High quality repair of osteochondral defects in rats using the extracellular matrix of antler stem cells

BACKGROUND Cartilage defects are some of the most common causes of arthritis.Cartilage lesions caused by inflammation,trauma or degenerative disease normally result in osteochondral defects.Previous studies have shown that decellularized extracellular matrix(ECM)derived from autologous,allogenic,or xenogeneic mesenchymal stromal cells(MSCs)can effectively restore osteochondral integrity.AIM To determine whether the decellularized ECM of antler reserve mesenchymal cells(RMCs),a xenogeneic material from antler stem cells,is superior to the currently available treatments for osteochondral defects.METHODS We isolated the RMCs from a 60-d-old sika deer antler and cultured them in vitro to 70%confluence;50 mg/mL L-ascorbic acid was then added to the medium to stimulate ECM deposition.Decellularized sheets of adipocyte-derived MSCs(aMSCs)and antlerogenic periosteal cells(another type of antler stem cells)were used as the controls.Three weeks after ascorbic acid stimulation,the ECM sheets were harvested and applied to the osteochondral defects in rat knee joints.RESULTS The defects were successfully repaired by applying the ECM-sheets.The highest quality of repair was achieved in the RMC-ECM group both in vitro(including cell attachment and proliferation),and in vivo(including the simultaneous regeneration of well-vascularized subchondral bone and avascular articular hyaline cartilage integrated with surrounding native tissues).Notably,the antler-stem-cell-derived ECM(xenogeneic)performed better than the aMSC-ECM(allogenic),while the ECM of the active antler stem cells was superior to that of the quiescent antler stem cells.CONCLUSION Decellularized xenogeneic ECM derived from the antler stem cell,particularly the active form(RMC-ECM),can achieve high quality repair/reconstruction of osteochondral defects,suggesting that selection of decellularized ECM for such repair should be focused more on bioactivity rather than kinship.

Isolation and Identification Methods for Oral Klebsiella pneumoniae Involved in Onset of Inflammatory Bowel Disease

Purpose: Recently, it was reported that Klebsiella pneumoniae is related to the onset of inflammatory bowel disease including the Crohn disease. It was frequently reported that K. pneumoniae was detected in human oral cavities. Regrettably, it currently remains unclear whether K. pneumoniae is part of the normal oral flora. The aim of this study was to establish the isolation and identification methods for K. pneumoniae from human oral cavities, and investigate its transmission pattern. Methods: A selective medium, OKPSM, for the isolation of K. pneumoniae from oral cavities was developed in this study. Also, PCR primer for the identification and detection at subspecies level of K. pneumoniae was designed. Results: OKPSM and PCR method using the primers designed in this study were useful for the isolation and identification of K. pneumoniae from human oral cavities. K. pneumoniae subsp. pneumoniae was detected at 10.0% in 30 saliva samples. On the other hand, K. pneumoniae subsp. ozaenae and K. pneumoniae subsp. rhinoscleromatis were detected from no sample. Moreover, K. pneumoniae subsp. pneumoniae isolates from same subject at 0 month and after 3 months showed same genotypes on AP-PCR using OPA-07 primer. Conclusion: These results indicated that human oral cavities were not suitable for the habitat of K. pneumoniae.

Study on Distribution of Four Pseudomonas Species in Living Environment Using Multiplex PCR

Purpose: The genus Pseudomonas is a ubiquitous microorganism frequently detected from immunocompromised patients. The inherent resistance to numerous antimicrobial agents contributes to the opportunistic character of this pathogen exhaustive monitoring of this pathogen is considered of critical importance to public health organizations. The reliable identification method able to distinguish genetic close Pseudomonas species is needed, because these organisms are difficult to differentiate by phenotypic or biochemical methods. The purpose of the present study was to design species-specific primers in order to identify and detect four Pseudomonas species which are frequently detected from the human oral cavities, and to investigate the distribution of these organisms in the living environment using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the rpoD gene of four Pseudomonas species. Swab samples were collected from fifty washstands, and the distribution of Pseudomonas species was investigated using a conventional PCR at genus level and a multiplex PCR at species level. Results: Multiplex PCR method developed in this study was able to distinguish four Pseudomonas species clearly. The genus Pseudomonas was detected from all samples (100%), whereas P. putida, P, aeruginosa, P. stutzeri and P. fluorescens were detected at 44%, 8%, 4% and 2% in fifty swab samples, respectively. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and works without requiring DNA extraction. It was indicated that washstands were the uninhabitable environment for P. putida, P, aeruginosa, P. stutzeri and P. fluorescens.

Craniofacial therapy:advanced local therapies from nano-engineered titanium implants to treat craniofacial conditions

Nano-engineering-based tissue regeneration and local therapeutic delivery strategies show significant potential to reduce the health and economic burden associated with craniofacial defects,including traumas and tumours.Critical to the success of such nano-engineered non-resorbable craniofacial implants include load-bearing functioning and survival in complex local trauma conditions.Further,race to invade between multiple cells and pathogens is an important criterion that dictates the fate of the implant.In this pioneering review,we compare the therapeutic efficacy of nano-engineered titanium-based craniofacial implants towards maximised local therapy addressing bone formation/resorption,soft-tissue integration,bacterial infection and cancers/tumours.We present the various strategies to engineer titanium-based craniofacial implants in the macro-,micro-and nano-scales,using topographical,chemical,electrochemical,biological and therapeutic modifications.A particular focus is electrochemically anodised titanium implants with controlled nanotopographies that enable tailored and enhanced bioactivity and local therapeutic release.Next,we review the clinical translation challenges associated with such implants.This review will inform the readers of the latest developments and challenges related to therapeutic nano-engineered craniofacial implants.

Specific RNA m6A modification sites in bone marrow mesenchymal stem cells from the jawbone marrow of type 2 diabetes patients with dental implant failure

The failure rate of dental implantation in patients with well-controlled type 2 diabetes mellitus(T2DM)is higher than that in nondiabetic patients.This due,in part,to the impaired function of bone marrow mesenchymal stem cells(BMSCs)from the jawbone marrow of T2DM patients(DM-BMSCs),limiting implant osseointegration.RNA N6-methyladenine(m6A)is important for BMSC function and diabetes regulation.However,it remains unclear how to best regulate m6A modifications in DM-BMSCs to enhance function.Based on the“m6A site methylation stoichiometry”of m6A single nucleotide arrays,we identified 834 differential m6Amethylated genes in DM-BMSCs compared with normal-BMSCs(N-BMSCs),including 43 and 790 m6A hypermethylated and hypomethylated genes,respectively,and 1 gene containing hyper-and hypomethylated m6A sites.Differential m6A hypermethylated sites were primarily distributed in the coding sequence,while hypomethylated sites were mainly in the 3’-untranslated region.The largest and smallest proportions of m6A-methylated genes were on chromosome 1 and 21,respectively.Maz F-PCR and real-time RTPCR results for the validation of erythrocyte membrane protein band 4.1 like 3,activity-dependent neuroprotector homeobox(ADNP),growth differentiation factor 11(GDF11),and regulator of G protein signalling 2 agree with m6A single nucleotide array results;ADNP and GDF11 m RNA expression decreased in DM-BMSCs.Furthermore,gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses suggested that most of these genes were enriched in metabolic processes.This study reveals the differential m6A sites of DM-BMSCs compared with N-BMSCs and identifies candidate target genes to enhance BMSC function and improve implantation success in T2DM patients.

Study on the Distribution at Species Level of Genus Candida in Human Oral Cavities, Using Culture and Multiplex PCR Methods

Purpose: Although the genus Candida is frequently isolated from human oral cavities, the distribution at the species level of these organisms has been little reported. The purpose of the present study was to assess the distribution at the species level of the genus Candida in human oral cavities. Methods: This study was performed using culture and Multiplex PCR methods. Moreover, the genotyping classification of C. albicans was analyzed with a PCR. Results: Of all subjects (n = 90), detection frequency of genus Candida was 42.2%. Genus Candida was not detected in the subjects between 0 to 9 years old, and there was no difference in the detection frequencies of this organism among each generation from 10s to 80s. C. albicans was the most dominant species, followed by C. parapsilosis, C. glabrata, and C. dubliniensis. Plural Candida species tended not to be detected in the individual sample. Genotype A was dominant in the C. albicans isolates. Conclusion: These results indicated that C. albicans of genotype A was dominant and that the genus Candida rarely coexists with other Candida species, in each individual oral cavity.

Origin of Candida albicans in Human Oral Cavity

Purpose: Candida albicans is regarded as a part of normal flora in the human oral cavity. However, it remains unclear whether the genus Candida, especially C. albicans, is an oral resident microorganism and causes marital infection or not. The purpose of the present study was to elucidate the origin of oral C. albicans by investigating the colonization and infection route to oral cavities of this organism with arbitrarily primed polymerase chain reaction (AP-PCR). Methods: After C. albicans was isolated from four subjects (average age: 42.2, range: 33 - 56), the isolations of this organism from them were performed six months later again. To investigate whether C. albicans is an oral resident microorganism, the genotype homology of each C. albicans isolates that were isolated twice from the same subjects was compared. Moreover, C. albicans was isolated from five pairs of married couples (average period of cohabitation: 12.4 years, range: 5 - 31). To investigate whether C. albicans causes marital infection, the genotype homology of C. albicans isolates that were isolated from each pair of married couples was compared. Results: AP-PCR patterns of C. albicans that were isolated from each subject at o month and after 6 months showed the identical genotypes among each individual. C. albicans isolates from five pairs of married couples showed the identical genotypes between a husband and wife of each pair on AP-PCR. Conclusion: These results indicated that C. albicans was an oral resident microorganism and caused the marital infection.

Five-year outcomes of immediate implant placement for mandibular molars with and without chronic apical periodontitis:A retrospective study

BACKGROUND Most physicians consider molars with chronic apical periodontitis(CAP)lesions as contraindications for immediate implant placement.At the patient’s request,we perform immediate implant placement of the mandibular molars with CAP in clinical practice.AIM To retrospectively analyze and compare the 5-year outcomes of immediate implant placement of the mandibular molars with CAP and those without obvious inflammation.METHODS The clinical data of patients with immediate implant placement of the mandibular molars in the Department of Oral and Maxillofacial Surgery,the Affiliated Hospital of Qingdao University,from June 2015 to June 2017 were collected.The patients were divided into CAP(n=52)and no-CAP(n=45)groups.Changes in bone mineral density and bone mass around implants were analyzed 5 years after implant restoration.RESULTS At 5 years after implantation,the peri-implant bone mineral density was 528.2±78.8 Hounsfield unit(HU)in the CAP group and 562.6±82.9 HU in the no-CAP group(P=0.126).Marginal bone resorption around implants did not differ significantly between the two groups,including buccal(P=0.268)or lingual(P=0.526)resorption in the vertical direction or buccal(P=0.428)or lingual(P=0.560)resorption in the horizontal direction.Changes in the peri-implant jump space did not differ significantly between the two groups,including the buccal(P=0.247)or lingual(P=0.604)space in the vertical direction or buccal(P=0.527)or lingual(P=0.707)space in the horizontal direction.The gray value of cone-beam computed tomography measured using Image J software can reflect the bone mineral density.In the CAP area,the gray values of the bone tissue immediately and 5 years after implant placement differed significantly from those of the surrounding bone tissue(P<0.01).CONCLUSION The results of this study suggest that immediate implant placement of the mandibular molars with CAP can achieve satisfactory 5-year clinical results,without significant differences in the complications,survival rate,or bone tissue condition from the no-CAP mandibular molars.

Investigation of Bacteria Species Most Involved in Peri-Implantitis

Purpose: Currently, bacteriological examinations of implant treatments target periodontopathic bacteria such as red complex bacteria, including Porphyromonas gingivalis, and detect them qualitatively or quantitatively. However, it seems that those examinations do not reflect the peri-implant tissue conditions precisely, because periodontopathic bacteria are also frequently detected from healthy peri-implant sites. The purpose of the present study was to investigate bacteria species most involved in peri-implantitis using a PCR method. Methods: Polymerase chain reaction (PCR) primers in this study were designed based on partial sequences of 16S rDNA of bacteria species involved in peri-implantitis that were described in numerous previous studies. Peri-implant sulcus fluid (PISF) samples were collected from thirty periodontally healthy patients with implants (HI) and thirty patients with peri-implantitis (PI). Each detection frequency of bacteria species in PISFs of both groups was investigated using a PCR method, and was compared using Fisher’s exact test. Results: In PI group, detection frequencies of Corynebacterium durum, Fretibacterium fastidiosum and Slackia exigua were significantly higher than those of HI group (p P. gingivalis and Tannerella forsythia belonging to red complex were frequently detected in the PISF samples of HI group (p > 0.05). Conclusion: It was suggested that monitoring C. durum and F. fastidiosum levels in PISF samples was useful as a clinical indicator for the evaluation of peri-implant tissue conditions.

Study on Distribution of Acinetobacter baumannii Complex in Dental Hospital Using Multiplex PCR

Purpose: In recent years, multidrug-resistant Acinetobacter baumannii has appeared and caused outbreaks of hospital infections all over the world. Close monitoring of this pathogen and other A. baumanii complex species is considered of critical importance to public health organizations. The reliable identification method able to distinguish A. baumannii from genetically close Acinetobacter species is needed, because these species are unable to be differentiated by phenotypic or biochemical methods. The purpose of the present study was to design species-specific primers in order to identify and detect A. baumanii complex species, and Acinetobacter lwoffii which is frequently detected from the human specimens, and to investigate the distribution of these organisms in dental hospital using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the 16S ribosomal RNA (16S rRNA) gene and DNA gyrase subunit B (gyrB) of each species of A. baumanii complex species. Swab samples were collected from ten dental spittoon units in dental hospital, and the distribution of A. baumanii complex species was investigated using a multiplex PCR. Results: These primers were able to distinguish each species of A. baumanii complex species clearly. A. baumanii and A. calcoaceticus were detected at 20.0% and 10.0% in ten swab samples, respectively. On the other hand, A. nosocomialis, A. lowffii, and A. pittii were detected from no sample. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and worked without requiring DNA extraction.

One Step Multiplex PCR for Identification of Candida haemulonii complex and Candia auris

Purpose: Recently, Candida haemulonii complex (Candida haemulonii, Candida duobushaemulonii and Candida haemulonii var. vulnera) and two genetically close species (Candida pseudohaemulonii and Candida auris) have emerged as an opportunistic fungal pathogen associated with various infectious diseases of humans, and most of those isolates have displayed antifungal resistance. Because it is difficult to differentiate these microorganisms by a current technique, unfortunately, it is important to establish a method for identifying them accurately. The purpose of the present study was to design species-specific primers in order to identify and detect C. auris, C. pseudohaemulonii, and C. haemulonii complex, i.e. , C. haemulonii, C. duobushaemulonii and C. haemulonii var. vulnera using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the 26S rRNA, 18S rRNA, and RPB1 genes and ITS region of five Candida species. Results: The multiplex PCR method developed in this study was able to distinguish five Candida species clearly. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and works without requiring DNA extraction.

Bone block from lateral window-correcting vertical and horizontal bone deficiency in maxilla posterior site:A case report

BACKGROUND Lateral window approach for sinus floor lift is commonly used for vertical bone augmentation in cases when the residual bone height is less than 5 mm.However,managing cases becomes more challenging when a maxillary sinus pseudocyst is present or when there is insufficient bone width.In this case,we utilized the bone window prepared during the lateral window sinus lift as a shell for horizontal bone augmentation.This allowed for simultaneous horizontal and vertical bone augmentation immediately after the removal of the maxillary sinus pseudocyst.CASE SUMMARY A 28-year-old female presented to our clinic with the chief complaint of missing upper left posterior teeth.Intraoral examination showed a horizontal deficiency of the alveolar ridge contour.The height of the alveolar bone was approximately 3.6 mm on cone beam computed tomography(CBCT).And a typical well-defined'dome-shaped'lesion in maxillary sinus was observed on CBCT imaging.The lateral bony window was prepared using a piezo-ultrasonic device,then the bony window was fixed to the buccal side of the 26 alveolar ridge using a titanium screw with a length of 10 mm and a diameter of 1.5 mm.The space between the bony window and the alveolar ridge was filled with Bio-Oss,covered with a Bio-Gide collagen membrane,and subsequently sutured.Nine months later,the patient’s bone width increased from 4.8 to 10.5 mm,and the bone height increased from 3.6 to 15.6 mm.Subsequently,a Straumann^(■)4.1 mm×10 mm implant was placed.The final all-ceramic crown restoration was completed four months later,and both clinical and radiographic examinations showed that the implant was successful,and the patient was satisfied with the results.CONCLUSION The bone block harvested from the lateral window sinus lift can be used for simultaneous horizontal bone augmentation acting as a shell for good two-dimensional bone augmentation.

AFF1 and AFF4 differentially regulate the osteogenic differentiation of human MSCs

AFF1 and AFF4 belong to the AFF （AF4/FMR2） family of proteins, which function as scaffolding proteins linking two different transcription elongation factors, positive elongation factor b （P-TEFb） and ELL1/2, in super elongation complexes （SECs）. Both AFF1 and AFF4 regulate gene transcription through elongation and chromatln remodeling. However, their function in the osteogenic differentiation of mesenchymal stem cells （MSCs） is unknown. In this study, we show that small interfering RNA （siRNA）-mediated depletion of AFF1 in human MSCs leads to increased alkaline phosphatase （ALP） activity, enhanced mineralization and upregulated expression of osteogenic-related genes. On the contrary, depletion of AFF4 significantly inhibits the osteogenic potential of MSCs. In addition, we confirm that overexpression of AFF1 and AFF4 differentially affects osteogenic differentiation in vitro and MSC-mediated bone formation in vivo. Mechanistically, we find that AFFI regulates the expression of DKK1 via binding to its promoter region. Depletion of DKK1 in HA-AFFl-overexpressing MSCs abrogates the impairment of osteogenic differentiation. Moreover, we detect that AFF4 is enriched in the promoter region of ID1. AFF4 knockdown blunts the BRE luciferase activity, SP7 expression and ALP activity induced by BMP2 treatment. In conclusion, our data indicate that AFF1 and AFF4 differentially regulate the osteogenic differentiation of human MSCs.AFF1 and AFF4 belong to the AFF （AF4/FMR2） family of proteins, which function as scaffolding proteins linking two different transcription elongation factors, positive elongation factor b （P-TEFb） and ELL1/2, in super elongation complexes （SECs）. Both AFFI and AFF4 regulate gene transcription through elongation and chromatln remodeling. However, their function in the osteogenic differentiation of mesenchymal stem cells （MSCs） is unknown. In this study, we show that small interfering RNA （siRNA）-mediated depletion of AFF1 in human MSCs leads to increased alkaline phosphatase （ALP） activity, enhanced mineralization and upregulated expression of osteogenic-related genes. On the contrary, depletion of AFF4 significantly inhibits the osteogenic potential of MSCs. In addition, we confirm that overexpression of AFF1 and AFF4 differentially affects osteogenic differentiation in vitro and MSC-mediated bone formation in vivo. Mechanistically, we find that AFFI regulates the expression of DKK1 via binding to its promoter region. Depletion of DKK1 in HA-AFFl-overexpressing MSCs abrogates the impairment of osteogenic differentiation. Moreover, we detect that AFF4 is enriched in the promoter region of ID1. AFF4 knockdown blunts the BRE luciferase activity, SP7 expression and ALP activity induced by BMP2 treatment. In conclusion, our data indicate that AFF1 and AFF4 differentially regulate the osteogenic differentiation of human MSCs.

Effect of Galla chinensis on the remineralization of two bovine root lesions morphous in vitro

The present study aims to evaluate the effect of Galla chinensis compounds on the remineralization of two artificial root lesions morphous in vitro. Sixty bovine dentine blocks were divided into two groups and individually treated with two levels of demineralization solutions to form erosive and subsurface artificial carious lesions in vitro. Each group was then divided into three subgroups, each of which were treated with a remineralization solution （positive control）, deionized water （negative control）, or 4 000 mg-L-1 aqueous solutions of Galla chinensis extract. The dentine blocks were then subjected to a pH-cycling regime for 7 days. During the first 4 days, the daily cycle included 21-h deal and 3-h demineralization applications. The dentine blocks were dealt with the entire day during the remaining 3 days. Two specimens from each of the treatment groups were selected and observed under a polarized light microscope. Data collected using a laser scanning confocal microscope were computerized and analyzed. Galla chinensis extract clearly enhanced the remineralization of both erosive lesion and subsurface lesion patterns in the specimens （P〈0.05）. The level of remineralization of the erosive lesion by Galla chinensis extract was lower than that of the subsurface lesion （P〈0.05）. In addition, the remineralization of the subsurface lesion by Galla chinensis extract was higher than that of the remineralization solution （P〈0.05）. No significant difference between the remineralization of erosive lesions by Galla chinensis extract and the remineralization solution was observed （P〉0.05）. So Galla chinensis extract has the potential to improve the remineralization of artificial root lesions under dynamic pH-cyclic conditions, indicating its potential use as a natural remineralization medicine.

Single-cell characterization of monolayer cultured human dental pulp stem cells with enhanced differentiation capacity

Human dental pulp stem cells(hDPSCs)are easily obtained multipotent cells,however,their potential value in regenerative medicine is hindered by the phenotypic and functional changes after conventional monolayer expansion.Here,we employed single-cell RNA sequencing(scRNA-seq)to comprehensively study the transcriptional difference between the freshly isolated and monolayer cultured DPSCs.The cell cluster analysis based on our scRNA-seq data showed that monolayer culture resulted in a significant cellular composition switch compared to the freshly isolated DPSCs.However,one subpopulation,characterized as MCAM(+)JAG(+)PDGFRA(−),maintained the most transcriptional characteristics compared to their freshly isolated counterparts.Notably,immunofluorescent staining revealed that the MCAM(+)JAG(+)PDGFRA(−)hDPSCs uniquely located in the perivascular region of human dental pulp tissue.Flow-cytometry analysis confirmed that their proportion remained relatively stable(~2%)regardless of physiological senescence or dental caries.Consistent with the annotation of scRNA-seq data,MCAM(+)JAG(+)PDGFRA(−)hDPSCs showed higher proliferation capacity and enhanced in vitro multilineage differentiation potentials(osteogenic,chondrogenic and adipogenic)compared with their counterparts PDGFRA(+)subpopulation.Furthermore,the MCAM(+)JAG(+)PDGFRA(−)hDPSCs showed enhanced bone tissue formation and adipose tissue formation after 4-week subcutaneous implantation in nude mice.Taken together,our study for the first time revealed the cellular composition switch of monolayer cultured hDPSCs compared to the freshly isolated hDPSCs.After in vitro expansion,the MCAM(+)JAG(+)PDGFRA(−)subpopulation resembled the most transcriptional characteristics of fresh hDPSCs which may be beneficial for further tissue regeneration applications.

Allogenic tooth transplantation using 3D printing: A case report and review of the literature

BACKGROUND The history of allogenic tooth transplantation can be traced back to the 16th century.Although there have been many successful cases,much needs to be better understood and researched prior to the technique being translated to everyday clinical practice.CASE SUMMARY In the present report,we describe a case of allogenic tooth transplantation between a mother and her daughter.The first left maxillary molar of the mother was diagnosed with residual root resorption and needed to be extracted.The 3rd molar of the daughter was used as a donor tooth.Prior to transplantation,a 3D printing system was introduced to fabricate an individualized reamer drill specifically designed utilizing the donor’s tooth as a template.The specific design of our 3D printed bur allowed for the recipient site to better match the donor tooth.With the ability to 3D print in layers,even the protuberance of the root can be matched and 3D printed,thereby minimizing unnecessary bone loss.CONCLUSION Our study is a pioneering case combining 3D printing with allogenic tooth transplantation,which could be able to minimize unnecessary bone loss and improve the implant stability.This article aims to enhance our understanding of allogenic tooth transplantation and 3D printing,and may potentially lead to tooth transplantation being utilized more frequently - especially since transplantations are so commonly utilized in many other fields of medicine with high success rates.

Application of exosome-derived noncoding RNAs in bone regeneration:Opportunities and challenges

With advances in the fields of regenerative medicine,cell-free therapy has received increased attention.Exosomes have a variety of endogenous properties that provide stability for molecular transport across biological barriers to cells,as a form of cell-to-cell communication that regulates function and phenotype.In addition,exosomes are an important component of paracrine signaling in stemcell-based therapy and can be used as a stand-alone therapy or as a drug delivery system.The remarkable potential of exosomes has paved the pathway for cell-free treatment in bone regeneration.Exosomes are enriched in distinct noncoding RNAs(ncRNAs),including microRNAs,long ncRNAs and circular RNAs.Different ncRNAs have multiple functions.Altered expression of ncRNA in exosomes is associated with the regenerative potential and development of various diseases,such as femoral head osteonecrosis,myocardial infarction,and cancer.Although there is increasing evidence that exosome-derived ncRNAs(exoncRNAs)have the potential for bone regeneration,the detailed mechanisms are not fully understood.Here,we review the biogenesis of exo-ncRNA and the effects of ncRNAs on angiogenesis and osteoblast-and osteoclast-related pathways in different diseases.However,there are still many unsolved problems and challenges in the clinical application of ncRNA;for instance,production,storage,targeted delivery and therapeutic potency assessment.Advancements in exo-ncRNA methods and design will promote the development of therapeutics,revolutionizing the present landscape.

Accelerated and enhanced osteointegration of MAO-treated implants:histological and histomorphometric evaluation in a rabbit model

Microarc oxidation（MAO） has become a promising technique for the surface modification of implants. Therefore, the aims of this study were to further quantitatively and qualitatively evaluate the osteointegration abilities of MAO-treated and smooth surface（SF） implants in vivo and to investigate the areas in which the superiority of MAO-treated implants are displayed. In a rabbit model,a comprehensive histomorphological, osteogenic, mineralizational, and integrative assessment was performed using light microscopy, fluorescence microscopy, confocal laser scanning microscopy, and radiographic analyses. Compared with the SF groups, the MAO-treated groups exhibited more active contact osteogenesis, as well as distant osteogenesis, under fluorescence examination, the mineral apposition rate was found to be greater for all of the MAO-treated implants, and the osteointegration index（OI） value was greater in the MAO-treated groups at different times. In conclusion, the calcium-rich amorphous layer created by MAO provided a better environment for osteointegration, with more active contact osteogenesis, a more rapid mineral apposition rate and greater OI values.

Analyses of oligodontia phenotypes and genetic etiologies

Oligodontia is the congenital absence of six or more teeth and comprises the more severe forms of tooth agenesis.Many genes have been implicated in the etiology of tooth agenesis,which is highly variable in its clinical presentation.The purpose of this study was to identify associations between genetic mutations and clinical features of oligodontia patients.An online systematic search of papers published from January 1992 to June 2021 identified 381 oligodontia cases meeting the eligibility criteria of causative gene mutation,phenotype description,and radiographic records.Additionally,ten families with oligodontia were recruited and their genetic etiologies were determined by whole-exome sequence analyses.We identified a novel mutation in WNT10A(c.99_105dup)and eight previously reported mutations in WNT10A(c.433 G>A;c.682 T>A;c.318 C>G;c.511.C>T;c.321 C>A),EDAR(c.581 C>T),and LRP6(c.1003 C>T,c.2747 G>T).Collectively,20 different causative genes were implicated among those 393 cases with oligodontia.For each causative gene,the mean number of missing teeth per case and the frequency of teeth missing at each position were calculated.Genotype–phenotype correlation analysis indicated that molars agenesis is more likely linked to PAX9 mutations,mandibular first premolar agenesis is least associated with PAX9 mutations.

Immunogenicity and Prediction of Epitopic Region of Antigen Ag Ⅰ/Ⅱ and Glucosyltransferase from Streptococcus mutans

The levels of Streptococcus（S.） mutans infections in saliva were evaluated and a comparison for specific antibody levels among children with different levels of S. mutans infection was made. The promising epitopic regions of antigen AgⅠ/ Ⅱ（PAc） and glucosyltransferase（GTF） for potential vaccine targets related to S. mutans adherence were screened. A total of 94 children aged 3–4 years were randomly selected, including 53 caries-negative and 41 caries-positive children. The values of S. mutans and those of salivary total secretory immunoglobulin A（s Ig A）, anti-PAc and anti-Glucan binding domain（anti-GLU） were compared to determine the correlation among them. It was found the level of s-Ig A against specific antigens did not increase with increasing severity of S. mutans infection, and the complete amino acid sequence of PAc and GTFB was analyzed using the DNAStar Protean system for developing specific anti-caries vaccines related to S. mutans adherence. A significantly positive correlation between the amount of S. mutans and children decayed, missing, and filled teeth index was observed. No significant difference was detected in specific s Ig A against PAc or GLU between any two groups. No significant correlation was found between such specific s Ig A and caries index. A total of 16 peptides from PAc as well as 13 peptides from GTFB were chosen for further investigation. S. mutans colonization contributed to early children caries as an important etiological factor. The level of s Ig A against specific antigens did not increase with increasing severity of S. mutans infection in children. The epitopes of PAc and GTF have been screened to develop the peptide-based or protein-based anti-caries vaccines.