Objective：Construction of erythropoietin (EPO) contained nanofibers by tissue engineering strategies and subsequent study of its pro-osteogenic effects. Methods：Biomimetic gelatin/hydroxyapatite (Gel/HA) nanofibrous scaffold was fabricated by electrostatic spinning method. Erythropoietin (EPO) loaded Gel/HA scaffold was prepared by freeze-dried method after crosslinking of Gel/HA scaffold. The release profile of EPO was detected by ELISA. The osteogenic effects of Gel/HA/EPO were detected by q-PCR. A 5mm diameter calvarial bone defect model was created in rat and scaffolds were implanted. Cone-Beam CT and histological technique were used to evaluate the bone repair effects. Results：SEM results showed that all the scaffolds were reticular and intertwined structure. There was some deposition of EPO protein on the surface of Gel/HA/EPO scaffold. The release profile of EPO showed an obvious burst release phenomenon, and only a small amount of EPO remnants kept releasing slowly. q-PCR results showed that EPO can promote osteoblast differentiation by promoting the expression of Col1 and Osterix. In vivo study showed that the Gel/HA/EPO group had a large amount of new bone formation both in 6-week and 12-week post implantation, which obviously was higher than those of the blank control group and Gel/HA group; Besides, the new bone formation in the Gel/HA group was also significantly higher than that of the blank control group. Conclusions：EPO loaded biomimetic Gelatin/HA nanofibers is an excellent scaffold material for bone tissue engineering.

Objective：Porphyromonas gingivalis could produce cyclic diguanylate (c-di-GMP) as a kind of virulence factor. This study investigated the effect of c-di-GMP on the regulation of M1 type macrophage polarization in vitro. Methods： In this study, mice derived RAW 264.7 macrophages were stimulated with different concentrations (0.25, 1, 4, 16, 64 μg/mL) c-di-GMP for different time (3, 6, 12 and 24 h). The gene and protein expression levels of IL-6, TNF-α, IL-23, IL-10 were detected by qPCR and ELISA. Flow cytometry was used to detect the expression of markers on the surface of macrophages. Results：qPCR results showed that after c-di-GMP stimulated RAW 264.7 cells, the mRNA expression of TNF-α, IL-6 and IL-23 was increased with the concentration and time of c-di-GMP in different degrees, which was dose and time dependent. The expression of IL-10 was not significantly increased in each group. ELISA results showed that the secretion of cytokines were according with the expression level of mRNA. Flow cytometry results showed that after c-di-GMP stimulated RAW 264.7 cells, the double positive rate of F4/80 and CD86, the single positive rate of CD86 and CD206 were significantly increased. Conclusions：C-di-GMP could stimulate the expression of TNF-α, IL-6 and IL-23, and induced M1 type macrophage polarization leading to the inflammatory reaction, suggesting that c-di- GMP may play an important role in the development and progression of periodontitis.

Objective：To investigate the effect of hydrogel loaded with erythropoietin (EPO) on the experimental periodontal disease (EPD) in rats.Methods：Hydrogel loaded with EPO was prepared, and measured for the gelation time and the effect on the growth and adhesion of BMSCs. Wistar rats were injected with 25 μL of the solution on the proximal and distal alveolar crest of the maxillary first molar immediately after EPD induction once every 3 days for 14 days. CBCT was used to detect the distance between cemento-enamel junction between the first molar and the second molar and alveolar bone crest; HE staining was used to observe the morphological changes of the tissue sections. The degree of inflammatory cell infiltration, alveolar bone and cementum integrity between the first molar and the second molar was evaluated by 0~3 grade. Results：The gelation time of hydrogels loaded with EPO was (220±15)s and BMSCs grew well with good adhesion. The results from the CBCT analysis demonstrated that animals treated with EPO showed no significant bone resorption and alveolar bone height was significantly greater than that of the control group, close to that of the blank group. Histopathological analysis confirmed the healthy bone structure for naive group animals, while the EPD group exhibited bone loss. In particular, the EPD+hydrogel loaded with EPO group showed significantly alveolar bone regeneration, The results of these groups were significantly different from the EPD+hydrogel loaded with EPO group (P<0.05). Conclusions：Our data suggests that hydrogel loaded with EPO was able to control periodontitis and promote alveolar bone regeneration, and may provide a new therapeutic strategy for periodontitis.

Objective：To explore the effect of knockout of Acvr1（activin A receptor type 1） on the formation of dentin. Methods： Acvr1 conditional knockout mice model was constructed using Cre-loxP system. The genotype Osterix-Cre; Acvr1fx/- was chosen as the experiment group and the genotype Osterix-Cre; Acvr1fx/+ was chosen as the control group in littermate. We observed the dentin formation and odontoblast morphology by imageology and HE staining. And we checked the expression of the odontoblast/osteoblast markers by immunohistochemical staining. Finally the expression of odontoblast/osteoblast genes was detected by qRT-PCR. Results：The mandibles of the experiment group showed more pores, the dentin of mandibular first molar was thinner, pre-dentin was thicker, and odontoblast looked irregular. The genes important for odontoblast differentiation such as Dspp, Nestin were downregulated. However, the incisor showed osteodentin, and the Dspp was down-regulated. IHC showed down-regulated DSPP in cKO molar. Conclusions：ACVR1 affected the formation of dentin by up-regulating odontoblast differentiation, and also had important role in odontoblast transition.

Objective： To investigate the source of cancer associated fibroblasts (CAFs). Methods：Normal fibroblasts (NFs) and CAFs were separated from normal gingival tissues and oral squamous carcinoma tissues, and then Cal27 medium was addedto NFs; we carried on the contrast research by immunohistochemistry, qRT-PCR technology, regarding NFs as negative control, CAFs as the positive control. Results：Compared with NFs, CAFs cell body was bigger, more disarranged, more rapid in growth, and with obviously different morphology. Cell immunohistochemistry showed: CAFs showed positive for vimentin, α-SMA was partly strongly positive, partly negative and FAP was strong positive, PDGFR-α was positive, CK (total) was negative. NFs was positive for vimentin, but the rest were negative. After culturing NFs with conditioned medium of Cal27, we found α-SMA was partly strongly positive, FAP was strong positive and PDGFR-α was positive. qRT-PCR showed: compared with NFs, α-SMA, FAP mRNA expression in CAFs and CM-NFs was significantly higher, and PDGFR-α mRNA expression was significantly lowered. Conclusions：Oral squamous cell cancer cells can activate the NFs to CAFs, NFs likely to be a source of CAFs.

Objective：To study the effects of activin receptor like kinase 2 (ALK2) on proliferation and differentiation of mouse pre-osteoblasts. Methods：The mouse pre-osteoblasts were cultured in vitro, silencing Alk2 gene by siRNA, detecting transfection by flow cytometry, using qRT-PCR to detect silence efficiency and the expression of genes related with osteogenesis. MTT method was used to detect cell proliferation and alizarin red staining was used to detect the ability of mineralization. Results：Mouse pre-osteoblasts were observed under inverted microscope. The cells were found to be irregular in shape, mostly triangular and polygonal, with more protuberances; flow cytometry and real-time RT-PCR showed that siRNA could reduce the expression of Alk2 in mouse pre-osteoblasts to 13%; MTT results showed that the number of cells increased after silencing Alk2 gene; the results of real-time RT-PCR showed that the expression of Runx2 decreased significantly(P<0.05) and the expression of Alp had downward trend after silencing Alk2 gene; alizarin red staining showed that the ability of mineralization decreased significantly after silencing Alk2 gene. Conclusions：ALK2 can inhibit the proliferation and promote the differentiation of mouse pre-osteoblasts.

Objective：To prepare light cured polyurethane acrylate adhesive for dentin bonding and we focus on its water absorption/solubility and in vitro cytotoxicity. Methods：Polyether and polyester blend polyurethane has been prepared with solution polymerization process, along with methyl methacrylate (MMA), butyl acrylate (BAM) as diluent preparation to obtain the polyurethane acrylate adhesive (PU) used in the dentin bonding. Then we evaluated water absorption/solubility and cytotoxicity of PU adhesive, and compared with two commercial adhesives（Single Bond2,Adper Easy One）. Results：water absorption: PU<Adper Easy One<Single Bond 2，water solubility: PU< Single Bond2 <Adper Easy One；In MTT assay，cytotoxicity of PU adhesive and two commercial adhesives exposed to human pulp cells was PU<Adper Easy One<Single Bond 2。Cytotoxicity was affected by the exposure time, 24 h<48 h<72 h. Conclusions：PU adhesive shows good resistance to hydrolysis performance and good biocompatibility compared with the traditional acrylic ester resin adhesive.

Objective: To set up a model and study the effect of gelatin, as organic matrix, on the process of biomineralization. Method: 30mM calcium solution and 5mM phosphate solution were added into the mineralization device respectively. Gelatin was injected into the reaction space between the cation-exchange membrane and dialysis membrane. The cation-exchange membranes were gained after 7 and 21days and the sediments on the membranes were prepared for SEM, XRD and EDX. Results: Crystals in the sediment become aciform or prismatical as the reaction time being prolonged. When 10% gelatin was added into the reaction space, the crystals became fasciculate. And the Ca/P proportion of the crystals raised when the gelatin’s concentration was changed to 20%, which meant the transformation of amorphous calcium phosphate to hydroxyapatite. Dentin-like sediments were formed on the ion-selective membrane with 20% gelatin added into the reaction space after 21 days. The crystals appeared typical hexagonal prism or hexagonal cone of around 100nm in diameter and 2um in height as they grew along c-axis direction. Analysis by energy dispersive x-ray diffraction showed that these precipitates were HA. Conclusions: Gelatin can promote the process of biomineralization and the transformation from amorphous calcium phosphate to hydroxyapatite.

Objective：To observe the effects of resveratrol on the proliferation and osteogenic differentiation of human dental pulp stem cells (hDPSCs) modified by SLA on oxidized stress. Methods：Enzyme digestion was used to isolate cultured human dental pulp derived hDPSCs in vitro. Oxidative stress was induced by hydrogen peroxide (200 μmol/L) for 24 hours. Application of 10 μmol/L resveratrol to hDPSCs was treated with hydrogen peroxide. The effects of resveratrol on the proliferation of hDPSCs after oxidative stress was observed by AlamerBlue method. The effects of resveratrol on the adhesion and morphology of hDPSCs on SLA titanium surface were observed by scanning electron microscopy. The effect of resveratrol on the osteogenic differentiation of hDPSCs on oxidized stress was observed by ALP staining. Results：The proliferation ability of hDPSCs of oxidative stress was significantly lower than that of the normal control group. Resveratrol could significantly increase the proliferation of hDPSCs of oxidative stress and promote the adhesion and morphology change of hDPSCs and the ALP expression on the surface of SLA modified. Conclusions：Resveratrol could promote the proliferation and bone differentiation ability of hDPSCs on the surface of SLA modified titanium.

Objective：To explore the anatomical relationship between the temporomandibular joint and its surrounding structures, and provide theoretical basis for avoiding the damage of important structure surrounding the temporomandibular joint. Methods：A total of 43 skull specimens of the temporomandibular joint were selected and the distance and angle between the temporomandibular joint and the important bone markers were measured with a vernier caliper and protractor. Results：The shortest distance from the squamotympanic fissure to the foramen Spinosum, foramen ovale, stylomastoid foramen was (22.69±2.02) mm, (27.23±2.07) mm, (17.67±1.76) mm. The shortest distance from the petrosquamous fissure to the foramen Spinosum, foramen ovale, stylomastoid foramen was(6.55±1.56) mm, (11.64±1.73) mm, (17.53±1.75) mm. The shortest distance from the sphenoidal crest to the foramen ovale, stylomastoid foramen was (9.53±1.39) mm, (20.08±2.14) mm. The shortest distance from the articular tubercle to the foramen Spinosum, foramen ovale, stylomastoid foramen was (27.33±2.14) mm,(30.33±2.29) mm,and(32.23±1.78) mm. The three angles of the triangle of articular tubercle, foramen Spinosum, stylomastoid foramen was (73.72±7.24)o, (55.58±6.49)o, (50.70±7.56)o. The three angles of the triangle of articular tubercle, foramen ovale, stylomastoid foramen was (62.41±6.88)o, (61.19±5.61)o, (56.40±7.60)o. Conclusions：Anatomical location measurements of the temporomandibular joint can provide a reference for temporomandibular joint surgery, thus reducing or avoiding some complications after surgery.

Objective：To measure the attached gingiva width(AGW) on buccal sides of healthy adults to provide the referencial range of AGW of healthy adults for the convenience of implanting, orthodontic, and prosthodontic treatment. Methods：144 healthy adults were enrolled, who were 20-30 years old, with healthy periodontal tissue and tooth structure, but without systemic diseases and any drug used in the past 6 months. The width of keratinized gingiva and depth of gingival sulcus were measured, then AGW were calculated and the data were statistically analyzed. Results：The WAG of lateral incisor was the most widest, ranging up to 5.2±1.1mm in maxillary, and the AGW of the first premolar was 1.8±0.8mm in mandibular which was the narrowest. The AGW in maxillary teeth were significantly greater than that in mandibular teeth, with statistically significant differences(P<0.05).There was no significant differences of the AGW between both genders (P>0.05). There was no significant difference of the AGW between the left and right sides (P>0.05). Conclusions：The AGW was the widest in lateral incisors, and narrowest in the first premolars. The AGW on maxillary arch is higher than that on madibular arch on the same position. The AGW is symmetrical on the same position of both sides and there is no difference of WAG on the same position between both genders.