Influence of the Transmucosal Surface of Dental Implants on the Soft Tissue Attachment Level and Marginal Bone Loss in Preclinical Studies: A Systematic Review.

PURPOSE: To determine the characteristics of dental implant transmucosal surfaces that influence soft tissue attachment and marginal bone loss (MBL). MATERIALS AND METHODS: The PubMed, Embase, and Cochrane Library electronic databases were searched based on predefined PICO eligibility criteria. Data from animal studies that compared junctional epithelium and connective tissue attachment and MBL from 4 days to 72 weeks were analyzed. The risk of bias was performed with the Systematic Review Centre for Laboratory Animal Experimentation tool. A rank analysis evaluation of data was performed, and the most frequently appearing materials/surfaces for each tissue compartment were identified. RESULTS: The search identified 3,549 studies, 28 of which were eligible for analysis, with an average risk of bias of 28% ± 10%. Machined, polished, etched, sandblasted, or coated titanium and zirconia materials/surfaces were most frequently examined. Several studies investigated lithium disilicate, polyether ether ketone (PEEK) or polyether ketone ketone (PEKK), aluminum oxide, and gold. Based on ranking and frequency of use at different time points, titanium grade IV (Ti-4) microthreads with a polished neck area most frequently supported natural tooth-like junctional epithelial attachment (≤ 1.5 mm), while machined Ti-4 and machined titanium grade V (Ti-5) most frequently supported connective tissue attachment (≤ 1.25 mm) and led to the least MBL (≤ 0.75 mm). CONCLUSIONS: Analyzed data suggest that Ti-4 microthreads with a polished neck area and machined Ti-4 and Ti-5 were the materials/surfaces of choice for the transmucosal part of implants. However, the extensive heterogeneity in reported studies precludes solid identification of the best materials/surfaces.

Effects of interleukin-1ß on human follicular dendritic cell-secreted protein gene expression in periodontal ligament cells.

Follicular dendritic cell-secreted protein (FDC-SP) is expressed in FDCs, human periodontal ligament (HPL) cells, and junctional epithelium. To evaluate the effects of interleukin-1 beta (IL-1ß) on FDC-SP gene expression in immortalized HPL cells, FDC-SP mRNA and protein levels in HPL cells following stimulation by IL-1ß were measured by real-time polymerase chain reaction and Western blotting. Luciferase (LUC), gel mobility shift, and chromatin immunoprecipitation (ChIP) analyses were performed to study the interaction between transcription factors and promoter regions in the human FDC-SP gene. IL-1ß (1 ng/mL) induced the expression of FDC-SP mRNA and protein levels at 3 h, and reached maximum levels at 12 h. IL-1ß increased LUC activities of constructs (-116FDCSP - -948FDCSP) including the FDC-SP gene promoter. Transcriptional inductions by IL-1ß were partially inhibited by 3-base-pair (3-bp) mutations in the Yin Yang 1 (YY1), GATA, CCAAT-enhancer-binding protein2 (C/EBP2), or C/EBP3 in the -345FDCSP. IL-1ß-induced -345FDCSP activities were inhibited by protein kinase A, tyrosine-kinase, mitogen-activated protein kinase (MEK)1/2, and PI3-kinase inhibitors. The results of gel shift and ChIP assays revealed that YY1, GATA, and C/EBP-ß interacted with the YY1, GATA, C/EBP2, and C/EBP3 elements that were increased by IL-1ß. These studies demonstrate that IL-1ß increases FDC-SP gene transcription in HPL cells by targeting YY1, GATA, C/EBP2, and C/EBP3 in the human FDC-SP gene promoter.

Consideraciones periodontales y protéticas para el tallado de piezas dentarias / Periodontal and prosthetic considerations for teeth grinding

Las preparaciones dentales para prótesis fija, involucran al tejido duro, además la proximidad que existe con la encía hace necesario la comprensión de su estructura y la función del aparato de protección, para que las restauracionesno invadan el ancho biológico constituido por el epitelio de unión y las fibras gingivales; basándonos en los estudios de Gargiulo en 1961, donde la inserción del tejido conjuntivo mide 1.07 mm y la adherencia epitelial 0.97 mm en promedio, sumados al espacio del surco gingival. El biotipo periodontal es de vital importancia para la elección de la altura de terminación del hombro de la preparación, donde el mismo deberá establecerse, previo sondaje óseo a no menos de 2.5 mm de la cresta según Kois (2008). La utilización de materiales y técnicas para la toma de impresiones definitivas deben ser amigables con los tejidos blandos a fin de no producir alteraciones y que las restauraciones sean predecibles, tanto funcional como estéticamente en el transcurso del tiempo

Dental preparations fixed prosthesis, involving the hard tissue, there is also proximity to the gum is necessary to understand the structure and function of the protection device, to restorations without invading the biologic width consisting of the epithelium junction and the gingival fibers; based on Gargiulo studies in 1961, where the insertion of the connective tissue and epithelial measured 1.07 mm 0.97 mm in average adhesion, coupled with sulcular space. The periodontal biotype is of vital importance for the choice of the termination of the shoulder height of the preparation, where it must be established prior tobone sounding no less than 2.5mm crest according Kois (2008). The use of materials and techniques for making final impressions should be friendly soft so as not to alter tissues and restorations predictable, both functionally and aesthetically over time.

Green tea extract inhibits the onset of periodontal destruction in rat experimental periodontitis.

BACKGROUND AND OBJECTIVE: Green tea extract exerts a variety of biological effects, including anti-inflammatory activities. However, there has been no report on the effect of green tea extract on loss of attachment, which is an important characteristic of periodontitis. Here, we examined the inhibitory effects of green tea extract on the onset of periodontitis in a rat model. MATERIAL AND METHODS: Rats were immunized intraperitoneally with Escherichia coli lipopolysaccharide (LPS). The LPS group (n = 12) received a topical application of LPS onto the palatal gingival sulcus every 24 h. The green tea extract group (n = 12) received a topical application of LPS mixed with green tea extract, sunphenon BG, every 24 h. The phosphate-buffered saline (PBS) group (n = 6) received a topical application of PBS every 24 h. The levels of anti-LPS immunoglobulin G (IgG) in serum were determined using ELISA. Rats in the LPS and green tea extract groups were killed after the 10th and 20th applications. Rats in the PBS group were killed after the 20th application. Loss of attachment, level of alveolar bone and inflammatory cell infiltration were investigated histopathologically and histometrically. RANKL-positive cells and the formation of immune complexes were evaluated immunohistologically. RESULTS: There was no significant difference in the serum levels of anti-LPS IgG between the LPS group and the green tea extract group. In contrast, loss of attachment, level of alveolar bone, inflammatory cell infiltration and RANKL expression in the green tea extract group were significantly decreased compared with those in the LPS group. CONCLUSION: These findings demonstrate that green tea extract suppresses the onset of loss of attachment and alveolar bone resorption in a rat model of experimental periodontitis.

Periodontal healing after application of enamel matrix derivative in surgical supra/infrabony periodontal defects in rats with streptozotocin-induced diabetes.

BACKGROUND AND OBJECTIVE: Epidemiologic and clinical studies have indicated that diabetes is a risk factor for periodontal disease progression and healing. The aim of the present study was to evaluate short-term healing after enamel matrix derivative (EMD) application in combined supra/infrabony periodontal defects in diabetic rats. MATERIAL AND METHODS: Thirty male Wistar rats were initially divided into two groups, one with streptozotocin-induced diabetes and another one with healthy (non-diabetic) animals. Bony defects were surgically created on the mesial root of the first maxillary molars. After root surface planing and EDTA conditioning, EMD was applied to the roots at one side of the maxillae, while those on the contralateral sides were left untreated. Animals were killed 3 wk after surgery, and block sections were prepared for histologic and histomorphometric analysis. RESULTS: There was statistically significant more gingival recession in diabetic animals than in non-diabetic animals. The length of the junctional epithelium was significantly shorter in the EMD-treated sites in both diabetic and normoglycemic rats. Sulcus depth and length of supracrestal soft connective tissue showed no statistically significant differences between groups. In all animals, new bone formation was observed. Although new bone occurred more frequently in healthy animals, the extent of new bone was not significantly different between groups. In none of the teeth, a layer of new cementum was detectable. EMD had no influence on bone or cementum regeneration. Adverse reactions such as excessive inflammation due to bacterial root colonization, ankylosis and bone fractures were exclusively observed in diabetic animals, irrespective of EMD treatment. CONCLUSION: Within the limits of the present study, it can be concluded that periodontal healing was impaired in streptozotocin-induced diabetic rats. EMD had no beneficial effects on new bone and cementum formation during short-term healing in this defect model and could not ameliorate the adverse effects in the systemically compromised animals.

Effects of electrical stimulation on periodontal tissue remodeling in rats.

BACKGROUND AND OBJECTIVE: Electric current is used to promote wound healing. However, it is unclear whether electrical stimulation contributes to gingival tissue remodeling. This study examined the effects of electrical stimulation on gingival tissue remodeling in a rat periodontitis model. MATERIAL AND METHODS: Male Wistar rats (n = 28, 8 wks of age) were divided into four groups of seven rats each. The control group did not receive any treatment for 6 wks. In the other groups, periodontitis was ligature-induced for 4 wks. After 4 wks, the rats with periodontitis were given daily electrical stimulation of 0, 50 or 100 µA for 2 wks. RESULTS: The periodontitis group stimulated with 0 µA showed a higher density of polymorphonuclear leukocytes and a lower density of collagen in gingival tissue compared with the control group (p < 0.05). The two remaining groups treated with 50 or 100 µA of electrical stimulation exhibited a lower density of polymorphonuclear leukocytes (p < 0.05) and a higher density of collagen than the group stimulated with 0 µA (p < 0.05). They also showed higher expression of fibroblast growth factor-2 than the group treated with 0 µA of electrical stimulation (p < 0.05). CONCLUSION: Electric stimulation may offer a novel approach to promote gingival tissue remodeling in periodontal lesions.

Supplementation of green tea catechins in dentifrices suppresses gingival oxidative stress and periodontal inflammation.

OBJECTIVE: this study examined the effects of a dentifrice containing green tea catechins on gingival oxidative stress and periodontal inflammation using a rat model. DESIGN: twenty-four male Wister rats were randomly divided into four groups. The first group (Control group) received no treatment for 8 weeks. Periodontal inflammation was induced in the second group for 8 weeks. Periodontal inflammation was induced in the last two groups for 8 weeks and dentifrices with or without green tea catechins were topically applied to the gingival sulcus daily for 4 weeks prior to the end of the experimental period. RESULTS: rats that had experimental periodontal inflammation showed apical migration of the junctional epithelium, alveolar bone loss and inflammatory cell infiltration in the connective tissue subjacent to the junctional epithelium at 8 weeks, whilst the control group showed no pathologic changes. Topical application of a green tea catechin-containing dentifrice reduced inflammatory cell infiltration in the periodontal lesions to a greater degree than the control dentifrice at 8 weeks. The gingiva in which green tea catechin-containing dentifrice was applied also showed a lower level of expression of hexanoyl-lysine (a marker of lipid peroxidation), nitrotyrosine (a marker of oxidative protein damage), and tumour necrosis factor-α (an indicator of pro-inflammatory cytokines) at 8 weeks compared to gingiva in which the control dentifrice was applied. CONCLUSIONS: adding green tea catechins to a dentifrice may contribute to prevention of periodontal inflammation by decreasing gingival oxidative stress and expression of pro-inflammatory cytokines.

Muscarinic acetylcholine receptor activation increases transcellular transport of macromolecules across mouse and human intestinal epithelium in vitro.

The intestinal epithelium acts as a barrier restricting uptake of luminal macromolecules such as dietary antigens and microbes. Here, we examined the role of cholinergic signalling in the regulation of permeability to macromolecules. Mouse jejunum was mounted in Ussing chambers and permeability was determined by measuring the flux of the antigen-sized protein, horseradish peroxidase (HRP), across the tissue. Baseline HRP permeability was significantly reduced by neural blockade with tetrodotoxin or cholinergic muscarinic antagonism with atropine, suggesting that ongoing release of endogenous acetylcholine from enteric nerves regulates barrier function. Exogenous addition of the muscarinic agonist bethanechol caused significant increases in both HRP flux and the area of HRP-containing endosomes in enterocytes. Bethanechol-enhanced HRP flux was abrogated by the M3 receptor antagonist, 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP), the phospholipase A(2) inhibitor quinacrine, and the cyclooxygenase inhibitor indomethacin. Complementary in vitro studies showed direct effects of bethanechol on T84 epithelial cells, where increased HRP uptake was associated with increased F-actin, and increased cytosolic phospholipase A(2) (cPLA(2)) phosphorylation. Taken together, these results provide evidence for cholinergic regulation of transepithelial transport of macromolecules, mainly mediated by activation of M3 receptors with subsequent involvement of phospholipase A(2) and cyclooxygenase products.

Effect of enamel matrix protein derivative on healing of surgical supra-infrabony periodontal defects in the rat molar: a histomorphometric study.

BACKGROUND: Enamel matrix protein derivative (EMD) has proven to enhance periodontal regeneration in human and animal studies. The present histomorphometric study evaluated healing of combined supra-infrabony periodontal defects with EMD. METHODS: The study comprised two groups of 10 Wistar rats each, 7 to 8 months old. Bony defects were created on the mesial aspect of the mesial root of the first maxillary molar. The root surface was planed and 24% EDTA gel applied for 2 minutes and then rinsed with water. In the study group, EMD was applied, and in the control group, only propylene glycol alginate was applied. Animals were sacrificed 12 weeks after surgery, and block sections were removed, demineralized, and embedded in paraffin. For histomorphometric analysis, three sections from the central area of the defect were selected. Root, surgical defect, epithelial attachment, sulcus, supracrestal connective tissue, ankylosis, and the length and area of new cementum and new bone were measured. RESULTS: No statistically significant differences between the two groups were found for root and defect measures. The remaining parameters were calculated as a percentage of the defect. In the study group, smaller gingival recession (P = 0.05), deeper gingival sulcus (P = 0.05), and shorter junctional epithelium (P = 0.01) were found. New cementum was observed in the study group only (P = 0.02). Ankylosis was six times larger in the control group but not statistically significant. New bone formation was similar in both groups. CONCLUSION: Enamel matrix protein derivative enhanced periodontal healing in this model by reducing gingival recession and junctional epithelium along the root surface and enhancing the formation of new cementum.

Effect of an oily calcium hydroxide suspension (Osteoinductal) on healing of intrabony periodontal defects. A pilot study in dogs.

The aim of the present study was to evaluate histologically in dogs the effect of treating intrabony defects with an oily calcium hydroxide suspension (OCHS). Intrabony defects were surgically created bilaterally at the distal aspects of the maxillary first premolars and at the mesial aspects of the third premolars in two mongrel dogs. Subsequently, the defects were randomly treated with (a) access flap surgery followed by the application of an OCHS or (b) access flap surgery alone. After 8 weeks of healing, the animals were killed. Dissected blocks containing the experimental specimens were fixed in formalin, decalcified in EDTA, and embedded in paraffin. The formation of new cementum and bone was assessed histomorphometrically. In the control group, healing was predominantly characterized by the formation of a long junctional epithelium along the root surface and limited periodontal regeneration at the most apical part of the defect. The OCHS-treated defects consistently revealed periodontal regeneration (i.e., new periodontal ligament, new cementum with inserting collagen fibers, and new bone). Within the limits of the present study, it can be concluded that OCHS may favor periodontal regeneration in acute-type intrabony periodontal defects.

Histological analysis of effects of 24% EDTA gel for nonsurgical treatment of periodontal tissues.

The aim of this study was to investigate, by means of histological and histomorphometric analysis, the effects of 24% ethylenediaminetetraacetic acid (EDTA) gel in periodontal tissue when used in combination with conventional periodontal treatment. Periodontitis was induced in the 2nd upper left permanent molars of 45 male Wistar rats by means of ligature. After 5 weeks, this was removed and debridement was performed. The animals were then randomly divided into 3 groups; group 1: mechanical treatment, group 2: mechanical treatment and EDTA gel application for 2 min, and group 3: mechanical treatment and placebo gel application for 2 min. After the treatment, rinsing was done with 0.9% saline solution for 1 min in all cases, followed by root notching in the deepest part of the pocket. After 4, 10, and 28 days the animals were sacrificed. The averages obtained were evaluated by means of test two-way analysis of variance (ANOVA) and Tukey statistical tests (P < 0.05). The results showed that with respect to the type of treatment employed, there were no statistically significant differences in the vitality of the periodontal tissue. It was concluded that 24% EDTA gel did not interfere with periodontal tissue repair when used in combination with conventional periodontal treatment.

A histopathologic investigation on the effects of electrical stimulation on periodontal tissue regeneration in experimental bony defects in dogs.

BACKGROUND: One endpoint of periodontal therapy is to regenerate the structure lost due to periodontal disease. In the periodontium, gingival epithelium is regenerated by oral epithelium. Underlying connective tissue, periodontal ligament, bone, and cementum are derived from connective tissue. Primitive connective tissue cells may develop into osteoblasts and cementoblasts, which form bone and cementum. Several procedural advances may support these regenerations; however, the regeneration of alveolar bone does not always occur. Therefore, bone stimulating factors are a main topic for periodontal reconstructive research. The present study was designed to examine histopathologically whether the application of an electrical field could demonstrate enhanced alveolar and cementum regeneration and modify tissue factors. METHODS: Seven beagle dogs were used for this experiment. Mandibular left and right sides served as control and experimental sides, respectively, and 4-walled intrabony defects were created bilaterally between the third and fourth premolars. The experimental side was treated with a capacitively coupled electrical field (CCEF) (sinusoidal wave, 60 kHz, and 5 V peak-to-peak), applied for 14 hours per day. The following measurements were performed on the microphotographs: 1) the distance from the cemento-enamel junction to the apical notch (CEJ-AN) and from the crest of newly formed bone (alveolar ridge) to the apical notch (AR-AN); 2) the thickness of new cementum in the apical notch region; and 3) the length of junctional epithelium. The following histopathologic parameters were assessed by a semiquantitative subjective method: 1) inflammatory cell infiltration (ICI); 2) cellular activity of the periodontal ligament; 3) number and morphology of osteoclasts; 4) resorption lacunae; and 5) osteoblastic activity. RESULTS: The results showed that the quantity of new bone fill and the mean value of the thickness of the cementum were significantly higher for the experimental side (P < 0.01). The location of the base of the pocket was positioned more coronally with respect to the apical point of the coronal notch in the experimental side (statistically significant P < 0.01). The length of the junctional epithelium and the number of osteoclasts were higher in the stimulated side than the coronal side; these findings were also statistically significant (P < 0.01). The comparison of the electrically stimulated versus non-stimulated mandibles with the semiquantitative subjective method demonstrated statistically significant differences in defined histopathologic parameters, except for osteoclast morphologies (P > 0.05). CONCLUSIONS: This study demonstrated that the CCEF method has the potential to produce reconstructive effects and bone deposits. Further investigations with respect to the theoretical determination of local field parameters of the periodontal tissue complex, such as permittivity, conductivity, strength of the field electrical stimulation applied to the periodontal field current density, wavelength, and signal frequency appropriate for this field, should be undertaken. Using different electromotive forces alone or in combination with bone graft materials, guided tissue regeneration techniques, and dental implants may achieve a new dimension in periodontal therapy in the near future.

Loaded custom-made zirconia and titanium implants show similar osseointegration: an animal experiment.

BACKGROUND: Zirconia might be an alternative material to titanium for dental implant fabrication. The aim of the present study was to investigate the histological behavior (osseointegration) of loaded zirconia implants in an animal model and to compare it with the behavior of titanium implants. METHODS: Five months after extraction of the upper anterior teeth, 12 custom-made titanium implants (control group) and 12 custom-made zirconia implants (test group) were inserted in the extraction sites in six monkeys. Before insertion, the titanium implant surfaces were sandblasted with Al2O3 and subsequently acid-etched. The zirconia implants were only sandblasted. Six months following implant insertion, impressions were taken for the fabrication of single crowns. A further 3 months later, nonprecious metal crowns were inserted. Five months after insertion of the crowns, the implants with the surrounding hard and soft tissues were harvested, histologically prepared, and evaluated under the light microscope regarding the peri-implant soft tissue dimensions and mineralized bone-to-implant contact. RESULTS: No implant was lost during the investigational period. The mean height of the soft peri-implant tissue cuff was 5 mm around the titanium implants and 4.5 mm around the zirconia implants. No statistically significant differences were found in the extent of the different soft tissue compartments. The mean mineralized bone-to-implant contact after 9 months of healing and 5 months of loading amounted to 72.9% (SD: 14%) for the titanium implants and to 67.4% (SD: 17%) for the zirconia implants. There was no statistically significant difference between the different implant materials. CONCLUSION: Within the limits of this animal experiment, it can be concluded that the custom-made zirconia implants osseointegrated to the same extent as custom-made titanium control implants and show the same peri-implant soft tissue dimensions.

Comparative study of enamel matrix derivative with or without GTR in the treatment of class II furcation lesions in dogs.

Chronic Class II furcation lesions were created in four dogs. After 21 days, group I remained as a control, group 2 was treated with membranes and enamel matrix derivative (EMD), and group 3 received EMD alone. Healing in group 1 was characterized by a long junctional epithelium and discrete bone formation; group 2 showed reduced bone formation; and group 3 showed significant bone regeneration (area of new bone = 67.36%+/-3.93%; distance from furcation roof to bone crest = 0.57+/-0.15 mm). The EMD led to significant regeneration of the furcation lesions, and the association with membranes was detrimental.

Healing of human intrabony defects following regenerative periodontal therapy with a bovine-derived xenograft and guided tissue regeneration.

The purpose of the present study was to histologically evaluate the healing of human intrabony defects following treatment with either a bovine-derived xenograft (BDX) and guided tissue regeneration (GTR) [BDX + GTR] or a bovine-derived xenograft mixed with collagen (BDX Coll) and GTR [BDX Coll + GTR]. Eight patients with chronic periodontitis and each with one very deep intrabony defect around a tooth scheduled for extraction were treated with either a combination of BDX + GTR (five patients) or with BDX Coll + GTR (three patients). The postoperative healing was uneventful in all eight cases. After a healing period of 6 months, the teeth or roots were extracted together with some of their surrounding soft and hard tissues and subsequently fixed in 10% buffered formalin. Following decalcification in EDTA, the specimens were embedded in paraffin and 8-microm histological sections were cut in the mesio-distal direction, parallel to the long axes of the teeth. The sections were alternatively stained with hematoxylin and eosin, van Giesson's connective tissue stain or with the Ladevig's connective tissue staining method and examined under the light microscope. Generally, formation of new cementum with inserting collagen fibers was found in seven out of the eight treated cases, whereas in the remaining case (treated with BDX + GTR) the healing was characterized by formation of a long junctional epithelium along the debrided root surface and no formation of cementum or bone. In the specimens demonstrating periodontal regeneration the new cementum was always of a cellular type. In most cases, the graft particles were surrounded by bone. In some areas, the bone tissue around the graft particles was connected by perpendicularly inserting collagen fibers to the newly formed cementum on the root surface. The epithelium downgrowth stopped always at the most coronal part of the newly formed cementum. No remnants of the membrane material were observed in any of the biopsies. Connective tissue encapsulation of the graft particles was rarely observed and was limited to the most coronal part of the defects. The findings of the present study provide evidence that treatment of intrabony defects with both BDX + GTR and BDX Coll + GTR may enhance periodontal regeneration in humans.

Fatty acids and epithelial permeability: effect of conjugated linoleic acid in Caco-2 cells.

Conjugated linoleic acid (CLA) is a collective term referring to the positional and geometric isomers of linoleic acid. This novel fatty acid has been shown to have a number of beneficial actions, including immunomodulatory, anticarcinogenic, and antiatherogenic effects. Tight junctions of epithelial cells determine epithelial membrane integrity and selective paracellular permeability to ions and macromolecules. Occludin and ZO-1 are integral structural components of the tight junction, which are involved in the biogenesis and functional integrity of the epithelial monolayer. This study investigated the effects of two isomers of CLA (cis-9 and trans-10 isomers) on Caco-2 cell transepithelial resistance (TER) development, paracellular epithelial permeability, and occludin and ZO-1 expression. Caco-2 cells were grown in media supplemented with 0.05 mM linoleic acid, cis-9 CLA, or trans-10 CLA for 21 days. The trans-10 CLA isomer delayed Caco-2 cell TER development, which is an in vitro measure of epithelial cell integrity, and increased paracellular epithelial permeability. Immunofluorescent staining of Caco-2 cell epithelial monolayers grown in media supplemented trans-10 CLA showed that the trans-10 CLA isomer altered distribution of occludin and ZO-1. The trans-10 CLA isomer delayed the acquisition of transepithelial resistance and altered the cellular distribution of occludin, which have important implications in relation to epithelial permeability.

Multi-layered periodontal pocket epithelium reconstituted in vitro: histology and cytokeratin profiles.

BACKGROUND: In order to study inter-individual differences in bacterial adhesion/invasion of periodontal tissues, an in vitro model for culturing multi-layered pocket epithelium without feeder layers or stromal equivalents (including the evaluation of their cytokeratin profiles) was developed. METHODS: Pocket epithelium was collected and grown until confluent in Falcon flasks using keratinocyte-serum free medium (KSFM), without a feeder layer. In the second passage, oral keratinocytes were re-grown in a 2 compartment system using either a clear polyester (transwell-clear [TCL]) or a collagen (transwell-col [TCO]) membrane as culture surface. After the first week, the calcium concentration was raised to 1.2 mM and in half the wells, the KSFM was supplemented with 10% fetal calf serum (FCS). Histology and immunohistochemistry were performed after 1, 2, and 3 weeks of additional growth. RESULTS: In general, all conditions resulted in a structured epithelium consisting of 3 to 5 layers, but important differences were observed between the membrane types and between the media. CK4 was rarely and only lightly expressed while CK18 and 19 (characteristic of junctional epithelium) were very strongly expressed in the older (2 and 3 weeks) cultures. CK13 and 14 (characteristic of any stratifiable epithelial cell) also tended to increase over time; CK13 seemed to be stronger in KSFM with FCS while the contrary was true for CK14. The multi-layer created by the combination TCL/KSFM + 10% FCS resembled a junctional epithelium most, while that grown on TCO without FCS mimicked the sulcular epithelium. CONCLUSIONS: It seems possible to create a histiotypic culture resembling either periodontal pocket or junctional epithelium without the use of stromal equivalents or feeder layers which make this approach more cumbersome. This multi-layered culture offers a model to investigate the permeability of pocket epithelium and the adhesion and penetration of bacteria under well-defined environmental conditions.

Effect of calcium sulphate on the healing of periodontal intrabony defects.

The purpose of this series of three studies was to evaluate the regenerative potential of calcium sulphate in the treatment of periodontal intrabony defects.

The mucosal attachment at different abutments. An experimental study in dogs.

The present experiment was performed to examine if the material used in the abutment part of an implant system influenced the quality of the mucosal barrier that formed following implant installation. 5 beagle dogs were included in the study. The mandibular premolars and the 1st, 2nd and 3rd maxillary premolars were extracted. Three fixtures of the Brånemark System were installed in each mandibular quadrant (a total of 6 fixtures per animal). Abutment connection was performed after 3 months of healing. In each dog the following types of abutments were used: 2 "control abutments" (c.p. titanium), 2 "ceramic abutments" (highly sintered Al2O3), 1 "gold abutment", and 1 "short titanium abutment". This "short titanium abutment" was provided with an outer structure made of dental porcelain fused to gold. Following abutment connection a plaque control program was initiated and maintained for 6 months. The animals were sacrificed and perfused with a fixative. The mandibles were removed and each implant region was dissected, demineralized in EDTA and embedded in EPON. Semithin sections representing the mesial, distal, buccal and lingual aspects of the peri-implant tissues were produced and subjected to histological examination. The findings from the analysis demonstrated that the material used in the abutment portion of the implant influenced the location and the quality of the attachment that occurred between the periimplant mucosa and the implant. Abutments made of c.p. titanium or ceramic allowed the formation of a mucosal attachment which included one epithelial and one connective tissue portion that were about 2 mm and 1-1.5 mm high, respectively. At sites where abutments made of gold alloy or dental porcelain were used, no proper attachment formed at the abutment level, but the soft tissue margin receded and bone resorption occurred. The abutment fixture junction was hereby occasionally exposed and the mucosal barrier became established to the fixture portion of the implant. It was suggested that the observed differences were the result of varying adhesive properties of the materials studied or by variations in their resistance to corrosion.