Abstract Objectives Self-etch adhesives are well adopted in general practice, obviously primarily thanks to their ease of use and fast application time. Nevertheless, phosphoric acid is still often ...recommended to beforehand etch enamel following a so-called ‘selective’ enamel-etch technique, this in particular when most cavity margins end in enamel. The purpose of this study was to test if a new one-step adhesive can be applied in a multi-mode manner, this following different, either ‘full’ or ‘selective’, self-etch and etch-and-rinse approaches. Specific research hypotheses tested were that prior phosphoric-acid etching did not affect the bonding effectiveness of the one-step adhesive to enamel and dentine, and that the bonding effectiveness to dentine was also not affected when the adhesive was applied either following a ‘dry-bonding’ or ‘wet-bonding’ etch-and-rinse technique. Methods The micro-tensile bond strength (μTBS) of the one-step self-etch adhesive G-Bond Plus (GC, Tokyo, Japan; 1-SEA) was measured when it was bonded to bur-cut enamel following either a ‘self-etch’ or an ‘etch-and-rinse’ adhesive protocol, and to bur-cut dentine when applied following either a ‘self-etch’, a ‘dry-bonding’ or a ‘wet-bonding’ etch-and-rinse adhesive protocol. Bond-strength testing was corroborated by ultra-structural analysis of the interfacial interaction at enamel and dentine using transmission electron microscopy (TEM). Results Prior phosphoric-acid etching significantly increased the bonding effectiveness of the 1-SEA to enamel. A clearly enhanced micro-retentive surface was revealed by TEM. To dentine, no statistically significant difference in bonding effectiveness was recorded when the 1-SEA was either applied following a self-etch or both etch-and-rinse approaches. The ‘dry-bonding’ etch-and-rinse protocol was significantly more effective than its ‘wet-bonding’ version. TEM however revealed indications of low-quality hybridisation following both etch-and-rinse approaches, in particular in the form of a porous and poorly resin-infiltrated collagen mesh. Conclusions While phosphoric-acid etching definitely improved bonding of the one-step self-etch adhesive to enamel, one should be more careful with additional phosphoric-acid etching of dentine. Although the bond strength was not reduced, the resultant adhesive interface appeared ultra-structurally more vulnerable to biodegradation.
MicroRNAs (miRNAs) are small RNA molecules of 21-25 nucleotides that regulate cell behavior through inhibition of translation from mRNA to protein, promotion of mRNA degradation and control of gene ...transcription. In this study, we investigated the miRNA expression signatures of cell cultures undergoing osteoblastic and osteocytic differentiation from mesenchymal stem cells (MSC) using mouse MSC line KUSA-A1 and human MSCs. Ninety types of miRNA were quantified during osteoblastic/osteocytic differentiation in KUSA-A1 cells utilizing miRNA PCR arrays. Coincidently with mRNA induction of the osteoblastic and osteocytic markers, the expression levels of several dozen miRNAs including miR-30 family, let-7 family, miR-21, miR-16, miR-155, miR-322 and Snord85 were changed during the differentiation process. These miRNAs were predicted to recognize osteogenic differentiation-, stemness-, epinegetics-, and cell cycle-related mRNAs, and were thus designated OstemiR. Among those OstemiR, the miR-30 family was classified into miR-30b/c and miR-30a/d/e groups on the basis of expression patterns during osteogenesis as well as mature miRNA structures. In silico prediction and subsequent qRT-PCR in stable miR-30d transfectants clarified that context-dependent targeting of miR-30d on known regulators of bone formation including osteopontin/spp1, lifr, ccn2/ctgf, ccn1/cyr61, runx2, sox9 as well as novel key factors including lin28a, hnrnpa3, hspa5/grp78, eed and pcgf5. In addition, knockdown of human OstemiR miR-541 increased Osteopontin/SPP1 expression and calcification in hMSC osteoblastic differentiation, indicating that miR-541 is a negative regulator of osteoblastic differentiation. These observations indicate stage-specific roles of OstemiR especially miR-541 and the miR-30 family on novel targets in osteogenesis.
In this study, we aimed to compare the long-term survival of vital teeth adjacent to bounded edentulous spaces rehabilitated using an implant-supported prosthesis (ISP), a resin-bonded fixed partial ...denture (RBFPD), or a conventional fixed partial denture (CFPD). The risk factors for complications in teeth adjacent to the edentulous space (TAES) were also investigated.
We followed-up a consecutive series of 514 patients who underwent rehabilitation of a single bounded edentulous space with vital TAES (ISP: 103; RBFPD: 216; and CFPD: 195) from 2008 to 2017. Cumulative survival rates of prosthesis and TAES, and complication-free rates of TAES, were evaluated using the Kaplan−Meier analysis and log-rank test. Risk factors were evaluated using a Cox proportional hazards model.
Cumulative complication-free rates of TAES showed no significant differences among the three groups. The cumulative survival rate of TAES in CFPD was significantly lower than that of ISP (p = 0.037); no significant differences were observed between ISP and RBFPD (p = 0.513), and RBFPD and CFPD (p = 0.076). Older age (p = 0.027) was the only independent significant risk factor for complications in TAES. Installation of CFPD (p = 0.019), ceramic prosthesis in edentulous space (p = 0.026), and deeper periodontal probing depth (p = 0.018) of TAES were significant risk factors for non-surviving TAES.
Rehabilitating a single bounded edentulous space with CFPD could increase the risk for TAES loss compared with ISP. Risk of TAES loss remained similar between ISP and RBFPD, which can minimize the loss of coronal tooth structure during tooth preparation.
Teeth adjacent to edentulous space show equivalent longevity when rehabilitating a single bounded edentulous space with resin-bonded fixed partial dentures or single standing implant-supported prosthesis, at least 10 years post-installation.
The importance of macrophages in tissue development and regeneration has been strongly emphasized. However, the specific roles of macrophage colony-stimulating factor (MCSF), the key regulator of ...macrophage differentiation, in glandular tissue development have been unexplored. Here, we disclose new macrophage-independent roles of MCSF in tissue development. We initially found that MCSF is markedly upregulated at embryonic day (E)13.5, at a stage preceding the colonization of macrophages (at E15.5), in mouse submandibular gland (SMG) tissue. Surprisingly, MCSF-induced branching morphogenesis was based on a direct effect on epithelial cells, as well as indirectly, by modulating the expression of major growth factors of SMG growth, FGF7 and FGF10, via the phosphoinositide 3-kinase (PI3K) pathway. Additionally, given the importance of neurons in SMG organogenesis, we found that MCSF-induced SMG growth was associated with regulation of neurturin expression and neuronal network development during early SMG development in an
organogenesis model as well as
These results indicate that MCSF plays pleiotropic roles and is an important regulator of early SMG morphogenesis.
Mesenchymal stem cells (MSCs) are known to promote tissue regeneration and suppress excessive inflammation caused by infection or trauma. Reported evidence indicates that various factors influence ...the expression of MSCs' endogenous immunomodulatory properties. However, the detailed interactions of MSCs with macrophages, which are key cells involved in tissue repair, and their regulatory mechanisms are not completely understood. We herein investigated how age-related immunomodulatory impairment of MSCs alters the interaction of MSCs with macrophages during bone healing using young (5-week old) and aged (50-week old) mice. To clarify the relationship between inflammatory macrophages (M1) and MSCs, their spatiotemporal localization at the bone healing site was investigated by immunostaining, and possible regulatory mechanisms were analyzed in vitro co-cultures. Histomorphometric analysis revealed an accumulation of M1 and a decrease in MSC number at the healing site in aged mice, which showed a delayed bone healing. In in vitro co-cultures, MSCs induced M1 apoptosis through cell-to-cell contact but suppressed the gene expression of pro-inflammatory cytokines by soluble factors secreted in the culture supernatant. Interestingly, interleukin 38 (
) expression was up-regulated in M1 after co-culture with MSCs. IL-38 suppressed the gene expression of inflammatory cytokines in M1 and promoted the expression of genes associated with M1 polarization to anti-inflammatory macrophages (M2). IL-38 also had an inhibitory effect on M1 apoptosis. These results suggest that MSCs may induce M1 apoptosis, suppress inflammatory cytokine production by M1, and induce their polarization toward M2. Nevertheless, in aged conditions, the decreased number and immunomodulatory function of MSCs could be associated with a delayed M1 clearance (i.e., apoptosis and/or polarization) and consequent delayed resolution of the inflammatory phase. Furthermore, M1-derived IL-38 may be associated with immunoregulation in the tissue regeneration site.
S-adenosylmethionine (SAM) is considered to be a useful therapeutic agent for degenerative cartilage diseases, although its mechanism is not clear. We previously found that polyamines stimulate the ...expression of differentiated phenotype of chondrocytes. We also found that the cellular communication network factor 2 (CCN2) played a huge role in the proliferation and differentiation of chondrocytes. Therefore, we hypothesized that polyamines and CCN2 could be involved in the chondroprotective action of SAM. In this study, we initially found that exogenous SAM enhanced proteoglycan production but not cell proliferation in human chondrocyte-like cell line-2/8 (HCS-2/8) cells. Moreover, SAM enhanced gene expression of cartilage-specific matrix (aggrecan and type II collagen), Sry-Box transcription factor 9 (SOX9), CCN2, and chondroitin sulfate biosynthetic enzymes. The blockade of the methionine adenosyltransferase 2A (MAT2A) enzyme catalyzing intracellular SAM biosynthesis restrained the effect of SAM on chondrocytes. The polyamine level in chondrocytes was higher in SAM-treated culture than control culture. Additionally, Alcian blue staining and RT-qPCR indicated that the effects of SAM on the production and gene expression of aggrecan were reduced by the inhibition of polyamine synthesis. These results suggest that the stimulation of polyamine synthesis and gene expression of chondrogenic differentiation factors, such as CCN2, account for the mechanism underlying the action of SAM on chondrocytes.
Development of human bodies, organs, and tissues contains numerous steps of cellular differentiation including an initial zygote, embryonic stem (ES) cells, three germ layers, and multiple expertized ...lineages of cells. Induced pluripotent stem (iPS) cells have been recently developed using defined reprogramming factors such as Nanog, Klf5, Oct3/4 (Pou5f1), Sox2, and Myc. This outstanding innovation is largely changing life science and medicine. Methods of direct reprogramming of cells into myocytes, neurons, chondrocytes, and osteoblasts have been further developed using modified combination of factors such as N-myc, L-myc, Sox9, and microRNAs in defined cell/tissue culture conditions. Mesenchymal stem cells (MSCs) and dental pulp stem cells (DPSCs) are also emerging multipotent stem cells with particular microRNA expression signatures. It was shown that miRNA-720 had a role in cellular reprogramming through targeting the pluripotency factor Nanog and induction of DNA methyltransferases (DNMTs). This review reports histories, topics, and idea of cellular reprogramming.
The antagonist-specific regulation in tissue engineering constitutes important attempts to achieve an improved and rapid bone regeneration by controlling the natural biological response of the ...natural body growth factors. L51P is molecularly engineered bone morphogentic protein-2 (BMP-2) variant with a substitution of the 51st leucine with a proline residue. L51P is deficient in BMP receptor binding, but maintains its structure and affinity for inhibitory proteins such as noggin, chordin, and gremlin. These modifications convert the BMP-2 variant L51P into a receptor-inactive inhibitor of BMP antagonists. This current approach may prevent the uncontrolled bone overgrowth using high concentration of BMPs and thus regulates the possible growth factor’s high-dose side effects. Exploring of L51P biological functions is required to broad our understanding of BMP mutant biological functions and their potential clinical applications. The progress of L51P researches would hopefully lead to the development of multiple applications for using the L51P in bone and fracture healing disorders.
Purpose: Dental implant therapy is a common clinical treatment for missing teeth. However, the esthetic result is not as satisfactory as expected in some cases, especially in the anterior maxillary ...area. Poor esthetic results are caused by inadequate preparation of the hard and soft tissues in this area before treatment. The socket shield technique may be an alternative for a desirable esthetic outcome in dental implant treatments.Study selection: In the present systematic review, PubMed-Medline, Google Scholar, and ScienceDirect were searched for clinical studies published from January 2000 to December 2018.Results: Twenty studies were included, comprising one randomized controlled trial, two cohort studies, 14 clinical human case reports, and three retrospective case series. In total, 288 patients treated with the socket shield technique with immediate implant placement and follow-up between 3–60 months after placement were included. A quality assessment showed that 12 of the 20 included studies were of good quality. Twenty-six of the 274 (9.5%) cases developed complications or adverse effects related to the socket shield technique. Most studies reported implant survival without the complications (90.5%); most of the cases that were followed up for more than 12 months after implant placement achieved a good esthetic appearance. The failure rate was low without the complications, although there were some failures due to failed implant osseointegration, socket shield mobility and infection, socket shield exposure, socket shield migration, and apical root resorption.Conclusions: The socket shield technique can be used in dental implant treatment, but it remains difficult to predict the long-term success of this technique until high-quality evidence becomes available.
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