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Current research on dental implants has mainly focused on the influence of surface roughness on the rate of osseointegration, while studies on the development of surfaces to also improve the interaction of peri-implant soft tissues are lacking. To this end, the first purpose of this study was to evaluate the response of human gingival fibroblasts (hGDFs) to titanium implant discs (Implacil De Bortoli, Brazil) having different micro and nano-topography: machined (Ti-M) versus sandblasted/double-etched (Ti-S). The secondary aim was to investigate the effect of the macrogeometry of the discs on cells: linear-like (Ti-L) versus wave-like (Ti-W) surfaces. The atomic force microscopy (AFM) and scanning electron microscopy (SEM) analysis showed that the Ti-S surfaces were characterized by a significantly higher micro and nano roughness and showed the 3D macrotopography of Ti-L and Ti-W surfaces. For in vitro analyses, the hGDFs were seeded into titanium discs and analyzed at 1, 3, and 5 days for adhesion and morphology (SEM) viability and proliferation (Cck-8 and MTT assays). The results showed that all tested surfaces were not cytotoxic for the hGDFs, rather the nano-micro and macro topography favored their proliferation in a time-dependent manner. Especially, at 3 and 5 days, the number of cells on Ti-L was higher than on other surfaces, including Ti-W surfaces. In conclusion, although further studies are needed, our in vitro data proved that the use of implant discs with Ti-S surfaces promotes the adhesion and proliferation of gingival fibroblasts, suggesting their use for in vivo applications.

This study investigates the role of NLRP3 inflammasome and its main effector Caspase-1 in inflammation and alveolar bone resorption associated with periodontitis. Heat-killed Aggregatibacter actinomycetemcomitans (Aa) was injected 3x/week (4 weeks) into gingival tissues of wild-type (WT), Nlrp3-KO and Caspase1-KO mice. Bone resorption was measured by µCT and osteoclast number was determined by tartrate-resistant acid phosphatase (TRAP) staining. Inflammation was assessed histologically (H/E staining and immunofluorescence of CD45 and Ly6G). In vitro studies determined the influence of Nlrp3 and Caspase-1 in Rankl-induced osteoclast differentiation and activity and on LPS-induced expression of inflammation-associated genes. Bone resorption was significantly reduced in Casp1-KO but not in Nlrp3-KO mice. Casp1-KO mice had increased in osteoclast numbers, whereas the inflammatory infiltrate or on gene expression were similar to those of WT and Nlrp3-KO mice. Strikingly, osteoclasts differentiated from Nlrp3-deficient macrophages had increased resorbing activity in vitro. LPS-induced expression of Il-10, Il-12 and Tnf-α was significantly reduced in Nlrp3- and Casp1-deficient macrophages. As an inceptive study, these results suggest that Nlrp3 inflammasome does not play a significant role in inflammation and bone resorption in vivo and that Caspase-1 has a pro-resorptive role in experimental periodontal disease.

Primary stem cells, after several cell divisions, enter into a senescence state, that is characterized by alterations to spindle-shape typical morphology. This concern is one of the main problems in the use of human mesenchymal stem cells (hMSCs) in clinical applications which demand cells in large numbers. Short peptides had geroprotective properties and stimulated stem cell differentiation. The aim of the study is to demonstrate the role of AEDG and KED peptides in maintaining oral hMSCs morphology and functions over long-term expansion. 2 types of hMSCs were investigated: human periodontal ligament stem cells (hPLSCs) and human gingival mesenchymal stem cells (hGMSCs). Cells at the 25th passage were divided into 3 groups: 1 - control (without adding peptide), 2 - treated with AEDG peptide, 3 - treated with KED peptide. Cell cultures were analyzed by an immunofluorescence method and RT-PCR on the p16 and p21 senescence markers expression. AEDG peptide decreased p16 and p21 mRNA expression by 1.56-2.44 times in comparison with the control group. KED peptide decreased p16 and p21 mRNA expression by 1.82-3.23 times in comparison with the control group. These results were confirmed by immunofluorescent visualization. AEDG and KED peptides could be used as supplementary substances in a culture medium to delay the expression of senescence markers in long term stem cell cultivation in order to promote the large-scale in vitro expansion necessarily required for stem cell therapy clinical application. The data obtained confirm the geroprotective effect of AEDG and KED peptide, which was shown early in animal and cells models.

Calcium signaling plays a central role in bone development and homeostasis. Store operated calcium entry (SOCE) is an important calcium influx pathway mediated by calcium release activated calcium (CRAC) channels in the plasma membrane. Stromal interaction molecule 1 (STIM1) is an endoplasmic reticulum calcium sensing protein important for SOCE. We generated a mouse model expressing the STIM1 R304W mutation, causing Stormorken syndrome in humans. Stim1 R304W/R304W mice showed perinatal lethality, and the only three animals that survived into adulthood presented with reduced growth, low body weight, and thoracic kyphosis. Radiographs revealed a reduced number of ribs in the Stim1 R304W/R304W mice. Microcomputed tomography data revealed decreased cortical bone thickness and increased trabecular bone volume fraction in Stim1 R304W/R304W mice, which had thinner and more compact bone compared to wild type mice. The Stim1 R304W/+ mice showed an intermediate phenotype. Histological analyses showed that the Stim1 R304W/R304W mice had abnormal bone architecture, with markedly increased number of trabeculae and reduced bone marrow cavity. Homozygous mice showed STIM1 positive osteocytes and osteoblasts. These findings highlight the critical role of the gain-of-function (GoF) STIM1 R304W protein in skeletal development and homeostasis in mice. Furthermore, the novel feature of bilateral subgingival hair growth on the lower incisors in the Stim1 R304W/R304W mice and 25 % of the heterozygous mice indicate that the GoF STIM1 R304W protein also induces an abnormal epithelial cell fate., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The phosphorylation of (+) alpha tocopherol produces adhesive nanostructures that interact with oral biofilms to restrict their growth. The aim of this work was to understand if these adhesive (+) alpha tocopheryl phosphate (α-TP) nanostructures could also control macrophage responses to the presence of oral bacteria. The (+) α-TP planar bilayer fragments (175 nm ± 21 nm) formed in a Trizma®/ethanol vehicle swelled when exposed to the cell lines (maximum stabilized size = 29 μm). The swelled (+) α-TP aggregates showed selective toxicity towards THP-1 macrophages (LD 50  = 304 μM) compared to human gingival fibroblasts (HGF-1 cells; LD 50  > 5 mM), and they inhibited heat killed bacteria stimulated MCP-1 production in both macrophages (control 57.3 ± 18.1 pg/mL vs (+) α-TP 6.5 ± 3.2 pg/mL) and HGF-1 cells (control 673.5 ± 133 pg/mL vs (+) α-TP - 463.9 ± 68.9 pg/mL)., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background The etiology of gingival overgrowth due to cyclosporine A (CsA) is still unknown. The aim of this study was to determine the possible role of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) on extra-cellular matrix (ECM) homeostasis when treated with different levels of CsA and its difference between fetal and adult human gingival fibroblasts (HGFs). Methods Each group of cells (adult and fetal) was cultured in 40 wells that consisted of four different CsA treatment concentrations. Every 10 wells were treated with 0, 50, 100, and 150 ng/mL of CsA which makes a total of 80 wells. Supernatants of every well were used to determine the concentration of MMPs and TIMPs using the Elisa kits from Boster, CA, USA. Results MMP-1 level increased with the treatment of CsA when treated with 50 and 150 ng/mL of CsA (p = 0.02 and p = 0.04) as TIMP-1 decreased (p < 0.0001) in adult group; while in the fetal group, TIMP-1 level increased with treatment of 150 ng/mL (p < 0.0001). MMP-2 level increased in both adult and fetal groups (p < 0.0001). MMP-3 level decreased in adult group (p < 0.0001) but went up in fetal HGFs (p = 0.01) when treated with 150 ng/mL CsA. TIMP-2 level increased in all wells significantly when treated with CsA (p < 0.0001). The study showed that CsA affects secretion of MMPs and TIMPs. MMP-1 increment and TIMP-1 decrement were observed, which indicate more degradation of ECM. This may be due to single donor use in this study. TIMP-2 and MMP-2 were both more active when treated with CsA which may be due to the gelatinase activity of them and that in CsA gingival overgrowth. There was more inflammation rather than fibrosis.

Human gingival fibroblasts (HGFs) are responsible for connective tissue repair and scarring, and are exposed to mechanical forces under physiological and pathological conditions. The exact mechanisms underlying gingival tissue reconstruction under mechanical forces remain unclear. The present study aimfed to investigate the effects of mechanical forces on the proliferation and extracellular matrix synthesis in HGFs by establishing a 3‑dimensional (3D) HGF culture model using poly(lactide‑co‑glycolide) (PLGA) scaffolds. HGFs were cultured in 3D PLGA scaffolds and a mechanical force of 0, 5, 15, 25 or 35 g/cm2 was applied to HGFs for 24 h. A mechanical force of 25 g/cm2 induced the highest proliferation rate, and thus was selected for subsequent experiments. Cell viability was determined using the MTT assay at 0, 24, 48 and 72 h. The expression levels of type I collagen (COL‑1) and matrix metallopeptidase (MMP)‑1 were examined by reverse transcription‑quantitative polymerase chain reaction and ELISA, and transforming growth factor (TGF)‑β expression was evaluated by ELISA. The application of mechanical force on HGFs cultured on the 3D PLGA scaffolds resulted in a significant increase in cell proliferation and COL‑1 expression, as well as a decrease in MMP‑1 expression. A TGF‑β1 inhibitor was also applied, which attenuated the effects of mechanical force on HGF proliferation, and COL‑1 and MMP‑1 expression, thus suggesting that TGF‑β signaling pathways may mediate the mechanical force‑induced alterations observed in HGFs. In conclusion, these findings helped to clarify the mechanisms underlying mechanical force‑induced HGF proliferation and ECM synthesis, which may promote the development of targeted therapeutics to treat various diseases, including gingival atrophy caused by orthodontic treatment.

Several human dental traits typical of modern humans appear to be associated with the prolonged period of development that is a key human attribute. Understanding when, and in which early hominins, these dental traits first appeared is thus of strong interest. Using x-ray multiresolution synchrotron phase-contrast microtomography, we quantify dental growth and development in an archaic Homo juvenile from the Xujiayao site in northern China dating to 161,000-224,000 years or 104,000-125,000 years before present. Despite the archaic morphology of Xujiayao hominins, most aspects of dental development of this juvenile fall within modern human ranges (e.g., prolonged crown formation time and delayed first molar eruption). For its estimated age-at-death (6.5 years), its state of dental development is comparable to that of equivalently aged modern children. These findings suggest that several facets of modern human dental growth and development evolved in East Asia before the appearance of fully modern human morphology.

Background: Periodontitis is an inflammatory disease that can result in destruction of the tooth attachment apparatus. Therefore, periodontal tissue regeneration is currently an important focus of research in the field. Approaches using stem cells and reprogrammed cells, such as induced pluripotent stem cells (iPSCs) or trans-differentiated cells, represent the cutting edge in periodontal regeneration, and have led to many trials for their clinical application. Objectives and Results: In this review, we consider all available stem cell sources, methods to obtain the cells, their capability to differentiate into the desired cells, and the extent of their utilization in periodontal regeneration. In addition, we introduce the new concepts of using iPSCs and transdifferentiated cells for periodontal regeneration. Finally, we discuss the promise of tissue engineering for improving cell therapy outcomes for periodontal regeneration., Conclusions: Despite their limitations, iPSCs and trans-differentiated cells may be promising cell sources for periodontal tissue regeneration. Further collaborative investigation is required for the effective and safe application of these cells in combination with tissue engineering elements, like scaffolds and biosignals., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Mesenchymal stem cells (MSCs) have been considered promising tools for tissue engineering and regenerative medicine. However, the optimal cell source for bone regeneration remains controversial. To better identify seed cells for bone tissue engineering, we compared MSCs from seven different tissues, including four from dental origins, dental pulp stem cells (DPSCs), periodontal ligament stem cells (PDLSCs), gingival MSCs (GMSCs), and dental follicle stem cells (DFSCs); two from somatic origins, bone marrow-derived MSCs (BM-MSCs) and adipose-derived stem cells (ADSCs); and one from birth-associated perinatal tissue umbilical cord (UCMSCs). We cultured the cells under a standardized culture condition and studied their biological characteristics. According to our results, these cells exhibited similar immunophenotype and had potential for multilineage differentiation. MSCs from dental and perinatal tissues proliferated more rapidly than those from somatic origins. Simultaneously, DPSCs and PDLSCs owned stronger antiapoptotic ability under the microenvironment of oxidative stress combined with serum deprivation. In respect to osteogenic differentiation, the two somatic MSCs, BM-MSCs and ADSCs, demonstrated the strongest ability for osteogenesis compared to PDLSCs and DFSCs, which were just a little bit weaker than the formers. However, GMSCs and UCMSCs were the most pertinacious ones to differentiate to osteoblasts. We also revealed that the canonical intracellular protein kinase-based cascade signaling pathways, including PI3K/AKT, MAPK/ERK, and p38 MAPK, possessed different levels of activation in different MSCs after osteoblast induction. Our conclusions suggest that PDLSCs might be a good potential alternative to BM-MSCs for bone tissue engineering.

Since the oral mucosa is continuously exposed to abundant microbes, one of its most important defense features is a highly proliferative, thick, stratified epithelium. The cellular mechanisms responsible for this are still unknown. The aim of this study was to determine whether multi-species oral biofilm contribute to the extensive stratification and primed antimicrobial defense in epithelium. Two in vitro models were used: 3D reconstructed human gingiva (RHG) and oral bacteria representative of multi-species commensal biofilm. The organotypic RHG consists of a reconstructed stratified gingiva epithelium on a gingiva fibroblast populated hydrogel (lamina propria). Biofilm was cultured from healthy human saliva, and consists of typical commensal genera Granulicatella and major oral microbiota genera Veillonella and Streptococcus. Biofilm was applied topically to RHG and host-microbiome interactions were studied over 7 days. Compared to unexposed RHG, biofilm exposed RHG showed increased epithelial thickness, more organized stratification and increased keratinocyte proliferation. Furthermore biofilm exposure increased production of RHG anti-microbial proteins Elafin, HBD2 and HBD3 but not HBD1, adrenomedullin or cathelicidin LL-37. Inflammatory and antimicrobial cytokine secretion (IL-6, CXCL8, CXCL1, CCL20) showed an immediate and sustained increase. In conclusion, exposure of RHG to commensal oral biofilm actively contributes to RHG epithelial barrier function.

Gingival overgrowth is a side effect of certain medications, including calcium channel blockers, cyclosporin A, and phenytoin. Phenytoin-induced gingival overgrowth is fibrotic. Lysyl oxidases are extracellular enzymes that are required for biosynthetic cross-linking of collagens, and members of this enzyme family are upregulated in fibrosis. Previous studies in humans and in a mouse model of phenytoin-induced gingival overgrowth have shown that LOXL2 is elevated in the epithelium and connective tissue in gingival overgrowth tissues and not in normal tissues. Here, using a novel LOXL2 isoform-selective inhibitor and knockdown studies in loss- and gain-of-function studies, we investigated roles for LOXL2 in promoting cultures of human gingival fibroblasts to proliferate and to accumulate collagen. Data indicate that LOXL2 stimulates gingival fibroblast proliferation, likely by a platelet-derived growth factor B receptor-mediated mechanism. Moreover, collagen accumulation was stimulated by LOXL2 enzyme and inhibited by LOXL2 inhibitor or gene knockdown. These studies suggest that LOXL2 could serve as a potential therapeutic target to address oral fibrotic conditions.

Tooth eruption is characterized by a concert of mechanisms that result in the emergence of teeth in the oral cavity. Genetic variants seem to regulate this process and the formation of a gingival sulcus around the teeth. Interindividual variability in the response to microbial triggers in the sulcus plays an important role in the onset and progression of periodontal diseases. Host genetic variants can influence this variability, affecting the response of the host to the subgingival biofilm. Genetic factors affecting tooth eruption could potentially influence susceptibility to periodontal diseases and, specifically, susceptibility to localized aggressive periodontitis. This review aims to discuss the evidence available for the role of host genetic variants in tooth eruption and to and to give some directions for prospective research in this topic., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: The aim of the study was to evaluate 2 types of collagen scaffold for gingival regeneration., Study Design: Two types of collagen scaffolds, CS-pH7.4 and CS-pH3.0, were prepared by processing atelocollagen at pH 7.4 or 3.0, respectively, followed by dehydrothermal treatment. Gingival wounds with sizes of 4 × 6 mm (rectangle) or 6 mm diameter (circle) were made with buccal incisions in beagle dogs. The defective area was surgically covered with the CS-pH7.4, CS-pH3.0, or no scaffold (control). Gingival regeneration was assessed by monitoring the differences in the lengths of the epithelial and submucosal tissues at the wound site and the normal site. Histopathologic assessments were performed by 4 evaluators independently; statistical significance was evaluated by using the Wald test., Results: Significantly higher recovery of epithelial and submucosal tissues, which, in turn, resulted in recovery of gum thickness, was observed in gingival wounds treated with the CS-pH7.4 compared with that in the control. CS-pH3.0 treatment also resulted in higher gingival regeneration compared with the control; however, the effects were more pronounced in wounds treated with the CS-pH7.4. CS-pH7.4-treated wounds showed better gingival regeneration compared with the control and CS-pH3.0-treated wounds, even after adjusting for interevaluator differences using a linear mixed model., Conclusions: CS-pH7.4 is a promising scaffold for gingival tissue regeneration., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Previous studies have demonstrated increased expression and raised levels of human β-defensin (hBD)-1 in gingival tissue and crevicular fluid of patients with chronic periodontitis and peri-implantitis, oral bone-resorbing diseases caused by enhanced osteoclastogenesis. Therefore, we aimed to investigate the effect of hBD-1 on osteoclast formation and function and to elucidate the involved signaling pathway in vitro. Human peripheral blood mononuclear cells (PBMCs) were first incubated with various doses of hBD-1 and cell viability was assayed by MTT. PBMCs were treated with macrophage-colony stimulating factor and receptor activator of nuclear factor kappa-B ligand (RANKL) in the presence or absence of non-toxic doses of hBD-1. In vitro osteoclastogenesis was analyzed by tartrate-resistant acid phosphatase (TRAP) staining, osteoclast-specific gene expression, and a resorption pit assay. Involvement of mitogen-activated protein kinases (MAPKs) was studied by immunoblotting and specific MAPK inhibitors. HBD-1 potentiated induction of in vitro osteoclastogenesis by RANKL, as shown by significantly increased number of TRAP-positive multinuclear cells and resorption areas on the dentin slices, and further up-regulated expressions of osteoclast-specific genes compared to those by RANKL treatment (p <0.05). However, hBD-1 treatment without RANKL failed to induce formation of osteoclast-like cells. A significant and further increase in transient phosphorylation of the p44/42 MAPKs was demonstrated by hBD-1 co-treatment (p<0.05), consistent with the inhibitory effect by pretreatment with U0126 or PD98059 on hBD-1-enhanced osteoclastogenesis. Collectively, hBD-1 potentiates the induction of in vitro osteoclastogenesis by RANKL via enhanced phosphorylation of the p44/42 MAPKs., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Objective: Premolar autotransplantation represents an effective therapeutic option for the treatment of juvenile dentition with either aquired or congenital hypodontia. The objective of this prospective clinical study was to quantitatively assess bone and soft tissue levels after autogenous premolar transplantation by clinical and radiographic parameters., Study Design: In the study, 26 premolars were transplanted in 20 patients after traumatic tooth loss (n = 16) or congenital aplasia (n = 10) in the anterior maxilla. Based on standardized photographic documentation, the relative soft tissue level was measured compared to the healthy adjacent teeth. Radiographic findings included evaluation of root resorption, pulp canal obliteration, and relative bone height., Results: Average survival rate of transplanted premolars (n = 26) was 100% over a follow-up period of 29 months (range 10-60 months). The relative soft tissue level significantly increased by +1.1 mm (P < .01). Radiographs showed a tendency toward vertical bone growth. Continuous root development and signs of pulpal healing were observed postoperatively in 18 transplants (69.2%)., Conclusions: Autogenous premolar transplantation represents a safe method to ensure functional and aesthetic rehabilitation in the anterior maxilla irrespective of the nature of tooth loss., (Copyright © 2017 Elsevier Inc. All rights reserved.)

While oral rinses used for cosmetic purposes only do not necessarily have to be antiseptic, antimicrobial activity is required for medical indications, including oral and periodontal surgery. So the question arises-is the antimicrobial activity of oral rinses associated with any destructive changes in cell viability in vitro? To answer this question, we examined twelve oral rinses with respect to their antimicrobial and cytotoxic activity. Antimicrobial activity was screened against five bacterial strains using disc diffusion. Cytotoxicity was determined by mitochondrial reductase activity with primary gingival fibroblasts, L929 cells, and HSC-2 epithelial cells. Phase contrast microscopy and trypan blue staining were then performed to reveal cell morphology. Cells remained vital after exposure to oral rinses that were only used for cosmetic purposes. Moderate cytotoxic effects were observed for oral rinses containing 0.05% chlorhexidine, ethanol, or pegylated hydrogenated castor oil and sodium dodecyl sulfate. Other oral rinses containing 0.2% chlorhexidine and cocamidopropyl betaine exhibited strong cytotoxic and antimicrobial activity. Strong cytotoxic but moderate antimicrobial activity was observed in oral rinses containing cetylpyridinium chloride. The in vitro data show that oral rinses are heterogeneous with respect to their cytotoxic and antimicrobial effects. Based on their respective properties, oral rinses can be selected either to reduce the microbial load or for cosmetic purposes.

Additive manufacturing is becoming increasingly important in dentistry for the production of surgical guides. The development of cost-effective desktop stereolithography (SLA) printing systems and the corresponding resins makes this novel technique accessible to dental offices and dental laboratories. The aim of the study was to reveal the response of soft tissue cells to Clear and Dental SG resins used in desktop SLA printing systems at different stages of processing. Cell activity of L929 cells and gingival fibroblasts (GF) in response to the materials was examined in indirect and direct monolayer culture models and a direct spheroid culture model based on MTT, resazurin-based toxicity assays, and live-dead staining. Overall we found that the impact of Clear and Dental SG resins on L929 and GF depends on the processing stage of the materials. Liquid Clear resin induced a stronger reduction of cell activity compared to Dental SG resin. Printing and postcuring reduced the impact on cell activity and viability. As in-house 3D printing for surgical guides is getting integrated in the digital workflow, our data suggest that careful adherence to processing guidelines-especially postcuring-is of clinical relevance.

The purposes of this case report were to describe a growing two-cm gingival mass that developed after natal teeth were extracted in a four-month-old female patient, present a review of the literature on the growth of a gingival mass after the extraction of natal teeth, and illustrate the clinical and histological features that differentiate this condition from other types of gingival masses in infants. Histological examination of the excised mass revealed that it contained tooth-like hard tissue (regular and irregular dentin) that intermingled with bone, dental pulp, and fibrous tissue. We found eight cases from 1962 to 2009 in which a soft-tissue mass with dentin-like hard tissue or a tooth-like structure had developed after the extraction of natal teeth. Based on clinical and histological findings, we deduced that the mass was the result of abnormal growth of a residual dental papilla, including mesenchymal stem cells. Consequently, dentists, obstetricians, gynecologists, and pediatricians should be aware of this potential complication and observe caution before they extract natal teeth.

Background and Objective: The gingiva is the first oral tissue directly exposed to cigarette smoke (CS). Exposure to CS compromises the structure and function of gingival tissue. Damaging or altering the gingival epithelium leads to a compromised protective barrier of the periodontium, resulting in several diseases. The aim of this study was to assess the effect of repeated exposure to CS on gingival epithelial cell growth and on expression of apoptotic protein and keratin., Material and Methods: Primary human gingival epithelial cells were seeded on a collagen scaffold for 5 d to allow growth and stratification. The cells were then exposed for 5 min to whole CS for 3, 6 and 9 d. At the end of each exposure period, cell proliferation [using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (MTT) and 5-bromo-2'-deoxyuridine (BrdU) assays], gene expression [by quantitative reverse transcription polymerase chain reaction (qRT-PCR)] and protein production (by western blot analysis) were investigated., Results: Higher metabolic activity was found in the CS-exposed cells than in the nonexposed cells, specifically after 3 and 6 d of exposure to CS. At 9 d there was no significant difference between CS-exposed and nonexposed cells. Metabolic activity was supported by the BrdU cell-proliferation analyses, which showed increased cell growth at 3 d compared with the control. However, at 6 and 9 d, cell proliferation in the CS-exposed culture was comparable to that in the nonexposed culture. Interestingly, the Bax/Bcl-2 protein ratios decreased with increased CS exposure, suggesting cell resistance. Moreover, protein analyses showed that CS decreased expression of keratin(K) 5 at 3, 6 and 9 d, and increased expression of K14 at 6 and 9 d. Finally, mRNA analyses showed significant decreases of K1, K6, K10 and K16 in CS-exposed cultures, correlating, at times, with a decrease of protein production., Conclusion: CS was shown to increase epithelial cell proliferation, which may involve cell resistance to apoptosis. This is supported by the modulation of expression of different keratin genes and proteins. Altogether, these data may explain the hyperplasia reported in gingival tissue, as well as periodontal disease, in smokers., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The dental morphology usually determines the shape and volume of the interdental space, which must be filled by a dense connective tissue covered by oral epithelium to achieve pleasant esthetics. When composite resin restorations are placed to solve esthetic problems, the restorative procedure must be designed to allow the formation of healthy interdental papilla. This case report discusses aspects that should be considered when composite resin restorations are proposed for diastema closure. A 23-year-old man sought treatment for variations of space in the anterior dentition after orthodontic treatment. Direct composite resin restorations were placed in a way that respected the emergence profile, even though "black triangles" were evident immediately after the procedure. At the 45-day follow-up, complete closure of the interdental spaces by healthy papillae was observed. The emergence profile should be identified and respected when restorations are placed to obtain diastema closure because healthy periodontal tissues and acceptable esthetics depend on it.

Gingival stem cells (GSCs) are a novel source of mesenchymal stem cells (MSCs) that are easily accessed from the oral cavity. GSCs were considered valuable autograft MSCs with particular characteristics. However, the limitation in the number of available GSCs remains an obstacle. Therefore, this study aimed to stimulate GSC proliferation by ascorbic acid (AA) and determined the effects of AA on GSC pluripotent potential-related gene expression. GSCs were isolated from gum tissue by explant culture and continuously subcultured before analysis of stemness and effects of AA on pluripotent-related gene expression. GSCs cultured with various concentrations of AA showed increased proliferation in a dose-dependent manner. AA-treated GSCs showed significantly higher expression of SSEA-3, Sox-2, Oct-3/4, Nanog, and TRA-1-60 compared with control cells. More importantly, GSCs also maintained their stemness with MSC phenotypes and failed to cause tumors in nude athymic mice. Our results show that AA is a suitable factor to stimulate GSC proliferation.

In this report we provide data on dental eruption and tooth germ maturation at birth in a large sample constituting the broadest array of non-human primates studied to date. Over 100 perinatal primates, obtained from natural captive deaths, were screened for characteristics indicating premature birth, and were subsequently studied using a combination of histology and micro-CT. Results reveal one probable unifying characteristic of living primates: relatively advanced maturation of deciduous teeth and M1 at birth. Beyond this, there is great diversity in the status of tooth eruption and maturation (dental stage) in the newborn primate. Contrasting strategies in producing a masticatory battery are already apparent at birth in strepsirrhines and anthropoids. Results show that dental maturation and eruption schedules are potentially independently co-opted as different strategies for attaining feeding independence. The most common strategy in strepsirrhines is accelerating eruption and the maturation of the permanent dentition, including replacement teeth. Anthropoids, with only few exceptions, accelerate mineralization of the deciduous teeth, while delaying development of all permanent teeth except M1. These results also show that no living primate resembles the altricial tree shrew (Tupaia) in dental development. Our preliminary observations suggest that ecological explanations, such as diet, provide an explanation for certain morphological variations at birth. These results confirm previous work on perinatal indriids indicating that these and other primates telegraph their feeding adaptations well before masticatory anatomy is functional. Quantitative analyses are required to decipher specific dietary and other influences on dental size and maturation in the newborn primate., (© 2015 Wiley Periodicals, Inc.)

This study was conducted to evaluate the usefulness of decellularized and lyophilized extracellular matrix, which was acquired from human amniotic membrane, for surgical closure of the mucogingival defects. Preliminarily, to create a gingival recession defect, silk ligature was applied on the gingival part of the upper incisor in the first (experimental) (n=20) and second (control) (n=20) groups. On the 14th day, the ligature was removed and the damaged gingival tissues were resected. The formed mucogingival defect, in the animals of the first group, was covered with acellular human amniotic three-dimensional scaffold with bone marrow stem cells. Animals with mucogingival defect of the second group were left untreated and served as controls. Unlike the animals from the control group, in animals from the experimental group the mucogingival defect already on the seventh day was completely closed and there was the newly formed epithelial lining, which in shape and color did not differ from the normal. Acellular human amniotic membrane as a three-dimensional scaffold boosts angiogenesis and increases the reparative regeneration of the damaged tissues; and it is well-tolerated by the gingival tissues. Hence, human amniotic membrane might be a suitable alternative to other conventional methods of treating gingival recession.

Knowledge of periodontal anatomy is essential when performing surgical and non-surgical procedures in the field of oral healthcare. Gingival thickness (GT) is often assessed for this purpose. A dental system prototype was recently developed for quantitative, non-invasive GT assessment by high-frequency (HF) ultrasound. Laboratory trials were conducted to validate system performance against a traditional method of assessment. A system with a 50 MHz broadband, spherically-focused transducer was used. The transducer was housed in a small, hand-held probe equipped with a continuous water supply. A-scans were obtained and thickness at each location was determined. For comparison, the traditional method of transgingival probing through tissue with an endodontic k-file needle was also implemented. Preliminary experiments were performed on phantoms simulating the anatomical and acoustic properties of human periodontal tissues. A porcine cadaver was obtained for further laboratory trials. The speed of sound through porcine gingiva was determined to be 1564 ± 21 m/s. Finally, a multiple-point experiment involved GT assessment in an array of locations on the buccal gingival surface in the fourth quadrant. Ultrasonic measurements were found to yield similar GT values to those obtained from invasive methods. Results obtained in this experiment validate the applicability of ultrasound as a diagnostic tool for assessing periodontal anatomy., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

Comprehensive health care in children and youth includes periodic oral examinations. The mucogingival complex undergoes significant changes during development. The factors that impact the width of attached and keratinized gingiva during this period of life are: tooth eruption phase, the position in the arch, the type of frena attachment and oral hygiene. Along with the child's development the width of attached and keratinized gingiva increases, except for the period of tooth replacement, when a temporary narrowing of attached gingiva of erupting permanent teeth is observed. The understanding of physiological processes of the mucogingival complex is prerequisite for diagnostics and treatment of gingival abnormalities in children and youth. Therefore close cooperation between paediatrician and dental specialists: paedodontist, orthodontist and periodontologist is essential.

Introduction: The cytotoxicity of 2 novel calcium silicate-containing endodontic sealers to human gingival fibroblasts was studied., Methods: EndoSequence BC (Brasseler, Savannah, GA), MTA Fillapex (Angelus Indústria de Produtos Odontológicos S/A, Londrina, PR, Brazil) and a control sealer (AH Plus; Dentsply DeTrey GmbH, Konstanz, Germany) were evaluated. Human gingival fibroblasts were incubated for 3 days both with the extracts from fresh and set materials in culture medium and cultured on the surface of the set materials in Dulbecco-modified Eagle medium. Fibroblasts cultured in Dulbecco-modified Eagle medium were used as a control group. Cytotoxicity was evaluated by flow cytometry, and the adhesion of the fibroblasts to the surface of the set materials was assessed using scanning electron microscopy. The data of cell cytotoxicity were analyzed statistically using a 1-way analysis of variance test at a significance level of P < .05., Results: Cells incubated with extracts from BC Sealer showed higher viabilities at all extract concentrations than cells incubated with extracts from freshly mixed AH Plus and fresh and set MTA Fillapex, esspecially for the high extract concentrations (1:2 and 1:8 dilutions). Extracts from set MTA Fillapex of 2 weeks and older were more cytotoxic than extracts from freshly mixed and 1-week-old cement. With extract concentrations of 1:32 and lower, MTA Fillapex was no longer cytotoxic. After setting, AH Plus was no longer cytotoxic, and the fibroblast cells grew on set AH Plus equally as well as on BC Sealer., Conclusions: BC Sealer and MTA Fillapex, the 2 calcium silicate-containing endodontic sealers, exhibited different cytotoxicity to human gingival fibroblasts., (Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

This paper reviews the effects that drugs may have on the gingival and periodontal tissues. Drug-induced gingival overgrowth has been recognised for over 70 years but is becoming a more prevalent occurrence with wider use of antihypertensive and immunosuppressant drugs. The anti-inflammatory steroids, non-steroidal drugs and anti-TNF-α agents might all be expected to exert a dampening effect on chronic periodontitis although the evidence is somewhat equivocal and none of these drugs has emerged as potentially valuable adjuncts to treat periodontal disease. Desquamative gingivitis is a clinical appearance of aggressive gingival inflammation with which a number of drugs have been associated and the oral contraceptives have also been implicated in the development of gingival inflammation. Patients who are prescribed bisphosphonates and anti-platelet drugs are at risk of serious side effects following more invasive dental procedures including extractions and surgical treatments although timely, conventional management of periodontal disease may be undertaken to reduce periodontal inflammation, prevent disease progression and ultimately the need for extractions.

Purpose: The aim of the present randomized study was to evaluate the efficacy of intermaxillary fixation screw (IMFS) versus eyelet interdental wiring for intermaxillary fixation (IMF) in minimally displaced mandibular fractures., Materials and Methods: A total of 50 patients with a minimally displaced mandibular fracture were enrolled, with 25 patients randomly selected for each group. In group I (study group, n = 25), the patients were treated using IMFS, and in group II (control group, n = 25), they received eyelet interdental wiring. Both techniques were assessed for the following parameters: time required for placement and removal of each type of IMF technique, time required for placement of IMF wires, postoperative occlusion, stability of the IMF wire, local anesthesia requirement during removal of each fixation type, oral hygiene status, glove perforation rate, and complications associated with both techniques. The collected data were analyzed using Student's unpaired t test or χ2 test. P < .05 was considered significant and the Statistical Package for Social Sciences software, version 10, was used for analysis., Results: The average time required for placement in groups I and II was 17.56 and 35.08 minutes, respectively (P = .000). The time required for placement of the IMF wire in group I was 2.1 minutes and in group II was 6 minutes. The oral hygiene status was assessed, and the mean plaque index score for groups I and II was 1.44 and 2.12, respectively (P = .00). The glove perforation rate was much less in group I than in group II. Finally, the most common complication in both groups was mucosal growth., Conclusions: The results established the supremacy of IMFS compared with eyelet interdental wiring. Thus, we have concluded that IMFS, in the present scenario, is a safe and time-saving technique. IMFS is a cost-effective, straightforward, and viable alternative to cumbersome eyelet interdental and other wiring techniques for providing IMF, with satisfactory occlusion during closed reduction or intraoperative open reduction internal fixation of fractures. In addition, oral hygiene can be maintained, and the glove perforation rate was very low using IMFS. The relatively small sample size and limited follow-up period were the study limitations., (Copyright © 2014 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.)

Orthodontic extrusion of nonrestorable teeth has been used for almost 20 years as an alternative to bone grafting in preparation for implant placement. Although this technique predictably creates bone and soft tissue, and improves the socket diameter and depth, most of the bone apposition occurs in the marginal alveolar and periapical areas of the extruded tooth. To create more labial bone, the standard orthodontic extrusion technique was modified to apply pressure on the hopeless tooth both coronally and palatally, which allowed bone at the site to develop apically and labially. Gingival thickness on the labial aspect was also increased, and the tissue biotype was improved. A clinical treatment is presented that illustrates the use of this technique., (Copyright © 2013 Editorial Council for the Journal of Prosthetic Dentistry. Published by Mosby, Inc. All rights reserved.)

The present study clarifies developmental organization of the oxytalan fiber system in the periodontal space of both the enamel (labial) and cementum (lingual) sides of rat incisors. The number of oxytalan fibers per unit area (μm(2)) was counted in rat incisors at stages of embryonic day 20 (E20) to postnatal day 35 (P35). Oxytalan fibers in the periodontal space of the enamel side were apt to decrease in number during the postnatal period, whereas their number remained almost unchanged on the cementum side during the developmental period. When the incisor emerged through the gum at P11, thinner oxytalan fibers distributed in the apical growing periodontium of the cementum side seemed to be fused with one another to become thicker fibers as has been reported for rat molars (Inoue et al., 2012). Thus, the oxytalan fiber system in the periodontal space represented significant differences in its distributional density between the enamel and cementum sides after E23. At the stage of P35, oxytalan fibers presented significantly denser distribution in all territories of the periodontal ligament of the cementum side versus the enamel side. The present findings claim that the oxytalan fiber system might bind the tooth to the periodontal ligament and provide equilibrium of vascular system and control of blood flow in the periodontal ligament of the cementum side, while it might exclusively regulate the high level of physiologically adapted vasculature in the periodontal space of the enamel side., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

We observed nonsurgical improvement of interdental papillae in a patient undergoing supportive periodontal therapy. The patient was a 47-year-old Japanese man presenting with widespread gingival recession at Daniele's papilla presence index level 3 and Miller Class I recession affecting the facial aspect of tooth number 42. Initial periodontal therapy for periodontitis was performed, included oral hygiene instruction, scaling and root planing, resulting in a reduction in inflammation. Use of an interdental brush was then suspended to allow the interdental papillae to recover. The type of toothbrush and tooth brushing method were checked repeatedly. Mechanical debridement was performed every 2 to 3 months. A gradual improvement was observed in recession of the interdental papillae over a period of several years together with coronal regrowth of the gingival margin.

The aim of this study was to investigate the effects of a single exposure to whole cigarette smoke on human gingival fibroblast behavior. Normal oral mucosa fibroblasts were exposed once to whole cigarette smoke for 5, 15, or 30 min, and then were used to analyze cell adhesion, β1-integrin expression, cell growth and viability, cell capacity to contract collagen gel, and cell migration following wound infliction. Our findings showed that when gingival fibroblasts were exposed once to whole cigarette smoke, this resulted in a significant inhibition of cell adhesion, a decrease in the number of β1-integrin-positive cells, increased LDH activity in the target cells, and reduced growth. The smoke-exposed fibroblasts were also not able to contract collagen gel matrix and migrate following insult. Overall results demonstrate that a single exposure to whole cigarette smoke produced significant morphological and functional deregulation in gingival fibroblasts. This may explain the higher predisposition of tobacco users to oral infections and diseases such as cancer.

Background: There are various types of oral mucosa specific to different parts of the mouth and each of these has a unique histological structure. The variations in the epithelial structure are consistently related to observable differences in the underlying microcirculation: i.e. differences in the course, conformation, and density of capillaries. The aim of this research has been to investigate oral microcirculatory differences between men and women as well as between various age groups, in order to map the oral mucosa, and to highlight changes occurring during aging., Methods: A total of 45 healthy subjects were enrolled for this study (12 men and 33 women; mean age 60.37; range 30-82). A complete in-vivo videocapillaroscopic mapping of the oral mucosa was done on each subject., Results: The capillaroscopic patterns of the various areas differ particularly in the course of the loops in relation to the mucosal surface. On the basis of statistical analysis of the collected data, it emerges that there are many differences in capillary loop density between men and women and between different age groups., Conclusion: This study demonstrates the necessity of approaching the investigation of patient microcirculation in different ways depending on sex or age.

Aim: To investigate the nisin Z innocuity using normal human gingival fibroblast and epithelial cell cultures, and its synergistic effect with these gingival cells against Candida albicans adhesion and transition from blastospore to hyphal form., Methods and Results: Cells were cultured to 80% confluence and infected with C. albicans in the absence or presence of various concentrations of nisin Z. Our results indicate that only high concentrations of nisin Z promoted gingival cell detachment and differentiation. Determination of the LD(50) showed that the fibroblasts were able to tolerate up to 80 microg ml(-1) for 24 h, dropping thereafter to 62 mug ml(-1) after 72 h of contact, compared to 160 microg ml(-1) after 24 h, and 80 microg ml(-1) after 72 h recorded by the gingival epithelial cells which displayed a greater resistance to nisin Z. The use of nisin Z even at low concentration (25 microg ml(-1)) at appropriate concentrations with gingival cells significantly reduced C. albicans adhesion to gingival monolayer cultures and inhibited the yeast's transition., Conclusion: These findings show that when used at non-toxic levels for human cells, nisin Z can be effective against C. albicans adhesion and transition and may synergistically interact with gingival cells for an efficient resistance against C. albicans., Significance and Impact of the Study: This study suggests the potential usefulness of nisin Z as an antifungal agent, when used in an appropriate range.

A high incidence of gingival overgrowth occurred in a group of New Zealand White rabbits receiving daily cyclosporine (15 mg/kg IM) while on a retinoblastoma study. Over the course of 2 mo, rabbits presented with clinical signs of ptyalism (4 of 18 rabbits), inappetence (3 of 18), or both (3 of 18); facial dermatitis and erythema occurred secondary to ptyalism. Reducing the dose of cyclosporine to 10 mg/kg led to complete resolution of clinical signs in all but 2 rabbits, which then received azithromycin (62.5 mg PO once daily for 7 d), a common treatment for cyclosporine-induced gingival overgrowth in other species. After dose reduction and azithromycin treatment, clinical signs resolved and did not reoccur for the remainder of the study. Fourteen rabbits were necropsied at the end of the study, and gingival width was measured. Although some rabbits were clinically normal, the gingiva in all rabbits was grossly thickened. Rabbits on cyclosporine had molar gingiva that was significantly thicker (4.8 mm) than controls (2.5 mm) not treated with cyclosporine. Histologic analysis of the gingiva revealed mild to moderate gingival epithelial hyperplasia, hyperkeratosis, and mild inflammation. Gingival overgrowth is a known side effect of cyclosporine administration in other species but, to our knowledge, this report is the first description of the condition in rabbits. Because rabbits frequently are used in studies that involve systemic cyclosporine administration, clinicians are advised to include this possibility in their differential list for cases involving hypersalivation, facial dermatitis, or inappetence in rabbits.

Follicular dendritic cell-secreted protein (FDC-SP) is a small secretory protein having structural similarities to statherin, a protein in saliva thought to play a role in calcium retention in saliva. In contrast, FDC-SP is thought to play a role in the immune system associated with germinal centers. We report here the very specific expression of FDC-SP in junctional epithelium at the gingival crevice. This region is very important for the host defense against pathogens and for periodontal protection. To be able to better understand the function of FDC-SP, we developed a novel gene expression system that exploited gene trapping and site-specific gene integration to introduce the protein into a mammalian cell culture system. Using this system we were able to express FDC-SP as a fusion protein with green fluorescent protein in an osteogenic progenitor cell line with long term stability, which we then used to find that the fusion protein specifically adsorbs onto mineral deposits and the surface of hydroxyapatite particles exogenously added to the culture. This adsorption was highly dependent on the structural integrity of FDC-SP. These results suggest that FDC-SP may play an important role, adsorbing onto the surface of cementum and alveolar bone adjacent to periodontal ligament and onto tooth surface at the gingival crevice.

Calcium (Ca2+) antagonists induce gingival overgrowth as a side effect but the pathogenic mechanism is still unknown. The Ca2+-channel activator Bay K 8644 elevates intracellular Ca2+ concentration ([Ca2+]i) and enhances the cell proliferation of gingival fibroblasts in a dose-dependent manner. Verapamil, an L-type Ca2+-channel blocker, also elevates [Ca2+]i in gingival fibroblasts, but it has no effect on other fibroblasts such as those of the lung, skin, and muscle. Moreover, verapamil enhances the proliferation of fibroblasts of the gingiva but has no effect on the proliferation of those of other tissues. These findings confirm that [Ca2+]i elevation induces the proliferation of gingival fibroblasts.

Matrix remodeling is regulated by matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). Periostin, originally identified in a mouse osteoblastic library, plays a role in cell adhesion and migration and in mechanical stress-induced matrix remodeling. In this study, we analyzed and compared the distribution patterns of TIMP-2 and periostin during mouse mandible development. Immunohistochemical staining for TIMP-2 and periostin was carried out on serial cryosections obtained from mice at embryonic days 13-16, postnatal day 2 (P2), P35, and 12 weeks of age. TIMP-2 and periostin exhibited a strikingly similar protein distribution during mandible development. From bud to early bell stages of molars, TIMP-2 and periostin were highly expressed on the lingual and anterior sides of the basement membrane and on the adjacent jaw mesenchyme. In pre- and postnatal incisors, the basement membrane of the apical loop and dental follicle was immunostained for TIMP-2 and periostin. At postnatal stages, TIMP-2 and periostin were prominently confined to the extracellular matrix (ECM) of gingival tissues, periodontal ligaments, and tendons (all recipients of mechanical strain). However, periostin was solely detected in the lower portion of the inner root sheath of hair follicles. Gingiva of P2 cultured in anti-TIMP-2 antibody-conditioned medium showed markedly reduced staining of periostin. We suggest that TIMP-2 and periostin are co-distributed on ECM exposed to mechanical forces and coordinately function as ECM modulators.

Purpose: This article describes different scenarios of facial osseous defects when the osseous-gingival relationship exceeds 3 mm and evaluates the effects of the morphology of the compromised facial bone on gingival dynamics after immediate tooth replacement and guided bone regeneration. The implant success rate and peri-implant bone change were also reported., Materials and Methods: Twenty-three patients treated consecutively with the mean age of 39.5 years (range, 25 to 63 years) underwent immediate tooth replacement and guided bone regeneration in sockets with facial bony defects exceeding 3 mm. Facial bony defects were categorized into V-, U-, and Ultra-U (UU)-shaped. The patients were evaluated clinically and radiographically at 1-year after implant placement., Results: At 1-year, the implant success rate was 100% (23/23). No marginal bone change of greater than 1 mm was observed. Greater than 1.5 mm of facial gingival recessions were noted in 8.3% (1/12) of V-shaped, 42.8% (3/7) of U-shaped, and 100% (4/4) of UU-shaped defects., Conclusions: U- and UU-shaped defects showed significantly higher frequency and magnitude of facial gingival recession (>1.5 mm) when compared with V-shaped defects 1-year after immediate tooth replacement and guided bone regeneration. It is important to identify the type of facial bony defect during diagnosis and treatment planning, so that appropriate treatment can be prescribed. The combination of delayed implant placement after staged reconstruction of unfavorable U- and UU-shaped labial extraction socket defects should be considered in areas of high esthetic concern.

Reformation of natural-appearing sulcular and papilla anatomy between adjacent implants in the esthetic zone presents a complex challenge for the implant team. Guidelines for implant placement of adjacent implants and soft tissue development are introduced that optimize esthetic results. The rule of 3x3x3 PIE is a mnemonic, based on biologically and prosthetically driven implant placement, which maximizes the potential for optimal esthetics between adjacent implants. The 4 interdependent principles of the 3x3x3 PIE rule are as follows: 1) the platforms of the implants should be located 3 mm apical to the zeniths of the predetermined facial-gingival margins of the planned restorations, 2) the centers of the implants should be placed at a distance at least 3 mm palatal to the anticipated facial margins, 3) interimplant spacing of 3 mm is required between adjacent implant platforms, and 4) the implants should emerge through the palatal incisal edge of the ensuing crown positions. After placement of adjacent implants by the surgeon according to the rule of 3x3x3 PIE, the restorative dentist must then develop the remaining soft tissue beginning with a provisional restoration. The reformed peri-implant gingiva is then supported by the subgingival contours of the definitive abutments and crowns. If ideal gingival esthetics are not achievable, pontics or gingival-colored porcelain can be acceptable alternatives. New technologies, including platform switching, are enhancing clinicians' abilities to deliver consistent esthetic results with adjacent implants.

With the introduction of in-office cone beam computed tomography (CT), improved access to conventional CT scanning, and dental implant treatment planning software allowing on-the-spot 3-dimensional evaluations of potential implant sites, the use of "flapless" implant surgery has gained popularity among surgeons. Although the flapless approach was initially suggested for and embraced by novice implant surgeons, the successful use of this approach often requires advanced clinical experience and surgical judgment. This article reviews the advantages and disadvantages of and indications and contraindications for flapless dental implant surgery, with special emphasis on requirements for establishing or maintaining long-term health and stability of the peri-implant soft tissues. Prerequisites for surgeons wishing to use the flapless tissue punch approach in dental implant surgery are outlined and put into perspective relative to conventional open-flap surgery techniques and other minimally invasive procedures currently used in implant surgery. Procedures for single- and multiple-tooth applications are illustrated.

Although the predictability of treating full or partial edentulism with dental implants has long been established, the choice of whether to use a fixed or removable prosthesis to restore such implants remains a complex one. Attention to such factors as the amount of vertical and horizontal bone loss that has occurred, the quality and quantity of the soft tissue, and the patient's restorative history can simplify treatment planning.

Introduction: We previously reported that, on average, maxillary molars erupt with buccal crown torque and upright with age, mandibular molars erupt with lingual crown torque and upright with age, and these molar crown torque changes are accompanied by concurrent increases in maxillary and mandibular intermolar widths. Our purpose in this study was to determine the transverse skeletal changes that accompany these molar movements during growth., Methods: Thirteen maxillary and mandibular transverse measurements were made by using casts and posteroanterior radiographs from 36 Class I untreated subjects from the Iowa Facial Growth Study at approximately ages 7.6, 10.3, 12.9, 16.5, and 26.4 years. Intermolar arch widths, maxillary basal bone widths (bijugale), and mandibular basal bone widths (bigonion, biantegonion) were recorded for each subject. Also recorded were the maxillary and mandibular cross-arch alveolar process widths measured at the molar gingival crest and the midalveolar level., Results: The results indicate a pattern of width changes in the maxilla, the maxillary alveolar process, the maxillary first molars, the mandibular first molars, and the mandibular alveolar process that occurs as a gradient in the vertical dimension. The greatest width change occurs superiorly (jugale point). The smallest width change occurs inferiorly (midalveolar point of the mandible). On average, the basal bone of the maxilla increased in width by 5.37 mm; maxillary cross-arch midalveolar process width increased by 3.20 mm (left buccal surface to right buccal surface) and 4.94 mm (left palatal surface to right palatal surface); maxillary intermolar width increased by 3.08 mm; mandibular intermolar width increased by 2.05 mm; and mandibular cross-arch crest level alveolar width increased by 1.60 mm (left buccal surface to right buccal surface) and 1.02 mm (left lingual surface to right lingual surface). The basal bone of the mandible increased in width by 14.54 mm., Conclusions: We concluded that transverse molar movements during growth mirror transverse maxillary basal bone width increases, maxillary cross-arch alveolar process width increases, and mandibular cross-arch alveolar process width increases. Furthermore, there is a pattern of width changes in the maxilla, the maxillary alveolar process, the maxillary first molars, the mandibular first molars, and the mandibular alveolar process that occurs as a gradient in the vertical dimension. On the other hand, transverse mandibular basal bone changes, measured as bigonion and biantegonion, do not fit this pattern.

Purpose: Gingival hyperplasia is a well known side effect of Cyclosporine A therapy. The aetiology of this is not totally understood and there is debate whether it is hyperplasia of the gingival epithelium or of the submucosal connective tissue, or both, and what roles play factors like age and gender of the patients, duration and dosage of the drug., Material and Methods: The influence of different Cyclosporine A concentrations (10(-6) g/ml; 5 x 10(-7) g/ml; 10(-9) g/ml) and of no medication (controls) on growth and proliferation of cultured human gingival keratinocytes was investigated after a culture period of 3, 6 and 9 days. Cell proliferation was assessed by counting anti Ki-67 stained nuclei, cell growth by counting total number of nuclei and by the EZ4U-assay., Results: There was no significant correlation of the cell proliferation rate and cellular growth with either gender (p > 0.568) or duration of medication (p > 0.876); but Cyclosporine A concentration showed a highly significant influence on cellular growth (p = 0.0001). Inhibition of cell growth was dependent on drug dosage, but a low concentration of 10(-9) g/ml even stimulated cell growth., Conclusions: There is evidence that Cyclosporine A in low concentrations (10(-9) g/ml as applied in long-term therapy) stimulates gingival keratinocyte growth and therefore might be related to hyperplasia of the gingiva. However, high Cyclosporine A concentrations may inhibit cell growths and factors like gender of the patient did not show any influence in-vitro.

The present work focused on the design of novel hydrogel microspheres based on both dextran- and gelatin-derived biomaterials, and discussed whether locally controlled delivery of IGF-I from dextran-co-gelatin hydrogel microspheres (DG-MP) was useful for periodontal regeneration enhancement. Microspheres were synthesized when gelatin was cooperating with glycidyl methacrylate (GMA) derivatized dextrans (Dex-GMA) and the resultant DG-MP with a hydrogel character of which the cross-linking density could be controlled by the degree of substitution (DS, the number of methacrylates per 100 glucopyranose residues) of Dex-GMA. In this study, three types of DG-MP (DG-MP4.7, DG-MP6.3 and DG-MP7.8) obtained from gelatin and Dex-GMA (differing in DS: 4.7, 6.3 and 7.8 respectively) were prepared and characterized by swelling and degradation properties, drug release kinetics and biological capability in promoting tissue regeneration. By swelling in aqueous positively charged IGF-I solutions, the protein could be encapsulated in DG-MP by polyionic complexation with negatively charged acidic gelatin. No obvious influence of Dex-GMA's DS on DG-MP's configuration and size was observed, and the release and degraded properties showed no significant difference between three types of DG-MP in PBS buffer either. However, high DS of Dex-GMA could lower microsphere's swelling, prolong its degraded time and minimize IGF-I burst release markedly in dextranase-containing PBS, where IGF-I release from a slow release type of microspheres (DG-MP7.8) could be maintained more than 28 days, and an effective protein release kinetics without a significant burst but a relevantly constant release after the initial burst was achieved. IGF-I in DG-MP resulted in more new bone formation in the periodontal defects within 4 or 8 weeks than IGF-I in blood clot directly did (P < 0.01). The observed newly formation of periodontal tissues including the height and percentage of new bone and new cementum on the denuded root surfaces of the furcation area in DG-MP7.8 group were more than that in other groups (P < 0.05). The adequate width of regenerative periodontal ligament (PDL), regular Sharpey's fibers and alveolar bone reconstruction could be observed only in DG-MP7.8 group. These combined results demonstrate that effective release kinetics can be realized by adjusting the DS of Dex-GMA and followed cross-linking density of DG-MP, and that locally controlled delivery of IGF-I from slow release type of DG-MP may serve as a novel therapeutic strategy for periodontal tissue regeneration.

Clinical research has long been testing techniques of integrating biomaterials with many external factors, such as simple proteins or more complicated devices, in order to achieve the restitutio ad integrum of periodontium. This study assessed the in vitro effectiveness of platelet derivate growth factor-BB (PDGF) and insulin growth factor I (IGF); the biocompatibility of materials like Paroguide, Oclastim membranes, Gingistat sponges, Surgiplaster, and Capset; and their efficacy as carriers for the platelet derivate growth factor-BB (PDGF) and insulin growth factor I (IGF). Fibroblasts from the human periodontal ligament were incubated with growth factors free or vehiculated. Mitogenic effect was evaluated by measuring the growth rate and biocompatibility by observing cell morphology at SEM. PDGF was the most effective in stimulating cell proliferation both in solution (p < 0.001) and vehiculated (p < 0.01). Surgiplaster and Capset were more biocompatible; however, final analysis to assess their efficacy as carriers failed to disclose significant differences between experimental findings and control.

Implant esthetics has been the focus of attention for the past decade, and one vital issue is the effect of interimplant distance on interimplant papilla formation and crestal bone loss. The aim of this study was to evaluate the effect of 1, 2, and 3 mm of interimplant distance on papilla formation and crestal resorption in submerged and nonsubmerged Ankylos implants after prosthetic restoration. Bilateral mandibular premolars of 7 dogs were extracted, and after 12 weeks each dog received 8 implants. Implants were placed so that 3 interimplant distances were created at 1 mm (group 1), 2 mm (group 2), and 3 mm (group 3). The sides and the position of the groups were randomly selected. Twelve weeks after placement, the implants received metallic prostheses that allowed 5 mm of space between the prosthetic contact point (CP) and the crestal bone (CB). After 8 weeks, the distance between the CP and the papilla (CP-P) and the gingival height at the distal proximal aspect of the prosthesis (CP-DE) was clinically measured. Radiographic images were obtained to measure the distance of the CP to the CB within the interimplant surfaces (CP-IP) and adjacent to the edentulous surfaces (CP-ED). The clinical measurement of CP-P for submerged and nonsubmerged implants was 3.57+/-1.17 mm and 3.10+/-0.82 mm for group 1, 3.57+/-0.78 mm and 3.16+/- 0.87 mm for group 2, and 3.35+/- 0.55 mm and 3.07+/-0.93 mm for group 3. The CP-DE was 3.25+/-0.77 mm for submerged and 2.78+/- 0.64 mm for nonsubmerged implants. The CP-IP for the submerged and nonsubmerged implants was 6.91+/-0.95 mm and 7.68+/-2.73 mm for group 1, 7.46+/-1.43 mm and 5.87+/-1.71 mm for group 2, and 7.72+/-0.81 mm and 7.59+/-1.33 mm for group 3. The CP-ED was 6.77+/-1.33 mm for submerged implants and 6.03+/-1.58 mm for nonsubmerged implants. There were no statistical significant differences for any of the measured parameters. We conclude that when the distance from the CP to the CB was 5 mm, interimplant distances of 1 to 3 mm did not affect papilla formation or crestal resorption of submerged or nonsubmerged implants in the dog model.

Owing to mathematical coupling, statistical analyses relating change to baseline values using correlation or regression are erroneous, where the statistical procedure of testing the null hypothesis becomes invalid. Alternatives, such as Oldham's method and the variance ratio test, have been advocated, although these are limited in the presence of measurement errors with non-constant variance. Furthermore, such methods prohibit the consideration of additional covariates (e.g., treatment group within trials) or confounders (e.g., age and gender). This study illustrates the more sophisticated approach of multilevel modelling (MLM) which overcomes these limitations and provides a comprehensive solution to the analysis of change with respect to baseline values. Although mathematical coupling is widespread throughout applied research, one particular area where several studies have suggested a strong relationship between baseline disease severity and treatment effect is guided tissue regeneration (GTR) within dental research. For illustration, we use GTR studies where the original data were found to be available in the literature for reanalysis. We contrast the results from an MLM approach and Oldham's method with the standard (incorrect) approach that suffers from mathematical coupling. MLM provides a robust solution when relating change to baseline and is capable of simultaneously dealing with complex error structures and additional covariates and/or potential confounders.