Potential Applications of Blautia wexlerae in the Regulation of Host Metabolism.

Blautia wexlerae (B. wexlerae) is a strong candidate with the potential to become a next-generation probiotics (NGPs) and has recently been shown for the first time to exhibit potential in modulating host metabolic levels and alleviating metabolic diseases. However, the factors affecting the change in abundance of B. wexlerae and the pattern of its abundance change in the associated indications remain to be further investigated. Here, we summarize information from published studies related to B. wexlerae; analyze the effects of food source factors such as prebiotics, probiotics, low protein foods, polyphenols, vitamins, and other factors on the abundance of B. wexlerae; and explore the patterns of changes in the abundance of B. wexlerae in metabolic diseases, neurological diseases, and other diseases. At the same time, the development potential of B. wexlerae was evaluated in the direction of functional foods and special medical foods.

A type I and type II chemical biology toolbox to overcome the hypoxic tumour microenvironment for photodynamic therapy.

Photodynamic therapy (PDT) is a minimally invasive therapeutic modality employed for the treatment of various types of cancers, localized infections, and other diseases. Upon illumination, the photo-excited photosensitizer generates singlet oxygen and other reactive species, thereby inducing cytotoxicity in the target cells. The hypoxic tumour microenvironment (TME), however, poses a limitation on the supply of oxygen in tumour tissues. Moreover, under such conditions, tumour metastasis and drug resistance frequently occur, further compromising the efficacy of PDT in combating tumours. Traditionally, type I photosensitizers with lower oxygen consumption demonstrate significant potential in overcoming hypoxic environments and play a crucial role in determining the therapeutic efficacy of PDT because type I photosensitizers can generate highly cytotoxic free radicals. In comparison, type II photosensitizers exhibit high oxygen dependence. The rate of reactive oxygen species (ROS) generation in the type II process is significantly higher than that in the type I process. Thus, the efficiency and selectivity of PDT depend on the properties of the photosensitizer. Here, the recent development and application of type I and type II photosensitizers, mainly in the past year, are summarized. The design methods, electronic structures, photophysical properties, lipophilic properties, electric charge, and other molecular characteristics of these photosensitizers are discussed in detail. These modifications alter the microstructure of photosensitizers and directly impact the results of PDT. The main content of this paper will have a positive promoting and inspiring effect on the future development of PDT.

Evaluating the Role of Postbiotics in the Modulation of Human Oral Microbiota: A Randomized Controlled Clinical Trial.

There is a lack of clinical data to support the effectiveness and safety of postbiotics in the modulation of human oral microbiota and oral health care. Here, volunteers were recruited and randomly assigned to two cohorts: a placebo group (n = 15) and a postbiotic group (n = 16). The placebo group used toothpaste that did not contain postbiotics, while the postbiotic group used toothpaste with postbiotics (3 × 1010 CFU inactivated Lactobacillus salivarius LS97, L. paracasei LC86, and L. acidophilus LA85). Saliva samples were collected at different time points and the immunoglobulin A (IgA) and short-chain fatty acid (SCFA) levels were determined, while the salivary microbiota was analyzed by 16S rRNA amplicon sequencing. The results showed that salivary IgA levels and acetic and propionic acid levels were notably higher in the postbiotic group (P < 0.05), accompanied by an increase in the level of alpha diversity of the salivary microbiota, and these indexes remained high 1 month after discontinuing the use of toothpaste with or without postbiotics. A notable decrease in the relative abundance of the unclassified_Enterobacteriaceae, Klebsiella, Escherichia, etc. in the postbiotic group was accompanied by a notable increase in Ruminofilibacter and Lactobacillus. However, both groups did not cause significant changes in the overall structure of the host salivary microbiota. In conclusion, postbiotics dramatically and consistently improved oral immunity levels and SCFA content in the host. In addition, postbiotics were able to increase the level of microbial alpha diversity and down-regulate the abundance of some harmful microbes without significantly altering the structure of the host salivary microbiota. Chinese Clinical Trial Registry (ChiCTR) ( www.chictr.org.cn ) under the registration number ChiCTR2300074088.

[An analysis of the wall width before and after root canal therapy in mandibular second molars with C-shaped canals].

PURPOSE: To measure the wall width in mandibular second molars with C-shaped canals before and after root canal therapy using cone-beam CT(CBCT). METHODS: A total of 55 mandibular second molars from 38 patients which met the criteria for inclusion at Nanjing Stomatological Hospital, Affiliated Stomatological Hospital of Medical School of Nanjing University from January 2020 to July 2021 were analyzed．From this sample, ten teeth had been treated, while another 45 of them not. CT images of the teeth were reestablished by Mimics software 20.0. Then we made a section every 1mm perpendicular to the long axis of the teeth from apex to pulp floor. The first slice from apex to pulp chamber was named the slice 1. Along the slice 1 to crown 1 mm was called slice 1, and so forth. The wall thickness at different locations of all the slices was measured. The data was entered into SPSS 20.0 software package for analysis. RESULTS: Regardless of whether the teeth were treated or not, both the mesial and distal canal walls' average width were thicker than 1mm in all slices. At the same time, the mesial and distal canal walls' width were thicker than the width of buccal and lingual canal walls in all the slices from C-shaped root canal, except slices which were near pulp chamber(P＜0.05). As for the C-shaped root canals without root canal therapy, the width of lingual wall in the slice 1 to 4, as well as apex third root, was thinner than 1 mm. The width of buccal canal wall was thicker than the width of lingual canal wall in all slices except slice 11 and 12. As for the C-shaped root canals with root canal therapy, the width of buccal canal wall in slice 1 to 5, equivalent of apex half root, and the width of lingual wall in the slice 1 to 7, amount to apex two-thirds of root, was thinner than 1 mm. The width of buccal canal wall was thicker than the width of lingual wall in all slices except slice 1 and 9. There was no significant difference between the distal canal walls' width of C-shaped canals with and without root canal therapy(P＞0.05) . There was significant difference between the buccal canal walls' width of C-shaped canals with and without root canal therapy, as same as the mesial canal walls' width and the width of lingual canal wall (P＜0.05). CONCLUSIONS: The lingual canal walls' width in apex third root of C-shaped root canal were thin before canal preparation. The buccal walls' width in apex half root and the lingual canal walls' in apex two-thirds of root of C-shaped root canal were thin after canal preparation.

New-designed 3D printed surgical guide promotes the accuracy of endodontic microsurgery: a study of 14 upper anterior teeth.

We aimed to design a novel three-dimensional (3D) printed surgical guide and evaluate its accuracy in assisting endodontic microsurgeries. A new 3D printed surgical guide was designed by computer-aided design and computer-aided manufacturing (CAD/CAM) technology and applied to 7 patients who underwent endodontic microsurgeries of upper anterior teeth from 2020.01 to 2020.12 as the experimental group. 7 patients who suffered from endodontic microsurgeries operated by the same surgeon without using the surgical guide from 2019.01 to 2019.12 were selected as the control group. Cone beam computed tomography (CBCT) was performed more than 12 months after operation, and the accuracy of apical resection was compared between the two groups. The accuracy of the microsurgery focused on the length and angle of the root apical resection. In the study, CBCT data and oral digital scanning data were used to reconstruct 3D models of periapical lesions with soft and hard tissue information, based on which we designed the new 3D printed surgical guides. The guides were successfully applied to the apectomy in endodontic microsurgeries. The deviation of the apical resection length of the experimental group (0.467 ± 0.146 mm) was better than that of the control group (1.743 ± 0.122 mm) (P < 0.0001), and the deviation of the apical resection angle of the experimental group (9.711 ± 3.593°) was significantly less than that of the control group (22.400 ± 3.362°) (P < 0.0001). The 3D-printed surgical guide could effectively guide endodontic microsurgery and improve its accuracy by fixing both the position and the angle of apectomy. The new type of surgical guide could accurately localize the root apex and guide the apical resection.

Lactobacillus gasseri LG08 and Leuconostoc mesenteroides LM58 exert preventive effect on the development of hyperuricemia by repairing antioxidant system and intestinal flora balance.

Introduction: Currently, hyperuricemia has shown a surprisingly rising trend, which attracts widespread attention due to potentially major health risks. Considering the inevitable side effects of long-term medicine, probiotics are emerging as potential therapeutics due to their ability to improve uric acid metabolism and superior safety. Methods: In our study, two strains of probiotics, Lactobacillus gasseri LG08 (LG08) and Leuconostoc mesenteroides LM58 (LM58) isolated from kimchi were evaluated for the prebiotic properties in vitro and uric-lowering effects in vivo. Here, hyperuricemia animal model and 16S rRNA gene amplicons analysis were further studied to investigate whether these probiotics exert different effects in prevention and treatment. Results: In vivo indicators and intestinal flora immunity revealed that both LG08 and LM58 significantly prevent the development and progression of hyperuricemia, repair the antioxidant system and maintain intestinal flora balance in healthy rats, especially LM58. After hyperuricemia was formed, although the effect of LG08 and LM58 could decrease the level of uric acid, the effect to reverse and repair antioxidant levels in the body was limited. Discussion: In our study, these findings have important implications for hyperuricemia prevention and therapy, and provided more mechanistic insights into the effect of probiotics in hyperuricemia.

[Effect of circRASA2 targeting miR-543/TRAF6 axis on LPS-induced periodontal ligament cell proliferation and osteogenic differentiation].

PURPOSE: To investigate the possible role of circRASA2 in periodontitis and its potential regulatory mechanism. METHODS: Periodontitis cell model was established by lipopolysaccharide(LPS)-induced periodontal ligament cells(PDLCs). Cell proliferation activity was detected by CCK-8 assay, cell migration ability was detected by Transwell chamber assay, and the expression of osteogenic differentiation-related proteins in cells was detected by Western blot. The target miRNA of circRASA2 and its downstream target genes were predicted using the databases circinteractome and starBase, respectively, and the targeting relationship between the target genes was verified by dual-luciferase reporter gene experiment. GraphPad Prism 8.0 software package was used to analyze the data. RESULTS: circRASA2 was highly expressed in LPS-treated PDLCs cells. LPS-induced PDLCs cell proliferation activity, migration ability and osteogenic differentiation ability decreased, while knockdown of circRASA2 promoted proliferation, migration and osteogenic differentiation ability of PDLCs under LPS treatment. circRASA2 targeted and negatively regulated the expression of miR-543, and overexpression of miR-543 promoted proliferation, migration and osteogenic differentiation of PDLCs under LPS treatment. TRAF6 was a downstream target gene of miR-543, knockdown of circRASA2 down-regulated the expression of TRAF6 through the sponge action of miR-543. Overexpression of TRAF6 reversed the promotion of circRASA2 knockdown on proliferation, migration and osteogenic differentiation of PDLCs. CONCLUSIONS: circRASA2 accelerated the pathological process of periodontitis in vitro through miR-543/TRAF6 axis, and might improve periodontitis by targeting down the expression of circRASA2.

Erythropoietin induces odontoblastic differentiation of human-derived pulp stem cells via EphB4-Mediated MAPK signaling pathway.

OBJECTIVES: Human-derived pulp stem cells play key roles during dentinogenesis. Erythropoietin is reportedly involved in osteoblastogenesis and facilitates bone formation. However, the mechanism is still unknown. This research was to study the potential of erythropoietin in enhancing odontoblastic differentiation of human-derived pulp stem cells and to determine the underlying mechanism. METHODS: The human-derived pulp stem cells were treated with erythropoietin, EphB4 inhibitor, and MAPK inhibitors, and the odontoblastic differentiation was measured by ALP staining, ALP activity assay, alizarin red S staining, and their quantitative analysis, and RT-qPCR of DSPP, DMP1, OCN, and RUNX2. The direct pulp capping model was established to evaluate the formation of tertiary dentin after treatment with erythropoietin. Western blot assay was conducted to assess relevant protein expressions in the phosphorylated EphB4 and MAPK pathway. RESULTS: The results showed that erythropoietin promoted odontoblastic differentiation of human-derived pulp stem cells at 20 U/ml. Erythropoietin induced tertiary dentin formation in vivo. The potential mechanism of this was upregulating phosphorylated EphB4 and phosphorylated MAPK; furthermore, this effect could be decreased by EphB4 inhibitors, which inhibited MAPK phosphorylation. Blockage of MAPK pathways attenuated human-derived pulp stem cells' odontoblastic differentiation, suggesting that MAPK pathways are involved. CONCLUSION: Erythropoietin induced tertiary dentin formation in vivo. And erythropoietin enhanced human-derived pulp stem cells' odontoblastic differentiation via the EphB4-mediated MAPK signaling pathway.

External apical root resorption in orthodontic tooth movement: the risk factors and clinical suggestions from experts' consensus.

External apical root resorption is among the most common risks of orthodontic treatment, and it cannot be completely avoided and predicted. Risk factors causing orthodontic root resorption can generally be divided into patient- and treatment-related factors. Root resorption that occurs during orthodontic treatment is usually detected by radiographical examination. Mild or moderate root absorption usually does no obvious harm, but close attention is required. When severe root resorption occurs, it is generally recommended to suspend the treatment for 3 months for the cementum to be restored. To unify the risk factors of orthodontic root resorption and its clinical suggestions, we summarized the theoretical knowledge and clinical experience of more than 20 authoritative experts in orthodontics and related fields in China. After discussion and summarization, this consensus was made to provide reference for orthodontic clinical practice.

Evaluation of the anti-biofilm effect of poloxamer-based thermoreversible gel of silver nanoparticles as a potential medication for root canal therapy.

The purpose of this study was to design silver nanoparticles (AgNPs) poloxamer thermoreversible gel (AgNPs-PL) and investigate whether this gel could provide sustained antibacterial activity against Enterococcus faecalis (E. faecalis) in the root canal. The gels fabricated were characterized in terms of gelatin temperature, particle size, in-vitro Ag+ release, and elemental content. Cytotoxicity of AgNPs-PL on primary human periodontal ligament fibroblasts (HPDLFs) was examined by CCK-8 assay. Characterization of AgNPs-PL gel revealed that it contained particles existing as large clumps/fused aggregates of different shapes, with a mean diameter of 21.624 ± 14.689 nm, exhibited sustained release of Ag+ for 9 days, and non-toxic to HPDLFs at a low dose (4-32 µg/mL) through 24, 48, and 72 h exposures. The antibacterial effect of 16 and 32 µg/mL concentrations of AgNPs-PL was compared with blank poloxamer gel (PL) and calcium hydroxide (CH) using three methods: (I) agar counting plate, (II) scanning electron microscope (SEM) observations, and (III) confocal laser scanning microscope (CLSM) analysis. AgNPs-PL at the two doses above was more effective than PL and CH in removing E. faecalis biofilm at 1, 3, 9 days. Thus, AgNPs-PL exhibits strong activity against E. faecalis and is easy to produce, with a continuous release profile of Ag+. AgNPs-PL gel may be a candidate for a new root canal disinfection.

Nanoscale metal-organic frameworks for tumor phototherapy.

Metal-Organic Frameworks (MOFs) are constructed from metal ions/cluster nodes and functional organic ligands through coordination bonds. Owing to the advantages of diverse synthetic methods, easy modification after synthesis, large adsorption capacity for heavy metals, and short equilibrium time, considerable attention has recently been paid to MOFs for tumor phototherapy. Through rational tuning of metal ions and ligands, MOFs present abundant properties for various applications. Light-triggered phototherapy, including photodynamic therapy (PDT) and photothermal therapy (PTT), is an emerging cancer treatment approach. Nanosized MOFs can be applied as phototherapeutic agents to accomplish phototherapy with excellent phototherapeutic efficacy. This review outlines the latest advances in the field of phototherapy with various metal ion-based MOFs.

[Guided endodontic access of calcified root canal by laser melting templates].

OBJECTIVE: The aim of this study is to treat calcified root canal and assess the accuracy of guided endodontics using laser melting templates. METHODS: Two cases with calcified anterior teeth were treated with laser melting templates. Cone beam computed tomography (CBCT) was used to scan the maxillary teeth of patients before surgery to establish the root canal system model. The maxillary impression of the patient was made and it was scanned by a 3D scanner. The CBCT scans were matched with surface scans of plaster model. Mimics 19.0 and 3-matic 11.0 software were used to design the virtual planning to access cavities. The templates were produced by laser melting 3D printing. Access cavity was prepared under the guidance of laser melting template. Then the deviations of planned and prepared cavities in three dimensions and angle were measured. RESULTS: The two teeth obtained satisfactory results. The first case had a angle deviation of 1.77°, a drilling base deviation of 0.403-0.497 mm, and a tip of 0.433-0.537 mm. The second case had a angle deviation of 3.26°, a drill base deviation of 0.18-0.347 mm, and a tip of 0.310-0.463 mm. CONCLUSIONS: Laser melting template-guided endodontics is an effective technique for the treatment of calcified root canal and can be used as a new strategy for the treatment of calcified canal.

Evaluation of the Effect of Implants Placed in Preserved Sockets Versus Fresh Sockets on Tissue Preservation and Esthetics: A Meta-analysis and Systematic Review.

OBJECTIVES: To compare the success rates, tissue preservation, and esthetics of implants placed in fresh and preserved sockets and to evaluate the factors influencing the outcomes. METHODS: Medline, Embase, CENTRAL, Wanfang, and China National Knowledge Infrastructure (CNKI) databases were searched electronically, and a manual search was conducted as well. Studies that compared the implant success rate, tissue preservation, and patient-related outcomes such as complications and esthetic outcomes of immediate implant placement (IIP) and alveolar ridge preservation (ARP) were included. A subgroup analysis according to the follow-up period, socket conditions, and regenerative strategies was performed to investigate how these factors influence the prognosis. RESULTS: A total of 12 studies with 588 implants, where 298 are implants after ARP and the remaining 290 are after IIP. The IIP was performed in 58.4% of 250 implants inserted in the sockets with an intact buccal wall, whereas the percentage declined to 41.9% when the buccal wall was defective. The implant success rate was similar between ARP and IIP for an intact buccal wall but different for a defective buccal wall (ARP 98.6% vs IIP 89.6%). Moreover, hard-tissue preservation and the Pink Esthetic Score (PES) of the ARP group were significantly better than those of the IIP group in the molar region (P < .05). Also short-term complications showed no significant differences in the ARP group (P = .06). In the anterior region, there appeared to be no significant difference in hard- and soft-tissue preservation PES and patient-related outcomes between the 2 protocols. CONCLUSION: An alveolar bone defect might reduce the success rate of IIP. Further studies on the tissue preservation and esthetics of implants placed by IIP and ARP are still needed.

In Vivo Detection of Subtle Vertical Root Fracture in Endodontically Treated Teeth by Cone-beam Computed Tomography.

INTRODUCTION: The purpose of this study was to evaluate the diagnostic accuracy of cone-beam computed tomographic (CBCT) imaging in the detection of subtle vertical root fractures (VRFs) in endodontically treated teeth in vivo and to discuss direct and indirect evidence for the diagnosis of subtle VRFs. METHODS: Twenty-nine endodontically treated teeth with suspected VRFs from 29 patients were examined using CBCT imaging. CBCT images were scored based on evaluation of the fracture line and vertical buccopalatal (lingual) bone loss. VRF was diagnosed only when a definite fracture line was detected on CBCT images, and findings of periodontal exploration or surgical extraction were considered the gold standard. The diagnostic sensitivity, specificity, and accuracy were assessed. RESULTS: Of the 29 teeth, 27 were positive and 2 were negative for VRF according to the gold standard. Sensitivity, specificity, and accuracy were 33.3%, 100%, and 37.9%, respectively, based on the direct finding of a fracture line on CBCT images. However, vertical buccopalatal (lingual) bone loss was found in 25 of 27 teeth with VRFs. CONCLUSIONS: Although the accuracy of CBCT imaging for the diagnosis of subtle VRFs in endodontically treated teeth in vivo was poor, vertical buccopalatal (lingual) bone loss is an important indirect sign for the diagnosis of VRFs, which can be found on CBCT images.

Bactericidal efficacy of three parameters of Nd:YAP laser irradiation against Enterococcus faecalis compared with NaOCl irrigation.

The success of endodontic treatment depends on the thorough removal of microorganisms from the root canal system. The search for new ways to eliminate the microorganisms is therefore justified. Nd:YAP is a laser that uses yttrium aluminum perovskite, doped with neodymium crystal, as active laser medium. We used the Nd:YAP laser in an in vitro experiment to evaluate the bactericidal effect of three parameters of Nd:YAP laser-activated irrigation on biofilms of Enterococcus faecalis in root canals. The canals of 45 extracted human single-root teeth were prepared on a #35 Mtwo instrument and contaminated with E. faecalis for 14 days. Forty infected single-root teeth were then randomly divided into four groups according to the irrigation agitation protocols as follows: 5.25% sodium hypochlorite (NaOCl), Nd:YAP laser (180 mJ) + NaOCl, Nd:YAP laser (280 mJ) + NaOCl, and Nd:YAP laser (360 mJ) + NaOCl. The remaining bacteria were counted immediately using the cell count method. Teeth were firstly spilt and one half examined by scanning electron microscopy (SEM). The other half involved examination of bacterial colonization in dentinal tubules using confocal laser scanning microscopy (CLSM). Nd:YAP laser (280 mJ) + NaOCl and Nd:YAP laser (360 mJ) + NaOCl completely removed the E. faecalis biofilms from the root canal walls and made it the cleanest among the treatment groups. Bacterial reductions in the treatment groups for dentinal tubules are presented in a descending order as follows: Nd:YAP laser (360 mJ) (53.7%), Nd:YAP laser (280 mJ) (51.5%) > Nd:YAP laser (180 mJ) (45.3%) > 5.25% NaOCl (31.9%) > control (19.3%) (p < 0.05). Nd:YAP laser of 280 mJ and 360 mJ showed effective bactericidal effect in removing E. faecalis biofilm from the root canal walls and dentinal tubules.

Facile Preparation of Nanosilver Particles with Excellent Antimicrobial Activity via Release of Ionic Silver.

Nanosilver particles (SNPs) have been widely exploited in various fields, including the medical sciences due to their excellent inhibitory and bactericidal effects. It is of great importance to prepare SNPs using green synthesis that has environmentally acceptable solvent systems and eco-friendly reducing agents. In the current study, gallic acid was employed as both a reducing agent and a stabilizing agent to synthesize SNPs at mild ambient conditions. The image of transmission electron microscopy (TEM) showed that SNPs exhibited approximately spherical shape with the average diameter of 13.81±2.21 nm. The absorbance peak of obtained SNPs was sharp with the maximum wavelength of 400.5 nm by ultraviolet-visible (UV-vis) spectroscopy, suggesting the formation of small and highly monodispersed SNPs. The antimicrobial potential of the SNPs was evaluated against multiple common pathogenic microbes. The results indicated that the microbial sensitivity to the SNPs was found to vary depending on the microbial species. Among them, the gram-negative bacteria exhibited more sensitive toward SNPs than the gram-positive bacteria. In addition, the N-acetyl-L-cysteine (NAC), a silver ion chelator, pretreatment could protect the E. coli and P. aeruginosa from the SNPs inhibition, while the pretreatment of the L-ascorbic acid, an antioxidant against oxidative stress, did not significantly influence the antibacterial effects of the SNPs. These data suggested that the ionic silver release, but not reactive oxygen species (ROS), played a key role in the antimicrobial effect of the SNPs. To sum up, this study provides an environmentally friendly technique for facile synthesis of SNPs with excellent antibacterial potential.

DSP-PP precursor protein cleavage by tolloid-related-1 protein and by bone morphogenetic protein-1.

Dentin sialoprotein (DSP) and phosphophoryn (PP), acidic proteins critical to dentin mineralization, are translated from a single transcript as a DSP-PP precursor that undergoes specific proteolytic processing to generate DSP and PP. The cleavage mechanism continues to be controversial, in part because of the difficulty of obtaining DSP-PP from mammalian cells and dentin matrix. We have infected Sf9 cells with a recombinant baculovirus to produce large amounts of secreted DSP-PP(240), a variant form of rat DSP-PP. Mass spectrometric analysis shows that DSP-PP(240) secreted by Sf9 cells undergoes specific cleavage at the site predicted from the N-terminal sequence of PP extracted from dentin matrix: SMQG(447)↓D(448)DPN. DSP-PP(240) is cleaved after secretion by a zinc-dependent activity secreted by Sf9 cells, generating DSP(430) and PP(240) products that are stable in the medium. DSP-PP processing activity is constitutively secreted by Sf9 cells, but secretion is diminished 3 days after infection. Using primers corresponding to the highly conserved catalytic domain of Drosophila melanogaster tolloid (a mammalian BMP1 homolog), we isolated a partial cDNA for a Spodopotera frugiperda tolloid-related-1 protein (TLR1) that is 78% identical to Drosophila TLR1 but only 65% identical to Drosophila tolloid. Tlr1 mRNA decreased rapidly in Sf9 cells after baculovirus infection and was undetectable 4d after infection, paralleling the observed decrease in secretion of the DSP-PP(240) processing activity after infection. Human BMP1 is more similar to Sf9 and Drosophila TLR1 than to tolloid, and Sf9 TLR1 is more similar to BMP1 than to other mammalian homologs. Recombinant human BMP1 correctly processed baculovirus-expressed DSP-PP(240) in a dose-dependent manner. Together, these data suggest that the physiologically accurate cleavage of mammalian DSP-PP(240) in the Sf9 cell system represents the action of a conserved processing enzyme and support the proposed role of BMP1 in processing DSP-PP in dentin matrix.

Expression of mineralized tissue associated proteins: dentin sialoprotein and phosphophoryn in rodent hair follicles.

BACKGROUND: Mammalian hair development and tooth development are controlled by a series of reciprocal epithelial-mesenchymal interactions. Similar growth factors and transcription factors, such as fibroblast growth factor (FGF), sonic hedgehog homolog (SHH), bone morphogenetic proteins (BMPs) and Wnt10a, were reported to be involved in both of these interactions. Dentin sialoprotein (DSP) and phosphophoryn (PP) are the two major non-collagenous proteins secreted by odontoblasts that participate in dentin mineralization during tooth development. Because of striking similarities between tooth development and hair follicle development, we investigated whether DSP and/or PP proteins may also play a role in hair follicle development. OBJECTIVE: In this study, we examined the presence and location of DSP/PP proteins during hair follicle development. METHODS: Rat PP proteins were detected using immunohistochemical/immunofluorescent staining. DSP-PP mRNAs were detected by in situ hybridization with riboprobes. LacZ expression was detected in mouse tissues using a DSP-PP promoter-driven LUC in transgenic mice. RESULTS: We found that PP proteins and DSP-PP mRNAs are present in rat hair follicles. We also demonstrate that an 8 kb DSP-PP promoter is able to drive lacZ expression in hair follicles. CONCLUSION: We have firmly established the presence of DSP/PP in mouse and rat hair follicles by immunohistochemical/immunofluorescent staining, in situ hybridization with riboprobes and transgenic mice studies. The expression of DSP/PP in hair follicles is the first demonstration that major mineralization proteins likely may also contribute to soft tissue development. This finding opens a new avenue for future investigations into the molecular-genetic management of soft tissue development.

[Effect of longterm lower Porphyromonas gingivalis infection on the progression of atherosclerosis in apolipoprotein E-knocked out mice].

OBJECTIVE: To assess the effect of longterm and lower oral inoculation with Porphyromonas gingivalis (Pg) on the progression of atherosclerosis in apolipoprotein E-knocked out (ApoE(-/-)) mice. METHODS: Six-week-old male ApoE(-/-) mice were inoculated orally with 0.1 ml live Pg(10(13)/L) or bouillon culture-medium quintic per week for 15 consecutive weeks, altogether 75 times of inoculations. The lesion area of atherosclerosis in the aortic tree was measured by en face quantification by red oil O staining method. The atherosclerotic lesion was examined by histopathology. The levels of total cholesterol and triglycerides were compared. RESULTS: At 22 weeks after inoculation, the mean atherosclerotic lesion area in inoculated mice was (98 363.68 ± 12 043.00) µm(2), which was significantly greater than that in noninoculated mice, which was (62 985.06 ± 7419.64) µm(2) (P = 0.035). CONCLUSIONS: Longterm lower oral inoculation of Pg can accelerate the progression of atherosclerosis in apolipoprotein E-knocked out mice.

[Effect of static magnetic field on bone morphogenetic protein 2 and collagen type I of osteoblast cell].

PURPOSE: To study the effect of static magnetic field on osteoblast,and to explore the possibility of osteogenesis of static magnetic field. METHODS: Rat calvarial osteoblasts were seeded on the 96-well culture plate, and exposed to 0.062T magnetic intensity field for 24 hours, 48 hours and 72 hours, respectively. Changes of BMP-2 and collagen type I in osteoblast were examined by Western blot. Immunohistochemical staining was carried out at different time periods. The data obtained were analyzed for ANOVA using SPSS 11.5 software package. RESULTS: The results of Western blot showed that magnetic static field affected expression of BMP-2 and collagen type I in osteoblast. The expression of BMP-2 in 48-hour group increased 2.1 fold and the expression of collagen type I in 72-hour group increased 1.6 fold compared to the control group (P<0.01). Moreover, the staining color of the collagen type I in cell of magnetic treated group was deeper than that of the control. CONCLUSIONS: Static magnetic field induced the expression of BMP-2 and stimulated secretion of collagen type I. The results indicate that osteoblasts are sensitive to 0.062T static magnetic field stimulation.