Differential Effects of Oral Doxercalciferol (Hectorol®) or Paricalcitol (Zemplar®) in the Cyp27b1-Null Mouse Model of Uremia

Authors

René St-Arnaud, Alice Arabian, Omar Akhouayri, Joyce C. Knutson, Stephen A. Strugnell

Abstract

Kidney disease patients experience declining calcitriol levels and develop secondary hyperparathyroidism (SHPT). Animal models of uremia based on 5/6 nephrectomy (NTX) do not consistently reproduce this calcitriol deficiency. We developed an animal model, the NTX Cyp27b1-null mouse, which completely lacks endogenous calcitriol, and examined the suitability of this model for evaluation of treatment with vitamin D analogs in uremia. Methods: NTX was performed at 2 months of age. One week post-NTX, animals were treated for 4 weeks with vehicle; doxercalciferol at 30, 100 or 300 pg/g body weight (b.w.); or paricalcitol at 100, 300 or 1,000 pg/g b.w. by gavage 3 times per week. Serum blood urea nitrogen and creatinine were elevated. Vehicle-treated NTX null mice had hypocalcemia and SHPT. Doxercalciferol at 100 or 300 pg/g b.w. normalized serum calcium and parathyroid hormone (PTH) levels. Paricalcitol at 300 or 1,000 pg/g normalized serum calcium, but PTH levels remained elevated. Osteomalacia was corrected by 100 pg/g b.w. of doxercalciferol or 1,000 pg/g b.w. of paricalcitol. The highest dose of doxercalciferol, but not of paricalcitol, significantly reduced osteitis fibrosa. Our results reveal the differential efficacy of doxercalciferol and paricalcitol in this novel animal model incorporating both calcitriol deficiency and renal insufficiency.

Link to Article

http://dx.doi.org/10.1159/000329663

Inhibition of β-catenin signaling in chondrocytes induces delayed fracture healing in mice

Authors

Yang Huang, Xiaoling Zhang, Kewei Du, Fei Yang, Yu Shi, Jingang Huang, Tingting Tang, Di Chen, Kerong Dai

Abstract

Appropriate and controlled chondrogenesis and endochondral ossification play fundamental roles in the fracture healing cascade, a regenerative process involved in highly coordinated biological events, including the Wnt/β-catenin signaling pathway. To examine the role and importance of this pathway in chondrocytes, we studied bone repair of closed tibias fractures in Col2a1-ICAT transgenic mice, in which the Wnt/β-catenin signaling pathway is specially inhibited in chondrocytes. Radiological, histological, and histomorphometric analyses at 7, 9, 12, 14, 21, and 28 days after fracture demonstrated the bone repairs were retarded in Col2a1-ICAT transgenic mice, due to reduced and delayed cartilage formation, chondrocyte hypertrophy, and bone generation. In addition, at 5 weeks, Col2a1-ICAT transgenic mice exhibited a weak mechanical tolerance to four-point bending. Furthermore, quantitative-PCR analysis revealed that the expression of genes associated specifically with cartilage extracellular matrix formation (collagen II, collagen X, and mmp13), bone remodeling (alp, collagen I, and osteocalcin), and vascular extravagation (vegf), and transcriptional activators involved in cartilage generation and ossification (sox9 and runx2) was decreased and delayed in the fracture sites of Col2a1-ICAT transgenic mice during healing. Collectively, these results suggest that Wnt/β-catenin signaling is critical for fracture healing, especially with respect to chondrogenesis and endochondral ossification. Thus, our study provides insight into the possible mechanisms of and therapeutic targets for improving normal facture repair and the healing of non-union fractures.

Link to Article

http://dx.doi.org/10.1002/jor.21505

Bone fragility and decline in stem cells in prematurely aging DNA repair deficient trichothiodystrophy mice

Authors

Karin E. M. Diderich, Claudia Nicolaije, Matthias Priemel, Jan H. Waarsing, Judd S. Day, Renata M. C. Brandt, Arndt F. Schilling, Sander M. Botter, Harrie Weinans and Gijsbertus T. J. van der Horst, et al.

Abstract

Trichothiodystrophy (TTD) is a rare, autosomal recessive nucleotide excision repair (NER) disorder caused by mutations in components of the dual functional NER/basal transcription factor TFIIH. TTD mice, carrying a patient-based point mutation in the Xpd gene, strikingly resemble many features of the human syndrome and exhibit signs of premature aging. To examine to which extent TTD mice resemble the normal process of aging, we thoroughly investigated the bone phenotype. Here, we show that female TTD mice exhibit accelerated bone aging from 39 weeks onwards as well as lack of periosteal apposition leading to reduced bone strength. Before 39 weeks have passed, bones of wild-type and TTD mice are identical excluding a developmental defect. Albeit that bone formation is decreased, osteoblasts in TTD mice retain bone-forming capacity as in vivo PTH treatment leads to increased cortical thickness. In vitro bone marrow cell cultures showed that TTD osteoprogenitors retain the capacity to differentiate into osteoblasts. However, after 13 weeks of age TTD females show decreased bone nodule formation. No increase in bone resorption or the number of osteoclasts was detected. In conclusion, TTD mice show premature bone aging, which is preceded by a decrease in mesenchymal stem cells/osteoprogenitors and a change in systemic factors, identifying DNA damage and repair as key determinants for bone fragility by influencing osteogenesis and bone metabolism.

Link to Article

http://dx.doi.org/10.1007/s11357-011-9291-8

The Effect of Bovine Bone Graft With or Without Platelet-Rich Plasma on Maxillary Sinus Floor Augmentation

Authors

Fatih Cabbar, Nurhan Güler, Mehmet Kürkcü, Ufuk Is̈eri, Kemal S̈ençift

Abstract

The purpose of this study was to compare Unilab Surgibone (USB) (Mississauga, Ontario, Canada), a bone xenograft (bovine), with platelet-rich plasma (PRP) and USB without PRP to augment the human maxillary sinus in preparation to receive dental implants. Patients who had bilateral pneumatized maxillary sinuses were included. Sinuses on one side were augmented with the USB-PRP combination and formed the study group, whereas the opposite-side sinuses were augmented with USB alone and served as controls. Bone biopsy specimens were taken during implant placement at 6.8 ± 0.9 months after maxillary sinus floor augmentation. Resonance frequency analysis measurements were performed at implant placement (first control) and before the prosthetic stage (second control), at 6.5 ± 0.7 months after implant surgery. There were 10 patients (7 men and 3 women; mean age, 53.7 ± 0.8 years). Integration between new bone and residual grafts was histologically observed in all samples. The volumes of soft tissue were 59.9% ± 7.5% and 57.8% ± 4.4% in the control and study groups, respectively; residual graft, 21.9% ± 6.6% and 23.6% ± 5.9%, respectively; new bone, 15.8% ± 4.8% and 16.0% ± 3.8%, respectively; and trabecular bone, 64.7% ± 22.5% and 69.1% ± 18.6%, respectively. A positive correlation was found between new bone volume and trabecular bone volume (P = .0001). The mean Implant Stability Quotient (ISQ) values were 71.7 ± 4.9 and 70.3 ± 5.7 in the control and study groups, respectively, at first control and 75.4 ± 6.4 and 74.4 ± 6.4, respectively, at second control. The mean ISQ values at second control in both groups were significantly higher than at first control (P = .043 and P = .028, respectively). No statistically significant differences were observed between groups (P > .05). The combination of USB and PRP does not have any effect on new bone formation and implant stabilization.

Link to Article

http://dx.doi.org/10.1016/j.joms.2011.03.040

Different effects on bone strength and cell differentiation in pre pubertal caloric restriction versus hypothalamic suppression

Authors

R.N. Joshi, F.F. Safadi, M. Barbe, Fe Del Carpio-Cano, S. Popoff and V.R. Yingling

Abstract

Hypothalamic amenorrhea and energy restriction during puberty affect peak bone mass accrual. One hypothesis suggests energy restriction alters hypothalamic function resulting in suppressed estradiol levels leading to bone loss. However, both positive and negative results have been reported regarding energy restriction and bone strength. Therefore, the purpose of this study was to investigate energy restriction and hypothalamic suppression during pubertal onset on bone mechanical strength and the osteogenic capacity of bone marrow-derived cells in two models: female rats treated with gonadotropin releasing hormone antagonists (GnRH-a) or 30% energy restriction. At 23 days of age, female Sprague Dawley rats were assigned to three groups: control group (C, n = 10), GnRH-a group (n = 10), and Energy Restriction (ER, n = 12) group. GnRH-a animals received daily injections for 27 days. The animals in the ER group received 70% of the control animals' intake. After sacrifice (50 days of age), body weight, uterine and muscle weights were measured. Bone marrow-derived stromal cells were cultured and assayed for proliferation and differentiation into osteoblasts. Outcome measures included bone strength, bone histomorphometry and architecture, serum IGF-1 and osteocalcin. GnRH-a suppressed uterine weight, decreased osteoblast proliferation, bone strength, trabecular bone volume and architecture compared to control. Elevated serum IGF-1 and osteocalcin levels and body weight were found. The ER model had an increase in osteoblast proliferation compared to the GnRH-a group, similar bone strength relative to body weight and increased trabecular bone volume in the lumbar spine compared to control. The ER animals were smaller but had developed bone strength sufficient for their size. In contrast, suppressed estradiol via hypothalamic suppression resulted in bone strength deficits and trabecular bone volume loss. In summary, our results support the hypothesis that during periods of nutritional stress the increased vertebral bone volume may be an adaptive mechanism to store mineral which differs from suppressed estradiol resulting from hypothalamic suppression.

Link to Article

http://dx.doi.org/10.1016/j.bone.2011.07.019

Conditional Inactivation of the CXCR4 Receptor in Osteoprecursors Reduces Postnatal Bone Formation Due to Impaired Osteoblast Development

Authors

Wei Zhu, Gang Liang, Zhiping Huang, Stephen B. Doty, Adele L. Boskey

Abstract

Cysteine (C)-X-C motif chemokine receptor 4 (CXCR4), the primary receptor for stromal cell-derived factor-1 (SDF-1), is involved in bone morphogenic protein 2 (BMP2)-induced osteogenic differentiation of mesenchymal progenitors. To target the in vivo function of CXCR4 in bone and explore the underlying mechanisms, we conditionally inactivated CXCR4 in osteoprecursors by crossing osterix (Osx)-Cre mice with floxed CXCR4 (CXCR4fl/fl) mice to generate knock-outs with CXCR4 deletion driven by the Osx promoter (Osx::CXCR4fl/fl). The Cre-mediated excision of CXCR4 occurred exclusively in bone of Osx::CXCR4fl/fl mice. When compared with littermate controls, Osx::CXCR4fl/fl mice developed smaller osteopenic skeletons as evidenced by reduced trabecular and cortical bone mass, lower bone mineral density, and a slower mineral apposition rate. In addition, Osx::CXCR4fl/fl mice displayed chondrocyte disorganization in the epiphyseal growth plate associated with decreased proliferation and collagen matrix syntheses. Moreover, mature osteoblast-related expression of type I collagen α1 and osteocalcin was reduced in bone of Osx::CXCR4fl/fl mice versus controls, suggesting that CXCR4 deficiency results in arrested osteoblast progression. Primary cultures for osteoblastic cells derived from Osx::CXCR4fl/fl mice also showed decreased proliferation and impaired osteoblast differentiation in response to BMP2 or BMP6 stimulation, and suppressed activation of intracellular BMP receptor-regulated Smads (R-Smads) and Erk1/2 was identified in CXCR4-deficient cells and bone tissues. These findings provide the first in vivo evidence that CXCR4 functions in postnatal bone development by regulating osteoblast development in cooperation with BMP signaling. Thus, CXCR4 acts as an endogenous signaling component necessary for bone formation.

Link to Article

http://dx.doi.org/10.1074/jbc.M111.250985