Johannes M. Rueger

Leptin Inhibits Bone Formation through a Hypothalamic Relay: A Central Control of Bone Mass

Gonadal failure induces bone loss while obesity prevents it. This raises the possibility that bone mass, body weight, and gonadal function are regulated by common pathways. To test this hypothesis, we studied leptin-deficient and leptin receptor-deficient mice that are obese and hypogonadic. Both mutant mice have an increased bone formation leading to high bone mass despite hypogonadism and hypercortisolism. This phenotype is dominant, independent of the presence of fat, and specific for the absence of leptin signaling. There is no leptin signaling in osteoblasts but intracerebroventricular infusion of leptin causes bone loss in leptin-deficient and wild-type mice. This study identifies leptin as a potent inhibitor of bone formation acting through the central nervous system and therefore describes the central nature of bone mass control and its disorders.

Rescue of the Skeletal Phenotype of Vitamin D Receptor- Ablated Mice in the Setting of Normal Mineral Ion Homeostasis: Formal Histomorphometric and Biomechanical Analyses*

1,25-Dihydroxyvitamin D3 has been shown to play an important role in vitro in regulating osteoblast gene transcription and promoting osteoclast differentiation. To address the role of the vitamin D receptor (VDR) in skeletal homeostasis, formal histomorphometric analyses were performed in VDR null mice in the setting of impaired mineral ion homeostasis as well as in VDR null mice in whom normal mineral ion homeostasis had been preserved. In hypocalcemic VDR null mice, there was an increase in bone volume as a result of a dramatic increase in osteoid. There was also an increase in the number of osteoblasts without a significant change in the number of osteoclasts. Examination of the growth plate revealed marked disorganization, with an increase in vascularity and matrix. Biomechanical parameters demonstrated increased bone fragility in the hypocalcemic VDR null mice. In the VDR ablated mice in whom normal mineral ion homeostasis had been preserved, none of these measurements was significantly different fro...

Genetic ablation of parathyroid glands reveals another source of parathyroid hormone.

The parathyroid glands are the only known source of circulating parathyroid hormone (PTH), which initiates an endocrine cascade that regulates serum calcium concentration. Glial cells missing2 (Gcm2), a mouse homologue of Drosophila Gcm, is the only transcription factor whose expression is restricted to the parathyroid glands. Here we show that Gcm2-deficient mice lack parathyroid glands and exhibit a biological hypoparathyroidism, identifying Gcm2 as a master regulatory gene of parathyroid gland development. Unlike PTH receptor-deficient mice, however, Gcm2-deficient mice are viable and fertile, and have only a mildly abnormal bone phenotype. Despite their lack of parathyroid glands, Gcm2-deficient mice have PTH serum levels identical to those of wild-type mice, as do parathyroidectomized wild-type animals. Expression and ablation studies identified the thymus, where Gcm1, another Gcm homologue, is expressed, as the additional, downregulatable source of PTH. Thus, Gcm2 deletion uncovers an auxiliary mechanism for the regulation of calcium homeostasis in the absence of parathyroid glands. We propose that this backup mechanism may be a general feature of endocrine regulation.

Increased bone mass is an unexpected phenotype associated with deletion of the calcitonin gene

Calcitonin (CT) is a known inhibitor of bone resorption. Calcitonin gene-related peptide-alpha (CGRPalpha), produced by alternative RNA processing of the CT/CGRP gene, has no clearly defined role in bone. To better understand the physiologic role of the CT/CGRP gene we created a mouse in which the coding sequences for both CT and CGRPalpha were deleted by homologous recombination. The CT/CGRP(-/-) knockout (KO) mice procreated normally, there were no identifiable developmental defects at birth, and they had normal baseline calcium-related chemistry values. However, KO animals were more responsive to exogenous human parathyroid hormone as evidenced by a greater increase of the serum calcium concentration and urine deoxypyridinoline crosslinks, an effect reversed by CT and mediated by a greater increase in bone resorption than in controls. Surprisingly, KO mice have significantly greater trabecular bone volume and a 1.5- to 2-fold increase in bone formation at 1 and 3 months of age. This effect appears to be mediated by increased bone formation. In addition, KO mice maintain bone mass following ovariectomy, whereas wild-type mice lose approximately one-third of their bone mass over 2 months. These findings argue for dual roles for CT/CGRP gene products: prevention of bone resorption in hypercalcemic states and a regulatory role in bone formation.

Biologically and chemically optimized composites of carbonated apatite and polyglycolide as bone substitution materials.

We report on the development and characterization of a new composite material consisting of amorphous carbonated apatite, Ca(5)(PO(4), CO(3))(3)(OH), and microstructured poly(hydroxyacetic acid), polyglycolide (PGA). This material is able to keep the pH of a surrounding solution within the physiological range (7.2-7.6). This was achieved by chemical fine-tuning of the counterplay between the acidic degradation of the polyester and the basic dissolution of calcium phosphate. Microporous samples with pore sizes of <1 microm and compact samples were prepared. The biological behavior was assayed in vitro by long-term osteoblast culture. Morphological and biochemical analyses of cell differentiation revealed excellent biocompatibility, leading to cell attachment, collagen and osteocalcin expression, and mineral deposition. This material could be of use as a biodegradable bone substitution material and as a scaffold for tissue engineering.

Impaired gastric acidification negatively affects calcium homeostasis and bone mass.

Activation of osteoclasts and their acidification-dependent resorption of bone is thought to maintain proper serum calcium levels. Here we show that osteoclast dysfunction alone does not generally affect calcium homeostasis. Indeed, mice deficient in Src, encoding a tyrosine kinase critical for osteoclast activity, show signs of osteopetrosis, but without hypocalcemia or defects in bone mineralization. Mice deficient in Cckbr, encoding a gastrin receptor that affects acid secretion by parietal cells, have the expected defects in gastric acidification but also secondary hyperparathyroidism and osteoporosis and modest hypocalcemia. These results suggest that alterations in calcium homeostasis can be driven by defects in gastric acidification, especially given that calcium gluconate supplementation fully rescues the phenotype of the Cckbr-mutant mice. Finally, mice deficient in Tcirg1, encoding a subunit of the vacuolar proton pump specifically expressed in both osteoclasts and parietal cells, show hypocalcemia and osteopetrorickets. Although neither Src- nor Cckbr-deficient mice have this latter phenotype, the combined deficiency of both genes results in osteopetrorickets. Thus, we find that osteopetrosis and osteopetrorickets are distinct phenotypes, depending on the site or sites of defective acidification (pages 610–612).

The treatment of intertrochanteric fractures: results using an intramedullary nail with integrated cephalocervical screws and linear compression.

Objective: A new device for the treatment of intertrochanteric fractures that uses 2 cephalocervical screws in an integrated mechanism allowing linear intraoperative compression and rotational stability of the head/neck fragment has been developed. The aim of this study was to describe the results using this device for the treatment of stable and unstable intertrochanteric fractures. Design: Prospective, consecutive. Setting: Academic Trauma Center. Methods: Between March 1, 2005, and July 31, 2006, 100 consecutive patients with an intertrochanteric fracture were treated with a new trochanteric antegrade nail (InterTan; Smith-Nephew, Memphis, TN). All living patients were followed up for a minimum of 1 year postoperatively (range 12-27 months). Clinical and radiographic examinations were performed until healing and at the 1-year anniversary of the index procedure. Healing, pain with ambulation, return to activities of daily living, the modified Harris hip score, and Barthel Index were used to evaluate outcomes. Results: The mean age of the patients was 81.2 (±11.3) years. Thirty-seven patients died, 12 were too infirmed for follow-up, and 3 could not be located, leaving 48 patients available for final evaluation. The average surgical time was 41 minutes (13-95 minutes). This rose significantly with the complexity of the fracture (OTA/AO classification: A1 versus A3, P = 0.016). All fractures healed within 16 weeks (range 10-16 weeks). Radiographic analysis at healing revealed no loss of reduction, no uncontrolled collapse of the neck, no nonunions, no femoral shaft fractures, and no implant failures. Two cases in the series were poorly reduced and settled into varus malalignment. There was no varus malposition seen in the remaining 46 fractures. The mean prefracture Harris hip score (75.1 ± 13.4) was significantly reduced at the time of follow-up (70.3 ± 14.5, P = 0.003); 58% of the patients recovered their prefracture status. No significant difference was seen for the Barthel Index. Conclusions: The InterTan device appears to be a reliable implant for the treatment of intertrochanteric femoral fractures. Its design provides for stability against rotation and minimizes neck malunions (shortening) through linear intraoperative compression of the head/neck segment to the shaft. As a result of the negligible complication rate and improved clinical outcomes, this implant is now the standard treatment for all intertrochanteric fractures at our institution.

Leptin Inhibits Bone Formation Not Only in Rodents, but Also in Sheep

This study examines the effect of long‐term ICV administration of leptin in ewes. We found that central application significantly decreased osteoblast activity as measured by serum analysis as well as by histomorphometry, resulting in decreased trabecular bone volume. These data provide additional evidence that bone formation and therefore bone remodeling is at least in part centrally controlled.

Control of bone formation by the serpentine receptor Frizzled-9.

Fzd9, induced upon osteoblast differentiation, is required for bone matrix mineralization in primary osteoblasts.

Increased Bone Formation in Mice Lacking Apolipoprotein E

ApoE is a plasma protein that plays a major role in lipoprotein metabolism. Here we describe that ApoE expression is strongly induced on mineralization of primary osteoblast cultures. ApoE‐deficient mice display an increased bone formation rate compared with wildtype controls, thereby showing that ApoE has a physiologic function in bone remodeling.