Histological Techniques for the Study of the Dentogingival Junction: A Scoping Review Using the Anatomical Quality Assurance Checklist (AQUA) / Técnicas Histológicas para el Estudio de la Unión Dentogingival: Una Revisión de Alcance Utilizando el Anatomical Quality Assurance Checklist (AQUA)

SUMMARY: The dentogingival junction (DGJ) is an adaptation of the oral mucosa composed of epithelial and connective tissues intimately related with the mineralised tissues of the tooth. The histological evidence available is mainly based on studies in animals, separate evaluations of hard and soft tissues, and studies using conventional histological techniques that eliminate the enamel from preparations. The aim of this study was to carry out a review of the existing evidence on histological techniques available for study of the tooth and periodontium in conjunction in humans. A scoping review was carried out of the available literature referring to study of the tooth and the periodontium in conjunction in humans, in the Web of Science (WoS), EMBASE, Scopus and SciELO databases, using the terms "Histological Techniques"[Mesh]) and "Epithelial Attachment"[Mesh]. One hundred and fifty-nine articles were found, of which 54 were selected for full- text reading. Ten were finally included in the qualitative synthesis, and we applied the Anatomical Quality Assurance (AQUA) checklist for analysis the methodological quality of the selected articles. The results showed that the only articles with a low risk of bias in all five domains according to the AQUA criteria corresponded to Silva et al. (2011) and Agustín-Panadero et al. (2020). Finally, we conclude that the quality of the histological sections to observe tissues that simultaneously contain the tooth and the periodontium, is conditioned by the selected technique and by the care required in certain specific tasks during the histological processing of the samples.

La unión dentogingival (DGJ) es una adaptación de la mucosa oral compuesta por tejidos epitelial y conectivo íntimamente relacionados con los tejidos mineralizados del diente. La evidencia histológica disponible se basa principalmente en estudios en animales, evaluaciones separadas de tejidos duros y blandos y estudios utilizando técnicas histológicas convencionales que eliminan el esmalte de las preparaciones. El objetivo de este estudio fue realizar una revisión de la evidencia existente sobre las técnicas histológicas disponibles para el estudio del diente y el periodonto en conjunto en humanos. Se realizó un scoping review de la literatura disponible referente al estudio del diente y el periodonto en conjunto en humanos, en las bases de datos Web of Science (WoS), EMBASE, Scopus y SciELO, utilizando los términos "Histological Techniques"[Mesh]) y "Epithelial Attachment"[Mesh]. Se encontraron 159 artículos, de los cuales 54 fueron seleccionados para lectura de texto completo. Diez fueron finalmente incluidos en la síntesis cualitativa, y se aplicó la lista de verificación Anatómica Quality Assurance (AQUA) para el análisis de la calidad metodológica de los artículos seleccionados. Los resultados mostraron que los únicos artículos con bajo riesgo de sesgo en los cinco dominios según los criterios AQUA correspondían a Silva et al. (2011) y Agustín-Panadero et al. (2020). Finalmente, concluimos que la calidad de los cortes histológicos para observar los tejidos que contienen simultáneamente el diente y el periodonto, está condicionada por la técnica seleccionada y por el cuidado requerido en ciertas tareas específicas durante el procesamiento histológico de las muestras.

Expression and distribution of calprotectin in healthy and inflamed periodontal tissues / 北京大学学报(医学版)

OBJECTIVE@#Calprotectin, the heterdimer of S100A8 and S100A9, is the major cytoplasmic protein of neutrophils, which is also expressed or induced in gingival epithelial cells, activated mononuclear macrophages and vascular endothelial cells. Calprotectin is intimately associated with the initiation and progression of periodontitis, but the in vivo expression patterns of calprotectin in healthy and inflamed periodontal tissue are not fully understood. To observe the expression, distribution and cellular localization of calprotectin in the samples of healthy periodontal tissues and experimental periodontitis tissues of Beagles and to explore their relationship with periodontal inflammation and possible effect.@*METHODS@#Experimental periodontitis model was established by ligation around the mandibular second molar of the Beagle dogs, while the contralateral teeth were healthy controls. Induction duration was 12 weeks, before the dogs were executed. Tissue specimens were demineralized and serial sections were made conventionally. The in vivo expression of calprotectin in the healthy and inflamed periodontal tissues were examined by immunohistochemistry. The in vitro expression of calprotectin in human primary gingival fibroblasts (GFs) and periodontal ligament (PDL) cells were detected by immunocytochemistry.@*RESULTS@#Immunohistochemistry analysis indicated that calprotectin was expressed in gingival epithelial cells and infiltrated neutrophils in the healthy periodontium within the gingival epithelium, S100A8/A9 was most strongly expressed in the junctional epithelium, followed by surface epithelium, and least expressed in the sulcular epithelium. The S100A8/A9 expression levels were sharply defined at the junction between the junctional epithelium and the sulcular epithelium. In periodontal inflammatory lesions, the expression level of calprotectin in sulcular epithelium and junctional epithelium was up-regulated than that in the healthy gingival epithelium. Calprotectin was inducibly expressed in fibroblast-like cells in gingival connective tissue and periodontal ligament tissue, microvascular endothelial cells (ECs) and bone marrow fibroblasts under inflammatory conditions. Additionally, the expression of calprotectin in primary human GFs and PDL cells was confirmed by immunnocytochemistry staining.@*CONCLUSION@#Constitutively expressed in neutrophils and gingival epithelial cells, and calprotectin might maintain the homeostasis and integrity of periodontium. Inflammation-induced expression of calprotectin in GFs, PDL cells, microvascular ECs and bone marrow fibroblasts might process anti-microbial function and promote leukocytes transmigration to defend the host against the microorganisms.

Effects of zinc during lactation on the junctional epithelium and inserted gum in rats / Efectos del zinc durante la lactancia sobre el epitelio de unión y la encía insertada en ratas

The purpose of this study was to evaluate the effects of zinc, during lactation, on the junctional epithelium and inserted gum of the first upper molar of rats. The study used one-day old male rats, divided into two groups: those whose mother had been treated with 300 mg zinc chloride (ZnCl2) in the drinker water (treated group), and those whose mothers did not receive ZnCl2 (control group). After 21 days, the rat pups were sacrificed. Using karyometrical techniques, the greater (D) and smaller (d) nuclear diameters of the different layers of the junctional and inserted gum epithelia were determined, and the mean geometric diameter, D/d ratio, perimeter, area, volume, volume/area ratio, eccentricity, shape coefficient, and the contour index were estimated. The 100-point Merz grid was used with the purpose of evaluating the citoplasmatic and celular volume, the nucleus/citoplasm relationship, number density, outer surface/basal layer ratio, the thickness of epithelial layers, and the surface density. The results were submitted to statistical analysis using the Wilcoxon-Mann-Whitney test. The nuclei of the studied structures were significantly smaller, and the stereological results demonstrated that there were smaller cells, hence meaning a greater number of cells per mm3 of tissue, in the treated group. Zinc caused changes on the studied epitheliums, according to morphometric and stereological evaluations.

El objetivo de este estudio fue evaluar los efectos del zinc durante la lactancia, sobre el epitelio de unión y la encía insertada del primer molar superior de ratas. Fueron utilizadas ratas macho de un día de edad, divididas en dos grupos: aquellas cuyas madres habían sido tratadas con 300 mg de cloruro de zinc (ZnCl2) con agua del bebedero (grupo tratado) y aquellas cuyas madres no recibieron ZnCl2 (grupo control). Las crías fueron sacrificadas después de 21 días. Utilizando técnicas cariométricas fueron medidos los diámetros mayor (D) y menor (d) de los núcleos de las células de los diferentes estratos del epitelio de unión y de la encía insertada, estimándose el diámetro geométrico medio, la relación D/d, perímetro, área, volumen, relación volumen/área, excentricidad, coeficiente de forma e índice de contorno. Fue usada la rejilla de Merz, de 100 puntos, con la finalidad de evaluar el volumen celular y citoplasmático, la relación núcleo/citoplasma, densidad numérica, relación superficie externa/superficie basal, espesor de las capas epiteliales y densidad de superficie. Los resultados fueron sometidos a análisis estadístico mediante el test de Wilcoxon-Mann-Whitney. En el grupo tratado los núcleos celulares de las estructuras estudiadas fueron significativamente menores y los resultados estereológicos demostraron que las células eran menores, por lo tanto, con mayor número por mm3 de tejido. De acuerdo a los resultados morfométricos y estereológicos, el zinc provocó cambios en los epitelios estudiados.

Effects of implant collar design on marginal bone and soft tissue / 대한치과보철학회지

PURPOSE: The purpose of this study was to investigate the effects of implant collar design on marginal bone change and soft tissue response by an animal test MATERIALS AND METHODS: Two types of Implant (Neobiotech Co. Seoul, Korea) that only differs in collar design were planted on two healthy Beagle dogs. The implants were divided into two groups, the first group with a beveled collar (Bevel Group) and the second group with "S" shaped collar (Bioseal group). Standardized intraoral radiographs were used to investigate the mesio-distal change of the marginal bone. Histological analysis was done to evaluate the bucco-lingual marginal bone resorption and the soft tissue response adjacent to the implant. Mann-Whitney test was done to compare the mesio-distal marginal bone change at equivalent time for taking the radiographs and the tissue measurements between the groups. RESULTS: Radiographic and histological analysis showed that there was no difference in marginal bone change between the two groups (P>.05). Histological analysis showed Bioseal group had more rigid connective tissue attachment than the Bevel group. There was no difference in biological width (P>.05). Bevel group showed significantly longer junctional epithelium attachment and Bioseal group showed longer connective tissue attachment (P<.05). CONCLUSION: For three months there were no differences in marginal bone change between the Bevel group and the Bioseal group. As for the soft tissue adjacent to the implant, Bioseal group showed longer connective tissue attachment while showing shorter junctional epithelium attachment. There were no differences in biologic width.

Efectos del cloruro de cobalto sobre el epitelio de unión y epitelio reducido del esmalte del primer molar superior de rata / Effects of cobalt chloride in junctional epithelium and reduced enamel epithelium of the rat maxillary first molar

El cobalto es uno de los principales componentes de las aleaciones metálicas fundidas, usadas frecuentemente en odontología. El metal es el constituyente de 45 a 70 por ciento de los trabajos protéticos. En virtud de la existencia de evidencias que elementos metálicos pueden causar toxicidad sistémica y local, este trabajo tuvo como objetivo evaluar los efectos del cobalto sobre el epitelio de unión y el epitelio del esmalte del primer molar superior de rata, durante la lactancia. Con esa finalidad fueron usadas ratas con 1 día de vida postnatal, cuyas madres recibieron 300 mg de cloruro de cobalto por litro de agua destilada en el bebedero, durante a la lactancia. Al cabo de 21 días, las crías fueron sacrificadas con sobredosis anestésica. Las cabezas fueron separadas, fijadas en "alfac", descalcificadas e incluidas en parafina. Fueron utilizados cortes frontales seriados teñidos con hematoxilina y eosina. Fueron estimados los siguientes parámetros nucleares: diámetros mayor, menor y geométrico medio, relación entre diámetros, perímetro, área, volumen, relación entre volumen y área, excentricidad, coeficiente de forma e índice de contorno. Mediante métodos estereológicos fueron evaluados: relación núcleo/citoplasma, volumen celular, densidad numérica celular, relación superficie externa/camada basal, espesor de las camadas epiteliales y densidad de superficie. Todos los datos obtenidos fueron sometidos a análisis estadístico mediante la prueba no-paramétrica de Wilcoxon-Mann-Whitney. Los núcleos de los tejidos estudiados mostraron valores menores para diámetros, perímetro, área, volumen y relación volumen/área. Estereológicamente, fue posible observar en el epitelio de unión y en el epitelio reducido del esmalte, células menores con citoplasma más escaso, lo que se refleja en mayor número de células por mm3 de tejido. En este estudio, el cobalto ocasionó un cuadro de atrofia epitelial, sugiriendo una acción directa sobre los epitelios de unión y del esmalte.

Cobalt is one of the main components of cast metal alloys broadly used in dentistry. It is the constituent of 45 to 70 percent of numerous prosthetic works. There are evidences that metal elements cause systemic and local toxicity. The purpose of the present study was to evaluate the effects of cobalt on the junctional epithelium and reduced enamel epithelium of the first superior molar in rats, during lactation. To do this, 1-day old rats were used, whose mothers received 300mg of cobalt chloride per liter of distilled water in the drinker, during lactation. After 21 days, the rat pups were killed with an anesthetic overdose. The heads were separated, fixed in "alfac", decalcified and embedded in paraffin. Frontal sections stained with hematoxylin and eosin were employed. Karyometric methods allowed to estimate the following parameters: biggest, smallest and mean diameters, D/d ratio, perimeter, area, volume, volume/area ratio, eccentricity, form coefficient and contour index. Stereologic methods allow to evaluate: cytoplasm/nucleus ratio, cell and cytoplasm volume, cell number density, external surface/basal membrane ratio, thickness of the epithelial layers and surface density. All the collected data were subjected to statistic analysis by the non-parametric Wilcoxon-Mann-Whitney test. The nuclei of the studied tissues showed smaller values after karyometry for: diameters; perimeter, area, volume and volume/area ratio. Stereologically, it was observed, in the junctional epithelium and in the reduced enamel epithelium, smaller cells with scarce cytoplasm, reflected in the greater number of cells per mm3 of tissue. In this study, cobalt caused epithelial atrophy, indicating a direct action on the junctional and enamel epithelium.

Ultrastructural changes of the junctional epithlium after dental preparation / 实用口腔医学杂志

Objective:To study the ultrastructural changes of the junctional epithelial after dental preparation. Methods: Junctional epithelium (JE) of left premaxillary teeth was detached by diamond bur in 8 rabbits (group 1) and by surgical knife in another 8 rabbits (group 2). JE of the right teeth without treatment was used as the controls. 4 animals in each group were sacrificed 1 and 2 weeks after operation respectively.Samples were prepared for transmission electron microscope observation.Results: In the two tested groups reattachment was observed 2 weeks after operation,degeneration of mitochondria in JE cells and destruction of basement membrane were observed. No significant difference was observed between group 1 and group 2.Conclusion: Dental preparation may result in ultrastructural changes of JE cells.

The Effects of calcium sulfate on healing of 1-wall intrabony defects in dogs / 대한치주과학회지

The main goal of periodontal therapy is the regeneration of periodontal tissue which has been lost due to destructive periodontal diseases. Although conventional forms of periodontal therapy show sound clinical results, the healing results in long junctional epithelium. There have been numerous materials and surgical techniques developed for new attachment and bone regeneration. Bone grafts can be catagorized into; autografts, allografts, xenografts and bone substitutes. Synthetic bone substitute materials include hydroxyapatite, tricalcium phosphate, calcium carbonate, and Plaster of Paris. Calcium sulfate has found its use in dental practice for the last 30 years. Recent animal studies suggest that periodontal regeneration in 3 wall intrabony defect may be enhanced by the presence of calcium sulfate. And it is well known that 2 wall & 1 wall defect have less osteogenic potential, So we need to study the effect of calcium sulfate in 1 wall intrabony defect in dogs. The present study evaluates the effects of calcium sulfate on the epithelial migration, alveolar bone regeneration and cementum formation in intrabony defects of dogs. Four millimeter-deep one-wall intrabony defects were surgically created in the mesial aspect of anterior teeth and mesial & distal aspects of premolars. The test group received calcium sulfate grafts with a flap procedure. The control underwent flap procedure only. Histologic analysis following 8 weeks of healing revealed the following results: 1. The lengths of junctional epithelium were; 2.52mm in the control, and 1.89mm in the test group. There was no statistical significance between the two groups. 2. Alveolar bone formation were; 0.61mm in the control, and 1.88mm in the test group. There was a statistically significant difference between the two groups (p<0.05). 3. Cementum formations were; 1.1mm in the control, and 2.46mm in the test group. There was a statistically significant difference between the two groups (p<0.05). 4. The length of CT adhesion were; 0.97mm in the control, and 0.17mm in the test group. There was no statistically significant differences between the two groups These results suggest that the use of calcium sulfate in intrabony defects has little effect on junctional epithelium migration, but has significant effects on new bone and new cementum formations.

Periodontal Wound Healing of the Experimental Subepithelial Connective Tissue Graft in Dogs / 대한치주과학회지

Several methods have been used for regeneration of tissue lost by periodontal disease. Subepithelial connective tissue graft technique, one of the technniques of mucogingival surgery, is used for the regeneration in esthetic problems such as recession, and denuded root coverage. This study is performed to evaluate the healing process and the regeneration and reattachment of periodontal tissue, including the reconstruction of junctional epithelium, and connective tissue. Alveolar defects in five adult dogs were treated with periodontal surgery and were attained by removing the marginal alveolar bone by 4x3mm from CEJ in the labial side of incisors, and root surfaces were planed. The experimental sites were divided into two groups as follows. 1. root planing alone(control group) 2. with connective tissue graft(Experimental Group) In the two groups flaps were positioned and sutured tightly, the healing processes were observed and were histologically compared with each other after 2days, 4days, 1week, 2weeks, 4weeks. The results were obtained as follows : 1. In the two groups blood clots were observed as early as 2 and 4 days, and were resorbed at 1 week. 2. In the two groups moderate inflammation was observed as early as 2 and 4 days, decreased at 1 and 2 weeks, and disappeared at 4 weeks. 3. Junctional Epithelium migration was more significant in the control group, and was restrained by graft materials in the experimental group. 4. Features of connective tissue fiber attachment partially showed the parallel pattern in the two groups from 2 weeks, and entirely from 4weeks. 5. Anastomosis, between graft and connective tissue, appeared from 4 days in the experimental group and the border between them was not discriminated at 4weeks.

The Effect of Calcium Sulfate on the Periodontal Healing of 2-Wall Intrabony Defects in Dogs / 대한치주과학회지

The purpose of this study is to evaluate the regenerative potential of calcium sulfate in the treatment of 2-wall intrabony defects as compared to the flap procedure alone. Periodontal healing of surgically created 2-wall intrabony defects grafted with calcium sulfate were evaluated in dogs. Experimental 2-wall intrabony defects of 4x4x4mm were surgically created in the upper anterior edentulous areas between the canines. The test sites include four 2-wall intrabony defects in 4 dogs treated with a calcium sulfate graft. Another four 2-wall intrabony defects in 4 dogs were treated with flap surgery alone as the control sites. Healing was evaluated after 8 weeks. Apical extention of junctional epithelium(JE) was 2.29mm in the control group and 0.50mm in the test group. The length of connective tissue adhesion(CTA) was 0.53mm in the control group and 1.16mm in the test group. The length of new cementum(NC) was 1.17mm for the control group and 2.55mm for the test group. The length of new bone(NB) was 1.02mm in the control group and 2.27mm for the test group. The test group showed statistically significant differences from the control group in junctional epithelium extension, new cementum and new bone formations (p<0.05). Within the limitations of the present study, the results suggest that calcium sulfate may be a safe and cost-effective bone graft material for the treatment of intrabony periodontal defects.

A Comparative Study of Initial Healing Process in White Rats after Gingivectomy using CO2 Laser of different watts / 대한치주과학회지

The use of laser in the treatment of soft tissue minimizes hemorrhage, provides better view of the operating field, and thereby minimizes operating time. Also, there will be far less post-operative swelling, pain and scar formation, and sterilizing effect are shown in some portions of the wound site. All these advantages of laser therapy contribute to its widespread use in the field of medicine and dentistry. Regarding such facts, we used CO2 laser of different watts in gingivectomy for white rats to compare initial healing process. For the control group, the least amount of output in performing gingivectomy(4watts) was offered, and for the experimental group, 6watts was given. Animals were sacrificed on the second, third days, 1 weeks, 2 weeks, and 3 weeks after operation, and their specimens were histologically analyzed. The following results were obtained: 1. Blood clot of small size was observed in both the control and experimental groups after two days, and no more thereafter. 2. In both the control and experimental groups, the inflammation zone size was the greatest after two days, and it decreased gradually to become almost invisble by the second week. The experimental group showed larger size of inflammation zone during second and third days; however, there was no difference after one week. 3. Granulation tissue in both the control and experimental groups showed gradual maturation with time, and by the second week, it was almost replaced by normal connective tissue. By the third week, complete healing pattern was observed. The experimental group showed larger granulation tissue than the control group until the third day, but there was no significant difference after one week. 4. In both the control and experimental groups, gingival epithelialization began on the second day. After one week, regeneration of rete peg and partial formation of junctional epithelium were observed; by the second week, keratinization of oral sulcular epithelium began, and it was completed by the third week.

The effects of horizontal orthodontic tooth movement on the periodontally diseased tissue in dogs / 대한치과교정학회지

Most adults, unlike growing children, have some periodontal problems which can influence the outcome of the orthodontic treatment. In cases where periodontal disease progression resulted in marked reduction of periodontium, orthodontic treatment could result in the worsening of the periodontal conditions, and therefore orthodontic treatment planning in such adult patients requires special considerations for the periodontal problems. This study investigates the effects of horizontal orthodontic tooth movement on the changes in the mesial, distal and furcation areas of the disease affected periodontium of adult dogs with advanced bone loss. Six adult dogs with healthy periodontium were selected, and mandibular 2nd premolars were extracted. In the mandibular 3rd premolars, angular bony defects in the mesial and distal sides, and horizontal bony defects in the furcation areas were created. Those that received the flap operation and plaque control were designated as the control, those that had horizontal tooth movement without plaque control after the flap operation as Experimental group I, those that had horizontal tooth movement under plaque control without the flap operation as Experimental group II, and those that had horizontal tooth movement under plaque control after the flap operation as Experimental group III. The control group was sacrificed 2 months postoperatively, and the experimental groups were sacrificed 5 months after the initiation of tooth movement. Specimens were histologically analyzed under light microscope. The results were as follows; 1. After the horizontal tooth movements, Experimental group I and II showed angular bony defects in the mesial sides of the roots and the distal side of the furcation areas, which correspond to the pressure sides. 2. After the horizontal tooth movements, Experimental group I and II showed decreased level of alveolar bone crest in the distal sides of the roots, which correspond to the tension sides. 3. Long junctional epithelium in the control group has not been replaced by periodontal connective tissue after the horizontal tooth movements. 4. Limited formation of new bone was observed in the angular bony defects in the mesial and distal aspects of the roots in the control group. 5. Inflammatory cell infiltration in the connective tissue was most severe in the Experimental group I, followed by Experimental group II, III, and the control group in that order. These results seem to indicate that plaque control was the most influencing factor in the alteration of the periodontal tissue after the horizontal tooth movements in the periodontal tissue with alveolar bone defects.