63, 95% CI: 1.35-1.96, P = 0.008, I 2 = 59.7%), but similar to that in postmenopausal ones (OR: 1.07, 95% CI: 0.95-1.20, P = 0.316, I 2 = 13.4%). The postmenopausal women were at a higher risk of moderate/severe MSP than the premenopausal ones (OR: 1.45, 95% CI: 1.21-1.75, P = 0.302, I 2 = 16.5%) or the perimenopausal ones (OR: 1.40, 95% CI: 1.09-1.79, P = 0.106, I 2 = 55.4%). In conclusion, the perimenopause is a state during which women are particularly predisposed to develop MSP. As to moderate to severe degrees of MSP, the odds increase linearly with age, from premenopause to peri- and then to postmenopause.

Vitamin D receptor Bsm I polymorphism and osteoporosis risk in postmenopausal women: a meta-analysis from 42 studies.

OBJECTIVE: This study aimed to quantitatively summarize the evidence for VDR BsmI gene polymorphism and osteoporosis risk in postmenopausal women. MATERIALS AND METHODS: The PubMed, EMBASE, Weipu, CNKI, and Wanfang databases were searched for eligible studies. Case-control studies containing available genotype frequencies of B/b were chosen, and odds ratio (OR) with 95% confidence interval (CI) was used to assess the strength of this association. RESULTS: 4485 osteoporosis and 5490 controls were identified in our meta-analysis. In the stratified analysis, a significant association was observed between VDR BsmI gene polymorphism and osteoporosis susceptibility in Caucasians (additive model: OR = 0.809, 95% CI 0.678~0.965, p = 0.019; recessive model: OR = 0.736, 95% CI 0.568~0.955, p = 0.021; and co-dominant model: bb vs. BB OR = 0.701, 95% CI 0.511~0.962 p = 0.028), and we failed to find any significant relationship in Asians. CONCLUSION: The present meta-analysis suggests that VDR BsmI genotype is associated with increased risk of postmenopausal osteoporosis in Caucasians but not in Asians. To draw comprehensive and true conclusions, further prospective studies with larger numbers of participants worldwide are needed to examine associations between VDR BsmI polymorphism and osteoporosis in postmenopausal women.

Preventive and Control Measures for the Coronavirus Pandemic in Clinical Dentistry.

A severe public health crisis has been declared worldwide since coronavirus disease 2019 (COVID-19) was classified as a pandemic of acute respiratory infectious disease by the World Health Organisation (WHO). China has taken strict measures to curb the spread of the disease to save lives, and has managed to control the outbreak. COVID-19 is mainly transmitted through respiratory droplets and close physical contact, so it is challenging to prevent nosocomial infection and possible spread during dental treatment. Since the initial phase of the COVID-19 outbreak, a disease prevention and control strategy based on the new concept of population risk classification and rational use of personal protective equipment has been implemented by the Peking University Hospital of Stomatology. Nosocomial infection prevention and control concepts and measures relating to dental diagnosis and treatment are critically checked in the hospital. Our experiences in handling this situation are shared here and may have wide-ranging implications for infection prevention and control (IPC) for COVID-19 in dental practices worldwide.

Clinical Efficiency and Patient Preference of Immediate Digital Impression after Implant Placement for Single Implant-Supported Crown.

OBJECTIVE: To evaluate the time efficiency and patient preference of three impression techniques by comparing immediate digital impression performed directly after implantation with regular digital impression and conventional implant impression performed 3 months after implantation. METHODS: Twenty consecutive patients with a missing single molar or premolar who received implant treatment were recruited into this prospective self-controlled clinical trial. Three different impression techniques were performed after implant surgery on all the participants: An intraoral scanning (IOS) impression performed immediately after implant placement (immediate digital impression) was compared with a regular digital impression and a classic polyether impression (conventional implant impression) performed 3 months after implant surgery. The operating time of each impression technique was recorded. Patients were asked to complete a visual analogue scale (VAS) questionnaire on their perception of the three techniques to describe their satisfaction and preference. Statistical analyses were performed with the Wilcoxon signed rank test. RESULTS: The clinical time of the immediate digital impression (10.97 ± 2.1 min) was significantly shorter than that of the conventional implant impression (14.45 ± 3.0 min) (P < 0.05). The mean time of the immediate digital impression (10.97 ± 2.1 min) was statistically the same as that of the regular digital impression (10.23 ± 2.7 min) (P > 0.05). Participants' subjective evaluation indicated higher satisfaction with the immediate digital impression than with the regular digital impression and the conventional implant impression. CONCLUSION: The immediate digital impression was more efficient than the conventional implant impression and had the same efficiency as the regular digital impression. Among the three impression techniques, the participants showed higher satisfaction with the immediate digital impression.

Origin and Clinical Applications of Neural Crest-Derived Dental Stem Cells.

Over the past few decades, scientific research into neural crest-derived stem cells has progressed rapidly. The migration and differentiation of neural crest-derived stem cells has been an interesting area of research. Stem cells within teeth originating from the embryonic neural crest have attracted increasing attention in clinical and scientific research because they are easy to obtain and have superb stemness. The stem cells within the teeth include dental pulp stem cells (DPSCs), dental follicle stem cells (DFSCs), stem cells from apical papilla (SCAPs), stem cells from human exfoliated deciduous teeth (SHEDs), and periodontal ligament stem cells (PDLSCs). To date, there have been several interesting studies focusing on dental pulp regeneration, neural regeneration and the revascularization for therapeutic applications.

Transcriptomics and Functional Analysis of Graphene-Guided Osteogenic Differentiation of Mesenchymal Stem Cells.

OBJECTIVE: To explore graphene's effects on the gene expression profile of mesenchymal stem cells, and to reveal the mechanisms of graphene-guided osteogenic differentiation. METHODS: Human adipose-derived mesenchymal stem cells (hASCs) were cultured on single-layer graphene-coated titanium disks or titanium disks in proliferation medium (control) or osteoinduction medium for 7 days before RNA extraction. After library construction and RNA sequencing, identification of differentially expressed genes was performed through Limma package of R platform, with a cut-off value of log fold change (logFC) > = |1|. Pathway and Gene ontology (GO) analyses were conducted on DAVID Bioinformatics Resources 6.8 (NIAID/NIH). Network analyses were performed by the Ingenuity Pathways Analysis (IPA). RESULTS: Signalling pathway analysis revealed the top five pathways - cytokine-cytokine receptor interaction, neuroactive-ligand receptor interaction, calcium signalling pathway, PI3K-Akt signalling pathway and cell adhesion molecules. GO analyses demonstrated significant changes on cell adhesion, calcium signalling, and epigenetic regulation. IPA network analyses demonstrated that inflammation-related pathways were influenced by graphene, while the downstream factors of histone H3 and H4 were also altered especially under the existence of osteoinduction medium. CONCLUSION: Graphene promotes osteogenic differentiation of hASCs mainly by influencing cell adhesion, cytokine-cytokine receptor interactions, inflammatory responses, and potentially influences histone H3 and H4 through epigenetic regulation.

[Promoted role of bone morphogenetic protein 2/7 heterodimer in the osteogenic differentiation of human adipose-derived stem cells].

OBJECTIVE: To investigate the role of bone morphogenetic protein 2/7 heterodimer (BMP-2/7) in the osteogenesis of human adipose-derived stem cells (hASCs). METHODS: hASCs were exposed to three different treatments in vitro: osteogenic medium with 150 µg/L BMP-2/7 (experimental group), osteogenic medium alone (OM group) and proliferation medium (PM group). After 1, 4 and 7 days of osteogenic induction, the amount of cellular DNA was measured to investigate the cytotoxicity. After 7 and 14 days, alkaline phosphatase (ALP) staining and quantification were performed to test the activity of ALP. After 21 and 28 days, the calcification deposition was determined by Alizarin Red S (ARS) staining and quantification. The expressions of the osteoblast-related genes were tested on days 1, 4, 7 and 14. In the in vivo study, 6 nude mice were used and 4 groups were set and implanted subcutaneously into the back of nude mice: (1) ß-TCP scaffold only (scaffold control group); (2) ß-TCP scaffold with hASCs cultured by PM in vitro for 1 week (PM control group); (3) ß-TCP scaffold with hASCs cultured by OM in vitro for 1 week (OM control group); (4) ß-TCP scaffold with hASCs cultured by OM with 150 µg/L BMP-2/7 in vitro for 1 week (test group). After 4 weeks of implantation, histological staining was performed to evaluate the in vivo osteogenesis of hASCs. RESULTS: After induction for 1 day, there was no significant difference between the experimental group and the PM group on the cellular DNA content (P>0.05). After 4 days, the cellular DNA content increased under the stimulation of BMP-2/7 (P<0.05). On day 7, there was no significant difference among the three groups (P>0.05). ALP activity was higher by the induction of BMP-2/7 than in OM alone and PM (P<0.05). More mineralization deposition and more expressions of osteoblast-related genes such as Runx2, ALP, COL-1A1 and OC were determined in the experimental group at different time points (P<0.05). HE staining showed that, in the test group and OM control group, the extracellular matrix (ECM) with eosinophilic staining were observed around hASCs, and newly-formed bone-like tissues could be found in ECM around the scaffold materials. Moreover, compared with the OM control group, more bone-like tissues could be observed in ECM with typical structure of bone tissue in the test group. Masson's trichrome staining showed that more expression of collagen could be observed in ECM in the test group compared with the other groups. There was small amount of expression of collagen in the OM and PM control groups. No obvious positive results were found in the scaffold group. CONCLUSION: BMP-2/7 heterodimer plays a significant role in the osteogenesis of hASCs and is able to enhance the osteogenic differentiation of hASCs in vitro and in vivo.

[Constructing 3-dimensional colorized digital dental model assisted by digital photography].

OBJECTIVE: To explore a method of constructing universal 3-dimensional (3D) colorized digital dental model which can be displayed and edited in common 3D software (such as Geomagic series), in order to improve the visual effect of digital dental model in 3D software. METHODS: The morphological data of teeth and gingivae were obtained by intra-oral scanning system (3Shape TRIOS), constructing 3D digital dental models. The 3D digital dental models were exported as STL files. Meanwhile, referring to the accredited photography guide of American Academy of Cosmetic Dentistry (AACD), five selected digital photographs of patients'teeth and gingivae were taken by digital single lens reflex camera (DSLR) with the same exposure parameters (except occlusal views) to capture the color data. In Geomagic Studio 2013, after STL file of 3D digital dental model being imported, digital photographs were projected on 3D digital dental model with corresponding position and angle. The junctions of different photos were carefully trimmed to get continuous and natural color transitions. Then the 3D colorized digital dental model was constructed, which was exported as OBJ file or WRP file which was a special file for software of Geomagic series. For the purpose of evaluating the visual effect of the 3D colorized digital model, a rating scale on color simulation effect in views of patients'evaluation was used. Sixteen patients were recruited and their scores on colored and non-colored digital dental models were recorded. The data were analyzed using McNemar-Bowker test in SPSS 20. RESULTS: Universal 3D colorized digital dental model with better color simulation was constructed based on intra-oral scanning and digital photography. For clinical application, the 3D colorized digital dental models, combined with 3D face images, were introduced into 3D smile design of aesthetic rehabilitation, which could improve the patients' cognition for the esthetic digital design and virtual prosthetic effect. CONCLUSION: Universal 3D colorized digital dental model with better color simulation can be constructed assisted by 3D dental scanning system and digital photography. In clinical practice, the communication between dentist and patients could be improved assisted by the better visual perception since the colorized 3D digital dental models with better color simulation effect.

[Establishing a luciferase reporter system to evaluate osteogenic differentiation potential of human adipose-derived stem cells].

OBJECTIVE: Human adipose-derived stem cells (hASCs) are a highly attractive source in bone tissue engineering. To generate a luciferase reporter system that could be used to quantitatively and rapidly examine osteogenic differentiation potential of human adipose-derived stem cells (hASCs) in vitro, and eventually make it possible to monitor the osteogenic differentiation of transplanted cells in vivo. METHODS: The genomic DNA harboring promotor regions of osteocalcin and DNA sequences encoding luciferase genes were amplified by PCR and cloned into the pLVX-pTRE-puro vector to generate the OC(pro)-Luc-Puro construct. Then, the OC(pro)-Luc-Puro construct together with three assistant vectors: pMDLg/pRRE, pRSV-REV, and pVSVG, were transiently transfected into HEK293T cells followed by viral supernatants collection, filtration and concentration. Next, the hASCs stably expressing luciferase reporter gene driven by osteocalcin promotor were created with the lentivirus carrying OC(pro)-Luc-Puro cassette under puromycin selection. The OC(pro)-Luc-hASCs were then cultured in the absence or presence of osteogenic differentiation medium. On the 7th and 14th days, after osteogenic induction, cellular extracts were collected and analyzed by luciferase reporter assay. Meanwhile, alizarin red staining and quantification as well as quantitative reverse transcription PCR (qRT-PCR) analysis of osteogenic associated genes osteocalcin (OC), runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP) were used to assess the osteogenic differentiation ability of OC(pro)-Luc-hASCs. RESULTS: OC(pro)-Luc-Puro plasmid and OCpro-Luc-hASCs were successfully generated. On the 7th and 14th days after osteogenic induction, the luciferase activity of the cellular extracts from OC(pro)-Luc-hASCs was dramatically increased. Consistently, the extracellular matrix mineralization, as shown by Alizarin red S (ARS) staining and quantification was also markedly intensified and a marked increase of the mRNA expression levels of OC, Runx2 and ALP, although to variable extent, was observed upon osteogenic differentiation. These results indicated that the observations from traditional experiments examining hASCs osteogenic differentiation were largely in agreement with that of our luciferase reporter assay in OC(pro)-Luc-hASCs. CONCLUSION: We established a luciferase reporter system that could be used to rapidly, quantitatively and specifically determine osteogenic differentiation ability of hASCs. Comparing with the traditional time-consuming methods, the system we generated here was highly effective. This system not only can be used to examine ostogenic differentiation of hASCs in a high throughput manner, but also provides a way to monitor ostogenic differentiation of cells in vivo.