Perinatal hypoxia and the risk of severe Molar-Incisor Hypomineralisation (MIH): a retrospective analysis of the pH value of umbilical arterial blood after birth.

PURPOSE: Molar-Incisor Hypomineralisation (MIH) remains a widespread developmental disorder of the teeth with a still largely unknown etiology. Perinatal events were blamed in previous studies for the development of MIH. The aim of the present study was to evaluate the influence of perinatal hypoxia-determined by the pH value of the umbilical cord blood-and to investigate its correlation with severe MIH retrospectively. In addition, cesarean section was recorded as differentiation variable. METHODS: A total number of 138 children (mean age 8.0 years ± 1.7), who were treated for severe MIH in a dental office in Berlin between the years 2008 and 2019, were included in the study. The control group was comprised of patients with the same date of birth (44 children, mean age 7.7 years ± 1.7). Information on the pH value of the arterial blood from the umbilical cord taken immediately after birth, whose recording is mandatory in Germany, was received from the parents by letter survey requesting the entries from the German Child Health Booklet. RESULTS: In the group of the male children born without cesarean section, the pH value of the control group was significantly lower (7.19 ± 0.09) than the pH value of the MIH group (7.27 ± 0.07, p = 0.0008). In female children born with or without cesarean section as well as in male children born by cesarean section there were no significant differences between the MIH and control group. CONCLUSIONS: No significant association between MIH and the pH value of the umbilical cord blood or birth by cesarean section could be found in the examined patient population.

Effect of Time and Temperature of Air Jet on the Mechanical and Biological Behavior of a Universal Adhesive System.

OBJECTIVES: To evaluate the influence of heat application on the degree of conversion (DC) of the 3M Single Bond Universal Adhesive System, as well as its transdentinal cytotoxicity and microtensile bond strength to dentin. METHODS: Experimental groups were established according to the time and temperature of the air jet: G1: 5 seconds-25°C; G2: 10 seconds-25°C; G3: 20 seconds-25°C; G4: 5 seconds-50°C; G5: 10 seconds-50°C; G6: 20 seconds-50°C. In control group (G7), no treatment was performed. The DC was assessed using the Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) technique. For the transdentinal cytotoxicity test, dentin discs fitted in artificial pulp chambers (APC) received the application of the adhesive system and the air jets. For the microtensile bond strength, healthy molars were restored and submitted to the microtensile test after 24 hours and 6 months, respectively. RESULTS: Significant reduction in viability of Mouse Dental Papilla Cell-23 (MDPC-23), which exhibited morphological changes, was observed in all experimental groups compared to control (p<0.05). Although all tested protocols resulted in transdentinal diffusion of 2-hydroxyethyl methacrylate (HEMA), the group G6 presented the highest degree of monomeric conversion and the lowest cytotoxic effect, with higher dentin bond strength values in comparison to group G1 (p<0.05). CONCLUSIONS: Applying an air blast at 50°C for 20 seconds increases the DC and microtensile bond strength of the 3M Single Bond Universal Adhesive System to dentin, as well as reduces the transdentinal cytotoxicity of the material to pulp cells.

Transcriptional Regulation of Jaw Osteoblasts: Development to Pathology.

Craniofacial and jaw bones have unique physiological specificities when compared to axial and appendicular bones. However, the molecular profile of the jaw osteoblast (OB) remains incomplete. The present study aimed to decipher the bone site-specific profiles of transcription factors (TFs) expressed in OBs in vivo. Using RNA sequencing analysis, we mapped the transcriptome of confirmed OBs from 2 different skeletal sites: mandible (Md) and tibia (Tb). The OB transcriptome contains 709 TF genes: 608 are similarly expressed in Md-OB and Tb-OB, referred to as "OB-core"; 54 TF genes are upregulated in Md-OB, referred to as "Md-set"; and 18 TF genes are upregulated in Tb-OB, referred to as "Tb-set." Notably, the expression of 29 additional TF genes depends on their RNA transcript variants. TF genes with no previously known role in OBs and bone were identified. Bioinformatics analysis combined with review of genetic disease databases and a comprehensive literature search showed a significant contribution of anatomical origin to the OB signatures. Md-set and Tb-set are enriched with site-specific TF genes associated with development and morphogenesis (neural crest vs. mesoderm), and this developmental imprint persists during growth and homeostasis. Jaw and tibia site-specific OB signatures are associated with craniofacial and appendicular skeletal disorders as well as neurocristopathies, dental disorders, and digit malformations. The present study demonstrates the feasibility of a new method to isolate pure OB populations and map their gene expression signature in the context of OB physiological environment, avoiding in vitro culture and its associated biases. Our results provide insights into the site-specific developmental pathways governing OBs and identify new major OB regulators of bone physiology. We also established the importance of the OB transcriptome as a prognostic tool for human rare bone diseases to explore the hidden pathophysiology of craniofacial malformations, among the most prevalent congenital defects in humans.

Efficacy of rotary instruments with greater taper in preparing oval root canals.

AIM: To compare the preparation quality of two rotary systems and NiTi-hand files in oval root canals, and to evaluate the effect of canal dimensions on the preparation. METHODOLOGY: Ninety roots with oval root canals were selected. The middle third was cross sectioned at two levels and photographed. The maximum and minimum diameters of the root and canal were recorded. Teeth were distributed in three groups (n = 30) using stratified randomization, and prepared under simulated clinical conditions with Mtwo, ProTaper, or NiTi-hand files. The pre- and post-preparation photographs were traced and superimposed, the thickness of dentine removed was measured and the ratio of prepared canal outline was calculated. The impact of preparation system and canal dimensions on the quality of the preparation was evaluated using regression analysis. RESULTS: With regards to the ratio of prepared canal outline, no statistical significant difference was found between Mtwo (0.75 [95%CI: 0.69; 0.81]) and ProTaper (0.75 [95%CI: 0.69; 0.80]), but both systems performed significantly better than NiTi-hand files (0.65 [95%CI: 0.60; 0.71]). In six root canals in Mtwo-group (20%), and eight root canals in ProTaper-group (27%), the minimal thickness of dentine-wall after preparation was less than 0.5 mm. In contrast to the maximum diameter of the root canal, the minimum diameter influenced the quality of the preparation (P = 0.0006). CONCLUSIONS: No instrumentation technique was able to circumferentially prepare the oval outline of root canals. Nevertheless, instruments with greater taper (ProTaper and Mtwo) were more efficient than NiTi- hand files, but this was, in some cases, at the expense of remaining dentine-wall thickness.

Ribavirin with either standard or pegylated interferon to treat recurrent hepatitis C after liver transplantation.

AIM: To investigate the efficacy of two anti-viral protocols in hepatitis C virus-reinfected liver transplant recipients. METHODS: In this prospective study, 26 liver transplant patients were treated with standard interferon-alpha2b for 12 months or standard interferon-alpha2b for 3 months followed by pegylated interferon-alpha2b for 9 months. Interferon was combined with ribavirin in all patients. The histological course of the study population was compared with an untreated historic control group (n = 38) with similar baseline characteristics. RESULTS: The sustained virological response rates in the standard interferon group and in the pegylated interferon group were 27.3% and 26.7%, respectively. Only 29% of patients with sustained virological response had end of treatment histological response, whereas 47% of viral non-responders showed end of treatment histological response. The percentage of patients with histological improvement was significantly higher in the study population when compared to the controls. Univariate analysis indicated that hepatitis C virus genotype non-1, high baseline alanine aminotransferase, the time interval between liver transplant and interferon therapy and the body mass index predicted sustained virological response. In the multivariate model, baseline alanine aminotransferase and the body mass index remained a significant predictor of sustained virological response. CONCLUSIONS: Both treatment regimens offer similar efficacy profiles. Failure to eradicate hepatitis C virus should not lead to treatment discontinuation if serial liver biopsies demonstrate histological response.

Surface biofunctionalization of ß-TCP blocks using aptamer 74 for bone tissue engineering.

Successful bone regeneration following oral and maxillofacial surgeries depends on efficient functionalization strategies that allow the recruitment of osteogenic progenitor cells at the tissue/implant interface. We have previously identified aptamer 74, which exhibited a binding affinity for osteogenically induced jaw periosteal cells (JPCs). In the present study, this aptamer was used for the surface biofunctionalization of ß-tricalcium phosphate (ß-TCP) blocks. Atomic force microscopy (AFM) measurements showed increased binding activity of aptamer 74 towards osteogenically induced JPCs compared to untreated controls. The immobilization efficiency of aptamer 74 was analyzed using the QuantiFluor ssDNA assay for 2D surfaces and by amino acid analysis for 3D ß-TCP constructs. Following the successful immobilization of aptamer 74 in 2D culture wells and on 3D constructs, in vitro assays showed no significant differences in cell proliferation compared to unmodified surfaces. Interestingly, JPC mineralization was significantly higher on the 2D surfaces and higher cell adhesion was detected on the 3D constructs with immobilized aptamer. Herein, we report an established, biocompatible ß-TCP matrix with surface immobilization of aptamer 74, which enhances properties such as cell adhesion on 3D constructs and mineralization on 2D surfaces. Further studies need to be performed to improve the immobilization efficiency and to develop a suitable approach for JPC mineralization growing within 3D ß-TCP constructs.

Coating of ß-tricalcium phosphate scaffolds-a comparison between graphene oxide and poly-lactic-co-glycolic acid.

Bone regeneration in critical size defects is a major challenge in oral and maxillofacial surgery, and the gold standard for bone reconstruction still requires the use of autologous tissue. To overcome the need for a second intervention and to minimize morbidity, the development of new biomaterials with osteoinductive features is the focus of current research. As a scaffolding material, ß-tricalcium phosphate (ß-TCP) is suitable for bone regeneration purposes, although it does not carry any functional groups for the covalent immobilization of molecules. The aim of the present study was to establish effective coating variants for ß-TCP constructs to enable the biofunctionalization of anorganic blocks with different osteogenic molecules in future studies. We established working protocols for thin surface coatings consisting of polylactic-co-glycolic acid (PLGA) and graphene oxide (GO) by varying parameters. Surface properties such as the angularity and topography of the developed scaffolds were analyzed. To examine biological functionality, the adhesion and proliferation behavior of jaw periosteal cells (JPCs) were tested on the coated constructs. Our results suggest that PLGA is the superior material for surface coating of ß-TCP matrices, leading to higher JPC proliferation rates and providing a more suitable basis for further biofunctionalization in the field of bone tissue engineering.

Degradable bisphosphonate-alkaline phosphatase-complexed hydroxyapatite implants in vitro.

Degradable hydroxyapatite (HA) implants complexed with the resorption inhibiting agent bisphosphonate (PCP) and the mineralizing agent alkaline phosphatase (ALP) can theoretically maintain alveolar bone mass directly after extraction of teeth. The present in vitro study investigated the surface properties of PCP-ALP-complexed HA implants in relation to the requirements of implant behavior and action. Adsorbed PCP (pH 3.49) resulted in a flattening and broadening of the phosphate peaks and the formation of carbonate peaks in the HA pattern of the implant indicating a chemical alteration of the HA surface. Adsorption of ALP onto PCP-altered HA surfaces was 26% lower than onto HA implant blank surfaces. PCP-ALP-complexed HA implants released the PCP and ALP steadily and continuously over observation periods of, respectively, 75 and 14 days. During these observation periods, the ceramic grains of the HA implant became smaller and intergrain boundaries became broader. These morphologic characteristics suggested preconditioning of the HA implant surface for future bonding and degradation in vivo. Individual grains were no longer bonded to other grains and detached from the implant which had become rounded in shape. From in vitro mice experiments we found that PCP concentrations between 10(-4) and 10(-3) M resulted in 45Ca-release from the bone HA. Our calculations showed, however, that only a total concentration of 1.4 x 10(-4) M PCP was gradually released over the whole observation period. In another experiment, it appeared that a PCP concentration in solution < 10(-3) M did not reduce ALP activity. It is concluded that release of PCP by the PCP-ALP-complexed implants is maintained at levels in the range to impair osteoclast bone resorption but not high enough to block osteoblast activity. The amount of ALP released can lead to induction of bone formation onto implant surfaces. pH-induced alterations in the microstructure and chemistry of the HA surface allow for controlled degradation of the HA implants in vitro. A PCP-ALP-complexed HA implant acting as temporary scaffolding for alveolar bone growth enhancement, mineralization, and maintenance seems to be a reasonable concept for preservation of the edentulous alveolus.

Synthesis, pharmacological and biophysical characterization, and membrane-interaction QSAR analysis of cationic amphiphilic model compounds.

Cationic amphiphilic drugs have a propensity to interact with biological interphases. This study was designed to gain more insight into the molecular properties of catamphiphilic drugs which govern this type of interaction. A series of phenylpropylamine model compounds were synthesized in which modifications were incorporated at the aromatic part of the molecule. The replacement of (45)Ca(2+) from phosphatidylserine monolayers served to monitor drug binding to the phospholipid. The influence on the phase-transition temperature of liposomes of dipalmitoylphosphatidic acid was measured to assess the perturbing action of the drugs on the structural organization of phospholipid assemblies. The antiarrhythmic activity of the compounds was determined in Langendorff preparations of guinea pig hearts to assess the membrane-stabilizing action. Quantitative structure-activity relationship (QSAR) models for these endpoints were developed using both intra- and intermolecular QSAR descriptors. Intermolecular membrane-interaction descriptors were derived from molecular dynamics simulations of the compounds in a model phospholipid monolayer. QSAR models were derived for all endpoints using partial least-squares regression (PLS) and a genetic algorithm tool, the genetic function approximation (GFA). Membrane-interaction descriptors appear to be of a particular importance in explaining the influence of the compounds on the phase-transition temperature of DPPA liposomes, while the other endpoints can be adequately modeled by intramolecular descriptors. The calcium-displacing activity at phosphatidylserine monolayers is governed by the electrostatic properties of the compounds. Measures of lipophilicity and molecular size are of particular importance for antiarrhythmic activity. Possible improvements to both the molecular modeling and the applied computational protocol of membrane-solute systems are identified and discussed.

Histological observations of coated and prestressed dental implants under flexible or rigid loading conditions.

Hydroxyapatite-coated and hydroxyapatite-prestressed titanium implants were implanted under load-free, flexible-loaded and rigid-loaded conditions in dog mandibula in vivo. The bone response, bone contact and the quality of the coating were evaluated histologically and histometrically. Hydroxyapatite coating on titanium implants enhanced the bone apposition. The bone contact can still be present while the coating is resorbed. Gingiva contact with the coating led to a faster degradation of the coating. There was no significant difference in bone contact between hydroxyapatite-coated and -prestressed implants with regard to the loading systems: rigid, flexible and load-free. All three implants showed the highest amount of bone contact apically. Only at the mesial side of the implant did the flexible element show rather high amounts of bone contact compared to the rigid and load-free implants. The hydroxyapatite coating more or less degraded during the implantation period. The degree of degradation varied in the implant itself, between the implants and between the animals.

Macroporous calcium phosphate bioceramics in dog femora: a histological study of interface and biodegradation.

This study examines the biodegradation behaviour of calcium phosphate macroporous bioceramics consisting of hydroxylapatite or beta-whitlockite implants. The implantations were performed in hard tissue of femora and muscular tissue of dogs for periods of 6 and 12 month. With microradiography and light microscopy, respectively, of ground and thin sections it was shown that hydroxylapatite remained unchanged and beta-whitlockite degraded within 6 month in bone tissue. In the muscular tissue, the biodegradation of beta-whitlockite was slower. In addition, beta-whitlockite gave rise to a cellular response of lymphocytes and plasma cells rather than hydroxylapatite in the soft tissue region. For tissue response hydroxylapatite is more suitable as an implant material than beta-whitlockite.

Macroporous calcium phosphate ceramics for bone substitution: a tracer study on biodegradation with 45Ca tracer.

The value of artificial materials in bone replacement depends highly on their biocompatibility and biostability. Porous calcium phosphate ceramics have a good compatibility with natural bone. To study the biodegradation process of calcium phosphate bioceramics, labelled [45Ca]-beta-whitlockite and [45Ca]-hydroxyapatite were implanted in the femurs of dogs. The effects accompanying the ingrowth of new bone into the pores of these bioceramics and its replacement by natural bone were investigated. In vivo degradation of the implant material by biochemical dissolution processes was observed. The beta-whitlockite implants showed a conspicuous decrease in radioactive calcium, in contrast to the hydroxyapatite implants. 45Ca was absent in adjacent bones and locally newly formed bone in both beta-whitlockite and hydroxyapatite suggesting a restricted availability of the 45Ca liberated in these processes. Indications of minor mechanical degradation of the materials were also found; in the lymph nodes a very small amount of 45Ca with a high specific activity was detectable. Urine, blood or faeces contained no detectable amounts of 45Ca activity.

A plastic composite of alginate with calcium phosphate granulate as implant material: an in vivo study.

A comparison of calcium phosphate granulate with a composite of alginate and calcium phosphate granulate, shows that the composite has less particle migration, less biodegradation and better handling properties. The tissue response to both materials is similar.

Permucosal implantation pilot study with HA-coated dental implant in dogs.

The histological response of transmucosal one-stage titanium dental implants coated with hydroxyapatite is described. The gingival adhesion to the implant was examined with regard to coated, partially coated or non-coated surfaces in the cervical region. From each coating type, 9 implants were inserted into dogs. Six months after the insertion, 19 implants could be evaluated, but 8 implants were lost. From these 19 implants, 6 implants showed severe pockets with inflammation up to the bony tissue. The 13 successful implants showed direct bone bonding with the hydroxyapatite coating and adhesion of submucosal connective tissue to the implant surface, with inflammation. The marginal gingiva showed slight inflammation. A totally coated implant will probably introduce inflammation by debris formation against the rough implant surface more easily. The hydroxyapatite coating often disappeared in the soft tissue or in the oral cavity. Bone which directly adapted to the coating seemed to prevent it from resorption.

Relationship between the degradation behaviour of calcium phosphate ceramics and their physical-chemical characteristics and ultrastructural geometry.

The neck geometry of different calcium phosphate materials was studied by scanning electron microscopy. The dissolution rate of these materials, especially of the necks, was determined by chemical analysis. The degradation of calcium phosphate materials in lactate buffer, related to the detachment of sintered particles, was found to be determined by neck dissolution rate and neck geometry. The latter factor depends on the crystallography and stoichiometry of the material and the sintering conditions.

Different calcium phosphate bioglass ceramics implanted in rabbit cortical bone. An interface study.

In order to study the interface of calcium phosphate bioglass ceramics, cylinders of standard size were implanted in the tibiae of rabbits. The materials were evaluated by radiography, light microscopy and microradiography. Bioceramics with hydroxyapatite surface give rise to a closer contact with new bone than calcium phosphate glass ceramics.

Long-term in vivo study of plasma-sprayed coatings on titanium alloys of tetracalcium phosphate, hydroxyapatite and alpha-tricalcium phosphate.

In order to study the interaction of calcium phosphate coatings with bone tissue, coated titanium plugs of standard size were implanted in dog femora. The bone bonding and bone formation of hydroxyapatite, alpha-tricalcium phosphate (alpha-TCP) and tetracalcium phosphate plasma-sprayed coatings were evaluated by mechanical push-out tests and histological observations after 3, 5, 15 and 28 months of implantation. During this time all coating types degraded. alpha-TCP showed the most significant degradation after 3 months of implantation. Hydroxyapatite and tetracalcium phosphate showed significant signs of degradation after about 5 months of implantation. All coatings showed a small increase in bone bonding after 5 months of implantation. In general, all types of implants showed similar bone response, some bone contact and several remodelling lacunae along the surfaces after long-term implantation.

Granuloma and plasma cell formation induced by the subcutaneous implantation of beta-whitlockite particles.

In this paper, the formation of a granuloma following subcutaneous implantation of beta-whitlockite powder is described. The implants were removed after 3, 5 and 7 wk and examined histologically. In the granuloma giant cells, macrophages, lymphocytes and plasma cells are present. The presence of plasma cells indicates that not only inflammation occurs, but that an immunoresponse also takes place. Resorption and phagocytosis of particles was observed and the particles were found in the regional lymph nodes after 7 wk. beta-Whitlockite powder induces an inflammation and both a cellular and a humoral immunological response, when implanted subcutaneously.

A comparative study of different beta-whitlockite ceramics in rabbit cortical bone with regard to their biodegradation behaviour.

Different beta-whitlockite ceramic cylinders of standard size were implanted in the tibiae of rabbits to study the influence of micropores and chemistry on the biodegradation rate. The materials were evaluated by radiography and light microscopy. Surface chemistry was varied by the addition of impurities, while different applied pressures before sintering and different sintering temperatures gave rise to different micropores. Both factors influenced the biodegradation rate.