Black bear parathyroid hormone has greater anabolic effects on trabecular bone in dystrophin-deficient mice than in wild type mice

Authors

Sarah K. Gray, Meghan E. McGee-Lawrence, Jennifer L. Sanders, Keith W. Condon, Chung-Jui Tsai, Seth W. Donahue

Abstract

Duchenne muscular dystrophy (DMD) is an X-linked neuromuscular disease that has deleterious consequences in muscle and bone, leading to decreased mobility, progressive osteoporosis, and premature death. Patients with DMD experience a higher-than-average fracture rate, particularly in the proximal and distal femur and proximal tibia. The dystrophin-deficient mdx mouse is a model of DMD that demonstrates muscle degeneration and fibrosis and osteoporosis. Parathyroid hormone, an effective anabolic agent for post-menopausal and glucocorticoid-induced osteoporosis, has not been explored for DMD. Black bear parathyroid hormone (bbPTH) has been implicated in the maintenance of bone properties during extended periods of disuse (hibernation). We cloned bbPTH and found 9 amino acid residue differences from human PTH. Apoptosis was mitigated and cAMP was activated by bbPTH in osteoblast cultures. We administered 24 nmol/kg of bbPTH 1–84 to 4-week old male mdx and wild type mice via daily (5×/week) subcutaneous injection for 6 weeks. Vehicle-treated mdx mice had 44% lower trabecular bone volume fraction than wild type mice. No changes were found in femoral cortical bone geometry or mechanical properties with bbPTH treatment in wild type mice, and only medio-lateral moment of inertia changed with bbPTH treatment in mdx femurs. However, μCT analyses of the trabecular regions of the distal femur and proximal tibia showed marked increases in bone volume fraction with bbPTH treatment, with a greater anabolic response (7-fold increase) in mdx mice than wild type mice (2-fold increase). Trabecular number increased in mdx long bone, but not wild type bone. Additionally, greater osteoblast area and decreased osteoclast area were observed with bbPTH treatment in mdx mice. The heightened response to PTH in mdx bone compared to wild type suggests a link between dystrophin deficiency, altered calcium signaling, and bone. These findings support further investigation of PTH as an anabolic treatment for DMD-induced osteoporosis.

Link to Article

http://dx.doi.org/10.1016/j.bone.2012.05.003

Microdamage of the cortical bone during mini-implant insertion with self-drilling and self-tapping techniques: A randomized controlled trial

Authors

Sumit Yadav, Madhur Upadhyay, Sean Liu, Eugene Roberts, William P. Neace, Ravindra Nanda

Abstract

The purpose of this research was to evaluate microdamage accumulation after mini-implant placement by self-drilling (without a pilot hole) and self-tapping (screwed into a pilot hole) insertion techniques. The null hypothesis was that the mini-implant insertion technique would have no influence on microcrack accumulation and propagation in the cortical bones of the maxillae and mandibles of adult hounds. Mini-implants (n = 162; diameter, 1.6 mm; length, 6 mm) were placed in the maxillae and mandibles of 9 hounds (12-14 months old) with self-drilling and self-tapping insertion techniques. The techniques were randomly assigned to the left or the right side of each jaw. Each hound received 18 mini-implants (10 in the mandible, 8 in the maxilla). Histomorphometric parameters including total crack length and crack surface density were measured. The null hypothesis was rejected in favor of an alternate hypothesis: that the self-drilling technique results in more microdamage (microcracks) accumulation in the adjacent cortical bone in both the maxilla and the mandible immediately after mini-implant placement. A cluster level analysis was used to analyze the data on the outcome measured. Since the measurements were clustered within dogs, a paired-samples t test was used to analyze the average differences between insertion methods at both jaw locations. A significance level of 0.05 was used for both analyses. The self-drilling technique resulted in greater total crack lengths in both the maxilla and the mandible (maxilla: mean difference, 18.70 ± 7.04 μm/mm2; CI, 13.29-24.11; mandible: mean difference, 22.98 ± 6.43 μm/mm2; CI, 18.04-27.93; P <0.05), higher crack surface density in both the maxilla and the mandible (maxilla: mean difference, 10.39 ± 9.16 μm/mm2; CI, 3.34-17.43; mandible: mean difference, 11.28 ± 3.41 μm/mm2; CI, 8.65-13.90; P <0.05). This study demonstrated greater microdamage in the cortical bones of adult hounds in both the maxilla and the mandible by the self-drilling insertion technique compared with the self-tapping technique.

Link to Article

http://dx.doi.org/10.1016/j.ajodo.2011.12.016

NELL-1 increases pre-osteoblast mineralization using both phosphate transporter Pit1 and Pit2

Authors

Catherine M. Cowan, Xinli Zhang, Aaron W. James, T. Mari Kim, Nichole Sun, Chia Soo, Benjamin Wu, Kang Ting

Abstract

NELL-1 is a potent osteoinductive molecule that enhances bone formation in multiple animal models through currently unidentified pathways. In the present manuscript, we hypothesized that NELL-1 may regulate osteogenic differentiation accompanied by alteration of inorganic phosphate (Pi) entry into the osteoblast via sodium dependent phosphate (NaPi) transporters. To determine this, MC3T3-E1 pre-osteoblasts were cultured in the presence of recombinant human (rh)NELL-1 or rhBMP-2. Analysis was performed for intracellular Pi levels through malachite green staining, Pit-1 and Pit-2 expression, and forced upregulation of Pit-1 and Pit-2. Results showed rhNELL-1 to increase MC3T3-E1 matrix mineralization and Pi influx associated with activation of both Pit-1 and Pit-2 channels, with significantly increased Pit-2 production. In contrast, Pi transport elicited by rhBMP-2 showed to be associated with increased Pit-1 production only. Next, neutralizing antibodies against Pit-1 and Pit-2 completely abrogated the Pi influx effect of rhNELL-1, suggesting rhNELL-1 is dependent on both transporters. These results identify one potential mechanism of action for rhNELL-1 induced osteogenesis and highlight a fundamental difference between NELL-1 and BMP-2 signaling.

Link to Article

http://dx.doi.org/10.1016/j.bbrc.2012.04.077

The use of autologous enriched bone marrow MSCs to enhance osteoporotic bone defect repair in long-term estrogen deficient goats

Authors

Lei Cao, Guangwang Liu, Yaokai Gan, Qiming Fan, Fei Yang, Xiaoling Zhang, Tingting Tang, Kerong Dai

Abstract

Bone defects are common in elderly patients suffering from osteoporosis. Current methods of bone defect treatment for osteoporosis are not always satisfactory. In this study, we demonstrated that bone marrow mesenchymal stem cells (MSCs) harvested from goats with long-term estrogen deficiencies exhibited a lower proliferation rate and decreased osteogenic capacity, which are critical obstacles for bone defect repair in the elderly. However, by combining autologous enriched bone marrow mesenchymal stem cells with porous β-TCP, we successfully repaired critical-sized bone defects in the medial femoral condyle of the osteoporotic goats. Both micro-CT images and histomorphometry analysis illustrated improved bone formation following the enriched MSC therapy. Thus, we proposed autologous enriched bone marrow mesenchymal stem cells as a quick, safe therapeutic strategy to treat osteoporotic bone defects.

Link to Article

http://dx.doi.org/10.1016/j.biomaterials.2012.03.069

Assessment of changes to the dental follicle in deep and partially impacted mandibular third molar

Author

Kamal, Abeer; Allam, Eman; Dehis, Mohammed; Zunt, Susan; Windsor, L. Jack

Abstract

Residual dental follicles surrounding the crowns of impacted teeth may represent a significant risk for their potential pathological changes. In this study, the expression levels of cell cycle proteins, proliferating cell nuclear antigen (PCNA), cyclin D1, and p21 were evaluated using immunohistochemistry. Twenty-two impacted mandibular third molars in 17 healthy individuals were evaluated in this study. They were divided into two groups on the basis of the depth of impaction: the partially impacted group and the fully impacted group. The impacted teeth were surgically removed and the surrounding follicle was curetted and examined by immunohistochemistry to evaluate whether there was a difference in cell cycle regulation between the two groups. PCNA was expressed in most of the epithelial cells with no significant difference between the groups. The p21 expression was significantly higher in the full bony impaction group. Cyclin D1 expression was expressed at low levels with no significant difference between the groups. Weak cyclin D1 expression was consistent with high p21, a negative regulator of the cell cycle. The high p21 expression could be a compensatory mechanism to the high PCNA levels. These results suggest the possibility that dental follicles associated with full bony impactions possess different cellular activities compared with partial bony/soft-tissue impactions.

Link to Article

http://dx.doi.org/10.1097/01.OMX.0000412880.68174.21

IL-17 mediates estrogen-deficient osteoporosis in an Act1-dependent manner

Authors

Carl J. DeSelm, Yoshifumi Takahata, Julia Warren, Jean C. Chappel, Taimur Khan, Xiaoxia Li, Caini Liu, Yongwon Choi, Youngmi Faith Kim, Wei Zou, Steven L. Teitelbaum

Abstract

Estrogen-deficient osteoporosis may be an inflammatory disorder and we therefore asked if IL-17 participates in its pathogenesis. Deletion of the principal IL-17 receptor (IL-17RA) protects mice from ovariectomy (OVX)-induced bone loss. Further supporting a central role of IL-17 in its pathogenesis, OVX-induced osteoporosis is prevented by a blocking antibody targeting the cytokine. IL-17 promotes osteoclastogenesis by stimulating RANK ligand (RANKL) expression by osteoblastic cells, mediated by the IL-17RA SEFIR/TILL domain. Estrogen deprivation, however does not enhance IL-17RA mRNA expression by osteoblasts or in bone, but augments that of Act1, an IL17RA-interacting protein and signaling mediator. Similar to IL-17RA-/- mice, those lacking Act1 are protected from OVX-induced bone loss. Also mirroring IL-17RA-deficiency, absence of Act1 in osteoblasts, but not osteoclasts, impairs osteoclastogenesis via dampened RANKL expression. Transduction of WT Act1 into Act1-/- osteoblasts substantially rescues their osteoclastogenic capacity. The same construct, however, lacking its E3 ligase U-box or its SEFIR domain, which interacts with its counterpart in IL-17RA, fails to do so. Estrogen deprivation, therefore, promotes RANKL expression and bone resorption in association with upregualtion of the IL-17 effector, Act1, supporting the concept that post-menopausal osteoporosis is a disorder of innate immunity.

Link to Article

http://dx.doi.org/10.1002/jcb.24165