Implementation of the "loaded implant" model in the rat using a miniaturized setup--description of the method and first results.

OBJECTIVE: To miniaturize the "loaded implant" model to permit its application to small rodents. In this model, two titanium implants are placed 8 mm apart with their heads protruding from the skin and are forced together by a dedicated actuator. To assess the effect of (i) the post-implantation healing period and the duration of stimulation and (ii) the intratissular strain level on the microtomographical bone parameters BV/TV, Tb.N., Tb.Th. and BIC. MATERIALS AND METHODS: Implants, 1 × 8 mm, were machined, inserted into the tibiae of rats and activated. A total of 123 animals were used. In series 1, the implants were left to heal for 2/4 weeks and then loaded to generate intratissular strains of 1125 ± 5% µÎµ for 4/8 weeks. Series 2 had their implants loaded to 750, 1500 and 2250 ± 5% µÎµ, respectively. RESULTS: Bone to implant contact increased upon loading. In series 1, no difference was observed regarding the duration of healing or the stimulation period. In series 2, at 750 µÎµ, the bone parameters did not differ from baseline. At 1500 µÎµ, all four parameters increased. At 2250 µÎµ, three of four parameters decreased relative to 1500 µÎµ. CONCLUSIONS: (i) The loaded implant model can be miniaturized to the millimeter range; (ii) in the present model, implant activation beyond 4 weeks did not affect the bone parameters; (iii) mechanical stimulation increased bone to implant contact by up to 20%; (iv) the results obtained are consistent with the concept of an anabolic effect from 750 to 1500 µÎµ and deleterious effects at strains in the 2250 µÎµ range; and (v) strains at 2250 µÎµ did not lead to implant dis-integration.

Systemic treatment with strontium ranelate accelerates the filling of a bone defect and improves the material level properties of the healing bone.

Rapid bone defect filling with normal bone is a challenge in orthopaedics and dentistry. Strontium ranelate (SrRan) has been shown to in vitro decrease bone resorption and increase bone formation, and represents a potential agent with the capacity to accelerate bone defect filling. In this study, bone tibial defects of 2.5 mm in diameter were created in 6-month-old female rats orally fed SrRan (625 mg/kg/d; 5/7 days) or vehicle for 4, 8, or 12 weeks (10 rats per group per time point) from the time of surgery. Tibias were removed. Micro-architecture was determined by micro-computed tomography (µCT) and material level properties by nanoindentation analysis. µCT analysis showed that SrRan administration significantly improved microarchitecture of trabecular bone growing into the defect after 8 and 12 weeks of treatment compared to vehicle. SrRan treatment also accelerated the growth of cortical bone over the defect, but with different kinetics compared to trabecular bone, as the effects were already significant after 4 weeks. Nanoindentation analysis demonstrated that SrRan treatment significantly increased material level properties of both trabecular bone and cortical bone filling the defect compared to vehicle. SrRan accelerates the filling of bone defect by improving cortical and trabecular bone microarchitecture both quantitatively and qualitatively.

Factors affecting subject-specific finite element models of implant-fitted rat bone specimens: critical analysis of a technical protocol.

The authors propose a protocol to derive finite element (FE) models from micro computer tomography scans of implanted rat bone. A semi-automatic procedure allows segmenting the images using specimen-specific bone mineral density (BMD) thresholds. An open-source FE model generator processes the segmented images to a quality tetrahedral mesh. The material properties assigned to each element are integrated from the BMD field. Piecewise, threshold-dependent density-elasticity relationships are implemented to limit the effects of metal artefacts. A detailed sensitivity study highlights the coherence of the generated models and quantifies the influence of the modelling parameters on the results. Two applications of the protocol are proposed. The stiffness of bare and implanted rat tibiae specimens is predicted by simulating three-point bending and inter-implant displacement, respectively. Results are compared with experimental tests. The mean value and the variability between the specimens are well captured in both tests.

External mechanical microstimuli modulate the osseointegration of titanium implants in rat tibiae.

PURPOSE: To assess the effect of external mechanical microstimuli of controlled magnitude on the microarchitecture of the peri-implant bone beds in rat tibiae. MATERIALS AND METHODS: Tibiae of forty rats were fitted with two transcutaneous titanium cylinders. After healing, the implants were loaded to 1 to 3 N, five days/week for four weeks. These force levels translated into intraosseous strains of 700 ± 200 µÎµ, 1400 ± 400 µÎµ, and 2100 ± 600 µÎµ. After sacrifice, the implants' pullout strength was assessed. Second, the bone's microarchitecture was analyzed by microcomputed tomography (µCT) in three discrete regions of interest (ROIs). Third, the effect of loading on bone material properties was determined by nanoindentation. RESULTS: The trabecular BV/TV significantly increased in an ROI of 0.98 mm away from the test implant in the 1 N versus the 3 N group with an opposite trend for cortical thickness. Pull-out strength significantly increased in the 2 N relatively to the nonstimulated group. Higher values of E-modulus and hardness were observed in the trabecular bone of the 2 N group. CONCLUSION: The in vivo mechanical loading of implants induces load-dependent modifications in bone microarchitecture and bone material properties in rat tibiae. In pull-out strength measurements, implant osseointegration was maximized at 2 N (1400 ± 400 µÎµ).

Hedgehog pathway activity is required for the lethality and intestinal phenotypes of mice with hyperactive Wnt signaling.

Several lines of evidence point to the central role of WNT signaling in the initiation of intestinal tumorigenesis, most often due to loss of APC, a negative regulator of the WNT-betaCATENIN/TCF pathway. Modeling human colon cancers in mice through loss of Apc has shown that inappropriate activation of Wnt signaling is sufficient to induce adenoma formation. More recent analyses have also demonstrated a key role for HEDGEHOG-GLI (HH-GLI) signaling in human colon cancers. However, how the WNT and HH pathways interact during intestinal development, homeostasis and cancer is not clear. Marker analyses suggest predominant paracrine signaling from rare Shh producing cells in the crypt's bottom to adjacent Gli1(+) mesenchymal cells in normal adult mice. Using conditional KO models, we show that inhibition of the function of the critical Hh mediator Smoothened (Smo) rescues the lethality and intestinal phenotypes of loss of Apc. The results uncover an essential role of the Hh pathway in tumors induced by hyperactive Wnt signaling, suggest the action of the Hh pathway in parallel or downstream of Wnt signaling, and validate this model for its use in preclinical work testing Hh pathway antagonists.

Interactions between HOXD and Gli3 genes control the limb apical ectodermal ridge via Fgf10.

The development of the vertebrate limb is dependent upon two signaling centers, the apical ectodermal ridge (AER), which provides the underlying mesenchyme with essential growth factors, and the zone of polarizing activity (ZPA), the source of the Sonic hedgehog (SHH) product. Recent work involving gain and loss of function of Hox genes has emphasized their impact both on AER maintenance and Shh transcriptional activation. Here, we describe antagonistic interactions between posterior Hoxd genes and Gli3, suggesting that the latter product protects the AER from the deleterious effect of the formers, and we present evidence that Fgf10 is the mediator of HOX-dependent AER expansion. Furthermore, the striking similarity between some of the hereby observed Hox/Gli3-dependent morphogenetic defects and those displayed by fetuses with severely altered retinoic acid metabolism suggests a tight connection between these various pathways. The nature of these potential interactions is discussed in the context of proximal-distal growth and patterning.

Hox gene function in vertebrate gut morphogenesis: the case of the caecum.

The digestive tract is made of different subdivisions with various functions. During embryonic development, the developing intestine expresses combinations of Hox genes along its anterior to posterior axis, suggesting a role for these genes in this regionalization process. In particular, the transition from small to large intestine is labelled by the transcription of all Hoxd genes except Hoxd12 and Hoxd13, the latter two genes being transcribed only near the anus. Here, we describe two lines of mice that express Hoxd12 ectopically within this morphological transition. As a consequence, budding of the caecum is impeded, leading to complete agenesis in homozygous individuals. This effect is concurrent with a dramatic reduction of both Fgf10 and Pitx1 expression. Furthermore, the interactions between ;anterior' Hox genes and ectopic Hoxd12 suggest a model whereby anterior and posterior Hox products compete in controlling Fgf10 signalling, which is required for the growth of this organ in mice. These results illuminate components of the genetic cascade necessary for the emergence of this gut segment, crucial for many vertebrates.