MCL-1 Inhibition Overcomes Anti-apoptotic Adaptation to Targeted Therapies in B-Cell Precursor Acute Lymphoblastic Leukemia.

Multiple targeted therapies are currently explored for pediatric and young adult B-cell precursor acute lymphoblastic leukemia (BCP-ALL) treatment. However, this new armamentarium of therapies faces an old problem: choosing the right treatment for each patient. The lack of predictive biomarkers is particularly worrying for pediatric patients since it impairs the implementation of new treatments in the clinic. In this study, we used the functional assay dynamic BH3 profiling (DBP) to evaluate two new treatments for BCP-ALL that could improve clinical outcome, especially for relapsed patients. We found that the MEK inhibitor trametinib and the multi-target tyrosine kinase inhibitor sunitinib exquisitely increased apoptotic priming in an NRAS-mutant and in a KMT2A-rearranged cell line presenting a high expression of FLT3, respectively. Following these observations, we sought to study potential adaptations to these treatments. Indeed, we identified with DBP anti-apoptotic changes in the BCL-2 family after treatment, particularly involving MCL-1 - a pro-survival strategy previously observed in adult cancers. To overcome this adaptation, we employed the BH3 mimetic S63845, a specific MCL-1 inhibitor, and evaluated its sequential addition to both kinase inhibitors to overcome resistance. We observed that the metronomic combination of both drugs with S63845 was synergistic and showed an increased efficacy compared to single agents. Similar observations were made in BCP-ALL KMT2A-rearranged PDX cells in response to sunitinib, showing an analogous DBP profile to the SEM cell line. These findings demonstrate that rational sequences of targeted agents with BH3 mimetics, now extensively explored in clinical trials, may improve treatment effectiveness by overcoming anti-apoptotic adaptations in BCP-ALL.

Tight regulation of FOXO1 is essential for maintenance of B-cell precursor acute lymphoblastic leukemia.

The FOXO1 transcription factor plays an essential role in the regulation of proliferation and survival programs at early stages of B-cell differentiation. Here, we show that tightly regulated FOXO1 activity is essential for maintenance of B-cell precursor acute lymphoblastic leukemia (BCP-ALL). Genetic and pharmacological inactivation of FOXO1 in BCP-ALL cell lines produced a strong antileukemic effect associated with CCND3 downregulation. Moreover, we demonstrated that CCND3 expression is critical for BCP-ALL survival and that overexpression of CCND3 protected BCP-ALL cell lines from growth arrest and apoptosis induced by FOXO1 inactivation. Most importantly, pharmacological inhibition of FOXO1 showed antileukemia activity on several primary, patient-derived, pediatric ALL xenografts with effective leukemia reduction in the hematopoietic, lymphoid, and central nervous system organ compartments, ultimately leading to prolonged survival without leukemia reoccurrence in a preclinical in vivo model of BCP-ALL. These results suggest that repression of FOXO1 might be a feasible approach for the treatment of BCP-ALL.

Bone Morphogenetic Protein 2 Coordinates Early Tooth Mineralization.

Formation of highly organized dental hard tissues is a complex process involving sequential and ordered deposition of an extracellular scaffold, followed by its mineralization. Odontoblast and ameloblast differentiation involves reciprocal and sequential epithelial-mesenchymal interactions. Similar to early tooth development, various Bmps are expressed during this process, although their functions have not been explored in detail. Here, we investigated the role of odontoblast-derived Bmp2 for tooth mineralization using Bmp2 conditional knockout mice. In developing molars, Bmp2LacZ reporter mice revealed restricted expression of Bmp2 in early polarized and functional odontoblasts while it was not expressed in mature odontoblasts. Loss of Bmp2 in neural crest cells, which includes all dental mesenchyme, caused a delay in dentin and enamel deposition. Immunohistochemistry for nestin and dentin sialoprotein (Dsp) revealed polarization defects in odontoblasts, indicative of a role for Bmp2 in odontoblast organization. Surprisingly, pSmad1/5/8, an indicator of Bmp signaling, was predominantly reduced in ameloblasts, with reduced expression of amelogenin ( Amlx), ameloblastin ( Ambn), and matrix metalloproteinase ( Mmp20). Quantitative real-time polymerase chain reaction (RT-qPCR) analysis and immunohistochemistry showed that loss of Bmp2 resulted in increased expression of the Wnt antagonists dickkopf 1 ( Dkk1) in the epithelium and sclerostin ( Sost) in mesenchyme and epithelium. Odontoblasts showed reduced Wnt signaling, which is important for odontoblast differentiation, and a strong reduction in dentin sialophosphoprotein ( Dspp) but not collagen 1 a1 ( Col1a1) expression. Mature Bmp2-deficient teeth, which were obtained by transplanting tooth germs from Bmp2-deficient embryos under a kidney capsule, showed a dentinogenesis imperfecta type II-like appearance. Micro-computed tomography and scanning electron microscopy revealed reduced dentin and enamel thickness, indistinguishable primary and secondary dentin, and deposition of ectopic osteodentin. This establishes that Bmp2 provides an early temporal, nonredundant signal for directed and organized tooth mineralization.

Central nervous system involvement in acute lymphoblastic leukemia is mediated by vascular endothelial growth factor.

In acute lymphoblastic leukemia (ALL), central nervous system (CNS) involvement is a major clinical concern. Despite nondetectable CNS leukemia in many cases, prophylactic CNS-directed conventional intrathecal chemotherapy is required for relapse-free survival, indicating subclinical CNS manifestation in most patients. However, CNS-directed therapy is associated with long-term sequelae, including neurocognitive deficits and secondary neoplasms. Therefore, molecular mechanisms and pathways mediating leukemia-cell entry into the CNS need to be understood to identify targets for prophylactic and therapeutic interventions and develop alternative CNS-directed treatment strategies. In this study, we analyzed leukemia-cell entry into the CNS using a primograft ALL mouse model. We found that primary ALL cells transplanted onto nonobese diabetic/severe combined immunodeficiency mice faithfully recapitulated clinical and pathological features of meningeal infiltration seen in patients with ALL. ALL cells that had entered the CNS and were infiltrating the meninges were characterized by high expression of vascular endothelial growth factor A (VEGF). Although cellular viability, growth, proliferation, and survival of ALL cells were found to be independent of VEGF, transendothelial migration through CNS microvascular endothelial cells was regulated by VEGF. The importance of VEGF produced by ALL cells in mediating leukemia-cell entry into the CNS and leptomeningeal infiltration was further demonstrated by specific reduction of CNS leukemia on in vivo VEGF capture by the anti-VEGF antibody bevacizumab. Thus, we identified a mechanism of ALL-cell entry into the CNS, which by targeting VEGF signaling may serve as a novel strategy to control CNS leukemia in patients, replacing conventional CNS-toxic treatment.

Childhood cancer predisposition syndromes-A concise review and recommendations by the Cancer Predisposition Working Group of the Society for Pediatric Oncology and Hematology.

Heritable predisposition is an important cause of cancer in children and adolescents. Although a large number of cancer predisposition genes and their associated syndromes and malignancies have already been described, it appears likely that there are more pediatric cancer patients in whom heritable cancer predisposition syndromes have yet to be recognized. In a consensus meeting in the beginning of 2016, we convened experts in Human Genetics and Pediatric Hematology/Oncology to review the available data, to categorize the large amount of information, and to develop recommendations regarding when a cancer predisposition syndrome should be suspected in a young oncology patient. This review summarizes the current knowledge of cancer predisposition syndromes in pediatric oncology and provides essential information on clinical situations in which a childhood cancer predisposition syndrome should be suspected.

Attenuated measles virus controls pediatric acute B-lineage lymphoblastic leukemia in NOD/SCID mice.

Novel therapies are needed for pediatric acute lymphoblastic leukemia resistant to conventional therapy. While emerging data suggest leukemias as possible targets of oncolytic attenuated measles virus, it is unknown whether measles virus can eradicate disseminated leukemia, in particular pediatric acute lymphoblastic leukemia. We evaluated the efficacy of attenuated measles virus against a large panel of pediatric xenografted and native primary acute lymphoblastic leukemias ex vivo, and against four different acute lymphoblastic leukemia xenografts of B-lineage in non-obese diabetic/severe combined immunodeficient mice. Ex vivo, attenuated measles virus readily spread among and effectively killed leukemia cells while sparing normal human blood cells and their progenitors. In immunodeficient mice with disseminated acute lymphoblastic leukemia a few intravenous injections of attenuated measles virus sufficed to eradicate leukemic blasts in the hematopoietic system and to control central nervous system disease resulting in long-term survival in three of the four xenografted B-lineage leukemias. Differential sensitivity of leukemia cells did not require increased expression of the measles entry receptors CD150 or CD46 nor absence of the anti-viral retinoic acid-inducible gene I/melanoma differentiation associated gene-5 /interferon pathway. Attenuated oncolytic measles virus is dramatically effective against pediatric B-lineage acute lymphoblastic leukemia in the pre-clinical setting warranting further investigations towards clinical translation.

Genetic basis for tooth malformations: from mice to men and back again.

Teeth arise from sequential and reciprocal interactions between the oral epithelium and the cranial neural crest-derived mesenchyme. Their formation involves a precisely orchestrated series of molecular and morphogenetic events. Numerous regulatory genes that have been primarily found in organisms such as Drosophila, zebrafish, xenopus and mouse are associated with all stages of tooth formation (patterning, morphogenesis, cytodifferentiation and mineralization). Most of these genes belong to evolutionary conserved signaling pathways that regulate communication between epithelium and mesenchyme during embryonic development. These signaling molecules together with specific transcription factors constitute a unique molecular imprint for odontogenesis and contribute to the generation of teeth with various and function-specific shapes. Mutations in several genes involved in tooth formation cause developmental absence and/or defects of teeth in mice. In humans, the odontogenic molecular program is not as well known as that of mice. However, some insight can be obtained from the study of mutations in regulatory genes, which lead to tooth agenesis and/or the formation of defective dental tissues.

Age estimation in an Indian population using pulp/tooth volume ratio of mandibular canines obtained from cone beam computed tomography.

The present study assessed the suitability of pulp/tooth volume ratio of mandibular canines for age prediction in an Indian population. Volumetric reconstruction of scanned images of mandibular canines from 140 individuals (aged ten - 70 years), using computed tomography was used to measure pulp and tooth volumes. Age calculated using a formula reported earlier for a Belgian sample, resulted in errors > ten years in almost 86% of the study population. The regression equation obtained for the Indian population: Age = 57.18 + (- 413.41 x pulp/tooth volume ratio), was applied to an independent control group (n = 48), and this resulted in mean absolute errors of 8.54 years which was significantly (p < 0.05) lower than those derived with the Belgian formula. The pulp/tooth volume ratio is a useful indicator of age, although correlations may vary in different populations and hence, specific formulae should be applied for the estimates.

Biodegradation, soft and hard tissue integration of various polyethylene glycol hydrogels: a histomorphometric study in rabbits.

OBJECTIVES: (i) To evaluate biodegradation, hard and soft tissue integration using various polyethylene glycol (PEG) hydrogels; (ii) to evaluate the influence of arginine-glycine-aspartic acid (RGD) on two types of PEG hydrogels. MATERIAL AND METHODS: In seven rabbits, six treatment modalities were randomly applied subperiosteally on the skull: (1) a dense network PEG hydrogel (PEG1), (2) PEG1 modified with RGD (PEG1-RGD), (3) a looser network PEG hydrogel (PEG2), (4) PEG2 modified with RGD (PEG2-RGD), (5) a collagen membrane, and (6) a polylactide/polyglycolide/trimethylene carbonate membrane. The animals were sacrificed at 14 days. Histomorphometric analyses were performed on undecalcified Epon sections using a standardized region of interest. For statistical analysis, paired t-test and signed rank test were applied. RESULTS: PEG1 and PEG1-RGD remained intact and maintained the shape. PEG2 and PEG2-RGD completely degraded and were replaced by connective tissue and bone. The largest amount of mineralized tissue was found for PEG2-RGD (21.4%), followed by PEG 2 (9.5%). The highest percentage of residual hydrogel/membrane was observed for PEG1-RGD (55.6%), followed by PEG1 (26.7%). CONCLUSIONS: Modifications of the physico-chemical properties of PEG hydrogels and the addition of RGD influenced soft and hard tissue integration and biodegradation. PEG1 showed an increased degradation time and maintained the shape. The soft tissue integration was enhanced by adding an RGD sequence. A high turn-over rate and extensive bone regeneration was observed using PEG2. The addition of RGD further improved bone formation and soft tissue integration.

Stem cells for tooth engineering.

Tooth development results from sequential and reciprocal interactions between the oral epithelium and the underlying neural crest-derived mesenchyme. The generation of dental structures and/or entire teeth in the laboratory depends upon the manipulation of stem cells and requires a synergy of all cellular and molecular events that finally lead to the formation of tooth-specific hard tissues, dentin and enamel. Although mesenchymal stem cells from different origins have been extensively studied in their capacity to form dentin in vitro, information is not yet available concerning the use of epithelial stem cells. The odontogenic potential resides in the oral epithelium and thus epithelial stem cells are necessary for both the initiation of tooth formation and enamel matrix production. This review focuses on the different sources of stem cells that have been used for making teeth in vitro and their relative efficiency. Embryonic, post-natal or even adult stem cells were assessed and proved to possess an enormous regenerative potential, but their application in dental practice is still problematic and limited due to various parameters that are not yet under control such as the high risk of rejection, cell behaviour, long tooth eruption period, appropriate crown morphology and suitable colour. Nevertheless, the development of biological approaches for dental reconstruction using stem cells is promising and remains one of the greatest challenges in the dental field for the years to come.

Intact apoptosis signaling in myeloid leukemia cells determines treatment outcome in childhood AML.

Recently we reported that intact apoptosis signaling is indicative of favorable outcome in childhood acute lymphoblastic leukemia. Here we addressed this issue in 45 pediatric acute myeloid leukemia patients analyzing 2 core apoptogenic events: cytochrome c release and caspase-3 activation. In patients with good prognosis cytochrome c release was clearly found to be caspasedependent and correlated with activated caspase-3, indicating that activation of initiator or amplifier caspases such as caspase-8 together with an intact apoptosome function are elementary for favorable outcome. The functional integrity of this apoptogenic checkpoint is reflected by the parameter caspase-dependent cytochrome c-related activation of caspase-3 (CRAC(dep)). Patients with positive CRAC(dep) values (intact signaling) exhibited superior survival compared with CRAC(dep) negative patients (deficient signaling). Thus, the propensity to undergo apoptosis of leukemia cells is an important feature for favorable treatment outcome and may serve as an additional stratification tool for pediatric AML patients. This trial was registered at www.ClinicalTrials.gov as #NCT00111345.

Histological, ultrastructural and quantitative investigations on the response of healthy human pulps to experimental capping with mineral trioxide aggregate: a randomized controlled trial.

AIM: To investigate the pulpal response to direct pulp capping in healthy human teeth with mineral trioxide aggregate (MTA) as against calcium hydroxide cement (Dycal) as control. METHODOLOGY: Twenty healthy human third molars had iatrogenic pulpotomy and direct pulp capping with MTA. Another 13 teeth were capped with Dycal as controls. The teeth were restored, with IRM, clinically reviewed and extracted after a number of pre-determined intervals (1 week, 1 month and 3 months). The specimens were fixed, decalcified, subdivided axially into two halves in the oro-buccal (lingual-buccal) plane, embedded in plastic, serial sectioned and evaluated qualitatively and quantitatively by correlative light and transmission electron microscopy with appropriate statistical evaluation of the quantitative data. RESULTS: Iatrogenic pulpal wounds treated with MTA were mostly free from inflammation after 1 week and became covered with a compact, hard tissue barrier of steadily increasing length and thickness within 3 months following capping. Control teeth treated with Dycal revealed distinctly less consistent formation of a hard tissue barrier that had numerous tunnel defects. The presence of pulpal inflammation up to the longest observation period (3 months) after capping, was a common feature in Dycal specimens. CONCLUSIONS: The MTA was clinically easier to use as a direct pulp-capping agent and resulted in less pulpal inflammation and more predictable hard tissue barrier formation than Dycal. Therefore, MTA or equivalent products should be the material of choice for direct pulp capping procedures instead of hard setting calcium hydroxide cements.

Effect of sodium hypochlorite on human root dentine--mechanical, chemical and structural evaluation.

AIM: To investigate the mechanical, chemical and structural alterations of human root dentine following exposure to ascending sodium hypochlorite concentrations. METHODOLOGY: Three-point bending tests were carried out on standardized root dentine bars (n = 8 per group, sectioned from sound extracted human third molar teeth) to evaluate their flexural strength and modulus of elasticity after immersion in 5 mL of water (control), 1% NaOCl, 5% NaOCl or 9% NaOCl at 37 degrees C for 1 h. Additional dentine specimens were studied using microelemental analysis, light microscopy following bulk staining with basic fuchsin, and scanning electron microscopy (SEM). Numerical data were compared using one-way ANOVA. Bonferroni's correction was applied for multiple testing. RESULTS: Immersion in 1% NaOCl did not cause a significant drop in elastic modulus or flexural strength values in comparison to water, whilst immersion in 5% and 9% hypochlorite reduced these values by half (P < 0.05). Both, carbon and nitrogen contents of the specimens were significantly (P < 0.05) reduced by 5% and 9% NaOCl, whilst 1% NaOCl had no such effect. Exposure to 5% NaOCl rendered the superficial 80-100 mum of the intertubular dentine permeable to basic fuchsin. Three-dimensional SEM reconstructions of partly demineralized specimens showed NaOCl concentration-dependent matrix deterioration. Backscattered electron micrographs revealed that hypochlorite at any of the tested concentrations left the inorganic dentine components intact. CONCLUSIONS: The current data link the concentration-dependent hypochlorite effect on the mechanical dentine properties with the dissolution of organic dentine components.

A comparative study on the disinfection potentials of bioactive glass S53P4 and calcium hydroxide in contra-lateral human premolars ex vivo.

AIM: To evaluate the effects of bioactive glass S53P4 versus calcium hydroxide when used as dressings in contra-lateral human premolars infected with Enterococcus faecalis ATCC 29212. METHODOLOGY: Pairs of contra-lateral premolar teeth plus single control premolars were obtained from 23 individuals aged 10-26 years undergoing orthodontic treatment. Root canals of teeth with fully formed apices (nine contra-lateral pairs, seven controls) were instrumented using a size 60 FlexoFiles 2 mm short of canal length. Canals with open apices (six contra-lateral pairs, four controls) were circumferentially instrumented using a FlexoFile. Root canals were rinsed with 1% sodium hypochlorite and 10% citric acid. Teeth were then suspended in tryptic soy broth (TSB) and autoclaved. Positive controls and study teeth were infected with E. faecalis ATCC 29212 for 2 weeks in TSB, while negative controls were kept in sterile TSB. Subsequently, contra-lateral premolars were dressed with bioactive glass S53P4 (BAG) or calcium hydroxide suspensions for 10 days. Dentine samples were obtained from teeth with fully formed apices using ISO-size 70, 80 and 90 FlexoFiles to working length and cultured. Teeth with open apices were fixed, fractured and examined using scanning electron microscopy (SEM). RESULTS: Calcium hydroxide had a strong antibacterial effect and was significantly more effective than BAG in preventing residual bacterial growth (P < 0.01). SEM analysis revealed apparent substance-specific modes of action. CONCLUSIONS: Calcium hydroxide was an effective disinfectant in human teeth.

Tubular sclerosis rather than the smear layer impedes dye penetration into the dentine of endodontically instrumented root canals.

AIM: To evaluate the effect of different root canal irrigating regimes on dentine penetration of Patent Blue dye. METHODOLOGY: Eighty extracted single-rooted human mandibular premolar teeth with narrow root canals were prepared using ProFile instruments. After each instrument, canals were irrigated with 1% sodium hypochlorite. Subsequently, teeth were randomly assigned to receive a 10 mL rinse of aqueous 17% (w/v) ethylenediaminetetraacetic acid or tap water for 2 or 10 min, followed by a final rinse with a 2% Patent Blue dye solution for 2 or 10 min (eight groups, n = 10 teeth per group). Teeth were then horizontally sectioned 3, 6 and 9 mm from the apex. Sections were digitally photographed and dye penetration was calculated as percentage of total dentine area using NIH Image J. Values were compared using one-way anova and Bonferroni correction with the alpha-type error set at <0.05. Representative tooth sections from all groups were further analysed using scanning electron microscopy. RESULTS: No significant impact of irrigating protocols on dye penetration was found. Dye penetration was significantly (P < 0.001) greater in the coronal than middle, and in middle than in apical root thirds. When observed microscopically, irrigant penetration was independent of the presence of a smear layer, but was rather a function of tubular sclerosis. CONCLUSIONS: Tubular sclerosis, a physiological phenomenon that starts in the third decade of life in the apical root region and advances coronally with age, was the main factor influencing penetrability of root dentine.

Observations on pulpal response to carbon dioxide laser drilling of dentine in healthy human third molars.

Preservation of pulpal health is the primary prerequisite for successful application of laser systems in the hard tissue management of vital teeth. The purpose of this study was to investigate the short and long-term pulpal effects to cavity preparations in healthy human teeth using carbon dioxide (CO2) laser. A total of seven, healthy, third molars that were scheduled to be removed due to space problems were used. After the laser drilling, the occlusal cavities were closed temporarily, and the teeth were extracted 7 days (n=5) and 3 months (n=2) after the operation. The specimens were fixed, decalcified, subdivided and processed for light and transmission electron microscopy. Seven days postoperatively all the five teeth that had been irradiated with the CO2 laser did not reveal any pathological changes in the pulpo-dentine complex. Three months postoperatively the two teeth that were prepared with the laser showed subtle but distinct apposition of tertiary dentine that was lined with intact odontoblasts. One of the specimens at 3 months revealed the presence of a mild, but very circumscribed, pulpal infiltration of chronic inflammatory cells subjacent to the cavity preparation. The latter is unlikely to be due to a direct effect of the laser irradiation but a possible consequence of microleakage of oral antigens and/or other tissue-irritating molecules through the temporary restoration and the remaining dentine thickness (RDT). Although these preliminary histological results suggest that the CO2 laser under investigation induced only minimal response of the dentine-pulp complex when used as a hard-tissue drilling tool, with specific energy settings, pulse duration within thermal relaxation time and emitting radiations at 9.6 microm of wavelength, larger clinical trials involving various types of teeth are necessary to reach definite conclusions for large-scale clinical application of the laser device.

Quantitative histological features and ultrastructure of opercula of human teeth showing normal and delayed eruption.

BACKGROUND: Failure of eruption of human permanent molars has been attributed to opercular lesions, although comparisons with specimens from normally erupting teeth are scarce. The aim of this study was to quantitatively analyse opercula associated with normal and delayed tooth eruption. METHOD: Twenty opercula covering permanent molars delayed in eruption were obtained from 13 patients aged 7.3-18.1 years. Six opercula from normally erupting molars of five 7.3-17.5-year-old subjects served as controls. Specimens were analysed light and electron microscopically and morphometrically. RESULTS: In addition to features recognized previously, prominent numbers of nerves, high endothelial-like venules and mast cells were observed. Ultrastructurally, large multinucleated cells did not reveal cell boundaries running between the nuclei, and mast cells seemed belonging to the MC(TC)-type. None of the features differed significantly between opercula from cases of delayed and normal tooth eruption. CONCLUSIONS: Disturbances of tooth eruption that are attributed to opercular lesions may represent retentions resulting from the failure of the eruption mechanism, rather than impactions because of a physical barrier.

Nature and attachment of cementum formed under guided conditions in human teeth. An electron microscopic study.

In an attempt at characterizing the nature and attachment of cementum formed under conditions of guided tissue regeneration (GTR) in humans, front teeth from 4 patients aged 42 to 72 years were examined at the electron microscopic level. All teeth were affected by complex periodontitis associated with advanced loss of periodontal support. Roots were surgically planed and notched, but not chemically conditioned. Either the mesial or distal surface of each tooth represented the experimental site and was covered with a biodegradable polyglactin 910 barrier, while the opposite approximal surface served as control. Following 3 months of healing, teeth were removed together with surrounding periodontal tissues including some alveolar bone. These blocks were fixed histologically, decalcified, embedded in epoxy, and sectioned for examination in the scanning (backscatter mode) and transmission electron microscope. Both experimental and control sites disclosed 2 types of regenerative cementum that seemed to be formed by cells resembling cementoblasts. The first type was characterized by a thin fringe of collagen fibrils which were arranged perpendicular to the root surface and appeared mineralized in a zone extending about 1 to 3 microm from the dentin. The second type occurred as thick patches which revealed scattered cementocytes and sheets of collagen fibrils oriented mainly parallel to the root surface, running both circularly and axially. In both situations, a continuous, thin, electrondense layer was interposed between newly formed cementum and preexisting radicular hard tissues. Interdigitation of collagen fibrils from cementum and dentin, such as observed along the natural cemento-dentinal junction, did not occur. Thus, regenerative cementum laid down in humans under guided conditions on previously diseased and planed, but not otherwise treated root surfaces shares some morphologic features with cementum formed during spontaneous repair of root resorptions. However, unlike in the course of such repair, a fibrous attachment of new cementum resembling the natural cemento-dentinal junction does not seem to be regenerated under guided conditions.

Age changes in the articular tissue of human mandibular condyles from adolescence to old age: a semiquantitative light microscopic study.

In a previous study (Luder, Anat. Rec., 1997;248:18-28), the articular tissue of the adult mandibular condyle was characterized semiquantitatively. However, questions about age changes of mature tissue were not answered, and the time course of tissue maturation from the end of condylar growth to the attainment of the adult appearance remained unknown. These issues are addressed in the present investigation. By using a light microscope, features of the superficial, intermediate, and deep articular tissue zones as well as of the subchondral bone were assessed at nine predetermined condylar sites. The frequencies of these features were recorded as scores from 0 (absent) to 10 (continuous) and were plotted against age. Analysis of covariance served for testing the significance of age and sex effects as well as intracondylar variability. Whereas almost all age-related changes in frequencies of tissue features were similar along the whole lateromedial dimension, changes at the putatively nonload-bearing, posterior slope differed significantly from those at the putatively load-bearing, anterior slope and zenith of the condyle. Two patterns of changes were noted. Frequencies of a first group of tissue features altered mainly during the age period from 15 years to 30 years and remained more or less stable thereafter. This course was characteristic for 1) a progressive cartilaginification of the superficial zone as well as 2) the disappearance of hypertrophic growth cartilage and 3) the appearance of grid-fibrous fibrocartilage in the deep zone, which were accompanied by 4) a decline in endochondral ossification and 5) the formation of a compact, subchondral bone plate. Frequencies of a second group of tissue features disclosed changes that continued up to middle and old age. This pattern was evident regarding 1) a decrease in the prominence associated with 2) a drop in cellularity and 3) progressive fibrosis or even cartilaginification of the intermediate zone. Among the age changes of condylar articular tissue, those affecting the superficial and deep zones as well as the subchondral bone are largely complete by about 30 years of age and seem to be related primarily to a gradual transition from growth to adulthood. In contrast, a second group of alterations, which progress to old age and involve mainly the intermediate zone, appears to be associated with continued maintenance and adaptive articular remodeling as well as possibly senescence. Both maturational and later age changes seem to depend markedly on articular load bearing.