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Objectives The peri-implant epithelium (PIE) plays an important role in the prevention against initial stage of inflammation. To minimize the risk of peri-implantitis, it is necessary to understand the biological characteristics of the PIE. The aim of this study was to investigate the characteristic gene expression profile of PIE as compared to junctional epithelium (JE) using laser microdissection and microarray analysis. Methods Left upper first molars of 4-week-old rat were extracted, and titanium alloy implants were placed. Four weeks after surgery, samples were harvested by laser microdissection, and total RNA samples were isolated. Comprehensive analyses of genes expressed in the JE and PIE were performed using microarray analysis. Confirmation of the differential expression of selected genes was performed by quantitative real-time polymerase chain reaction and immunohistochemistry. Results The microarray analysis showed that 712 genes were more than twofold change upregulated in the PIE compared with the JE. Genes Scgb1a1 were significantly upregulated more than 19.1-fold, Lpo more than 19.0-fold, and Gbp2 more than 8.9-fold, in the PIE (P

Aim To apply a novel human model to evaluate the morphogenesis of the mucosal attachment to implants. Material and methods Twenty one patients receiving implant-supported single-tooth replacement were enrolled in this study. After implant installation, a custom-designed experimental abutment was connected to the implant. Soft tissue biopsies representing 2, 4, 8 or 12 weeks of healing were collected by the use of a circular cutting device and prepared for histological analysis. Results The soft tissue biopsies were retrieved, preserved and processed with a technique that was safe and reproducible. The results from the histological analysis in regards to dimensional and qualitative changes in the mucosa over time were consistent with those reported from animal experiments. At 8 weeks, the soft tissue dimension was about 3.6 mm and included a barrier epithelium of 1.9 mm and a connective tissue portion of 1.7 mm. Similar dimensions were found at 12 weeks. Conclusion It is suggested that the new human model provides advantages in terms of cost-effectiveness in research as well as from ethical aspects and should be considered as an alternative to pre-clinical in vivo studies in animals.

Objectives This study was undertaken to evaluate the influences of concave, machined and concave-roughened profiles of transmucosal implant designs on early peri-implant tissue responses. Materials and Methods Implants were used and classified by transmucosal profile and surface type as straight-machined implants (SM), concave-machined implants (CM), or concave-roughened implants (CR). A total of 30 implants (10 per each type) with matching transmucosal profiles were placed directly on alveolar crests in randomized order in the edentulous mandibular ridges of three beagle dogs. Healing abutments were connected 4 weeks after implant placement, and prostheses were connected 8 weeks after implant placement and functionally loaded. All animals were sacrificed at 16 weeks. Peri-apical radiographs were obtained and measured to evaluate peri-implant marginal bone levels. Histological specimens were prepared to measure bone resorption, connective tissue contact, epithelial tissue height, biological width, and length of implant-abutment junction to the apical portion of junctional epithelium. Results Radiographic and histometric analysis showed that least bone resorption occurred around CM implants and greatest bone resorption around SM implants (P

AIM To validate the "platform switching" concept at oral implants with respect to the preservation of the alveolar crestal bone levels in an animal model. MATERIAL & METHODS Five minipigs received three implants each with a 0.25 mm implant/abutment mismatch and were placed flush (T(0)), 1 mm below (T(1)) and 1 mm above (T(+1)) the alveolar bony crest, and as a control, one conventionally restored implant placed at the bone level. The implants were randomly inserted flapless into the mandible. Four months after implant insertion, the animals were sacrificed, and undecalcified block sections were obtained and used for histological analyses. RESULTS The mean values for peri-implant bone resorption were 1.09 ± 0.59 mm (Control), 0.51 (± 0.27 mm, T(0)), 0.50 (± 0.46 mm, T(+1)) and 1.30 (± 0.21 mm, T(-1)), respectively. Statistically significant differences (P

Objective: Implant designs that can stimulate and integrate with an epithelial wound-healing process may significantly enhance the efficacy of dental implants. Here, we evaluated the potential of “step-type” implant systems to improve the sealing between the peri-implant epithelium (PIE) and the implant surface, and investigated the effect of implant structure on PIE down-growth. Materials and methods: Right maxillary first molars were extirpated from rats and implanted with either a straight-type or a step-type implant varying in step height and/or width (Ns: 0.8 mm height, 0.1 mm width; Ws: 0.8 mm height, 0.2 mm width; Hs: 0.4 mm height, 0.1 mm width). Maxillae were harvested at various time points over 16 weeks to evaluate laminin-5 distribution as an indicator of wound healing and PIE formation, horse-radish peroxidase (HRP) penetration as a measurement of epithelial sealing, and PIE down-growth formation. Results: In all implant models, the PIE formed from the oral sulcular epithelium and spread apically along the implant surface. In the Ws group, HRP penetration was detected only in the coronal region of the PIE at 4 weeks, whereas in the straight-type, it was observed in the apical region and the connective tissue. At 16 weeks, the Ws implants exhibited markedly less PIE down-growth than the Con, Ns or Hs implants, and were equivalent to that observed in natural teeth. Conclusion: The step-type implant system may have the potential for improving epithelial sealing at the tissue–implant interface, as well as reducing apical PIE down-growth, thus enhancing dental implant efficacy. To cite this article: Atsuta I, Ayukawa Y, Ogino Y, Moriyama Y, Jinno Y, Koyano K. Evaluations of epithelial sealing and peri-implant epithelial down-growth around “step-type” implants. Clin. Oral Impl. Res. 23, 2012 459–466 doi: 10.1111/j.1600-0501.2011.02163.x

Objectives: The aim of this study was to describe the ultrastructure of the dentogingival border at replanted teeth and implants. Material and methods: Wistar rats (8 weeks old) were divided into groups for replantation and implantation experiments. In the former, the upper right first molars were extracted and then immediately replanted. In the latter, pure titanium implants were used. All tissues were fixed, demineralized and embedded in epoxy resin for ultrastructural observations. Results: One week after replantation, the junctional epithelium was lost, and the oral sulcular epithelium covered the enamel surface. The amount of the epithelium increased in 2 weeks, and resembled the junctional epithelium, and the internal basal lamina and hemidesmosomes were formed in 4 weeks. One week after implantation, peri-implant epithelium was formed, and in 2 and 4 weeks, this epithelium with aggregated connective tissue cells were observed. In 8 weeks, the peri-implant epithelium receded, and aligned special cells with surrounding elongated fibroblasts and bundles of collagen fibers appeared to seal the implant interface. Conclusion: In replantation of the tooth, the internal basal lamina remained at the surface of the enamel of the replanted tooth, which is likely to be related to regeneration of the junctional epithelium and the attachment apparatus at the epithelium–tooth interface. Following implantation, a layer of cells with characteristics of connective tissue cells, but no junctional epithelium and attachment apparatus, was formed to seal the site of the implant.

The aim of the present study was to evaluate histometric changes around dental implants inserted at different levels in relation to the crestal bone, under different loading conditions.Thirty-six implants were inserted in the edentulous mandible of six mongrel dogs. Each implant was assigned to an experimental group according to the distance from the top of the implant to the crestal bone: Bone Level (at the crestal bone level), Minus 1 (1 mm below the crestal bone) or Minus 2 group (2 mm below the crestal bone). Each hemimandible was submitted to a loading protocol: conventional or immediate restoration. After 90 days, the animals were killed. Specimens were processed, and measurements were performed concerning the length of soft and hard peri-implant tissues. Data were analyzed using ANOVA and Student's t test (alpha=5%).Among conventionally restored sites, the distance from the most coronal position of soft tissue margin (PSTM) and first bone-implant contact (fBIC) was greater for Minus 2 than for Bone Level and Minus 1 sites (P=0.03), but significant differences were not observed among immediately restored sites. Differences among groups were not observed concerning the PSTM, and the distance from the implant-abutment junction to fBIC. Greater amounts of lateral bone loss were observed for conventionally than for immediately restored sites (P=0.006).These findings suggest that the apical positioning of the top of the implant may not jeopardize the position of soft peri-implant tissues, and that immediate restoration can be beneficial to minimize lateral bone loss. Further studies are suggested to evaluate the clinical significance of these results in longer healing periods.

PURPOSE: The objective of the present experiment was to study the morphogenesis of the mucosal attachment to implants made of c.p. titanium. MATERIAL AND METHODS: All mandibular premolars were extracted in 20 Labrador dogs. After a healing period of 3 months, four implants (ITI Dental Implant System) were placed in the right and left sides of the mandible. A non-submerged implant installation technique was used and the mucosal tissues were secured to the conical marginal portion of the implants with interrupted sutures. The sutures were removed after 2 weeks and a plaque control program including daily cleaning of the remaining teeth and the implants was initiated. The animals were sacrificed and biopsies were obtained at various intervals to provide healing periods extending from Day 0 (2 h) to 12 weeks. The mandibles were removed and placed in the fixative. The implant sites were dissected using a diamond saw and processed for histological analysis. RESULTS: Large numbers of neutrophils infiltrated and degraded the coagulum that occupied the compartment between the mucosa and the implant during the initial phase of healing. At 2 weeks after surgery, fibroblasts were the dominating cell population in the connective tissue interface but at 4 weeks the density of fibroblasts had decreased. Furthermore, the first signs of epithelial proliferation were observed in specimens representing 1-2 weeks of healing and a mature barrier epithelium occurred after 6-8 weeks of healing. The collagen fibers of the mucosa were organized after 4-6 weeks of healing. CONCLUSION: It is suggested that the soft-tissue attachment to implants placed using a non-submerged installation procedure is properly established after several weeks following surgery.

Oral implants pierce through the mucosa thus establishing a connection between the oral environment and underlying tissues. The soft-tissue connection to the transmucosal part is of crucial importance as it relates to the stability of the peri-implant tissues and the prevention of peri-implant infection with subsequent destruction of the peri-implant structures. The adhesion of soft tissue to the material surface is one prerequisite for the longterm success. This adhesion is depending upon the topography and chemical composition of the implanted biomaterial. As regards to surface topography of implants we refer to the definitions proposed by Albrektsson & Wennerberg (2004a, 2004b), where minimally rough surface has been defined in the range of Sa 0.5–1 mm (e.g., turned implants), moderately rough Sa 1–2 mm (e.g., acid etched, sandblasted or anodized) and rough Sa 42mm (e.g., plasma sprayed).

The purpose of this study was to determine the peri-implant soft tissue dimension (PSD) and peri-implant bone level (BL) of dental implants with different designs and surface modifications after functional loading without oral hygiene. Three types of dental implants were placed in the posterior jaws of adult baboons, three of the same design per quadrant, and fitted with fixed partial dentures. After 1.5 years of functional loading and plaque accumulation, all implants showed severe peri-implant mucositis and comparatively high BL. A histomorphometric evaluation of the sulcus depth (SD), the dimension of the junctional epithelium (JE) and the connective tissue contact (CTC) resulted in no significant differences between the three implant designs, neither in the maxilla nor in the mandible (P>0.05). The sum of SD, JE and CTC forming the PSD was nearly the same in the maxilla (commercially pure titanium, CpTi: 3.5 mm 2.9/4.1 confidence interval (CI); titanium plasma sprayed (TPS): 3.5 mm 2.9/4.2 CI; sand blasted acid-etched (GBAE): 3.2 mm 2.7/3.9 CI) and in the mandible (CpTi: 3.2 mm 2.6/3.8 CI; TPS: 3.2 mm 2.6/3.8 CI; GBAE: 3.2 mm 2.7/3.9 CI; P>0.05). There was no difference in BL around the three implant designs (maxilla: CpTi: 0.9 mm 0.5/1.6 CI; TPS: 0.9 mm 0.5/1.5CI; GBAE: 0.9 mm 0.5/1.6 CI; mandible: CpTi: 0.8 mm 0.5/1.2 CI; TPS: 0.6 mm 0.4/0.9 CI; GBAE: 0.7 mm 0.5/1.1 CI; P>0.05). Overall, the data presented did not show any significant differences in peri-implant soft tissue conditions in baboons. Moreover, plaque accumulation and propagation of peri-implant mucositis after 1.5 years of functional loading was not influenced by implant design and surface modifications in baboons.

Today, implant-supported prostheses are widely accepted as a reliable treatment modality, but failures in longitudinal studies have been shown. In some cases, peri-implantitis with a progressive periodontal bone loss takes place, and mechanical or load factors and biological or plaque-induced lesions have been claimed as main etiologic factors. We compared five cases of peri-implantitis, with five cases of healthy peri-implant tissues and five cases of aggressive periodontitis in order to give new findings on the osseointegration loss process. Biopsy specimens from the peri-implant tissues including oral (O), sulcular, and junctional epithelium and the underlying and supracrestal connective tissue, were taken in all cases for histological and immunohistochemical analysis. T lymphocytes were the most prominent cell in the peri-implantitis (PG) and aggressive periodontitis (AG) groups, but not in the peri-implant healthy group (HG). CD1a-positive cells (Langerhans and immature dendritic cells) were observed more frequently in the O than in the sulcular-junctional (S-J) epithelium: they were located in the basal and parabasal layers, without any differences between the three groups. Vascular proliferation analysed by immunoreactivity for CD34, Factor VIII, and vascular endothelial growth factor was more prominent in the PG comparing with HG and AG in the S-J area. Apoptosis, analysed by bcl2 and p53 immunoreactivity, was similar in the three groups. In conclusion, we suggest that the osseointegration loss process is due to an inflammatory process similar to that observed in aggressive periodontitis according to the number of T lymphocytes, but not to the vascular proliferation.

Background/aims Clinical probing of the soft tissues around oral implants has become a frequently used parameter for clinical monitoring. However, the healing of the disruption of the soft tissue seal as a result of probing has not yet been studied. The purpose of this study was to evaluate the healing events in the periimplant mucosal tissues following standardized clinical probing. Materials and methods In three foxhounds the mandibular premolars were extracted and, after 3 months of healing, TPS screw-shaped implants (ITO Dental Implant System) were installed. A plaque control regimen was performed throughout the duration of the experiment. After 3 months of healing of the transmucosal implants, clinical implant stability and healthy periimplant mucosal tissues with mostly absence of bleeding on probing were obtained. Clinical probing of the mesial and distal implant sites was performed immediately before 1, 2, 3, 5 and 7 days prior to the sacrifice of the animals. A specially manufactured titanium cylinder with a mesial and distal groove was fixed to the implants to standardize the insertion geometry of a pressure-sensitive probe (0.2-0.25 N, tip diameter 0.45 mm). The distances from the alveolar crest to the coronal border of the connective tissue adaptation to the implant and the length of the epithelial attachment were measured histomorphometrically in nondecalcified ground sections. The buccal and lingual aspects of the implants were used to determine the components of the biological width in unprobed control sites. Results The probe caused a separation between the surface of the implant and the junctional epithelium, but not within the connective tissue adaptation. In general, the probe tip was located at the most coronal level of the supracrestal connective tissue as determined histologically. By 1 day after separation of the periimplant mucosal tissue by probing, an epithelial attachment of approximately 0.5 mm in the apico-coronal direction was observed. The length of the epithelial adaptation showed a tendency to increase over time (day 2: 1.15 mm, day 3: 1.52 mm), and was complete at day 5 (1.92 mm). At the unprobed sites the epithelial attachment showed a mean length of 1.69 mm. Inflammatory infiltrates were practically nonexistent, indicating the absence of tissue trauma or infection as a result of probing. Conclusions Clinical probing around osseointegrated implants does not appear to have detrimental effects on the soft tissue seal and, hence, does not seem to jeopardize the longevity of oral implants. The 'healing of the epithelial attachment' seems to be complete 5 days after clinical probing.

Research in implant dentistry has mainly focused on hard tissue integration with much less data available with regards to soft tissue integration involving epithelium and connective tissue. In the present study, the implantogingival junction of unloaded and loaded non-submerged titanium implants has been analyzed histometrically in the canine mandible. In 6 foxhounds, 69 implants were placed. Dogs in the unloaded group were sacrificed 3 months after implant placement. Loaded implants were restored with gold crowns and those dogs were sacrificed after 3 months and 12 months of loading. Non-decalcified histologic sections were analyzed histometrically measuring the dimensions of the Sulcus Depth (SD), the Junctional Epithelium (JE), and the Connective Tissue Contact (CTC). Histometric evaluation revealed that significant changes within tissue compartments (SD, JE, CTC) occurred over time (P0.05). Sulcus Depth had a mean of 0.49 mm and 0.50 mm after 3 months and 6 months of healing, but after 15 months was 0.16 mm which was significantly different. Similarly, the length of the Junctional Epithelium after 3 months and 6 months of healing was 1.16 mm and 1.44 mm, respectively, and these values were significantly different from measurements taken after 15 months (1.88 mm). The area of Connective Tissue Contact showed a different pattern of change in that after 3 months of healing (1.36 mm) it was significantly different from the same area after 6 months and 15 months which were 1.01 mm and 1.05 mm, respectively. Interestingly, the sum of SD, JE, and CTC, forming the Biologic Width, did not change over the observation period (P0.05). These data indicate that the Biologic Width is a physiologically formed and stable structure over time in the case of non-submerged, one-piece titanium implants as evaluated histometrically under unloaded and loaded conditions. Dynamic changes did occur, however, within the overall Biologic Width dimension. Thus, the use of non-submerged, one-piece implants allow for stable overall peri-implant soft tissues as evaluated under loaded conditions for up to 12 months.

Objectives To study early soft tissue healing of immediately placed implants with or without immediate loading in the dog. Material and methods Forty-eight implants were placed in the distal sockets of Pm3 and Pm4 in the lower jaw of 12 beagle dogs immediately after tooth extraction. In the control group, no loading was applied. In the test group, an immediate loading restoration with occlusal contacts was performed. Dogs were sacrificed at 2, 4, and 8 weeks for histological analysis. Results At the end of the study, there was a 100% implant and prosthesis survival. The biological width dimension was similar in both groups at all the studied healing periods. This dimension tended to decrease from week 2 to 8 in both groups, on both the buccal and lingual side. The barrier epithelium tended to stop at the implant–abutment interface in both groups and also decreased in length from week 2 to 8, on the buccal and the lingual side. Soft tissue recession remained low and occurred mainly in the test group. Conclusions The characteristics, dimension, and healing pattern of the peri-implant soft tissues were similar around immediate implants with or without immediate loading.

This study details the structural and ultrastructural features of the interfaces between titanium implants and their surrounding tissues. The material stemmed from an experiment in dogs in which guided tissue regeneration with Gore-Tex membranes was used to treat peri-implant, ligature-induced tissue breakdown around submerged and nonsubmerged commercially pure titanium implants. Specimens from the nonsubmerged group were evaluated under light microscopy and scanning and transmission electron microscopy. A healthy gingiva and a gingival sulcus were formed around the implant necks. A regenerated junctional epithelium provided the epithelial union between implant and gingiva. The supracrestal connective tissue was characterized by a 3-dimensional network of collagen fibers, fibroblasts and blood vessels. Near the implant surface the collagen fibers ran parallel to the titanium surface or were orientated perpendicular to the implant. The connective tissue-implant interface was characterized by a fine fibrillar material interposed between the implant surface and the connective tissue. An unidentified material was also observed between the endings of functionally orientated collagen fibrils and the metallic surface. The apical portions of the implants were anchored in compact bone. At the bone-implant interface, either mineralized bone matrix was intimately adapted to the titanium surface without any intervening space or a 0.5-micron wide unmineralized layer was interposed. These findings indicate that a perimucosal seal was formed around the implants consisting of a junctional epithelium-implant union coronally and supported by the connective tissue-implant junction apically. The implants were integrated in connective tissue, but only tightly adapted to bone.

To validate the "platform switching" concept at oral implants with respect to the preservation of the alveolar crestal bone levels in an animal model.Five minipigs received three implants each with a 0.25 mm implant/abutment mismatch and were placed flush (T(0)), 1 mm below (T(1)) and 1 mm above (T(+1)) the alveolar bony crest, and as a control, one conventionally restored implant placed at the bone level. The implants were randomly inserted flapless into the mandible. Four months after implant insertion, the animals were sacrificed, and undecalcified block sections were obtained and used for histological analyses.The mean values for peri-implant bone resorption were 1.09 ± 0.59 mm (Control), 0.51 (± 0.27 mm, T(0)), 0.50 (± 0.46 mm, T(+1)) and 1.30 (± 0.21 mm, T(-1)), respectively. Statistically significant differences (P0.05) were found among the test (T(0), T(-1)) and the control sites. Control implants presented an average biologic width length of 3.20 mm (± 0.33), with a connective tissue adaptation compartment of 1.29 mm (± 0.53) and an epithelial attachment of 1.91 mm (± 0.71). T(0), T(+1) and T(-1) implants presented with a mean biologic width of 1.97 mm (± 1.20), 2.70 mm (± 1.36) and 2.84 mm (± 0.90), respectively, with a connective tissue adaptation compartment of 1.21 mm (± 0.97), 1.21 mm (± 0.65) and 1.50 mm (± 0.70) and an epithelial attachment of 0.84 mm (± 0.93), 1.66 mm (± 0.88) and 1.35 mm (± 0.44), respectively. Differences between the configurations were mainly associated with the length of the epithelial attachment. The epithelial attachment was significantly longer in the C sites than in T(0) (P=0.014). However, no other differences between configurations were detected.If the implants are positioned at the level of the alveolar bony crest, the platform-switching concept may have a minor impact on the length of the epithelial attachment (0.84 vs. 1.91 mm), while the connective tissue adaptation compartment remains relatively unaffected. Moreover, platform switching resulted in less resorption of the alveolar crest (0.58 mm).

The aim of the present study was to evaluate the peri-implant soft tissue dimensions in flapless immediate implants with and without immediate loading.This study was carried out on six beagle dogs. Four implants were placed (two per side) immediately after tooth extraction (third and fourth premolars). Flapless immediate implant placement was performed in one hemimandible (control). The same procedure was performed in the contralateral side and immediate prosthesis was connected (test). After 3 months of healing, the dogs were sacrificed.None of the implants and prosthesis were lost. Barrier epithelium in the loaded group was 2.51 mm at the buccal and 2.34 mm at the lingual aspect. In the no loaded group, the results were similar, 2.54 and 2.2 mm at the buccal and lingual side, respectively. Connective tissue in the loaded group was 1.38 mm at the buccal and 0.65 mm at the lingual aspect, and in the no loaded group 1.48 mm at the buccal and 0.53 mm at the lingual side. Biological width dimensions were 3.9 mm at the buccal and 2.95 mm at the lingual aspect for the loaded group, and 4.01 and 2.64 mm at the buccal and lingual aspect for the no loaded group.The results of the present study suggested that soft tissues dimensions around immediate implants with immediate loading were similar to immediate implants without loading.

Implant designs that can stimulate and integrate with an epithelial wound-healing process may significantly enhance the efficacy of dental implants. Here, we evaluated the potential of "step-type" implant systems to improve the sealing between the peri-implant epithelium (PIE) and the implant surface, and investigated the effect of implant structure on PIE down-growth.Right maxillary first molars were extirpated from rats and implanted with either a straight-type or a step-type implant varying in step height and/or width (N(s): 0.8 mm height, 0.1 mm width; W(s): 0.8 mm height, 0.2 mm width; H(s): 0.4 mm height, 0.1 mm width). Maxillae were harvested at various time points over 16 weeks to evaluate laminin-5 distribution as an indicator of wound healing and PIE formation, horse-radish peroxidase (HRP) penetration as a measurement of epithelial sealing, and PIE down-growth formation.In all implant models, the PIE formed from the oral sulcular epithelium and spread apically along the implant surface. In the W(s) group, HRP penetration was detected only in the coronal region of the PIE at 4 weeks, whereas in the straight-type, it was observed in the apical region and the connective tissue. At 16 weeks, the W(s) implants exhibited markedly less PIE down-growth than the Con, N(s) or H(s) implants, and were equivalent to that observed in natural teeth.The step-type implant system may have the potential for improving epithelial sealing at the tissue-implant interface, as well as reducing apical PIE down-growth, thus enhancing dental implant efficacy.

Connective tissue in contact to transgingival/-dermal implants presents itself as tight scar formation. Although rough surfaces support the attachment they increase bacterial colonisation as well. In contrast to surface roughness, little is known about the influence of surface wettability on soft-tissue healing in vivo. We therefore investigated the influence of different surface wettabilities on connective tissue healing at polished implant surfaces in vivo.Three polished experimental groups (titanium, titanium coated with hydrophobic nano-crystalline diamond (H-NCD) and titanium coated with hydrophilic nano-crystalline diamond (O-NCD) were inserted into the subcutaneous connective tissue of the abdominal wall of 24 rats. Animals were sacrificed after 1 and 4 weeks resulting in eight specimen per group per time point. Specimen were subjected to histological evaluation (van Giesson's staining) and immunohistochemistry staining for proliferating cell nuclear antigen (PCNA), fibronectin and tumour necrosis factor-alpha (TNF-α).Histological evaluation revealed dense scar formation at the titanium and H-NCD surfaces. In contrast, the connective tissue was loose at the O-NCD surface with a significantly higher number of cells after 4 weeks. O-NCD demonstrated a strong expression of PCNA and fibronectin but a weak expression of TNF-α. In contrast, the PCNA and fibronectin expression was low at the titanium and H-NCD, with a strong signal of TNF-α at the H-NCD surface.Hydrophilicity influences the connective tissue healing at polished implant surfaces in vivo positively. The attachment of connective tissue and the number of cells in contact to the surface were increased. Moreover, the inflammatory response is decreased at the hydrophilic surface.

To assess the marginal soft tissue healing process after flap or flapless surgery in immediate implant placement in a dog model.This study was carried out on five Beagle dogs. Four implants were placed in the lower jaw in each dog immediately after tooth extraction. Flap surgery was performed before the extraction on one side (control) and flapless on the other (test). After 3 months of healing, the dogs were sacrificed and prepared for histological analysis.Ten implants were placed in each group. Two failed (one of each group). The length of the junctional epithelium in the flapless group was 2.54 mm (buccal) and 2.11 mm (lingual). In the flap group, the results were very similar: 2.59 mm (buccal) and 2.07 mm (lingual), with no significant differences observed between the groups. The length of the connective tissue in the flapless group was 0.68 mm (buccal) and 0.54 mm (lingual), and 1.09 mm at the buccal and 0.91 mm at the lingual aspect in the flap group, with no significant differences between groups. The difference between the mean distance from the peri-implant mucosa margin to the first bone-implant contact at the buccal aspect was significant between both groups (3.02 mm-flapless and 3.69 mm flap group). However, this difference was mostly due to the Pm3 group (flapless: 2.95/flap: 3.76) because no difference could be detected in the Pm4 group. Both groups showed minimal recession, with no significant differences between groups (flapless group - 0.6 mm buccal and 0.42 mm lingual; flap group - 0.67 and 0.13 mm).The clinical evaluation of immediate implant placement after 3 months of healing indicated that buccal soft tissue retraction was lower in the flapless group than in the flap group, without significant differences. The mean values of the biological width longitudinal dimension at the buccal aspect were higher in the flap group than in the flapless group, this difference being mostly due to the Pm3, probably because of a thinner biotype in this region.

To compare the remodeling of the alveolar process at implants installed immediately into extraction sockets by applying a flap or a "flapless" surgical approach in a dog model.Implants were installed immediately into the distal alveoli of the second mandibular premolars of six Labrador dogs. In one side of the mandible, a full-thickness mucoperiosteal flap was elevated (control site), while contra-laterally, the mucosa was gently dislocated, but not elevated (test site) to disclose the alveolar crest. After 4 months of healing, the animals were sacrificed, ground sections were obtained and a histomorphometric analysis was performed.After 4 months of healing, all implants were integrated (n=6). Both at the test and at the control sites, bone resorption occurred with similar outcomes. The buccal bony crest resorption was 1.7 and 1.5 mm at the control and the test sites, respectively."Flapless" implant placement into extraction sockets did not result in the prevention of alveolar bone resorption and did not affect the dimensional changes of the alveolar process following tooth extraction when compared with the usual placement of implants raising mucoperiosteal flaps.

Objectives: The objective of this study was to evaluate peri-implant soft tissue attachment and alveolar bone height on nanoporous TiO2 thin film on commercial titanium dental implants compared with unmodified standard implants. Material and methods: In six adult beagle dogs, the mandibular premolars P2–P4 were extracted bilaterally. Sol–gel-derived nanoporous TiO2 thin film was produced on smooth coronal part of standard ITI® Straumann implants (4.1 mm × 8.0 mm) by dip coating method. After 3 months healing period of the extraction sockets modified (n=24) and unmodified (n=11) control implants were placed bilaterally. The animals were killed after 8 weeks and the samples were retrieved and processed for histologic/histomorfometric and TEM/SEM evaluations. Results: Histological examination showed mild or absent inflammatory reaction in peri-implant connective tissues around the surface modified implants. Further, junctional epithelium (JE)/connective tissue (CT) appeared to be in immediate contact with the experimental implants. Of the experimental implants, 22% were judged to be detached from the implant surface while 45% of the untreated control implants were detached. Dense plaques of hemidesmosomes were found in TEM evaluation of the JE cell membrane facing the surface-treated implants. In the histomorfometric analysis, the distance between the implant margin and alveolar bone crest was significantly shorter in surface-treated implants than in the control implants (P

The response of nerve fibres in the peri-implant epithelium to titanium implantation was investigated with an experimental model using rat maxilla and immunohistochemical techniques. The latter employed antibodies to protein gene product 9.5 (PGP9.5), and to calcitonin gene-related peptide (CGRP). In control rats without an implantation, a dense innervation of PGP9.5- and CGRP-positive nerve fibres was recognized throughout the junctional epithelium, as has been previously reported. A titanium-implantation induced a remarkable inflammatory reaction, as well as the destruction of covering epithelial cells. By 3-5 days post-implantation, inflammatory reaction showed a tendency to disappear, and the peri-implant epithelium showed proliferation and down-growth along the implant. At this stage, no nerve fibres were found around the peri-implant epithelium. At 10 days, a few nerve fibres reached the basal cell layers of the peri-implant epithelium, and entered it 15 days after implantation when the peri-implant epithelial cells showed morphological features roughly resembling those of normal junctional epithelial cells. At the complete osseointegration stage (days 20-30), the PGP9.5- and CGRP-positive nerve fibres, thin and beaded in appearance, were found distributed in the peri-implant epithelium. After 20 days, the numerical density of the intraepithelial nerves in the peri-implant epithelium appeared the same as, or less than, that in the normal junctional epithelium. These findings indicate that the peri-implant epithelium shows the same innervation as that in normal junctional epithelium, and that the intraepithelial nerve fibres in the peri-implant epithelium might have diverse functions, which have been suggested in the literature.

Horseradish peroxidase (HRP) tracer was applied to the gingival sulcus of implants or natural teeth at 5, 25, or 50 mg/ml to investigate the sealing capacities of the peri-implant epithelium (PIE) and junctional epithelium (JE); the extent of HRP penetration was observed under electron microscopy. A Ti-6Al-4V implant was inserted either immediately (immediate implantation) or 2 weeks (delayed implantation) after extraction of the maxillary left first molar of rats. The JE of the right molar was used as a control. Although the whole PIE of undecalcified sections appeared to be attached to the implant surface, electron microscopically, the internal basement lamina (IBL) and hemidesmosomes were deficient in the coronal-middle region of the PIE. There were extensive extracellular deposits of HRP in the intercellular spaces between PIE cells, and HRP was blocked to some extent by the lamina lucida and lamina densa of the external basal lamina and basal cell junction. HRP was detected in the connective tissue under the PIE, but was not found in the connective tissue under the JE. Intracellularly, HRP was found in the vesicles and granules of PIE cells and JE cells. These were fewer in number in PIE cells than in JE cells. There were no differences between the findings for immediate and delayed implantation. The results indicate that a deficiency in the IBL permitted penetration of HRP from the gingival sulcus into the connective tissue under the PIE, and suggest that the endocytotic capacity of PIE cells is inferior to that of JE cells.

The aim of this study was to evaluate the periodontal parameters of osseointegrated dental implants. The condition of the peri-implant mucosa was assessed using periodontal parameters, i.e., of plaque index, bleeding on probing, probing pocket depth, probing attachment level and Periotest scores as well as a radiographic parameter, over a 4-year follow-up period. 32 non-submerged ITI dental implants, all placed in the mandible, were studied in 12 patients who had good oral hygiene. All patients were regularly recalled at 6-month intervals. The overall implant success rate was 100%. None of the implants showed any signs of inflammation, radiographic bone loss or any detectable mobility during the follow-up period. Methods similar to those used to evaluate the natural dentition were effectively employed to assess the clinical status of the dental implants. The diagnostic value of these parameters could not, however, be determined from this study due to the absence of any peri-implant tissue complications. The results indicated that some periodontal parameters of healthy peri-implant mucosa might be slightly different from healthy periodontal tissue.

In 5 monkeys a total of 30 transmucosal endosseous dental implants were inserted in edentulous areas of the mandible with presence or absence of keratinized mucosa. After a healing period of 3 months with optimal plaque control, all implants were exposed to plaque accumulation for periods up to 9 months. To secure abundant plaque accumulation on half the number of the implants, cotton wool ligatures were placed around the implants at the entrance to the peri-implant sulcus. Attachment loss was measured clinically and histometrically, and tissue recession was measured clinically. Ligated implants without keratinized mucosa demonstrated significantly more recession and slightly more attachment loss than the other implants. The results of this study suggest that the absence of keratinized mucosa around dental endosseous implants increases the susceptibility of the peri-implant region to plaque-induced tissue destruction.

This study details the structural and ultrastructural features of the interfaces between titanium implants and their surrounding tissues. The material stemmed from an experiment in dogs in which guided tissue regeneration with Gore-Tex membranes was used to treat peri-implant, ligature-induced tissue breakdown around submerged and nonsubmerged commercially pure titanium implants. Specimens from the nonsubmerged group were evaluated under light microscopy and scanning and transmission electron microscopy. A healthy gingiva and a gingival sulcus were formed around the implant necks. A regenerated junctional epithelium provided the epithelial union between implant and gingiva. The supracrestal connective tissue was characterized by a 3-dimensional network of collagen fibers, fibroblasts and blood vessels. Near the implant surface the collagen fibers ran parallel to the titanium surface or were orientated perpendicular to the implant. The connective tissue-implant interface was characterized by a fine fibrillar material interposed between the implant surface and the connective tissue. An unidentified material was also observed between the endings of functionally orientated collagen fibrils and the metallic surface. The apical portions of the implants were anchored in compact bone. At the bone-implant interface, either mineralized bone matrix was intimately adapted to the titanium surface without any intervening space or a 0.5-micron wide unmineralized layer was interposed. These findings indicate that a perimucosal seal was formed around the implants consisting of a junctional epithelium-implant union coronally and supported by the connective tissue-implant junction apically. The implants were integrated in connective tissue, but only tightly adapted to bone.

The installment of endosseous dental implants has become an accepted treatment procedure, and the long-term clinical results appear excellent. The composition of the soft tissue environment, however, is different from that around natural teeth. One characteristic of original junctional epithelium is its association with plasminogen activator (PA) activity. In 11 patients with a total of 30 ITI hollow-screw titanium dental implants, 16 biopsies were taken. Histologic cryostat sections were assayed for the presence of PA in the junctional epithelium. The results demonstrated that junctional epithelium around titanium implants yields PA activity in a manner very similar to that of natural teeth. The ability to produce this enzyme activity is not related to the developmental origin of the junctional cells, but to their position and function at the base of the gingival sulcus.

This investigation is the result of a multicenter research project between the Catholic University of Leuven and the University of Madrid with the purpose of studying histometrically and ultrastructurally periimplant marginal tissue reactions in mendium-term Brånemark implants. 12 patients were selected and distributed according to predefined clinical and radiological parameters into a periimplant infection group (A) and a non-periimplant infection group (B). Biopsies were taken from these patients and analyzed by means of microscopy (LM), histometry (HM) and transmission electron microscopy (TEM). With LM group A showed pathological features consistent with an advanced gingivitis lesion, while group B showed signs of healthy gingiva in the connective tissue and vauolae formation in the epithelium. With HM, group A showed significantly higher transmigration of inflammatory cells in the epithelium than group B. In the connective tissue, group A showed a higher % of inflammatory infiltrate with significantly higher numbers of plasma cells and mononuclear cells than group B. With TEM, group A showed an ultrastructural picture consistent with plaque-induced periodontitis, while group B showed a rather healthy connective tissue and vacuolae formation in the epithelium. These results suggest that supracrestal marginal tissues around osseointegrated implants react to plaque bacteria similarly to periodontal tissues around teeth by chronic inflammation.

In the present animal experiment, analyses and comparisons were made between the structure and composition of clinically healthy supraalveolar soft tissues adjacent to implants and teeth. 5 beagle dogs were used. The right mandibular premolar region was selected in each dog for placement of titanium implants, while the left mandibular premolar region served as control. Extractions of the mandibular premolars were preformed, healing allowed, following which titanium fixtures were installed in the edentolous premolar region. Abutment connection was carried out 3 months later. After another 2 months of healing, plaque control was initiated and maintained for 8 weeks. At the end of the plaque control period, clinical examinations were performed and biopsies harvested from the implant site and the contralateral premolar tooth region. Following fixation and decalcification, all tissue samples were embedded in EPON and examined by histometric and morphometric means. The result from the analyses demonstrated that the periimplant mucosa which formed at titanium implants following abutment connection had many features in common with gingival tissue at teeth. Thus, like the gingiva, the peri-implant mucosa established a cuff-like barrier which adhered to the surface of the titanium abutment. Further, both the gingiva and the peri-implant mucosa had a well-keratinized oral epithelium which was continuous with a junctional epithelium that faced the enamel or the titanium surface. In the periimplant mucosa, the collagen fibers appeared to commence at the marginal bone and were parallel with the abutment surface. All gingival and periimplant units examined were free from infiltrates of inflammatory cells. It was suggested that under the conditions of study, both types of soft tissues, gingiva and periimplant mucosa, have a proper potential to prevent subgingival plaque formation.