Development, Establishment, and Validation of a Model for the Mineralization of Periodontium Remodelling Cells: Cementoblasts.

Chronic kidney disease (CKD) patients undergoing dialysis are at high risk of bone fractures. CKD-induced mineral and bone disorder is extended to periodontal disease due to changes in the ionic composition of saliva in CKD patients, dysregulating mineralization, hindering regeneration and thereby promoting the progression of dental complications. Despite the importance of cementum for overall oral health, the mechanisms that regulate its development and regeneration are not well comprehended, and a lack of sufficient in vitro experimental models has hindered research progress. In this study, the impact of experimental conditions on the calcification of cementoblasts was systematically investigated, aimed at establishing a standardized and validated model for the calcification of cementoblasts. The effects of phosphate, calcium, ascorbic acid, ß-glycerolphosphate, dexamethasone, and fetal calf serum on the calcification process of cementoblasts were analyzed over a wide range of concentrations and time points by investigating calcium content, cell viability, gene expression and kinase activity. Cementoblasts calcified in a concentration- and time-dependent manner with higher concentrations of supplements cause a higher degree of calcification but decreased cell viability. Phosphate and calcium have a significantly stronger effect on cementoblast calcification processes compared to osteogenic supplements: ascorbic acid, ß-glycerolphosphate, and dexamethasone induce calcification over a wide range of osteogenic signalling pathways, with osteopontin being a central target of gene regulation. Conversely, treatment with ascorbic acid, ß-glycerolphosphate, and dexamethasone leads to activating only selected pathways, especially promoting bone sialoprotein expression. The developed and validated cementoblast calcification protocol, incubating up to 60% confluent cementoblasts with 1.9 mmol L-1 of phosphate supplementation for a reasonable, multi-pathway calcification induction and 10 mmol L-1 ß-glycerolphosphate, 75 µmol L-1 ascorbic acid and 10 nmol L-1 dexamethasone for a reasonable osteogenic differentiation-based calcification induction, provides standard in vitro experimental models for better understanding cementoblast function and regeneration.

Efficacy and safety of P11-4 for the treatment of periodontal defects in dogs.

OBJECTIVES: This study's aim was to investigate the safety and performance of a self-assembling peptide matrix (SAPM) P11-4 for the treatment of periodontal disease in a controlled pre-clinical study. MATERIALS AND METHODS: Acute buccal bony dehiscence defects (LxW: 5 × 3 mm) were surgically created on the distal root of four teeth on one mandible side of 7 beagle dogs followed by another identical surgery 8 weeks later on the contralateral side. SAPM P11-4 (with and without root conditioning with 24% EDTA (T1, T2)), Emdogain® (C) and a sham intervention (S) were randomly applied on the four defects at each time point. Four weeks after the second surgery and treatment, the animals were sacrificed, the mandibles measured by micro-computed tomography (µ-CT) and sections of the tissue were stained and evaluated histologically. RESULTS: Clinically and histologically, no safety concerns or pathological issues due to the treatments were observed in any of the study groups at any time point. All groups showed overall similar results after 4 and 12 weeks of healing regarding new cementum, functionality of newly formed periodontal ligament and recovery of height and volume of the new alveolar bone and mineral density. CONCLUSION: A controlled clinical study in humans should be performed in a next step as no adverse effects or safety issues, which might affect clinical usage of the product, were observed. CLINICAL RELEVANCE: The synthetic SAPM P11-4 may offer an alternative to the animal-derived product Emdogain® in the future.

Dental cementum examination with SEM/EDX for unraveling details along the lifeline to assist identification of a female individual.

In this work, we present a novel approach of forensic examination of the dental cementum by Scanning Electron Microscopy (SEM) as an imaging tool, complemented by semiquantitative elemental analysis of the incremental lines with an Energy Dispersion X-ray analyzer. Tooth samples were collected from female donors who are known to have given single or multiple childbirths. This research is aimed to prove the following hypothesis - if pregnancy (possibly followed by breastfeeding) affects the calcium and phosphorus levels in female metabolism, then increased levels of Ca and P should be detected in the dental cementum region corresponding to the age of the pregnancy. For this purpose, EDX spectra were taken from sampling points, evenly aligned along the direction of the cementum growth. The most intensive X-ray peaks were analyzed employing qualitative and semiquantitative elemental analysis. Results showed that the changes in the peak values of Ca and P levels in the cementum along the lifeline could be related to the biological age of pregnancy, possibly followed by breastfeeding. The observed variations in other elements could also suggest some other physiological conditions or nutrition patterns. These preliminary results could trigger more profound and extensive research involving more sophisticated techniques for elemental analysis, such as PIXE (Proton Induced X-ray Emission). Correlation of the element levels to certain conditions may bring a new paradigm of the unique forensic value of the dental cementum, on which the life story of a person is literally "written in a stone". This method could serve pure forensic purposes and archeological examinations of human findings in support of historical facts.

Effect of photobiomodulation on external root resorption during orthodontic tooth movement - a randomized controlled trial.

OBJECTIVE: The objective of the study was to compare external root resorption during orthodontic tooth movement between patients who were subjected to photobiomodulation and those who were not. MATERIALS AND METHODS: The study was designed as a split-mouth, single-blind randomized controlled trial conducted on twenty-two orthodontic patients requiring extraction of maxillary first premolars (age group 13-30years). A buccal force was applied to all the premolars using a 0.019″×0.025″ beta-titanium cantilever spring for 28days. The test group premolars were irradiated by indium-gallium-arsenide (InGaAs) diode laser (980nm, 100mW) 0, 3, 7, 11, 15 and 28days after force application. Each premolar was irradiated at ten locations on the buccal and palatal gingiva resulting in a total energy of 10J delivered per tooth. After 28days, the root surface of all the premolars were studied for the number and volume of resorption craters using three-dimensional optical profilometry by a blinded assessor. One-sample Kolmogorov-Smirnov test was used to ascertain normality and Mann-Whitney U test was applied for data analysis. RESULTS: All twenty-two patients enrolled completed the study. The root surface analysis showed formation of 32.78% less number of resorption craters (P<0.05) and 39.49% less volume of resorption craters (P<0.05) in the test group premolars relative to the control group premolars. CONCLUSIONS: The results of the present study, therefore, suggest that photobiomodulation reduces external root resorption during orthodontic tooth movement. TRIAL REGISTRATION/REGISTRATION NUMBER: CTRI/2018/04/013520.

The effect of hydrochloric acid (HCl) on permanent molars: A scanning electron microscope (SEM) and energy dispersive X-ray spectroscopy (EDS) study.

It is well established that acid disposal is a potentially effective method used by criminal syndicates to hinder the identification of victims. This study documents the effects of continuous immersion in hydrochloric acid (HCl, 37%) on molars using macroscopic analysis, scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDS). The goal of this study is to aid in distinguishing visually unrecognizable fragments of dental remains when drastic changes in morphology have occurred as a result of acid exposure. Macroscopic, SEM, and EDS analysis were conducted on seven maxillary molars before and after HCl treatment. Molars reduced in weight relative to the length of time immersed in HCl and the dissolution time was over 40 hours longer than reported in previous studies, at just over 66 hours. SEM and EDS analysis showed acid-treated teeth exhibited morphological patterns such as cracking and layering visible at high magnification. Calcium/phosphorous ratios fell within the expected range of 1.6-2.5, indicating that HCl-treated teeth are still identifiable as osseous or dental tissue even when not visually identifiable as teeth. This is the first study to present SEM images of molar cementum before and after immersion in HCl and to present EDS results. This information can assist researchers and investigators in determining the presence of dental tissue in a forensic context associated with acid disposal.

Irisin promotes cementoblast differentiation via p38 MAPK pathway.

OBJECTIVE: Irisin is a newly identified exercise-induced myokine which can affect glucose metabolism and cortical bone mass and strength. However, the influence of irisin on cementoblasts remains largely unknown. MATERIAL AND METHODS: An immortalized mouse cementoblast cell line OCCM-30 was used in this study. Cementoblast differentiation markers and PGC-1α in cells cultured with mineral induction medium were evaluated by qRT-PCR. Cementoblast mineralization was evaluated by alizarin red staining. Differentiation markers and the activity of p38 MAPK pathway under irisin stimulation were assessed by qRT-PCR or Western blot analysis. p38 MAPK pathway inhibitor SB203580 or p38 siRNA was used to further identify the regulatory mechanism. Cell proliferation treated with irisin was examined by CCK-8 method. RESULTS: The expression of Runx2, osterix, ALP, and PGC-1α was up-regulated consistently under mineral induction. The formation of mineralized nodules was increased by irisin. Runx2, osterix, ALP, and osteocalcin were obviously up-regulated under irisin stimulation as well as the activity of p38 MAPK pathway. When pretreated with SB203580 or p38 siRNA before irisin stimulation, the irisin-induced differentiation was distinctly suppressed. OCCM-30 cell proliferation was enhanced when treated with high-dose irisin for long time. CONCLUSION: Irisin can promote the differentiation of cementoblasts via p38 MAPK pathway.

Calcium sulfate-based bioactive cement for periodontal regeneration: An In Vitro study.

BACKGROUND AND OBJECTIVE: Various types of osteoconductive graft materials are used for the management of alveolar bone defects arising out of periodontal disease. Inorganic, self-setting, bioactive bone cements are suggested to be most appropriate because they can conformally fill the bone defect and resorb progressively along with the regeneration of the host site. A new calcium sulfate-based bioactive bone cement (BioCaS) is developed, having simplicity and effectiveness for bone grafting applications. The response of primary human periodontal ligament (hPDL) cells to this material is investigated through in vitro cell culture model so as to qualify it for the repair of periodontal infrabony defects. METHOD: The BioCaS was designed as powder-liquid combination with in-house synthesized high purity calcium sulfate hemihydrate incorporating hydrogen orthophosphate ions. hPDL cells were isolated, cultured and characterized using optimized primary cell culture techniques. The cytotoxicity and cytocompatibility of the BioCaS samples were evaluated using the hPDL cells, with hydroxyapatite ceramic material as control. Osteogenic differentiation of the hPDL cells in presence of BioCaS was also evaluated using Alizarin red staining, Alizarin red assay, Von Kossa staining and Masson's trichrome staining. RESULTS: The primary cell culture techniques yielded a healthy population of periodontal ligament cells, with fibroblast morphology and characteristic marker expressions. The hPDL cells exhibited good viability, adhesion and spreading to the BioCaS cement in comparison to sintered hydroxyapatite. In addition, the cells differentiated to osteogenic lineage in the presence of the BioCaS cement, without extraneous osteogenic supplements, confirming the inherent bioactivity of the cement. CONCLUSION: The new BioCaS cement is a potential candidate for the repair of periodontal defects.

Physical properties of root cementum: Part 27. Effect of low-level laser therapy on the repair of orthodontically induced inflammatory root resorption: A double-blind, split-mouth, randomized controlled clinical trial.

INTRODUCTION: The purpose of this 2-arm-parallel split-mouth trial was to investigate the effect of low-level laser therapy (LLLT) on the repair of orthodontically induced inflammatory root resorption (OIIRR). METHODS: Twenty patients were included in this study, with 1 side randomly assigned to receive LLLT, and the other side served as a sham. Eligibility criteria included need for bilateral maxillary first premolar extractions as part of fixed appliance treatment. OIIRR was generated by applying 150 g of buccal tipping force on the maxillary first premolars for 4 weeks. After the active force was removed, the teeth were retained for 6 weeks. LLLT commenced with weekly laser applications using a continuous beam 660-nm, 75-mW aluminum-gallium-indium-phosphorus laser with 1/e2 spot size of 0.260 cm2, power density of 0.245 W/cm2, and fluence of 3.6 J/cm2. Contact application was used at 8 points buccally and palatally above the mucosa over each tooth root for 15 seconds with a total treatment time of 2 minutes. After 6 weeks, the maxillary first premolars were extracted and scanned with microcomputed tomography for primary outcome OIIRR calculations. Subgroup analysis included assessment per root surface, per vertical third, and sites of heaviest compressive forces (buccal-cervical and palato-apical). Randomization was generated using www.randomization.com, and allocation was concealed in sequentially numbered, opaque, sealed envelopes. Blinding was used for treatment and outcome assessments. Two-tailed paired t tests were used to determine whether there were any statistically significant differences in total crater volumes of the laser vs the sham treated teeth. RESULTS: Total crater volumes were 0.746 mm3 for the laser treated teeth and 0.779 mm3 for the sham. There was a mean difference of 0.033 ± 0.39 mm3 (95% CI, -0.21 to 0.148 mm3) greater resorption crater volume in the sham group compared with the laser group; this was not statistically significant (P = 0.705). No harm was observed. CONCLUSIONS: No significant difference was found between LLLT and sham control groups in OIIRR repair.

Immunohistopathologic evaluation of Drynaria fortunei rhizome extract in the management of Class II furcation defects in a canine model.

BACKGROUND: Management of furcation defects is still a challenging subject in periodontal therapy. Drynaria fortunei (Df) is a common type of traditional Chinese herb in the area of orthopedics and traumatology. In vitro and tissue engineering studies have shown that Df induces osteoblastic proliferation and promotes the differentiation of human periodontal ligament cells. This study investigated the management of Class II furcation defects in dogs using guided tissue regeneration (GTR) and Df granules mixed with ß-tricalcium phosphate (ß- TCP) alloplast. METHODS: Sixteen Class II critical-sized furcation defects were surgically created in four mongrel dogs: Eight defects were treated with GTR and Df granules mixed with (ß-TCP) alloplast served as the experimental group, while the other eight were managed with GTR and alloplast, served as control. Dogs were sacrificed at 4 and 8 weeks and the premolars were processed for the evaluation of treatment outcome including; osteoblastic count (OC), cementum layer thickness (CLT), percentage of collagen in bone matrix (CBM), and alkaline phosphatase (ALP) immunoreaction. RESULTS: Experimental group treated with Df showed a significant increase (P < 0.001) in the values of OC, CLT, CBM, and ALP immunoreactivity when compared with control at 4 and 8 weeks after treatment. CONCLUSION: Drynaria fortunei demonstrated increased regeneration and bone formation when used in the treatment of furcation defects in a canine model.

Relationship between root resorption and individual variation in the calcium/phosphorous ratio of cementum.

INTRODUCTION: The purpose of this study was to investigate whether individual variation in the hardness and chemical composition of the cementum in the root apex affects the degree of root resorption. METHODS: In a previous study, we evaluated the Vickers hardness scale of 50 extracted teeth. For this study, we classified the 50 extracted teeth into soft, moderate, and hard groups according to the Vickers hardness scale. Then, we randomly selected 7 teeth from each group and measured the resorbed areas of the apical cementum in vitro using human osteoclast precursor cells. We also investigated the calcium/phosphorous (Ca/P) and magnesium/calcium ratios of these 21 extracted teeth using energy-dispersive x-ray microanalysis studies to determine the chemical composition of the cementum in the root apex. RESULTS: In the pit formation assay, the resorbed area in the soft group showed a greater extent than it did in the moderate and hard groups (P < 0.01). A correlation was noted between the Vickers hardness and the resorbed area of the cementum in the root apex (r = -0.714; P < 0.01). The Ca/P ratios in the soft and moderate groups were lower than the ratio in the hard group (P < 0.01 and P < 0.05, respectively). A correlation was noted between the Vickers hardness and the Ca/P ratio of the cementum in the root apex (r = 0.741; P < 0.01). CONCLUSIONS: These results suggest that the hardness and Ca/P ratio of the cementum may be involved in root resorption caused by orthodontic forces.

Biostimulation with diode laser positively regulates cementoblast functions, in vitro.

The aim of this study was to evaluate the effects of diode laser biostimulation on cementoblasts (OCCM.30). A total of 40 root plates were obtained from healthy third molar teeth and assigned to the following two groups: (1) control group and (2) laser-treated group. Root plates were placed into the cell culture inserts, and OCCM.30 cells were seeded onto root plates. Cells were irradiated with a low level of diode laser (power: 0.3 W in continuous wave, 60 s/cm2). Proliferation and mineralized tissue-associated gene's and BMP's messenger RNA (mRNA) expressions of cementoblasts were evaluated. Total RNAs were isolated on day 3 and integrin-binding sialoprotein (Ibsp), bone gamma-carboxyglutamate protein (Bglap), Type I collagen (Col1a1), osteoblastic transcription factor, runt-related transcription factor (Runx2), and Bone Morphogenetic Protein (BMP)-2, 3, 4, 6, and 7 mRNA expressions were determined using quantitative RT-PCR. von Kossa staining was performed to evaluate biomineralization of OCCM.30 cells. In the proliferation experiment, while there was no significant difference until 96 h, laser irradiation retarded the decrease in cell proliferation trend after 96 h compared to the untreated control group. Statistically significant increase in Ibsp, Bglap, and BMP-2,3,6,7 mRNA expressions were noted in the laser groups when compared to the untreated control group (p < 0.05). Laser irradiation induced mineralized nodule formation of cementoblasts. The results of this study reveal that the biostimulation setting of diode laser modulates the behavior of cementoblasts inducing mineralized tissue-associated gene's mRNA expressions and mineralization. Therefore, biostimulation can be used during regenerative periodontal therapies to trigger cells with periodontal attachment apparatus.

Mineralising and antibacterial effects of modified calcium phosphate treatment on human root cementum.

BACKGROUND: Aging population will lead to the increase of incidence of root caries globally. The clinical management of root caries is challenging due to the difficulty in moisture isolation. The root caries is caused by the release of organic acids from cariogenic bacteria which results in the dissolution of cementum and dentin of the root. The purpose of this study is to study the efficacy of modified saturated calcium phosphate solution (CaP) supplement with zinc (Zn(2+)) and/or fluoride (F(-)) in providing root cementum surfaces less susceptible to acid dissolution and bacterial colonization. METHODS: Human root cementum sections from extracted premolars were treated with three modified calcium phosphate solutions (M/A-CaPs) respectively: (A) CaP-F/Zn, supplemented with F(-) and Zn(2+); (B) CaP-F, supplemented with F(-) only; (C) CaP-Zn, supplemented with Zn(2+) only. The surface characteristics of treated cementum sections were investigated using scanning electron microscopy (SEM) and fourier transform infrared spectroscopy (FT-IR). Following the acid attack and Streptococcus mutans challenge, M/A-CaPs treated cementum surfaces were analysed using inductive coupled plasma (ICP) and SEM respectively. RESULTS: Compared with the control group, M/A-CaPs treated cementum presented significant improvements in resistance to acid dissolution and bacterial colonization. Among M/A-CaPs, the CaP-F/Zn treated cementum surfaces released the lowest amount of Ca(2+) ions (2.11 ± 0.51 ppm) upon acid challenge (n = 3, p < 0.01) and also presented the most significant inhibiting effect against the colonization of S. mutans (n = 180, p < 0.05). CONCLUSIONS: Saturated calcium phosphate solution CaP supplemented with both F(-) and Zn(2+) could be applied as an effective coating material providing acid resistance and antibacterial property on cementum surfaces. The modified calcium phosphate-based solution could be a new treatment strategy to prevent the development of root caries and arrest the further progression of root caries.

Cementochronology and sex: A reappraisal of sex-associated differences in survival in past French societies.

The objective of the present study is to test our general knowledge of sex-specific survival differences in past northern France societies by implementing the tooth cementum annulations method of age estimation (i.e., cementochronology) to bio-archaeological series. 1255 individual estimated ages at death covering a millennium from the 3rd c. AD to the 15th c. AD matched different patterns of sex mortality from the late Antiquity to the Late Middle Age. Female survival curves are consistently inferior to those of their male counterparts. Maternal mortality is clearly visible in survival curves between 20 and 50 years of age in individual sites and pooled samples. Variations of sex mortalities also affected sites with peculiar recruitment, such as religious communities, pathological samples, leprosaria, and migrants. Whisker plots of median ages at death variations confirmed in both sex that populations within the Early Middle Ages were better off compared to Late Antiquity and Late Medieval Ages when group inequalities prevailed. Due to its sensitivity and applicability to small samples, cementochronology should be extended to other series.

Consideraciones metodológicas sobre el estudio de la adaptación del cemento sellador a las paredes del conducto: comparación entre el microscopio electrónico de barrido y el confocal láser / Methodological considerations on the study of endodontic sealer adaptation to root canal walls: comparison between scanning electronic and laser confocal microscopes

Objetivo: Evaluar la correlación entre las observaciones con el microscopio electrónico de barrido (MEB) y con el microscopio confocal láser (MCL) para el estudio de la adaptación a las paredes del conducto de TopSeal. Material y métodos: Diez dientes unirradiculares fueron instrumentados con el sistema de limas rotatorias MTwo hasta el 35/.04. Se irrigó con 5 ml de NaOCl (5,25%) entre limas y con 5 ml de EDTA (17%) como irrigante final. La obturación se realizó con condensación lateral utilizando conos de gutapercha y cemento TopSeal marcado con un fluorocromo. Se obtuvieron secciones transversales a 3, 5 y 8 milímetros del ápice, que fueron analizadas con MCL y luego metalizadas y observadas con MEB. Se estableció el grado de adaptación al perímetro del conducto en cada sección con ambos procedimientos. En el análisis estadístico se utilizaron el coeficiente de correlación de Pearson para la evaluación de la interfase cemento-dentina entre ambos métodos y el ANOVA para el análisis de la penetración intratubular del cemento evaluada con MCL. Resultados: El coeficiente de correlación de Pearson fue de 0,75 (p = 0,001) para los porcentajes medios de adaptación. Medias: MCL 71,40 y MEB 83,67. Los coeficientes para los tercios apical, medio y coronal fueron 0,940, 0,772 y 0,877 respectivamente, en todos los casos significativos. Respecto a la penetración del cemento sellador, no se encontraron diferencias significativas en los niveles estudiados. Conclusiones: Ambas técnicas microscópicas son complementarias, mientras que el MEB proporciona mayor seguridad en el estudio de la interfase, el MCL permite analizar la penetración intratubular del material

Aim: To compare two different microscopic techniques (SEM and CLSM) in studying filling materials adaptation to canal walls. Material and Methods: Ten sound extracted single-rooted teeth were selected, instrumented and filled with gutta-percha and TopSeal using lateral condensation technique. Following root filling, teeth were sectioned transversally at 3, 5 and 8 mm from the apex and analyzed using Confocal Laser Scanning Microscope and Scanning Electronic Microscope. Percentage of root canal perimeter with sealer penetration to the dentinal tubules and depth of penetration were compared between groups and statistically analyzed using a Pearson correlation analysis (p=0.001) and ANOVA test for intratubular penetration. Pearson’s correlation coefficient for the whole sample was 0.75 (p=0.001). Results: When comparing sealer adaptation to dentinal walls between thirds, apical medium and coronal, mean values were statistically significant: 0.940, 0.772 and 0.877 respectively. For intratubular penetration no significant difference between root canal thirds was found. Conclusions: Both techniques are complementary in the study of endodontic filling materials performance inside root Canals

PTH and Vitamin D Repress DMP1 in Cementoblasts.

A complex feedback mechanism between parathyroid hormone (PTH), 1,25(OH)2D3 (1,25D), and fibroblast growth factor 23 (FGF-23) maintains mineral homeostasis, in part by regulating calcium and phosphate absorption/reabsorption. Previously, we showed that 1,25D regulates mineral homeostasis by repressing dentin matrix protein 1 (DMP1) via the vitamin D receptor pathway. Similar to 1,25D, PTH may modulate DMP1, but the underlying mechanism remains unknown. Immortalized murine cementoblasts (OCCM.30), similar to osteoblasts and known to express DMP1, were treated with PTH (1-34). Real-time quantitative polymerase chain reaction (PCR) and Western blot revealed that PTH decreased DMP1 gene transcription (85%) and protein expression (30%), respectively. PTH mediated the downregulation of DMP1 via the cAMP/protein kinase A (PKA) pathway. Immunohistochemistry confirmed the decreased localization of DMP1 in vivo in cellular cementum and alveolar bone of mice treated with a single dose (50 µg/kg) of PTH (1-34). RNA-seq was employed to further identify patterns of gene expression shared by PTH and 1,25D in regulating DMP1, as well as other factors involved in mineral homeostasis. PTH and 1,25D mutually upregulated 36 genes and mutually downregulated 27 genes by ≥2-fold expression (P ≤ 0.05). Many identified genes were linked with the regulation of bone/tooth homeostasis, cell growth and differentiation, calcium signaling, and DMP1 transcription. Validation of RNA-seq results via PCR array confirmed a similar gene expression pattern in response to PTH and 1,25D treatment. Collectively, these results suggest that PTH and 1,25D share complementary effects in maintaining mineral homeostasis by mutual regulation of genes/proteins associated with calcium and phosphate metabolism while also exerting distinct roles on factors modulating mineral metabolism. Furthermore, PTH may modulate phosphate homeostasis by downregulating DMP1 expression via the cAMP/PKA pathway. Targeting genes/proteins mutually governed by PTH and 1,25D may be a viable approach for designing new therapies for preserving mineralized tissue health.

Mineral density volume gradients in normal and diseased human tissues.

Clinical computed tomography provides a single mineral density (MD) value for heterogeneous calcified tissues containing early and late stage pathologic formations. The novel aspect of this study is that, it extends current quantitative methods of mapping mineral density gradients to three dimensions, discretizes early and late mineralized stages, identifies elemental distribution in discretized volumes, and correlates measured MD with respective calcium (Ca) to phosphorus (P) and Ca to zinc (Zn) elemental ratios. To accomplish this, MD variations identified using polychromatic radiation from a high resolution micro-computed tomography (micro-CT) benchtop unit were correlated with elemental mapping obtained from a microprobe X-ray fluorescence (XRF) using synchrotron monochromatic radiation. Digital segmentation of tomograms from normal and diseased tissues (N=5 per group; 40-60 year old males) contained significant mineral density variations (enamel: 2820-3095 mg/cc, bone: 570-1415 mg/cc, cementum: 1240-1340 mg/cc, dentin: 1480-1590 mg/cc, cementum affected by periodontitis: 1100-1220 mg/cc, hypomineralized carious dentin: 345-1450 mg/cc, hypermineralized carious dentin: 1815-2740 mg/cc, and dental calculus: 1290-1770 mg/cc). A plausible linear correlation between segmented MD volumes and elemental ratios within these volumes was established, and Ca/P ratios for dentin (1.49), hypomineralized dentin (0.32-0.46), cementum (1.51), and bone (1.68) were observed. Furthermore, varying Ca/Zn ratios were distinguished in adapted compared to normal tissues, such as in bone (855-2765) and in cementum (595-990), highlighting Zn as an influential element in prompting observed adaptive properties. Hence, results provide insights on mineral density gradients with elemental concentrations and elemental footprints that in turn could aid in elucidating mechanistic processes for pathologic formations.

Effect of low-level laser therapy on dental root cementum remodeling in rats.

OBJECTIVE: To investigate the amount of the cementum layer formed over the rat's dental root surfaces by daily application of low-level laser therapy (LLLT) for 2 weeks. METHODS: Twelve female Sprague-Dawley (SD) rats were divided into two groups: six rats received daily LLLT (Ga-Al-As, 830 nm), and six rats received no treatment (control). The treatment lasted 2 weeks. In vivo Micro-CT imaging analyzed the root's hard tissue volumetric changes. The cementum thickness was evaluated histologically. RESULTS: Total cementum thicknesses in the LLLT group increased significantly (p = 0.015) compared to the control group. This significant increase in the cementum thickness, verified histologically, was not detectable during in vivo Micro-CT imaging, which showed no significant difference between the groups regarding the root hard tissues volumetric changes over the 2-week evaluation period. CONCLUSION: Two weeks of daily application of LLLT significantly increased rat's dental root cementum thickness as determined histologically. However, in vivo Micro-CT imaging failed to accurately reveal this cementum growth as it was not possible to differentiate dentinal changes.

Topical application of herbal formula for the treatment of ligature-induced periodontitis

PURPOSE: The aim of this study was to investigate the therapeutic effects of a herbal formula, PerioH-035, containing Angelica sinensis, steamed Rehmannia glutinosa, Angelica dahurica, Cimicifuga heracleifolia, and Zanthoxylum piperitum on the periodontal breakdown in a well-established ligature-induced periodontitis model in rats. METHODS: Sprague-Dawley rats were randomly assigned to 1 of 4 groups: NL (non-ligatured), L (ligatured), P1 (ligatured and treated with 1 mg/mL PerioH-035), P100 (ligatured and treated with 100 mg/mL PerioH-035). Periodontitis was induced by placing a ligature around the mandibular first molars. PerioH-035 was topically applied to both sides of the first molar for 2 weeks. The right side of the mandibles was retrieved for micro-computed tomography (CT) and methylene blue staining to analyze alveolar bone loss. The left side of the mandibles was histologically analyzed by TRAP and H&E staining. The MMP-9 mRNA level in gingival tissue was investigated by RT-PCR. RESULTS: Alveolar bone resorption was significantly reduced in the PerioH-035-treated groups. The number of dense multi-nucleated cells found to be TRAP-positive by staining in the ligatured rats was markedly decreased by PerioH-035 application. In addition, periodontal tissue destruction, especially cementum demineralization, was ameliorated in the P1 and P100 groups. Moreover, gingival tissue from the PerioH-035-treated group showed a decrease in the MMP-9 mRNA level, resulting in recovery of collagen degradation. CONCLUSIONS: These results suggest that PerioH-035 has therapeutic effects on periodontitis, and thus, PerioH-035 shows promise as a treatment for periodontitis. GRAPHICAL ABSTRACT:

Histologic comparison between platelet-rich plasma and blood clot in regenerative endodontic treatment: an animal study.

INTRODUCTION: In regenerative endodontic treatment (RET) for immature permanent tooth, better treatment results could be obtained by applying platelet-rich plasma (PRP) as the scaffold rather than the blood clot. The goal of this study was to compare the histologic differences between using PRP and blood clot in RET. METHODS: Three 6-month-old beagles each carrying 9 premolars with double root canals were randomly assigned to the PRP group, blood clot group, or negative control group. All experimental teeth suffered apical periodontitis, and RET was performed. In the blood clot group, bleeding was induced from the periapical tissues to fill the canal space. In the PRP group, autologous PRP was injected into each root canal. The animals were sacrificed 3 months later. Histologic sections were stained with hematoxylin-eosin. Statistical analysis was performed by the Fisher exact test, with the significance set at 0.05. RESULTS: With the ingrowth of cellular cementumlike tissues, the canal wall was thickened, and the apical apex was closed in both the PRP and blood clot groups. Cementocytelike cells were present in the newly formed tissues. Meanwhile, no statistical difference was found in both experimental groups for the average percentage of apical closure, new tissue formation, and pulplike tissue formation. Noticeably, a large number of inflammatory cells were present in some root canals in both groups although the postoperative radiograph revealed the disappearance of periapical radiolucency. CONCLUSIONS: PRP application could be an option for clinical cases in which little or no bleeding were found when irritating the apical tissue during RET.

Acemannan sponges stimulate alveolar bone, cementum and periodontal ligament regeneration in a canine class II furcation defect model.

BACKGROUND AND OBJECTIVE: Periodontal disease is a common infectious disease, found worldwide, causing the destruction of the periodontium. The periodontium is a complex structure composed of both soft and hard tissues, thus an agent applied to regenerate the periodontium must be able to stimulate periodontal ligament, cementum and alveolar bone regeneration. Recent studies demonstrated that acemannan, a polysaccharide extracted from Aloe vera gel, stimulated both soft and hard tissue healing. This study investigated effect of acemannan as a bioactive molecule and scaffold for periodontal tissue regeneration. MATERIAL AND METHODS: Primary human periodontal ligament cells were treated with acemannan in vitro. New DNA synthesis, expression of growth/differentiation factor 5 and runt-related transcription factor 2, expression of vascular endothelial growth factor, bone morphogenetic protein-2 and type I collagen, alkaline phosphatase activity, and mineralized nodule formation were determined using [(3)H]-thymidine incorporation, reverse transcription-polymerase chain reaction, enzyme-linked immunoabsorbent assay, biochemical assay and alizarin red staining, respectively. In our in vivo study, premolar class II furcation defects were made in four mongrel dogs. Acemannan sponges were applied into the defects. Untreated defects were used as a negative control group. The amount of new bone, cementum and periodontal ligament formation were evaluated 30 and 60 d after the operation. RESULTS: Acemannan significantly increased periodontal ligament cell proliferation, upregulation of growth/differentiation factor 5, runt-related transcription factor 2, vascular endothelial growth factor, bone morphogenetic protein 2, type I collagen and alkaline phosphatase activity, and mineral deposition as compared with the untreated control group in vitro. Moreover, acemannan significantly accelerated new alveolar bone, cementum and periodontal ligament formation in class II furcation defects. CONCLUSION: Our data suggest that acemannan could be a candidate biomolecule for periodontal tissue regeneration.