Role of LRP5/6/GSK-3ß/ß-catenin in the differences in exenatide- and insulin-promoted T2D osteogenesis and osteomodulation.

BACKGROUND AND PURPOSE: Insulin and exenatide are two hypoglycaemic agents that exhibit different osteogenic effects. This study compared the differences between exenatide and insulin in osseointegration in a rat model of Type 2 diabetes (T2D) and explored the mechanisms promoting osteogenesis in this model of T2D. EXPERIMENTAL APPROACH: In vivo, micro-CT was used to detect differences in the peri-implant bone microstructure in vivo. Histology, dual-fluorescent labelling, immunofluorescence and immunohistochemistry were used to detect differences in tissue, cell and protein expression around the implants. In vitro, RT-PCR and western blotting were used to measure the expression of osteogenesis- and Wnt signalling-related genes and proteins in bone marrow mesenchymal stromal cells (BMSCs) from rats with T2D (TBMSCs) after PBS, insulin and exenatide treatment. RT-PCR was used to detect the expression of Wnt bypass cascade reactions under Wnt inactivation. KEY RESULTS: Micro-CT and section staining showed exenatide extensively promoted peri-implant osseointegration. Both in vivo and in vitro experiments showed exenatide substantially increased the expression of osteogenesis-related and activated the LRP5/6/GSK-3ß/ß-catenin-related Wnt pathway. Furthermore, exenatide suppressed expression of Bmpr1a to inhibit lipogenesis and promoted expression of Btrc to suppress inflammation. CONCLUSION AND IMPLICATIONS: Compared to insulin, exenatide significantly improved osteogenesis in T2D rats and TBMSCs. In addition to its dependence on LRP5/6/GSK-3ß/ß-catenin signalling for osteogenic differentiation, exenatide-mediated osteomodulation also involves inhibition of inflammation and adipogenesis by BMPR1A and ß-TrCP, respectively.

Inhibition of PTEN promotes osteointegration of titanium implants in type 2 diabetes by enhancing anti-inflammation and osteogenic capacity of adipose-derived stem cells.

Background: The low osteogenic differentiation potential and attenuated anti-inflammatory effect of adipose-derived stem cells (ADSCs) from animals with type 2 diabetes mellitus (T2DM) limits osseointegration of the implant. However, the underlying mechanisms are not fully understood. Methods: Western blotting and qRT-PCR analyses were performed to investigate the effects of PTEN on the osteogenic capacity of ADSCs of T2DM rats (TADSCs). We conducted animal experiments in T2DM-Sprague Dawley (SD) rats to evaluate the osteogenic capacity of modified TADSC sheets in vivo. New bone formation was assessed by micro-CT and histological analyses. Results: In this study, adipose-derived stem cells of T2DM rats exhibited an impaired osteogenic capacity. RNA-seq analysis showed that PTEN mRNA expression was upregulated in TADSCs, which attenuated the osteogenic capacity of TADSCs by inhibiting the AKT/mTOR/HIF-1α signaling pathway. miR-140-3p, which inhibits PTEN, was suppressed in TADSCs. Overexpression or inhibition of PTEN could correspondingly reduce or enhance the osteogenic ability of TADSCs by regulating the AKT/mTOR/HIF-1α signaling pathway. TADSCs transfected with PTEN siRNA resulted in higher and lower expressions of genes encoded in M2 macrophages (Arg1) and M1 macrophages (iNOS), respectively. In the T2DM rat model, PTEN inhibition in TADSC sheets promoted macrophage polarization toward the M2 phenotype, attenuated inflammation, and enhanced osseointegration around implants. Conclusion: Upregulation of PTEN, which was partially due to the inhibition of miR-140-3p, is important for the attenuated osteogenesis by TADSCs owing to the inhibition of the AKT/mTOR/HIF-1α signaling pathway. Inhibition of PTEN significantly improves the anti-inflammatory effect and osteogenic capacity of TADSCs, thus promoting peri-implant bone formation in T2DM rats. Our findings offer a potential therapeutic approach for modifying stem cells derived from patients with T2DM to enhance osseointegration.

Synchronous stabilization of Pb, Zn, Cd, and As in lead smelting slag by industrial solid waste.

In order to prevent heavy metal (HM) pollution from lead smelting slag (LSS) to the surrounding environment, this work investigated the feasibility, influencing factors, and mechanisms of using industrial solid waste such as fly ash (FA), oil sludge pyrolysis residue (PR), and steel slag (SS) as remediation amendments. The results demonstrated that the stabilization process was influenced by the material dosage, water content, and LSS particle size. Compared to single materials, the combination amendment PR2FA1 (with a mass ratio of PR to FA as 2:1) exhibited the best stabilization effect, simultaneously reducing the leaching concentrations of As, Zn, Cd, and Pb in LSS to 0.032, 0.034, 0.002, and 0.014 mg/L, respectively. The pH value of the leachate remained between 8 and 9, which met the requirements of surface water quality class IV (GB3838-2002). Through morphological analysis, microscopic characterization, and simulated solution adsorption experiments, it was determined that the stabilization process of HMs was controlled by various mechanisms, including electrostatic attraction, physical adsorption, ion exchange, and chemical precipitation. PR2FA1 had more active components, and its fine-porous structure provided more active sites, resulting in good stabilization performance for As, Zn, Cd, and Pb. Furthermore, cost analysis showed that PR2FA1, as an environmentally friendly material, could generate profits of 157.2 ¥/ton. In conclusion, the prepared PR2FA1 not only addressed the HMs pollution from lead smelting slag to the surrounding environment but also achieved the safe and resourceful disposal of hazardous waste-oil sludge. Its excellent performance in stabilizing HMs and cost-effectiveness suggested promising commercial applications.

HIF-1α increases the osteogenic capacity of ADSCs by coupling angiogenesis and osteogenesis via the HIF-1α/VEGF/AKT/mTOR signaling pathway.

BACKGROUND: Stabilization and increased activity of hypoxia-inducible factor 1-α (HIF-1α) can directly increase cancellous bone formation and play an essential role in bone modeling and remodeling. However, whether an increased HIF-1α expression in adipose-derived stem cells (ADSCs) increases osteogenic capacity and promotes bone regeneration is not known. RESULTS: In this study, ADSCs transfected with small interfering RNA and HIF-1α overexpression plasmid were established to investigate the proliferation, migration, adhesion, and osteogenic capacity of ADSCs and the angiogenic ability of human umbilical vein endothelial cells (HUVECs). Overexpression of HIF-1α could promote the biological functions of ADSCs, and the angiogenic ability of HUVECs. Western blotting showed that the protein levels of osteogenesis-related factors were increased when HIF-1α was overexpressed. Furthermore, the influence of upregulation of HIF-1α in ADSC sheets on osseointegration was evaluated using a Sprague-Dawley (SD) rats implant model, in which the bone mass and osteoid mineralization speed were evaluated by radiological and histological analysis. The overexpression of HIF-1α in ADSCs enhanced bone remodeling and osseointegration around titanium implants. However, transfecting the small interfering RNA (siRNA) of HIF-1α in ADSCs attenuated their osteogenic and angiogenic capacity. Finally, it was confirmed in vitro that HIF-1α promotes osteogenic differentiation and the biological functions in ADSCs via the VEGF/AKT/mTOR pathway. CONCLUSIONS: This study demonstrates that HIF-1α has a critical ability to promote osteogenic differentiation in ADSCs by coupling osteogenesis and angiogenesis via the VEGF/AKT/mTOR signaling pathway, which in turn increases osteointegration and bone formation around titanium implants.

TRAF6 promotes osteogenesis in ADSCs through Raf-Erk-Merk-Hif1-a pathway.

Critical-size defects (CSDs) are challenging oral clinical issues that need to be solved. Adipose-derived mesenchymal stem cells (ADSCs) and gene therapy offer a new target to solve these issues. Consequently, ADSCs attract more and more attention because of advantages such as easy obtainability and no ethical concerns. TNF receptor-associated factor 6 (TRAF6) is a significant binding protein both of tumour necrosis factor superfamily and of the toll/interleukin-1 receptor superfamily. Evidence is accumulating that TRAF6 inhibited osteoclast formation and promoted the proliferation of multiple myeloma cell lines and bone resorption. Here, we reported that overexpression of TRAF6 enhanced the proliferation, migration and osteogenesis of ADSCs through Raf-Erk-Merk-Hif1a pathway. Cell sheet of ADSCs combined with TRAF6 accelerated the healing of CSDs. In a word, TRAF6 enhanced osteogenesis, migration and proliferation through Raf-Erk-Merk-Hif1a pathway.

Predictors of peri-implant bone remodeling outcomes after the osteotome sinus floor elevation: a retrospective study.

BACKGROUND: This study aimed to evaluate the radiographic outcomes of implants after osteotome sinus floor elevation (OSFE), and further identify the separate predictors for these radiographic outcomes. METHODS: In this retrospective cohort study, a total of 187 implants were inserted into 138 patients using the OSFE technique. Seventy-four patients in the grafted group, and 64 patients in the non-grafted group completed this study. The vertical bone gain (VBG) and marginal bone loss (MBL) at 3 years following surgery were assessed as outcome variables. Based on extensive literature results, variables considered potential predictors of outcome variables included sex, age, tooth position, implant length, implant diameter, with or without grafting materials, residual bone height, sinus width, bone density, and sinus membrane thickness. Subsequently, the binary logistic regression analysis was applied with VBG and MBL as dependent variables, respectively. The receiver operating characteristic curve (ROC) with its area under the curve (AUC) was performed to further determine the predictive value of these predictors. RESULTS: One hundred and six implants in grafted group and 81 implants in the non-grafted group were analyzed. The average VBG was 2.12 ± 1.94 mm for the grafted group and 0.44 ± 1.01 mm for the non-grafted group at 3 years (P < 0.05). The mean MBL was 1.54 ± 1.42 mm for the grafted group and 1.13 ± 1.69 mm for the non-grafted group at 3 years (P > 0.05). After the adjustment for confounders, logistic regression analysis demonstrated that implant length, grafting, residual bone height, and sinus membrane thickness were predictors of VBG. The odds ratio for VBG was 3.90, 4.04, 4.13 and 2.62, respectively. Furthermore, grafting exhibited the largest AUC at 0.80. While tooth position and implant length were predictors of MBL, the odds ratio for MBL was 3.27 and 7.85, respectively. Meanwhile, implant length exhibited the largest AUC at 0.72. CONCLUSIONS: OSFE with or without simultaneous grafting materials both showed predictable clinical outcomes. Additionally, the present study is the first quantitative and significant verification that VBG has a significant association with sinus membrane thickness, as well as residual bone height, implant length and grafting. Whereas tooth position and implant length are markedly associated with MBL.

The Proangiogenic Potential of Rat Adipose-Derived Stromal Cells with and without Cell-Sheet Induction: A Comparative Study.

A functional vasculature for survival remains a challenge for tissue regeneration, which is indispensable for oxygen and nutrient supply. Utilizing mesenchymal stromal cells (MSCs) to alleviate tissue ischemia and repair dysfunctional or damaged endothelium is a promising strategy. Compared to other populations of MSCs, adipose-derived stromal cells (ASCs) possess a more significant proangiogenic potential and are abundantly available. Cell sheet technology has recently been widely utilized in bone engineering. Compared to conventional methods of seeding seed cell suspension onto biological scaffolds, cell sheet technology prevents cell loss and preserves the extracellular matrix (ECM). Nevertheless, the proangiogenic potential of ASC sheets remains unknown. In this study, rat ASC sheets were constructed, and their macro- and microstructures were examined. In addition, we investigated the effects of ASCs and ASC sheets on the biological properties and angiogenic capacity of endothelial cells (ECs). The results demonstrated that the ASC sheets gradually thickened as the number of cells and ECM increased over time and that the cells were in an active state of secretion. Similar to ASC-CM, the conditioned medium (CM) of ASC sheets could significantly enhance the proliferative capacity of ECs. ASC sheet-CM has significant advantages over ASC-CM in promoting the migration and angiogenesis of ECs, where the exosomes secreted by ASC sheets play an essential role. Therefore, using ASC sheets for therapeutic tissue and organ regeneration angiogenesis may be a valuable strategy.

Exosomes derived from reparative M2-like macrophages prevent bone loss in murine periodontitis models via IL-10 mRNA.

BACKGROUND: Periodontitis is characterized by progressive inflammation and alveolar bone loss resulting in tooth loss finally. Macrophages including pro-inflammatory M1-like macrophages and reparative M2-like macrophages play a vital role in inflammation and tissue homeostasis in periodontitis. Among them, reparative M2-like macrophages have been shown to promote tissue repair and prevent bone loss. However, the mechanism of reparative M2 macrophages-induced osteoprotective effect remains elusive. RESULTS: Exosomes from reparative M2-like macrophages (M2-Exos) were isolated and identified successfully. M2-Exos could promote bone marrow stromal cells (BMSCs) osteogenic differentiation while suppressing bone marrow derived macrophage (BMDM) osteoclast formation, and prohibit pathological alveolar bone resorption because of the intercellular communication via exosomes. High expression level of IL-10 mRNA was detected not only in reparative M2-like macrophages but also in M2-Exos. Meanwhile, IL-10 expression level in BMSCs or BMDM was also upregulated significantly after co-culturing with M2-Exos in a concentration-dependent manner. In vitro, recombinant IL-10 proteins had the ability to selectively promote osteogenic differentiation of BMSCs and hinder osteoclast differentiation of BMDM. Moreover, after treatment with M2-Exos and IL-10R antibody together, the capacity of promoting osteogenesis and suppressing osteoclastogenesis of M2-Exos was significantly reversed. In vivo experiments further showed that M2-Exos reduced alveolar bone resorption in mice with periodontitis via IL-10/IL-10R pathway. CONCLUSION: In conclusion, our results demonstrate that the reparative M2-like macrophages could promote osteogenesis while inhibiting osteoclastogenesis in vitro as well as protect alveolar bone against resorption in vivo significantly. M2-Exos could upregulate the IL-10 cytokines expression of BMSCs and BMDM via delivering exosomal IL-10 mRNA to cells directly, leading to activation of the cellular IL-10/IL-10R pathway to regulate cells differentiation and bone metabolism. These results might partly account for the mechanism of osteoprotective effect of reparative M2-like macrophages and provide a novel perspective and a potential therapeutic approach on improving alveolar resorption by M2-Exos.

Construction and performance of exendin-4-loaded chitosan-PLGA microspheres for enhancing implant osseointegration in type 2 diabetic rats.

The updating and optimization of drug delivery systems is critical for better in vivo behaviors of drugs, as well as for improving impaired implant osseointegration in diabetes. Numerous studies have reported the benefits of exendin-4 on diabetic bone, with the potential to enhance osseointegration in diabetes. To construct an appropriate sustained-release system of exendin-4 targeting implant osseointegration in diabetes, this study fabricated exendin-4-loaded microspheres using poly(lactic-co-glycolic acid) (PLGA) and chitosan. The morphology, size, encapsulation efficiency, and drug release behavior of microspheres were investigated. The bioactivity of drug-loaded microspheres on cell proliferation and osteogenic differentiation of diabetic BMSCs was investigated to examine the pharmacologic action of exendin-4 loaded into chitosan-PLGA microspheres. Further, the influence of microspheres on osseointegration was evaluated using type 2 diabetes mellitus (T2DM) rat implant model. After 4 weeks, the samples were evaluated by radiological and histological analysis. The results of in vitro experiments showed that the prepared exendin-4-loaded chitosan-PLGA microspheres have good properties as a drug delivery system, and the chitosan could improve the encapsulation efficiency and drug release of PLGA microspheres. In addition, exendin-4-loaded microspheres could enhance the proliferation and osteogenic differentiation of diabetic BMSCs. The results of in vivo experiments showed the exendin-4-loaded microspheres significantly improved the impaired osseointegration and bone formation around implants in T2DM rats without affecting blood glucose levels. Thus, the local application of exendin-4-loaded chitosan-PLGA microspheres might be a promising therapeutic strategy for improving the efficacy of dental implants in T2DM individuals.

Expert consensus on the bone augmentation surgery for alveolar bone defects / 口腔疾病防治

@#Alveolar bone is an important anatomic basis for implant-supported denture restoration, and its different degrees of defects determine the choices of bone augmentation surgeries. Therefore, the reconstruction of alveolar bone defects is an important technology in the clinical practice of implant restoration. However, the final reconstructive effect of bone quality, bone quantity and bone morphology is affected by many factors. Clinicians need to master the standardized diagnosis and treatment principles and methods to improve the treatment effect and achieve the goal of both aesthetic and functional reconstruction of both jaws. Based on the current clinical experience of domestic experts and the relevant academic guidelines of foreign counterparts, this expert consensus systematically and comprehensively summarized the augmentation strategies of alveolar bone defects from two aspects: the classification of alveolar bone defects and the appropriate selection of bone augmentation surgeries. The following consensus are reached: alveolar bone defects can be divided into five types (Ⅰ-0, Ⅰ-Ⅰ, Ⅱ-0, Ⅱ-Ⅰ and Ⅱ-Ⅱ) according to the relationship between alveolar bone defects and the expected position of dental implants. A typeⅠ-0 bone defect is a bone defect on one side of the alveolar bone that does not exceed 50% of the expected implant length, and there is no obvious defect on the other side; guided bone regeneration with simultaneous implant implantation is preferred. Type Ⅰ-Ⅰ bone defects refer to bone defects on both sides of alveolar bone those do not exceed 50% of the expected implant length; the first choice is autologous bone block onlay grafting for bone increments with staged implant placement or transcrestal sinus floor elevation with simultaneous implant implantation. Type Ⅱ-0 bone defects show that the bone defect on one side of alveolar bone exceeds 50% of the expected implant length, and there’s no obvious defect on the other side; autologous bone block onlay grafting (thickness ≤ 4 mm) or alveolar ridge splitting (thickness > 4 mm) is preferred for bone augmentation with staged implant placement. Type Ⅱ-Ⅰ bone defects indicate that the bone plate defect on one side exceeds 50% of the expected implant length and the bone defect on the other side does not exceed 50% of the expected implant length; autologous bone block onlay grafting or tenting techniques is preferred for bone increments with staged implant implantation. Type Ⅱ-Ⅱ bone defects are bone plates on both sides of alveolar bone those exceed 50% of the expected implant length; guided bone regeneration with rigid mesh or maxillary sinus floor elevation or cortical autologous bone tenting is preferred for bone increments with staged implant implantation. This consensus will provide clinical physicians with appropriate augmentation strategies for alveolar bone defects.

Clinical and radiographic variables related to implants with simultaneous grafts among type 2 diabetic patients treated with different hypoglycemic medications: a retrospective study.

BACKGROUND: The influence of different hypoglycemic agents on peri-implant variables among type 2 diabetes mellitus patients is still unclear. Therefore, the aim of this study was to assess the radiographic marginal bone loss and clinical parameters around implants in patients using different hypoglycemic agents. METHODS: In this retrospective cohort study, the dental implant records of type 2 diabetes mellitus patients who met the inclusion criteria were collected. The patients using only single medication as follows: insulin, metformin, or glucagon-like peptide-1 (GLP-1) drugs, were grouped according to their medication. These patients received implant placement with the same initial status, and all the prosthesis restorations were cement-retained ceramic crowns. The peri-implant marginal bone levels were evaluated by periapical radiographs immediately after implant placement and at 1 and 2-year follow-up visits. The baseline characteristics were compared among groups. The peri-implant radiographic marginal bone loss and clinical parameters were preliminarily compared using the Kruskal-Wallis test, and then the covariates were controlled by covariance analysis. Bonferroni post hoc adjustment test was performed for the multiple comparisons. RESULTS: After a review of more than 7000 medical records, a total of 150 patients with 308 implants at 1-year follow-up were assessed. The peri-implant marginal bone loss in the GLP-1 drug group was significantly smaller than the insulin group and metformin group (P < 0.01). The radiographic bone loss in the metformin group was higher than the insulin group (P < 0.05). Some of these included patients were lost to follow-up. Only 74 patients with 129 implants completed the 2-year follow-up. The radiographic bone loss in the metformin group was still higher than the insulin group (P < 0.05) and GLP-1 group (P < 0.01). There was no significant difference in the BOP (+) and the mean PD among groups (P > 0.05). CONCLUSIONS: The radiographic variables were not exactly the same among the patients with different hypoglycemic agents at both the 1 and 2-year follow-ups. After ensuring consistency in baseline characteristics, the positive effect of GLP-1 drugs on peri-implant bone remodeling may be no less than insulin or metformin. More studies are needed to verify the direct effect of these drugs on peri-implant bone. Clinical trial registration number ChiCTR2000034211 (retrospectively registered).

The effect of hypoglycemic drugs on bone metabolism and dental implantation in type 2 diabetes mellitus patients / 口腔疾病防治

@#Patients with type 2 diabetes mellitus (T2DM) have a large demand for dental implants, but the pathologic state of T2DM patients could compromise the efficacy of implant treatment. Glycemic control can improve the success rate of implants in the T2DM population, but the early osseointegration of individuals still needs to be improved. Strengthening early osseointegration in patients with T2DM is one of the urgent problems for clinicians. The pharmacological mechanisms of hypoglycemic drugs on the market for bone metabolism are different and may require different interventions on the bone around the implant, but there is a lack of direct clinical evidence of the protective effect of hypoglycemic drugs. This review integrated the bone metabolic effect of drugs in clinical medical research and dental implant research. The aim was to provide medication guidance for T2DM patients who require implant surgery, and it is recommended to avoid the use of drugs with negative effects on bone as far as possible without violating the clinical medication guidelines, including SGLT-2 inhibitors and thiazolidinediones. Instead, they should choose glucose-lowering drugs that are beneficial to bone metabolism, such as insulin, metformin and GLP-1 receptor agonists. However, the comparative clinical effects of these drugs on periimplant bone need to be further elucidated. The researcher should select appropriate drugs (incretin drugs) to enhance the early osseointegration of implants in patients with T2DM.

[Insights into peri-implantitis and its prevention].

Peri-implantitis is one of the most common complications in dental implant treatment. Peri-implantitis is a crucial implication of implant failure, which is characterized by high morbidity and intractability. Thus, how to understand peri-implantitis correctly and deeply, and how to prevent its occurrence, are important problems that every dental implant surgeon has to face.

A preliminary study on submariners with xerostomia after a 3-month deployment.

To observe the clinical manifestations and salivary secretion of xerostomia patients in submariners who engaged in a three-month deployment. The general conditions and clinical examination of the 136 submariners were evaluated, by which the patients with xerostomia were screened out and their clinical manifestations were recorded. Besides, the flow rate of unstimulated saliva and stimulated saliva was measured and calculated. Subsequently, the related factors of xerostomia were quantitatively classified and statistically analyzed. In all the involved submariners, 42 were diagnosed to have xerostomia by physical examination after they returned from the task, among which 71.4% showed a decrease in unstimulated salivary flow rate and it was significantly correlated with the accompanying symptoms and their general conditions. Therefore, it was concluded that the occurrence of xerostomia could be related to the service life and job responsibilities of the submariners. The main manifestations were the reduction of unstimulated salivary secretion and the accompanying clinical symptoms such as cheilosis and angular cheilitis. Noticeably, the high psychological pressure and harsh living conditions need to be concerned, and further study should place more concentrations on these comprehensive influence factors and preventive actions of xerostomia.

Inhibition of GDF11 could promote bone healing in the tooth extraction socket and facilitate mesenchymal stem cell osteogenic differentiation in T2DM pigs.

BACKGROUND: Growth differentiation factor 11 (GDF11) might be a key factor responsible for the weakening of mesenchymal stem cell (MSC) osteogenic differentiation in tooth extraction sockets in patients with type 2 diabetes mellitus (T2DM). This study aimed to confirm that inhibition of GDF11 could promote bone healing in tooth extraction sockets and facilitate MSC osteogenic differentiation under T2DM conditions. METHODS: Three streptozotocin-induced T2DM pig models and two control pig models were established. The T2DM pigs were treated with an intrasocket injection of GDF11 inhibitor in the left mandible, whereas the right side was maintained for natural healing. The postextraction socket healing of the T2DM pigs was compared with that of nondiabetic controls. Healing was quantitatively verified by microcomputed tomography, and the GDF11 expression level was detected. MSCs from T2DM pig sockets were cultured and treated with a GDF11 inhibitor. The osteogenic differentiation ability of MSCs was also compared among groups. RESULTS: The expression of GDF11 in the tooth extraction sockets from T2DM pigs increased significantly post extraction. Bone healing was promoted by periodic injection of the GDF11 inhibitor into the extraction sockets of T2DM pigs. Furthermore, the osteogenic differentiation ability of T2DM-MSCs was improved in pigs treated with the GDF11 inhibitor. CONCLUSIONS: GDF11 inhibition could promote bone healing in the tooth extraction socket and facilitate MSC osteogenic differentiation under T2DM conditions. GDF11 could be a potential therapeutic target for undesirable alveolar bone healing in T2DM patients.

Hypermethylation in Calca Promoter Inhibited ASC Osteogenic Differentiation in Rats with Type 2 Diabetic Mellitus.

The abnormal environment of type 2 diabetes mellitus (T2DM) leads to a substantial decrease in osteogenic function of stem cells. However, the gene sequence does not vary before and after disease for the patient. This phenomenon may be related to changes in osteogenesis-related gene expression caused by DNA methylation. In this study, we established T2DM models to extract adipose-derived stem cells (ASCs) for different gene identifications through DNA methylation sequencing. Specific fragments of methylation changes in the target gene (Calca) were identified by IGV analysis. CGRP was applied to compare the effects on ASCs-T2DM morphology via phalloidin staining, proliferation through CCK-8 assay, and osteogenic differentiation with osteogenic staining, qPCR, and repair of calvarial defect. Furthermore, 5-azacytidine (5-az) was used to intervene ASCs-T2DM to verify the relationship between the methylation level of the target fragment and expression of Calca. We found that the DNA methylation level of target fragment of Calca in ASCs-T2DM was higher than that in ASCs-C. CGRP intervention showed that it did not change the morphology of ASCs-T2DM but could improve proliferation within a certain range. Meanwhile, it could significantly enhance the formation of ALP and calcium nodules in ASCs-T2DM, increase the expression of osteogenesis-related genes in vitro, and promote the healing of calvarial defects of T2DM rat in a concentration-dependent manner. 5-az intervention indicated that the reduction of the methylation level in Calca target fragment of ASCs-T2DM indeed escalated the gene expression, which may be related to DNMT1. Taken together, the environment of T2DM could upregulate the methylation level in the promoter region of Calca and then decrease the Calca expression. The coding product of Calca revealed a promoting role for osteogenic differentiation of ASCs-T2DM. This result provides an implication for us to understand the mechanism of the decreased osteogenic ability of ASCs-T2DM and improve its osteogenic capacity.

Selective laser melted titanium implants play a positive role in early osseointegration in type 2 diabetes mellitus rats.

Success of dental implant is associated with the surface modification. To evaluate whether the selective laser melting-superfinished titanium (Ti6Al4V) implants have a better early osseointegration in type 2 diabetes mellitus rats compare to pure titanium implants and acid-etched treated (SLA) implants, we designed a screw-shaped implant which was batch-fabricated by selective laser melting (SLM). Then the implants were randomly inserted in tibias of rats with type 2 diabetes mellitus (T2DM). After surgical operation, the SLM group showed the best bone formation around the implants with the highest bone-implant contact rate among the three groups.Removal torque tests and histomorphological analyses all revealed a stronger connection between the bone because its good surface characteristics and mechanical properties. SLM implant may be a novel implant for T2DM patients.

Research progress of superhydrophilic implants / 口腔疾病防治

@#Through a review of the literature on surface treatment of superhydrophilic implants and its clinical application, this paper discusses the shortening of load time, the improvement of the planting success rate and its long-term effect. Additionally, attention should be paid to the nonindication of superhydrophilic implants and issues requiring attention. The literature review showed that healthy patients could carry out an early load 21 days after implantation of superhydrophilic implants, and the load could be completed as soon as 6 weeks after implantation with superhydrophilic short implants when the residual alveolar bone height of the posterior dental area was repaired. Even if the residual alveolar bone density of the patient is low, the application of superhydrophilic implants can shorten the healing period to 8 weeks. Notably, some studies have reported that superhydrophilic implants have no significant effect on patients with a history of radiotherapy and the use of anticoagulants. Because the adhesion of the superhydrophilic implant to the bacteria is also improved to some extent, it is very important to prevent the use of antibiotics when using the superhydrophilic implant. Finally, this paper discusses and anticipates the future research direction of superhydrophilic implants: longer periodic follow-up and more in-depth molecular mechanism studies.

Exendin­4 promotes osteogenic differentiation of adipose­derived stem cells and facilitates bone repair.

Inflammation­related bone defects pose a heavy burden on patients and orthopedic surgeons. Although stem­cell­based bone repair has developed rapidly, it is of great significance to characterize bio­active molecules that facilitate bone regeneration. It is reported that a glucagon­like peptide 1 receptor agonist, exendin­4, promoted bone regeneration mediated by the transplantation of adipose­derived stem cells in a metaphyseal defect mouse model of femur injury. However, the underlying mechanism is unclear. Bone imaging, immunohistochemistry real­time PCR and western blot analysis were used in the present study, and the results revealed that exendin­4 increased the transcription of the osteogenic differentiation­related genes and induced osteogenic differentiation in situ. Furthermore, the present data obtained from sorted adipose­derived stem cells revealed that exendin­4 promoted osteogenic differentiation and inhibited adipogenic differentiation in vitro. These findings indicated that exendin­4 facilitates osteogenic differentiation of transplanted adipose­derived stem cells for bone repair and illuminated clinical prospects of both adipose­derived stem cells and exendin­4 in stem­cell­based bone defect repair.

Local Application of Semaphorin 3A Combined with Adipose-Derived Stem Cell Sheet and Anorganic Bovine Bone Granules Enhances Bone Regeneration in Type 2 Diabetes Mellitus Rats.

Bone tissue regeneration is considered to be the optimal solution for bone loss. However, diabetic patients have a greater risk of poor bone healing or bone grafting failure than nondiabetics. The purpose of this study was to investigate the influence of the complexes of an adipose-derived stem cell sheet (ASC sheet) and Bio-Oss® bone granules on bone healing in type 2 diabetes mellitus (T2DM) rats with the addition of semaphorin 3A (Sema3A). The rat ASC sheets showed stronger osteogenic ability than ASCs in vitro, as indicated by the extracellular matrix mineralization and the expression of osteogenesis-related genes at mRNA level. An ASC sheet combined with Bio-Oss® bone granules promoted bone formation in T2DM rats as indicated by microcomputed tomography (micro-CT) and histological analysis. In addition, Sema3A promoted the osteogenic differentiation of ASC sheets in vitro and local injection of Sema3A promoted T2DM rats' calvarial bone regeneration based on ASC sheet and Bio-Oss® bone granule complex treatment. In conclusion, the local injection of Sema3A and the complexes of ASC sheet and Bio-Oss® bone granules could promote osseous healing and are potentially useful to improve bone healing for T2DM patients.