[Show abstract][Hide abstract] ABSTRACT: Periodontitis is a kind of chronic inflammatory disease that affects the tooth-supporting tissues. ET-1 is related to periodontitis and involved in the regulation of cytokines, but the mechanisms remain unclear. The aim of this study is to investigate how ET-1 affects proinflammatory cytokine expression and differentiation in human periodontal ligament stem cells (PDLSCs). PDLSCs were isolated from the periodontal ligament tissues of periodontitis patients and then treated with ET-1 (1, 10, or 100 nM) for 12 h, 24 h, or 72 h. The osteogenic potential of PDLSCs was tested using ALP staining. TNF-
, and IL-6 levels were evaluated by ELISA and western blot. Runx2, OCN, and COL1 mRNA and western levels were detected by RT-PCR and western blot, respectively. To examine the signaling pathways and molecular mechanisms involved in ET-1-mediated cytokine expression and osteogenic differentiation, ETR pathway, MAPKs pathway, Wnt/
-catenin pathway, and Wnt/Ca
pathway were detected by RT-PCR and western blot, respectively. ET-1 promoted differentiation of PDLSCs into osteoblasts by increasing secretion of TNF-
, and IL-6 in a dose- and time-dependent manner. ET-1 also increased expression of Runx2, OCN, and COL1. ET-1 promotes differentiation of PDLSCs into osteoblasts through ETR, MAPK, and Wnt/
-catenin signaling pathways under inflammatory microenvironment.
Preview · Article · Jan 2016 · Mediators of Inflammation
[Show abstract][Hide abstract] ABSTRACT: Bone is a self-renewing tissue. Bone marrow mesenchymal stromal cells (BMSCs) are located in the adult skeleton and are believed to be involved in the maintenance of skeletal homeostasis throughout life. With increasing age, the ability of the skeleton to repair itself decreases, possibly due to the reduced functional capacity of BMSCs. Recent evidence has suggested the existence of at least two populations of BMSCs with different embryonic origins that cannot be interchanged during stem cell recruitment: craniofacial BMSCs (neural crest origin) and appendicular BMSCs (mesoderm origin). Questions arise as to whether the site-specific characteristics alter the effect of aging on the skeleton. In this study, the effects of biological aging on human BMSCs were compared with BMSCs derived from craniofacial bone vs. BMSCs derived from the appendicular skeleton. The phenotype, proliferation and functional characteristics (osteogenic differentiation, cytokine secretion and bone formation in vivo) of the BMSCs were investigated. The results demonstrated that the proliferative capacity and osteogenic differentiation of the BMSCs decrease significantly with age both in vitro and in vivo. For age-matched groups, the osteogenic differentiation capacity of alveolar BMSCs was higher than that of femoral BMSCs in the middle-aged and old groups while there was no significant difference for the young groups. Compared with old alveolar BMSCs, old femoral BMSCs had a significantly longer population doubling time, a smaller colony-forming population and less bone formation in vivo while there was no significant difference for the young and middle-aged groups. Distinct differences in the expression of cytokine factors were also found. In conclusion, human BMSCs display an age-related decrease in functional capacity, and embryonic origins may play a critical role in mediating the aging rate of BMSCs. These data provide novel insights into the skeletal site-specific characteristics of aged BMSCs.
No preview · Article · Dec 2015 · Rejuvenation Research
[Show abstract][Hide abstract] ABSTRACT: Introduction:
This study was to investigate whether there was statistical difference between the bilateral temporomandibular joint (TMJ) in patients with unilateral TMJ pain or joint sounds, using cone beam computed tomography (CBCT).
TMJ CBCT images of 123 cases were used to preliminarily determine the indicators suitable for the measuring method. TMJ CBCT image reconstruction was performed and 19 indicators were measured. Thirty-six cases without TMJ complaint served as controls. The comparison of bilateral TMJs was analyzed by paired t-test to find out the indicators without statistical significance. Twenty-nine patients with unilateral TMJ pain or joint sounds who underwent CBCT at the hospital were enrolled for the comparative study. The measured values were analyzed by paired t-test to determine the indicators with statistical difference.
In the control group, only radius value of bilateral TMJ was different statistically (P < 0.05). In the TMJ complaint group, the vertical 60° joint space of the bilateral TMJ was statistically different (P < 0.05) and the rest of the measured values showed no statistical difference.
In the patients with unilateral TMJ pain or joint sounds, the vertical 60° joint space of the symptomatic side was significantly increased comparing with the asymptomatic side.
No preview · Article · Sep 2015 · International Journal of Clinical and Experimental Medicine
[Show abstract][Hide abstract] ABSTRACT: NIN1/RPN12 binding protein 1 homolog (NOB1) facilitates the maturation of the 20S proteasome and is then degraded by 26S proteasome to complete 26S proteasome biogenesis. It also accompanies the pre-40S ribosomes during nuclear export and is cleaved at D-site of 20S pre-rRNA to form mature 18S rRNA in growing cells. NOB1 was reported to be involved in the development of several types of cancer. However, the role of NOB1 in oral squamous cell carcinoma (OSCC) has not been addressed. In the present study, the expression of NOB1 in 50 OSCC patients with different genders, ages, TNM and pathological grades was detected using immunohistochemistry and western blotting. A loss-of‑function study was carried out by the lentivirus‑mediated siRNA knockdown of NOB1 in the CAL27 and TCA-8113 OSCC cell lines. The results showed that, NOB1 expression increased with pathological grades. In the CAL27 and TCA-8113 cell lines, knockdown of NOB1 in OSCC cells decreased cell proliferation, colony formation, increased cell apoptosis and also induced cell cycle arrest in the S phase. The results suggested that NOB1 is important in OSCC development and serves as a candidate indicator of aggressiveness and a therapeutic target of OSCC.
No preview · Article · Sep 2015 · Oncology Reports
[Show abstract][Hide abstract] ABSTRACT: Objective:
. A novel injectable magnesium/calcium sulfate hemihydrate (Mg/CSH) composite with improved properties was reported here.
Composition, setting time, injectability, compressive strength, and bioactivity in simulated body fluid (SBF) of the Mg/CSH composite were evaluated. Furthermore, the cellular responses of canine bone marrow stromal cells (cBMSCs) and bone formation capacity after the implantation of Mg/CSH in tibia defects of canine were investigated.
Mg/CSH possessed a prolonged setting time and markedly improved injectability and mechanical property (p < 0.05). Mg/CSH samples showed better degradability than CSH in SBF after 21 days of soaking (p < 0.05). Moreover, the degrees of cell attachment, proliferation, and capability of osteogenic differentiation on the Mg/CSH specimens were higher than those on CSH, without significant cytotoxicity and with the increased proliferation index, ALP activity, and expression levels of integrin β1 and Coll I in cBMSCs (p < 0.05). Mg/CSH enhanced the efficiency of new bone formation at the tibia defect area, including the significantly elevated bone mineral density, bone area fraction, and Coll I expression level (p < 0.05).
The results implied that this new injectable bone scaffold exhibited promising prospects for bone repair and had a great potential in bone tissue engineering.
[Show abstract][Hide abstract] ABSTRACT: To analyze angulations of anterior teeth with reference to the alveolar bone.
Cone-beam computed tomographic (CBCT) images of 105 participants were taken with the same machine (ProMax 3D Max CBCT), showing the intact anterior teeth. The angulations formed between the long axis of the anterior teeth and the alveolus were measured using cross-sectional images. The thicknesses of alveolar bone on different area of root surfaces were also measured.
Maxillary anterior teeth were found to be close to the buccal alveolar surface in apical level with the angulations referred to alveolar bone in center incisor, lateral incisor, and canine were 17.65 ± 6.8, 18.79 ± 7.4, and 23.82 ± 6.96 degrees, respectively. Means of angulations of mandibular anterior teeth were less than 8 degree. The thicknesses of buccal bone at mid-root level in 77% to 90% maxillary anterior teeth were less than 1 mm. The determinations provided high intrarater/interrater reliability.
The application of CBCT is reliable for a complete calculation of angulations and thicknesses in a Chinese Han population. As a result, adequate planning combined with image examination preoperatively would contribute to a favorable outcome.
No preview · Article · Jun 2015 · Implant dentistry
[Show abstract][Hide abstract] ABSTRACT: Free radical hypothesis which is one of the most acknowledged aging theories was developed into oxidative stress hypothesis. Protein carbonylation is by far one of the most widely used markers of protein oxidation. We studied the role of age and gender in protein carbonyl content of saliva and plasma among 273 Chinese healthy subjects (137 females and 136 males aged between 20 and 79) and discussed the correlation between protein carbonyl content of saliva and plasma. Protein carbonyl content of saliva and plasma were, respectively, 2.391 ± 0.639 and 0.838 ± 0.274 nmol/mg. Variations of saliva and plasma different age groups all reached significant differences in both male and female (all p < 0.05) while both saliva and plasma protein carbonyls were found to be significantly correlated with age (r = 0.6582 and r = 0.5176, all p < 0.001). Gender was discovered to be unrelated to saliva and plasma protein carbonyl levels (all p > 0.05). Saliva and plasma protein carbonyls were positively related (r = 0.4405, p < 0.001). Surprisingly, saliva and plasma protein carbonyls/ferric reducing ability of plasma (FRAP) ratios were proved to be significantly correlated with age (r = 0.7796 and r = 0.6938, all p < 0.001) while saliva protein carbonyls/FRAP ratio and plasma protein carbonyls/FRAP ratio were also correlated (r = 0.5573, p < 0.001). We concluded that saliva protein carbonyls seem to be an alternative biomarker of aging while the mechanisms of protein carbonylation and oxidative stress and the relationship between saliva protein carbonyls and diseases need to be further investigated.
[Show abstract][Hide abstract] ABSTRACT: Bone mesenchymal stem cells (BMSCs) have multiple potentials to differentiate into osteoblasts and adipocytes, and methods to enhance their osteogenic differentiation are gaining increasing attention. MicroRNAs are critical regulation factors during the process of the osteogenic induction in BMSCs, and mir-205 has been substantiated to be involved in the osteogenic process, but the underlying mechanisms remain unclear. The purpose of this article is to investigate the role of mir-205 in the osteogenic differentiation of BMSCs. We found that mir-205 expression was down-regulated in a time-dependent manner during BMSC osteo-induction. Inhibition of mir-205 enhanced osteogenic abilities by up-regulating bone sialoprotein (BSP) and osteopontin (OPN) protein levels and increasing alkaline phosphatase (ALP) activity and osteocalcin secretion. Furthermore, we found that mir-205 could regulate protein expression of special AT-rich sequence-binding protein 2 (SATB2) and runt-related transcription factor 2 (Runx2), and over-expression of SATB2 activated Runx2 and reversed the negative effects of mir-205 on osteoblastic differentiation. Furthermore, we examined the extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (p38 MAPK) pathways during osteogenic induction and our data indicates that mir-205 might exert negative functions on the osteogenic differentiation in BMSCs at least partly via altering phosphorylation of ERK and p38 MAPK. These results shed new light on the molecular mechanisms of microRNAs in governing differentiation of BMSCs.
Preview · Article · May 2015 · International Journal of Molecular Sciences
[Show abstract][Hide abstract] ABSTRACT: Diabetes mellitus involves
metabolic changes that can impair bone repair. Bone mesenchymal stem cells (BMSCs) play an important role in bone regeneration. However, the bone regeneration ability of BMSCs is inhibited in high glucose microenvironments. It can be speculated that this effect is due to changes in BMSCs' proliferation and migration ability, because the recruitment of factors with an adequate number of MSCs and the microenvironment around the site of bone injury are required for effective bone repair. Recent genetic evidence has shown that the Cyclin D1 and the CXC receptor 4 (CXCR-4) play important roles in the proliferation and migration of BMSCs. In this study we determined the specific role of glycogen synthase kinase-3β (GSK3β) in the proliferation and migration of BMSCs in high glucose microenvironments. The proliferation and migration ability of BMSCs were suppressed under high glucose conditions. We showed that high glucose activates GSK3β but suppresses CXCR-4, β-catenin, LEF-1, and cyclin D1. Inhibition of GSK3β by LiCl led to increased levels of β-catenin, LEF-1, cyclin D1, and CXCR-4 expression. Our data indicate that GSK3β plays an important role in regulating the proliferation and migration of BMSCs by inhibiting cyclin D1 and CXCR-4 under high glucose conditions.
No preview · Article · Apr 2015 · Journal of Bone and Mineral Metabolism
[Show abstract][Hide abstract] ABSTRACT: An injectable bone cement, nHAC/CSH, which consists of nano-hydroxyapatite/collagen (nHAC) and calcium sulphate hemihydrate (CaSO4.½H2O; CSH) was investigated as a tissue-engineered scaffold material with blood-acquired mesenchymal progenitor cells (BMPCs) as seeding cells. An in vitro study on the cytocompatability of nHAC/CSH and an in vivo study on the ectopic bone formation of nHAC/CSH loaded with dBMPCs were both conducted. The dBMPCs morphology, proliferation, differentiation and apoptosis assays were conducted using the direct contact and extract method. The cells tests exhibited normal growth and bioactive function in vitro. Studies in vivo showed that this injectable tissue engineered bone (ITB) formed bone structure in the heterotopic site of nude mice. These findings indicate that the ITB composed of nHAC/CSH and dBMPCs may represent a useful strategy for clinical reconstruction of irregular bone defects.
No preview · Article · Jan 2015 · Journal of Materials Science Materials in Medicine
[Show abstract][Hide abstract] ABSTRACT: Our previous studies demonstrated that glimepiride enhanced the proliferation and differentiation of osteoblasts and led to activation of the PI3K/Akt pathway. Recent genetic evidence shows that endothelial nitric oxide synthase (eNOS) plays an important role in bone homeostasis. In this study, we further elucidated the roles of eNOS, PI3K and Akt in bone formation by osteoblasts induced by glimepiride in a high glucose microenvironment. We demonstrated that high glucose (16.5 mM) inhibits the osteogenic differentiation potential and proliferation of rat osteoblasts. Glimepiride activated eNOS expression in rat osteoblasts cultured with two different concentrations of glucose. High glucose-induced osteogenic differentiation was significantly enhanced by glimepiride. Down-regulation of PI3K P85 levels by treatment with LY294002 (a PI3K inhibitor) led to suppression of P-eNOS and P-AKT expression levels, which in turn resulted in inhibition of RUNX2, OCN and ALP mRNA expression in osteoblasts induced by glimepiride at both glucose concentrations. ALP activity was partially inhibited by 10 µM LY294002. Taken together, our results demonstrate that glimepiride-induced osteogenic differentiation of osteoblasts occurs via eNOS activation and is dependent on the PI3K/Akt signaling pathway in a high glucose microenvironment.
[Show abstract][Hide abstract] ABSTRACT: Zinc finger protein, X-linked (ZFX) has been identified as a transcriptional factor and is implicated in the development of variant types of cancer. Furthermore, it has been reported that ZFX is essential for the survival and self-renewal of embryonic stem cells. To investigate the involvement of ZFX in squamous cell carcinoma of the tongue, in the present study, we explored the expression of ZFX in clinical specimens from patients with squamous cell carcinoma of the tongue and the correlation between ZFX expression and multiple clinical pathological parameters. We further evaluated the impact of ZFX knockdown on the proliferation, colony formation ability, cell cycle distribution and survival of two human tongue squamous cell carcinoma cell lines to explore its critical role in the development of squamous cell carcinoma of the tongue. Our results showed that ZFX expression was aberrantly higher in samples from patients with squamous cell carcinoma of the tongue and revealed that ZFX expression is positively correlated with tumor grade and stage. Consistent with these findings, we further found that ZFX knockdown impaired cell proliferation and colony formation ability and induced cell apoptosis and cell cycle arrest in two human tongue squamous cell carcinoma cell lines. Our results indicate that ZFX is essential for the development and progression of squamous cell carcinoma of the tongue and represents a potential target for the development of effective therapy.
No preview · Article · Oct 2014 · Oncology Reports
[Show abstract][Hide abstract] ABSTRACT: Aim:
To evaluate the effects of autologous blood vessels and nerves on vascularization.
A dog model of tissue-engineered bone vascularization was established by constructing inferior alveolar neurovascular bundles through the mandibular canal. Sixteen 12-month-old healthy beagles were randomly divided into two groups (n=8). Group A retained inferior alveolar neurovascular bundles, and Group B retained inferior alveolar nerves. Bone marrow mesenchymal stem cells were injected into β-tricalcium phosphate to prepare internal tissue-engineered bone scaffold. A personalized titanium mesh was then prepared by rapid prototyping and fixed by external titanium scaffold. Two dogs in each group were sacrificed on the 30th, 45th, 60th and 90th postoperative days respectively. The bone was visually examined, scanned by CT, and subjected to HE staining, immunohistochemical staining, vascular casting and PCR to detect the changes in osteogenesis and vascularization.
The two groups had similar outcomes in regard to osteogenesis and vascularization (P>0.05): both showed remarkable regenerative capacities.
The model of tissue-engineered bone vascularization is potentially applicable in clinical practice to allow satisfactory osteogenesis and vascularization.
Preview · Article · Oct 2014 · Biomedical papers of the Medical Faculty of the University Palacky, Olomouc, Czechoslovakia
[Show abstract][Hide abstract] ABSTRACT: Background:
The pain caused by orthodontic treatment has been considered as tough problems in orthodontic practice. Danggui-shaoyao-san (DSS) is a traditional Chinese medicine (TCM) prescription which has long been used for pain treatment and possesses antioxidative, cognitive enhancing and antidepressant effects. We raise the hypothesis that DSS exerts analgesic effect for orthodontic pain via inhibiting the activations of neuron and microglia.
DSS was given twice a day from day 5 prior to experimental tooth movement (ETM). Directed face grooming and vacuous chewing movements (VCM) were evaluated. Immunofluorescent histochemistry and Western blot analysis were used to quantify the Iba-1 (microglia activation) and Fos (neuronal activation) expression levels in the trigeminal spinal nucleus caudalis (Vc).
ETM significantly increased directed face grooming and VCM which reached the peak at post-operative day (POD) 1 and gradually decreased to the baseline at POD 7. However, a drastic peak increase of Fos expression in Vc was observed at 4 hours and gradually decreased to baseline at POD 7; while the increased Iba-1 level reached the peak at POD 1 and gradually decreased to baseline at POD 7. Furthermore, pre-treatment with DSS significantly attenuated the ETM induced directed face grooming and VCM as well as the Fos and Iba-1 levels at POD 1.
Treatment with DSS had significant analgesic effects on ETM-induced pain, which was accompanied with inhibition of both neuronal and microglial activation.
No preview · Article · Oct 2014 · Chinese medical journal
[Show abstract][Hide abstract] ABSTRACT: Objective:
To observe the activity change of astrocyte in related nucleus caused by acute pulpitis in rats.
Rat acute pulpitis model was induced by lipopolysaccharides (LPS). And, according to processing time, a total of 30 rats were divided into 5 groups of control, 6, 12, 24 and 48 h. Immunohistochemistry and Western blot were employed to detect the dynamic expression of glial fibrillary acidic protein (GFAP) in spinal nucleus of trigeminal nerve (Vc).
The relative gray value of ipsilateral Vc GFAP expression in experimental groups was 153 ± 11 at 12 h. And it significantly increased versus the control group (100 ± 4)(P < 0.05). The relative gray value of GFAP was 198 ± 12 at 24 h and was 231 ± 9 at 48 h. And they significantly increased versus the control group (both P < 0.01). Their expressions were time-dependent.
In this acute pulpitis model, activated glial cells are probably involved in the processes of pulpitis and hyperalgesia.
No preview · Article · Apr 2014 · Zhonghua yi xue za zhi
[Show abstract][Hide abstract] ABSTRACT: Objective:
To explore the effect of glimepiride on the glucose uptake as well as glucose transporter (GLUT)-1 and GLUT-3 expression levels of rat mandibular osteoblasts in hyperglycemia.
Primary osteoblasts were isolated and cultured. Then, the cells were placed in an osteogenic medium containing two glucose concentrations (5.5 and 16.5 mmol X L(-1)), with or without glimepiride (10 micromol x L(-1)). Glucose uptake was determined by employing 18F-deoxyglucose (18F-FDG) in the cells, and GLUT-1 and GLUT-3 expression levels were evaluated by Western blot analysis.
Glucose at 16.5 mmol x L(-1) significantly inhibited 18F-FDG uptake and downregulated GLUT-3 protein expression in osteoblasts. Hyperglycemia increased GLUT-1 protein expression. Glimepiride significantly increased glucose uptake and upregulated GLUT-1 and GLUT-3.
Glimepiride enhance the glucose transporter in rat osteoblasts at two different glucose concentrations.
No preview · Article · Apr 2014 · Hua xi kou qiang yi xue za zhi = Huaxi kouqiang yixue zazhi = West China journal of stomatology
[Show abstract][Hide abstract] ABSTRACT: Objective
To investigate the antiosteoporotic effects of L-Arginine on ovariectomied rats.
Forty twelve-week-old female Sprague-Dawley rats were randomly divided into four groups of bilaterally ovariectomy and one group of Sham animals, with 8 animals in each group. Twelve additional weeks elapsed before initiation of the treatment with L-Arginine in order to induce significant bone loss in the ovariectomied animals. A 4-week daily treatment (5 days a week, Monday–Friday) at doses of 5 mg/kg/d, 10 mg/kg/d, 20 mg/kg/d of L-Arginine or vehicle only was administered by s.c. injection to the ovariectomied groups respectively. At the end of the treatment period, blood samples from all animals were collected for biochemical analysis. Micro-computerized tomography analysis, histopathological study and biomechanical test were performed on the femur of each animal.
Serum alkaline phosphatase and osteocalcin were reduced in ovariectomied rats after the administration of different dosage of L-Arginine. L-Arginine of 10 mg/kg showed the most significant effect. Micro-computerized tomography 3-D images revealed that the increase of bone mass in 5 mg/kg and 10 mg/kg groups were significant greater than that in Sham group. The biomechanical parameters of femur were improved significantly in L-Arginine 10 mg/kg group when compared to untreated ovariectomied rats.
L-Arginine (10 mg/kg) contributes significantly to the treatment of the bone loss induced by ovariectomy on rats, demonstrated by increased bone mass, improved bone structure and recovery of bone biomechanical activity.
No preview · Article · Apr 2014 · Biomedicine and Aging Pathology
[Show abstract][Hide abstract] ABSTRACT: To study the relationship between vascular endothelial growth factor (VEGF) and formation and repair of engineering bone, second-generation bone marrow stromal cells (BMSCs) of New Zealand white rabbits that were separated in vitro were transfected with VEGF 165 gene vectors by adenovirus to detect gene expressions. Transfected BMSCs and β-tricalcium phosphate material were complexed and implanted at the femoral injury sites of the study group (n = 12), and the control group (n = 12) were implanted with engineering bones that were not transfected with VEGF. Femoral recoveries of the two groups were observed on the 15th, 30th, 45th and 60th days, and their vascularization and ossification statuses were observed by immunohistochemical methods. The BMSCs transfected with VEGF highly expressed VEGF genes and excreted VEGF. The two groups both experienced increased vascularization and bone volume after implantation (t = 7.92, P<0.05), and the increases of the study group were significantly higher than those of the control group (t = 6.92, P<0.05). VEGF is clinically applicable because it can accelerate the formation and repair of engineering bone by promoting vascularization and ossification.
[Show abstract][Hide abstract] ABSTRACT: Osteocyte generation can be used in bone defect repair; the generation efficiency needs to be further improved. This study aims to evaluate the role of ubiquitin A20 (A20) in facilitating the expression of osteocalcin in adipose-derived stem cells (ADSCs). In this study, adipose tissue was obtained from 10 healthy human subjects; ADSCs were isolated from the adipose samples. The ADSCs were transfected with core binding factor alpha 1 (Cbfa1) and/or insulin-like growth factor-1 receptor (IGF-1R). Expression of osteocalcin, A20 in ADSCs was assessed by quantitative RT-PCR (qRT-PCR) and Western blotting. Apoptosis of ADSCs was analyzed by flow cytometry. The results showed that after the gene transfection and stimulation of insulin, the ADSCs expressed high levels of osteocalcin. However, apoptotic ADSCs were induced by the activation of IGF-1R. Exposure to insulin down-regulated the expression of Bcl-xL and A20, and increased Bax, in ADSCs. The addition of exogenous A20 prevented the ADSC apoptosis. We conclude that activation of IGF-1R can induce apoptosis in ADSCs, which can be prevented by addition of exogenous A20.
No preview · Article · Dec 2013 · Biochemistry and Cell Biology