Yifei Du

Expression of cytosolic phospholipase A2 and cyclooxygenase 2 and their significance in human oral mucosae, dysplasias and squamous cell carcinomas.

Prostaglandin E2 (PGE2) plays an important role in promoting carcinogenesis. Cytosolic phospholipase A2 (cPLA2) and cyclooxygenase 2 (COX-2) are both key enzymes for PGE2 biosynthesis. Recent evidence suggests that the coordinated function of cPLA2 and COX-2 in the arachidonic acid pathway may contribute to the process of carcinogenesis in various tissue types. However, the concomitant effect of these enzymes on oral carcinogenesis remains unclear. In this study,we evaluated the expression of cPLA2 and COX-2 in normal oral mucosa, dysplastic oral mucosa and squamous carcinoma (SCC) using immunohistochemistry. In an in vitro assay, Tca8113 and KB oral cancer cells were treatedwith NS-398 (a selective inhibitor of COX-2) for varying time intervals: 6, 12, 24, 48 and 72 h. The levels of cPLA2 and COX-2 expression were evaluated by Western blot, and PGE2 production was analyzed by radioimmunoassay. We found that cPLA2 and COX-2 were expressed at higher levels in oral dysplasia and SCC than in normal oral mucosa. cPLA2 expression was also found to correlate closely with COX-2 expression. Moreover, the enzymatic activities of cPLA2 and COX-2 were gradually downregulated with longer durations of treatment with NS-398, as demonstrated by a reduction in the amount of PGE2 production over time. Our data suggest that the coordinated activity of cPLA2 and COX-2 contribute to the process of oral carcinogenesis, thus identifying cPLA2 as a potential target for the prevention and treatment of oral cancers.

SATB2-Nanog axis links age-related intrinsic changes of mesenchymal stem cells from craniofacial bone

Bone mesenchymal stem cells (BMSCs) senescence contributes to age-related bone loss. The alveolar bone in jaws originates from neural crest cells and possesses significant site- and age-related properties. However, such intrinsic characteristics of BMSCs from alveolar bone (AB-BMSCs) and the underlying regulatory mechanisms still remain unknown. Here, we found that the expression of special AT-rich binding protein 2 (SATB2) in human AB-BMSCs significantly decreased with aging. SATB2 knockdown on AB-BMSCs from young donors displayed these aging-related phenotypes in vitro. Meanwhile, enforced SATB2 overexpression could rejuvenate AB-BMSCs from older donors. Importantly, satb2 gene- modified BMSCs therapy could prevent the alveolar bone loss during the aging of rats. Mechanistically, the stemness regulator Nanog was identified as the direct transcriptional target of SATB2 in BMSCs and functioned as a downstream mediator of SATB2. Collectively, our data reveal that SATB2 in AB-BMSCs associates with their age-related properties, and prevents AB-BMSCs senescence via maintaining Nanog expression. These findings highlight the translational potential of transcriptional factor-based cellular reprogramming for anti-aging therapy.

Estrogen regulates stemness and senescence of bone marrow stromal cells to prevent osteoporosis via ERβ-SATB2 pathway†

Decline of pluripotency in bone marrow stromal cells (BMSCs) associated with estrogen deficiency leads to a bone formation defect in osteoporosis. Special AT‐rich sequence binding protein 2 (SATB2) is crucial for maintaining stemness and osteogenic differentiation of BMSCs. However, whether SATB2 is involved in estrogen‐deficiency associated‐osteoporosis is largely unknown. In this study, we found that estrogen mediated pluripotency and senescence of BMSCs, primarily through estrogen receptor beta (ERβ). BMSCs from the OVX rats displayed increased senescence and weaker SATB2 expression, stemness, and osteogenic differentiation, while estrogen could rescue these phenotypes. Inhibition of ERβ or ERα confirmed that SATB2 was associated with ERβ in estrogen‐mediated pluripotency and senescence of BMSCs. Furthermore, estrogen mediated the upregulation of SATB2 through the induction of ERβ binding to estrogen response elements (ERE) located at −488 of the SATB2 gene. SATB2 overexpression alleviated senescence and enhanced stemness and osteogenic differentiation of OVX‐BMSCs. SATB2‐modified BMSCs transplantation could prevent trabecular bone loss in an ovariectomized rat model. Collectively, our study revealed the role of SATB2 in stemness, senescence, and osteogenesis of OVX‐BMSCs. These results indicate that estrogen prevents osteoporosis by promoting stemness and osteogenesis, and inhibiting senescence of BMSCs through an ERβ‐SATB2 pathway. Therefore, SATB2 is a novel anti‐osteoporosis target gene.

The angiogenic variation of skeletal site-specific human BMSCs from same alveolar cleft patients: a comparative study

Tissue engineering strategies hold great potential for alveolar cleft reconstruction. Bone marrow stromal cells (BMSCs) from iliac crest and craniofacial regions are candidate seeding cells with site-specific characteristics and bone-repairing properties. Craniofacial BMSCs seem to possess stronger multipotency and osteogenic capabilities than BMSCs isolated from iliac crest. However, the angiogenic capabilities of these two type cell is rarely reported. We obtained human BMSCs (hBMSCs) of maxilla (M-hBMSCs) and iliac crest (I-hBMSCs) from same alveolar cleft patients to investigate the agiogenic variations using co-culture system with human umbilical vein endothelial cells (HUVECs). From in vitro comparison, M-hBMSCs allowed HUVECs to form more tube-like structures and sprouting angiogenesis by tube formation assays and 3D fibrin vasculogenic assay, respectively. By transplantation in vivo, M-hBMSCs enhanced larger size vessel like structures distributed the entire implants compared with I-hBMSCs. Western blotting was used to assess the angiogenesis related factors including hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). The results showed a significant higher expression of bFGF protein in M-hBMSCs and HUVECs co-culture system both in vitro and in vivo. As bFGF could promote migration and proliferation of endothelial cells, scratch wound healing and transwell migration assays were performed as well as MTT assays and cell cycle analysis. The data suggested the effect of M-hBMSCs on HUVECs was stronger than I-hBMSCs. Taken together, these results indicated that craniofacial BMSCs seemed to have greater angiogenesis capability than iliac crest BMSCs and this might be associated with the different levels of bFGF protein expression in co-culture system.

Association between BRCA1 P871L polymorphism and cancer risk: evidence from a meta-analysis

Breast cancer 1 (BRCA1) gene makes great contributions to the repair of DNA. The association between BRCA1 P871L polymorphism and cancer risk has been investigated in a growing number of studies, but the conclusions are not conclusive. To obtain a comprehensive conclusion, we performed a meta-analysis of 24 studies with 13762 cases and 22388 controls. The pooled results indicated that BRCA1 gene P871L variant decreased risk of overall cancer (homozygous model: odds ratio (OR) = 0.89, 95%confidence interval (CI) = 0.79-1.00; recessive model: OR = 0.89, 95% CI = 0.80-0.99). The stratified analysis observed decreased risk associated with BRCA1 P871L in subgroups among Asians and high score studies, but not Caucasians or low score studies. In conclusion, despite several limitations, this meta-analysis suggested that BRCA1 P871L genetic variation may be associated with decreased susceptibility to cancer.

Accurate Evaluation of Cone-Beam Computed Tomography to Volumetrically Assess Bone Grafting in Alveolar Cleft Patients.

Objective: The aim of this study was to investigate the accuracy of cone-beam computed tomography (CBCT) to assess the volume of bone graft in alveolar cleft patients. Materials and Methods: Twelve patients of unilateral alveolar cleft were included in this study. All patients were taken CBCT preoperative and 1 week postoperative. The digital imaging and communications in medicine (DICOM) files were imported into Simplant software and three-dimensional (3D) reconstruction of the alveolar defect was achieved. With 3D volumetric measurements module, the volume of alveolar cleft was calculated preoperatively. During operation, the syringe compression method was adopted to calculate the actual amount of bone graft. One week postoperative, CBCT scan was performed again to measure the bone volume grafted to the defect. The volumetric ratio of the syringe compression method to preoperative CBCT assessment and the volume difference between syringe compression method and postoperative CBCT assessment were analyzed to evaluate the accuracy of CBCT measurement. Results: The 3D structure of the alveolar cleft and the boundary of bone graft was clear from CBCT images. The estimated volume of alveolar cleft by preoperative CBCT scans was 1.06 ± 0.09 cm3, and the actual amount of bone graft determined by the syringe compression method was 1.51 ± 0.12 cm3. The ratio between the latter to the former was 1.43 ± 0.07. The calculated volume of bone graft by 1-week postoperative CBCT scans was 1.53 ± 0.11 cm3, with no significant difference compared with the actual amount of bone graft (P > 0.05). Conclusions: CBCT was an accurate measurement to calculate the volume of alveolar defect and bone graft in alveolar cleft patients. Preoperative scans could aid in quantitatively determining the bone amount needed to adequately fill the bone defect, and the postoperative scans give accurate follow-up evaluation after surgery.

A new design of facial artery perforator flaps for the reconstruction of small- to medium-sized intraoral defects.

Aim The aim of this study was to investigate the reliability and outcomes of a new design of facial artery perforator flaps, extending from the angle of the mouth to the border of the mandible, for the reconstruction of small- to medium-sized intraoral defects. Patients and Methods The technique had been used in 23 patients between February 2009 and August 2012. The locations of intraoral defects included the tongue, the floor of the mouth, the inferior gingival mucosa, and the sublingual gland. All flaps were monitored for complications including skin loss and ischemia. The functions of appearance, swallowing, and speech were assessed 6 months after operation with the University of Washington Quality of Life Questionnaire. Results All flaps presented with satisfactory results except for one, which demonstrated superficial tip necrosis that settled after conservative treatment. The follow-up period ranged from 6 to 12 months, and 1 patient died as a result of pulmonary metastasis, and 3 patients underwent second surgery because of local tumor recurrence (2patients) and cervical recurrence (1 patient). Nineteen patients were assessed with the University of Washington Quality of Life Questionnaire, and the mean (SD) scores of appearance, swallowing, and speech were 57.89 (14.45), 83.68 (19.98), and 81.58 (23.16), respectively. Conclusions This design of facial artery perforator flaps could provide an efficient and cost-effective method for reconstruction of small- to medium-sized intraoral defects with a low surgical morbidity and satisfactory levels of quality of life.

Assessment of aging characteristics of female condylar trabecular structure by cone-beam computed tomography

ObjectivesThis study was performed to analyze the aging-related changes of the female condylar bone mineral density (BMD) and trabecular structure by cone-beam computed tomography (CBCT), and determine whether the condylar structure shows obvious changes after menopause.MethodsThe CBCT images of 160 female patients who met the inclusion criteria for the study were collected and divided into four groups by age (20–29 years, 30–39 years, premenopausal, and postmenopausal groups). Computer processing software CT-Analyser (Version 1.15.2.2+; SkyScan, Antwerp, Belgium) was used to measure the condylar BMD and related indexes, namely the bone volume/tissue volume ratio (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), trabecular structure model index (SMI), and bone surface area/volume ratio (BS/BV). SPSS 12.0 (SPSS Inc., Chicago, IL, USA) was used to analyze the radiographic findings and statistical differences.ResultsNo significant differences were found between the bilateral condyles in each group (P > 0.05). BV/TV, Tb.N, and Tb.Th of the condyle decreased with age, and the postmenopausal group showed significantly different values for each index compared with the other groups (P < 0.01). Tb.Sp, SMI, and BS/BV of the condyle increased with age, and the postmenopausal group showed significantly different values for each index compared with the other groups (P < 0.01).ConclusionsWith increasing age, the female condylar bone volume decreases, the Tb.N and Tb.Th decrease, the gap between the trabecular bone increases, and plate-like trabecular bone gradually transforms into a rod-like form. These changes are much more obvious in postmenopausal women.

Block iliac bone grafting enhances osseous healing of alveolar reconstruction in older cleft patients: A radiological and histological evaluation

Background Older alveolar cleft patients (&12 years old) often have wide bone defect as well as teeth loss, resulting in poor osseous healing with conventional alveolar bone grafting (ABG). In this study, we investigated a surgical technique of block iliac bone grafting for the alveolar cleft reconstruction and evaluated the clinical and radiological outcomes of these cleft patients. Material and Methods Fifteen patients were included in this study. All cases received preoperative cone bean computed tomography (CBCT) scans for the alveolar cleft evaluation. Osseous outcomes of block iliac bone grafting were assessed at 1 week, 3- and 6-month postoperatively. Volume changes and bone resorption rates were calculated using the measurement modules of Simplant software. Bone samples from one patient undergoing dental implantation were assessed by micro-CT and histological examination. The morbidities of donor-site were analyzed by clinical examination and questionnaire survey. Results The average age of the case series was 18.53±2.50 years. The intraoral incision of thirteen cases healed well. However, two cases had oronasal fistula and graft exposure at 1-week postoperatively. The results of follow-up CBCT scans showed significant resistance to radiation on both sides of the bone graft, suggesting a good osseous healing and new bone formation. The mean residual bone volume was 1.68±0.26 cm3, 1.29±0.23 cm3 and 1.15±0.23 cm3 at 1-week, 3- and 6-month postoperatively. Correspondingly, the mean bone resorption rates in 3- and 6-month postoperative were 21.78±6.88% and 30.66±8.97%, respectively. From micro-CT and HE examinations, the block bone samples exhibited a cancellous structure in which mature bone trabecula and functional blood vessels appeared. The average scores of donor-site morbidities were drastically decreased at 3- and 6-month postoperatively compared with those at 1-week postoperatively. Conclusions Our results demonstrated that block iliac bone grafting could achieve satisfying osseous outcomes in older alveolar cleft patients, and this technique provided favorable bony condition for further treatments, especially dental implantation. Key words:Alveolar bone grafting, Block bone grafting, Osseous healing, CBCT.

A functional polymorphism in the pre-miR-146a gene is associated with the risk of nonsyndromic orofacial cleft

microRNAs (miRNAs) are widely involved in craniofacial development, and genetic variants of miRNAs may be associated with the risk of nonsyndromic orofacial cleft (NSOC). Here, we systematically selected five single nucleotide polymorphisms (SNPs) of miRNAs and investigated the associations between these variants and NSOC susceptibility in a two‐stage case–control study including 1,406 NSOC patients and 1,578 controls from the Chinese population. We found that compared with the C allele, the rs2910164 G allele of pre‐miR‐146a was associated with an increased risk of NSOC (additive model: odds ratio [OR] = 1.17, 95% confidence interval [CI]: 1.06–1.30, P = 0.002), including both cleft lip with or without cleft palate (CL/P) and cleft palate only (CPO). Bioinformatic prediction and functional assays revealed that the C allele of rs2910164 was significantly associated with inhibited HEK‐293 and HEPM cell proliferation and decreased abundance of TRAF6. Both miR‐146a and TRAF6 were expressed in the lip tissue samples of NSOC patients, and a moderate inverse correlation was observed between them. Taken together, these results demonstrated that miR‐146a/rs2910164 is associated with susceptibility to NSOC, providing novel insights into the genetic etiology and underlying biology of NSOC.