Sclerostin-Antibody Treatment Decreases Fracture Rates in Axial Skeleton and Improves the Skeletal Phenotype in Growing oim/oim Mice.

In osteogenesis imperfecta (OI), vertebrae brittleness causes thorax deformations and leads to cardiopulmonary failure. As sclerostin-neutralizing antibodies increase bone mass and strength in animal models of osteoporosis, their administration in two murine models of severe OI enhanced the strength of vertebrae in growing female Crtap-/- mice but not in growing male Col1a1Jrt/+ mice. However, these two studies ignored the impact of antibodies on spine growth, fracture rates, and compressive mechanical properties. Here, we conducted a randomized controlled trial in oim/oim mice, an established model of human severe OI type III due to a mutation in Col1a2. Five-week-old female WT and oim/oim mice received either PBS or sclerostin antibody (Scl-Ab) for 9 weeks. Analyses included radiography, histomorphometry, pQCT, microcomputed tomography, and biomechanical testing. Though it did not modify vertebral axial growth, Scl-Ab treatment markedly reduced the fracture prevalence in the pelvis and caudal vertebrae, enhanced osteoblast activity (L4), increased cervico-sacral spine BMD, and improved the lumbosacral spine bone cross-sectional area. Scl-Ab did not impact vertebral height and body size but enhanced the cortical thickness and trabecular bone volume significantly in the two Scl-Ab groups. At lumbar vertebrae and tibial metaphysis, the absolute increase in cortical and trabecular bone mass was higher in Scl-Ab WT than in Scl-Ab oim/oim. The effects on trabecular bone mass were mainly due to changes in trabecular number at vertebrae and in trabecular thickness at metaphyses. Additionally, Scl-Ab did not restore a standard trabecular network, but improved bone compressive ultimate load with more robust effects at vertebrae than at metaphysis. Overall, Scl-Ab treatment may be beneficial for reducing vertebral fractures and spine deformities in patients with severe OI.

Isolated bilateral transverse agenesis of the distal segments of the lower limbs at the level of the knee joint in a human fetus.

Congenital limb anomalies occur in Europe with a prevalence of 3.81/1,000 births and can have a major impact on patients and their families. The present study concerned a female fetus aborted at 23 weeks of gestation because she was affected by non-syndromic bilateral absence of the zeugopod (leg) and autopod (foot). Autopsy of the aborted fetus, X-ray imaging, MRI, and histochemical analysis showed that the distal extremity of both femurs was continued by a cartilage-like mass, without joint cavitation. Karyotype was normal. Moreover, no damaging variant was detected by exome sequencing. The limb characteristics of the fetus, which to our knowledge have not yet been reported in humans, suggest a developmental arrest similar to anomalies described in chicks following surgical experiments on the apical ectodermal ridge of the lower limbs.

In vitro evaluation of peri-implantitis treatment modalities on Saos-2osteoblasts.

OBJECTIVES: Mechanical treatment of the implant surface through surgical approach is recommended to control peri-implantitis. Few conclusive data exist about the physical and chemical properties of treated titanium surfaces and their biocompatibility towards osteoblasts. This in vitro study aimed to evaluate four clinical procedures: plastic curette, air-abrasive device (Perio-Flow(®) ), titanium brush (Ti-Brush(®) ) and implantoplasty in terms of biocompatibility and osteogenic effect when cultured with Saos2. MATERIALS AND METHODS: Titanium disks were treated with plastic curette, air-abrasive device (Perio-Flow(®) ), titanium brush (Ti-Brush(®) ) and implantoplasty. Their surface microtopography (SEM), chemical composition (EDX) and wettability were evaluated. After seeding with Saos-2, cell morphology (1 h, 24 h), viability (three and 6 days) and alkaline phosphatase (ALP), osteoprotegerin (OPG) and osteocalcin (OCN) production (7 days) were analyzed. RESULTS: Control, plastic curette, Perio-Flow(®) and Ti-Brush(®) groups presented complex microstructures including craters and micropits, whereas the implantoplasty group appeared much smoother (SEM). Titanium, oxygen, aluminium and carbon were identified as the main components in all disks with a decrease in the percentage of oxygen, carbon and an increase in the percentage of titanium in the implantoplasty group (EDX). Implantoplasty disks were also significantly more hydrophilic than the other ones, whose surfaces appeared hydrophobic. Saos-2 showed no morphological difference at 1 h. At 24 h, they appeared round shaped in all groups, except the implantoplasty group where the cells appeared stretched and elongated. Viability was similar in all groups, but significantly higher in the Perio-Flow(®) than the control group at day six. ALP, OPG and OCN protein expression at 7 days was similar in all groups. CONCLUSIONS: Although implantoplasty was the only modality to modify the titanium surface morphology, composition and wettability, all treatment modalities promoted ALP, OPG and OCN production and appeared as valid approaches in terms of biocompatibility.

Microradiographic and histological evaluation of the bone-screw and bone-plate interface of orthodontic miniplates in patients.

OBJECTIVES: To describe the tissue reactions at the bone-titanium interface of orthodontic miniplates in humans. MATERIALS AND METHODS: Forty-two samples, consisting of tissue fragments attached or not to miniplates or their fixation screws, were collected from 24 orthodontic patients treated with miniplate anchorage, at the time of removal of their miniplates. The samples were embedded in methylmethacrylate and cut into undecalcified sections which were submitted to microradiographic analysis. The sections were also stained and examined under ordinary light. RESULTS: Three types of reactions were observed both on the histological sections and on the microradiographs. 1. The majority of the stable miniplates were easy to remove (34/42). The tissue samples collected consisted mainly in mature lamellar bone with some medullary spaces containing blood vessels, 2. two screws were highly osseointegrated and required the surgeon to remove them by trephining (2/42). They were surrounded by bone tissue which extended to the miniplate. The histological features were similar to the previous group, though the bone-screw contact was higher, and 3. in six samples obtained after unstable miniplate removal during the treatment, we observed either some woven bone trabeculae or loose connective tissue, without any histological sign of inflammation. LIMITATIONS AND CONCLUSION: For evident ethical reasons, our data were limited by the size of the tissue fragments and the limited number of patients and variety of clinical presentations. The healing reactions consisted mainly in mature lamellar bone tissue sparsely in contact with the screw or the miniplate, with signs of a moderate remodelling activity.

Postnatal growth defect in mice upon persistent Hoxa2 expression in the chondrogenic cell lineage.

Hoxa2 is a homeotic transcription factor, which is downregulated once chondrogenic differentiation is initiated. We previously generated a transgenic mouse model, which turns Hoxa2 on in cells expressing Collagen II A1, i.e. in cells entering chondrogenesis. As a consequence, mice display a general embryonic delay of ossification and then a postnatal growth defect. Col2a1-Cre mice were crossed with an inducible ß-actin driven Hoxa2 transgene. Spines, vertebrae and limbs were measured and skeletal elements were studied by X-ray, microCT, pQCT, TEM, western-blotting, histomorphometry and immunohistochemistry. Mice expressing Hoxa2 in chondrogenic cells feature a proportionate short stature phenotype with a severe lordosis, which appeared significant from postnatal day 4. Analysis of both cartilage and bone development in affected embryos and mice from birth till P35 did not reveal any major defect in histogenesis, except a reduced number of chondrocytes in the vertebral anlage at E13.5. In conclusion, the sustained expression of Hoxa2 in the chondrocyte lineage is characterized by a proportionate short stature resulting from skeletal growth defect. The indepth analysis of cartilage and bone histogenesis points towards an initial deficit in cell mobilization to enter chondrogenesis.

Molecular study of a Hoxa2 gain-of-function in chondrogenesis: a model of idiopathic proportionate short stature.

In a previous study using transgenic mice ectopically expressing Hoxa2 during chondrogenesis, we associated the animal phenotype to human idiopathic proportionate short stature. Our analysis showed that this overall size reduction was correlated with a negative influence of Hoxa2 at the first step of endochondral ossification. However, the molecular pathways leading to such phenotype are still unknown. Using protein immunodetection and histological techniques comparing transgenic mice to controls, we show here that the persistent expression of Hoxa2 in chondrogenic territories provokes a general down-regulation of the main factors controlling the differentiation cascade, such as Bapx1, Bmp7, Bmpr1a, Ihh, Msx1, Pax9, Sox6, Sox9 and Wnt5a. These data confirm the impairment of chondrogenic differentiation by Hoxa2 overexpression. They also show a selective effect of Hoxa2 on endochondral ossification processes since Gdf5 and Gdf10, and Bmp4 or PthrP were up-regulated and unmodified, respectively. Since Hoxa2 deregulation in mice induces a proportionate short stature phenotype mimicking human idiopathic conditions, our results give an insight into understanding proportionate short stature pathogenesis by highlighting molecular factors whose combined deregulation may be involved in such a disease.

A cadaveric model of anterior compartment leg syndrome: Subcutaneous minimally invasive fasciotomy versus open fasciotomy.

OBJECTIVE: Because of disabling sequelae of open fasciotomy in anterior compartment syndrome (ACS) of the leg, we wanted to describe and validate a cadaveric model of ACS. We hypothesized that, first, anterior compartment syndrome (ACS) could be reproduced in cadaveric leg and, second, fasciotomy without complete skin incision could lower the intramuscular pressure (IMP) in an equivalent range to complete dermatofasciotomy. MATERIALS AND METHODS: Lower limb ACS was reproduced by progressive injection of physiologic serum in the anterior compartment of 23 fresh frozen cadaveric legs with monitoring of IMP, in order to reach a maximal stabilised IMP higher than 30mmHg. Subcutaneous minimally invasive fasciotomy was performed on 14 legs through 5 transversal mini-incisions of the skin (2cm) along the axis from the tibial tuberosity to the posterior aspect of the lateral malleolus. Standard open fasciotomy of the anterior compartment was performed on the remaining 9 legs as control. IMP was measured after the skin incisions and after every fasciotomy through skin incisions in the first group and after skin and fascia incisions in the control group. RESULTS: A maximal IMP of 43±2mmHg was obtained by injection of 177±9ml physiologic serum into the anterior compartment of the leg. In the control open fasciotomy group, the skin incision alone did not lower IMP significantly, whereas fasciotomy lowered IMP to 10±1mmHg, which is statistically different from maximal IMP (p<0.001). In the subcutaneous fasciotomy group, complete fasciotomy lowered significantly the IMP to 11±4mmHg (p<0.001), without statistical difference with the control group. DISCUSSION: This cadaveric model is effective to reproduce the hyperpressure encountered in ACS. In this model, IMP release after fasciotomy is as efficient through minimally invasive subcutaneous incision as with control open fasciotomy. This in vitro technique appears as an attractive alternative treatment in anterior compartment syndrome of the leg. It should be tested in the other compartments of the leg and its in vivo feasibility in acute conditions has to be clarified. LEVEL OF EVIDENCE: III, control laboratory study.

Root repair after damage due to screw insertion for orthodontic miniplate placement.

BACKGROUND: The aim of this investigation was to describe the healing reactions following root damage caused by placement of a miniplate anchorage system. MATERIAL AND METHODS: In 4 beagle dogs, 4 titanium miniplates (2 self-tapping screws per miniplate) were placed in each maxilla, after drilling of pilot-holes. Six fixation screws were unintentionally inserted damaging the root of maxillary canines. Two weeks later, half of the miniplates were loaded with a coil spring. Two dogs were euthanized 7 weeks after placement of the miniplates, while the remaining two after 29 weeks. Histological sections were prepared, microradiographed, observed under U.V. light, then stained and analysed under ordinary light. RESULTS: Four screws caused direct root damage; one was damaged during the drilling process; one caused damage to the periodontal ligament only. Among these 6 screws, 2 were mobile and 4 were stable at sacrifice. Limited root damage showed some repair after 29 weeks, consisting in a thick layer of mineralized cementum including anchoring periodontal fibres. Tissue repair was not related to screw stability or loading status. CONCLUSIONS: Limited root damage has shown potential to heal, while extensive root damage has not. Precise position of insertion of the miniplates is thus of utmost importance. Key words:Temporary anchorage devices, animal studies, root resorption.

Sclerostin antibody reduces long bone fractures in the oim/oim model of osteogenesis imperfecta.

Osteogenesis imperfecta type III (OI) is a serious genetic condition with poor bone quality and a high fracture rate in children. In a previous study, it was shown that a monoclonal antibody neutralizing sclerostin (Scl-Ab) increases strength and vertebral bone mass while reducing the number of axial fractures in oim/oim, a mouse model of OI type III. Here, we analyze the impact of Scl-Ab on long bones in OI mice. After 9 weeks of treatment, Scl-Ab significantly reduced long bone fractures (3.6 ±â€¯0.3 versus 2.1 ±â€¯0.8 per mouse, p < 0.001). In addition, the cortical thickness of the tibial midshaft was increased (+42%, p < 0.001), as well as BMD (+28%, p < 0.001), ultimate load (+86%, p < 0.05), plastic energy (+184%; p < 0.05) and stiffness (+172%; p < 0.01) in OI Scl-Ab mice compared to OI vehicle controls. Similar effects of Scl-Ab were observed in Wild type (Wt) mice. The plastic energy, which reflects the fragility of the tissue, was lower in the OI than in the Wt and significantly improved with the Scl-Ab treatment. At the tissue level by nanoindentation, Scl-Ab slightly increased the elastic modulus in bones of both OI and Wt, while moderately increasing tissue hardness (+13% compared to the vehicle; p < 0.05) in Wt bones, but not in OI bones. Although it did not change the properties of the OI bone matrix material, Scl-Ab reduced the fracture rate of the long bones by improving its bone mass, density, geometry, and biomechanical strength. These results suggest that Scl-Ab can reduce long-bone fractures in patients with OI.

Patients' and orthodontists' perceptions of miniplates used for temporary skeletal anchorage: a prospective study.

INTRODUCTION: Temporary skeletal anchorage is a relatively recent addition to orthodontic treatment. Surgical miniplates, modified with intraoral attachments, provide an alternative to miniscrews for skeletal anchorage. In this study, we wanted to determine patients' and providers' perceptions of miniplate use during orthodontic treatment. METHODS: Consecutive patients having miniplates placed as part of their treatment completed questionnaires about their experiences during surgery and orthodontic treatment. A total of 200 miniplates were placed for 97 patients. The 30 orthodontists treating these patients also completed questionnaires concerning miniplate success, handling complexity, and whether these devices simplified treatment. RESULTS: The success rate was 92.5%. The devices were well tolerated by the patients. After a year, 72% of the patients reported that they did not mind having the implant, and 82% said that the surgical experience was better than expected, with little or no pain. The most frequent problems were postsurgical swelling, lasting 5 days on average, and cheek irritation experienced initially by more than a third of the patients, but it lessened over time. The clinicians reported that these devices were easy to use and greatly simplified orthodontic treatment. CONCLUSIONS: Miniplates are well accepted by patients and providers and are a safe and effective adjunct for complex orthodontic treatments.

Anatomical bases for the radiological delineation of lymph node areas. Upper limbs, chest and abdomen.

Cancer spreads locally through direct infiltration into soft tissues, or at distance by invading vascular structures, then migrating through the lymphatic or blood flow. Although cancer cells carried in the blood can end in virtually any corner of the body, lymphatic migration is usually stepwise, through successive nodal stops, which can temporarily delay further progression. In radiotherapy, irradiation of lymphatic paths relevant to the localisation of the primary has been common practice for decades. Similarly, excision of cancer is often completed by lymphatic dissection. Both in radiotherapy and in surgery, advanced knowledge of the lymphatic pathways relevant to any tumour location is an important information for treatment preparation and execution. This second part describes the lymphatics of the upper limb, of the thorax and of the upper abdomen. Providing anatomical bases for the radiological delineation of lymph nodes areas in the axilla, in the chest and in the abdomen, it also offers a simplified classification for labeling the mediastinal and intra-abdominal nodal levels, grouped in each location inside three major functional areas (called I, II and III) which are all divided into three sublevels (named a, b or c).

Oxidative Stress and Upregulation of Antioxidant Proteins, Including Adiponectin, in Extraocular Muscular Cells, Orbital Adipocytes, and Thyrocytes in Graves' Disease Associated with Orbitopathy.

BACKGROUND: Graves' orbitopathy (GO) is the main extrathyroidal manifestation associated with Graves' disease (GD). It is characterized by reduced eye motility due to an increased volume of orbital fat and/or of extraocular muscles (EOMs) infiltrated by fibrosis and adipose tissue. The pathogenetic mechanisms leading to fibrosis and adipogenesis are mainly based on the interaction between orbital fibroblasts and immune cells (lymphocytes and mast cells) infiltrating the GO EOMs. METHODS: Analysis of the morphological status, oxidative stress (OS), and antioxidant defenses in the orbital muscular cells and adipocytes in GO patients compared with controls was conducted. RESULTS: Both cell types are affected by OS, as shown by the increased expression of 4-hydroxynonenal, which leads to apoptosis in muscular cells. However, the EOMs and the adipocytes possess antioxidant defenses (peroxiredoxin 5 and catalase) against the OS, which are also upregulated in thyrocytes in GD. The expression of adiponectin (ApN) and proliferator-activated receptor gamma (PPARγ) is also increased in GO muscular cells and adipocytes. OS and antioxidant proteins expression are correlated to the level of blood antithyrotropin receptor antibodies (TSHR-Ab). CONCLUSION: Even when TSHR-Ab level is normalized, OS and antioxidant protein expression is high in EOM muscular cells and adipocytes in GO compared with controls. This justifies a supplementation with antioxidants in active as well as chronic GO patients. Orbital muscular cells are also the sources of PPARγ and ApN, which have direct or indirect local protective effects against OS. Modulation of these proteins could be considered as a future therapeutic approach for GO.

Gingiva laser welding: preliminary study on an ex vivo porcine model.

OBJECTIVE: The use of lasers to fuse different tissues has been studied for 50 years. As none of these experiments concerned the oral soft tissues, our objective was to assess the feasibility of laser gingiva welding. MATERIALS AND METHODS: Porcine full-thickness gingival flaps served to prepare calibrated samples in the middle of which a 2 cm long incision was closed, either by conventional suture or by laser tissue welding (LTW). To determine the irradiation conditions yielding the best tensile strength, 13 output power values, from 0.5 to 5 W, delivered either at 10 Hz or in continuous wave mode, were tested on six indocyanine green (ICG) concentrations, from 8% to 13% (588 samples). Then, some samples served to compare the tensile strength between the laser welded and the sutured gingiva; the other samples were histologically processed in order to evaluate the thermal damage extent. The temperature rise during the LTW was measured by thermocouples. Another group of 12 samples was used to measure the temperature elevation by thermal camera. RESULTS: In the laser welding groups, the best tensile strength (p<0.05) was yielded by the 9% ICG saline solution (117 mM) at 4.5 W, 10 Hz, and a fluence of 31.3 kJ/cm(2). The apposition strength revealed no statistically significant difference (p<0.05) between the sutured and the laser welded gingiva at 4.5 W, 10 Hz, and 9% ICG solution. The mean temperature was 74±5.4°C at the upper surface and 42±8.9°C at the lower surface. The damaged zone averaged 333 µm at the upper surface. CONCLUSIONS: The 808 nm diode laser associated with ICG can achieve oral mucosa LTW, which is conceivable as a promising technique of gingival repair.

Influence of sodium hypochlorite on Er:YAG laser-irradiated dentin and its effect on the quality of adaptation of the composite restoration margins.

OBJECTIVE: The aims of this in vitro study were to evaluate: (1) the influence of 5% NaOCl application on Er:YAG-irradiated dentin; and (2) its effect on the quality of adaptation of the composite restoration margins. BACKGROUND DATA: Previous research has shown that Er:YAG dentin irradiation produces a thermally affected tissue layer that results in lower bond strength than that of nonirradiated dentin. The removal of this thermally-affected layer may enhance the quality of dentin bonding. MATERIALS AND METHODS: Forty-nine caries-free extracted human molars were transversely sectioned in order to totally expose the dentin. Four standardized cavities were created on the dentinal surface of each molar. First, two cavities were irradiated with Er:YAG laser (2.94 nm): 150 mJ, 10 Hz, variable square pulse (VSP) mode (100 µsec), beam diameter=0.9 mm, speed of irradiation=1 mm/sec, 20% air and 20% water. Then, one of irradiated cavities and one of nonirradiated cavities were treated for 30 sec with 5% NaOCl solution. Finally, they went through a standard bonding treatment for composite restoration, etching, bonding, and composite filling. We obtained four groups of cavities: (1) one control group of nonirradiated cavities not pretreated with NaOCl; (2) one group of nonirradiated cavities, pretreated with NaOCl; (3) one group of irradiated cavities, not pretreated with NaOCl; and (4) one group of irradiated cavities, pretreated with NaOCl. All samples were subjected to thermocycling. Every cavity was immersed into a 0.5% solution of methylene blue. The percentage of dye penetration (microleakage) in the composite-dentin interface was evaluated. Six molars were analyzed by scanning electron microscope. RESULTS: Dye infiltration depth was significantly reduced in irradiated cavities treated with 5% NaOCl solution. CONCLUSIONS: The application of a 5% NaOCl solution on Er:YAG irradiated cavities can significantly improve the marginal quality of composite bonding.

Implant osseointegration in the irradiated mandible. A comparative study in dogs with a microradiographic and histologic assessment.

This research focuses on the effects of radiotherapy on the osseointegration of dental implants placed before or after radiotherapy in 11 male beagles. After the extraction of all mandibular premolars 1st and 2nd molars, three dogs were implanted without radiotherapy (Control group), four dogs were irradiated 4 weeks after implantation (IrA group) and four dogs were irradiated 8 weeks before implantation (IrB group). Eight implants were placed in each dog, in an alternating pattern: four nonsubmerged ITI Bonefit titanium plasma spray-coated and four submerged Steri-Oss hydroxyapatite-coated. The irradiated dogs received 4.3 Gy daily for 10 days. Two different fluorescent markers were administered at the time of implantation and of irradiation. The dogs were sacrificed 6 months after implantation, i.e. 5 months after radiotherapy for the IrA group and 8 months for the IrB group. Each mandible was submitted to histological and microradiographic analysis. Bone formation occurred around 85 of the 88 implants and consisted mostly of the successive deposit of woven and lamellar bone. Both irradiated groups showed obvious bone remodeling in alveolar bone as well as in the basilar part of the mandible. Nevertheless, in the IrA group, the resorption phenomena predominated over osteogenesis. The balance between these two opposite processes seemed to be restored 8 months after the end of radiotherapy (IrB group). In spite of focal lesions of radiation-specific bone destruction emphasized in some irradiated dogs, we conclude from our results that osseointegration of dental implants is possible in irradiated bone tissue.

Perforations of cortical bone allografts improve their incorporation.

The incorporation of perforated cortical bone allografts was compared with non-perforated allografts. A 5-cm circumferential defect in the middiaphysis at the tibia was created in adult sheep. A frozen tibial allograft was implanted and fixed with a locked nail for 6 months. There was no postoperative immobilization. Group I consisted of eight sheep with non-perforated allografts, whereas Group II was comprised of 10 sheep with perforated allografts. Union was evaluated radiographically, whereas the central part of the allograft had a densitometric evaluation. Creeping substitution was assessed on microradiographs from cross-sections of the central 3 cm of graft by measurement of porosity and percentage of new and old bone area within the confines of the graft. The width of periosteal and endosteal callus also was determined. There was no statistical difference between both groups for the union score and bone density. However, the cortical bone graft porosity and the amount of new bone within the cortical bone differed significantly between the perforated allografts and the non-perforated ones. Periosteal callus was similar in both groups, whereas endosteal callus was significantly more wide and extended in the perforated allografts. Perforation of a cortical bone substantially improved the amount of newly formed bone by the host when compared with a non-perforated bone. The creation of channels seemed to increase the interface between living soft tissues of the host and the allografted bone with a resulting enhanced incorporation.