Diverse Osteoclastogenesis of Bone Marrow From Mandible Versus Long Bone

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141128/1/jper0829.pd

Nanodiamond-Gutta Percha Composite Biomaterials for Root Canal Therapy.

Root canal therapy (RCT) represents a standard of treatment that addresses infected pulp tissue in teeth and protects against future infection. RCT involves removing dental pulp comprising blood vessels and nerve tissue, decontaminating residually infected tissue through biomechanical instrumentation, and root canal obturation using a filler material to replace the space that was previously composed of dental pulp. Gutta percha (GP) is typically used as the filler material, as it is malleable, inert, and biocompatible. While filling the root canal space with GP is the standard of care for endodontic therapies, it has exhibited limitations including leakage, root canal reinfection, and poor mechanical properties. To address these challenges, clinicians have explored the use of alternative root filling materials other than GP. Among the classes of materials that are being explored as novel endodontic therapy platforms, nanodiamonds (NDs) may offer unique advantages due to their favorable properties, particularly for dental applications. These include versatile faceted surface chemistry, biocompatibility, and their role in improving mechanical properties, among others. This study developed a ND-embedded GP (NDGP) that was functionalized with amoxicillin, a broad-spectrum antibiotic commonly used for endodontic infection. Comprehensive materials characterization confirmed improved mechanical properties of NDGP over unmodified GP. In addition, digital radiography and microcomputed tomography imaging demonstrated that obturation of root canals with NDGP could be achieved using clinically relevant techniques. Furthermore, bacterial growth inhibition assays confirmed drug functionality of NDGP functionalized with amoxicillin. This study demonstrates a promising path toward NDGP implementation in future endodontic therapy for improved treatment outcomes

Osteogenic Oxysterol, 20(S)-Hydroxycholesterol, Induces Notch Target Gene Expression in Bone Marrow Stromal Cells

We previously reported that specific oxysterols stimulate osteogenic differentiation of pluripotent bone marrow stromal cells (MSCs) through activation of hedgehog (Hh) signaling and may serve as potential future therapies for intervention in osteopenia and osteoporosis. In this study we report that the osteogenic oxysterol 20(S)-hydroxycholesterol (20S) induces the expression of genes associated with Notch signaling. Using M2-10B4 (M2) MSCs, we found that 20S significantly induced HES-1, HEY-1, and HEY-2 mRNA expression compared with untreated cells, with maximal induction after 48 hours, whereas the nonosteogenic oxysterols did not. Similar observations were made when M2 cells were treated with sonic hedgehog (Shh), and the specific Hh pathway inhibitor cyclopamine blocked 20S-induced Notch target gene expression. 20S did not induce Notch target genes in Smo−/− mouse embryonic fibroblasts, further confirming the role of Hh signaling in 20S-induced expression of Notch target genes. Despite the inability of liver X-receptor (LXR) synthetic ligand TO901317 to induce Notch target genes in M2 cells, LXR knockdown studies using siRNA showed inhibition of 20S-induced HEY-1 but not HES-1 expression, suggesting the partial role of LXR signaling in MSC responses to 20S. Moreover, 20S-induced Notch target gene expression was independent of canonical Notch signaling because neither 20S nor Shh induced CBF1 luciferase reporter activity or NICD protein accumulation in the nucleus, which are hallmarks of canonical Notch signaling activation. Finally, HES-1 and HEY-1 siRNA transfection significantly inhibited 20S-induced osteogenic genes, suggesting that the pro-osteogenic effects of 20S are regulated in part by HES-1 and HEY-1. © 2010 American Society for Bone and Mineral Researc

The effect of three hemostatic agents on early bone healing in an animal model

<p>Abstract</p> <p>Background</p> <p>Resorbable bone hemostasis materials, oxidized regenerated cellulose (ORC) and microfibrillar collagen (MFC), remain at the site of application for up to 8 weeks and may impair osteogenesis. Our experimental study compared the effect of a water-soluble alkylene oxide copolymer (AOC) to ORC and MFC versus no hemostatic material on early bone healing.</p> <p>Methods</p> <p>Two circular 2.7 mm non-critical defects were made in each tibia of 12 rabbits. Sufficient AOC, ORC or MFC was applied to achieve hemostasis, and effectiveness recorded. An autologous blood clot was applied to control defects. Rabbits were sacrificed at 17 days, tibiae excised and fixed. Bone healing was quantitatively measured by micro-computed tomography (micro-CT) expressed as fractional bone volume, and qualitatively assessed by histological examination of decalcified sections.</p> <p>Results</p> <p>Hemostasis was immediate after application of MFC and AOC, after 1-2 minutes with ORC, and >5 minutes for control. At 17 days post-surgery, micro-CT analysis showed near-complete healing in control and AOC groups, partial healing in the ORC group and minimal healing in the MFC group. Fractional bone volume was 8 fold greater in the control and AOC groups than in the MFC group (0.42 ± 0.06, 0.40 ± 0.03 vs 0.05 ± 0.01, <it>P </it>< 0.001) and over 1.5-fold greater than in the ORC group (0.25 ± 0.03, <it>P </it>< 0.05). By histology, MFC remained at the application site with minimal healing at the defect margins and early fibrotic tissue within the defect. ORC-treated defects showed partial healing but with early fibrotic tissue in the marrow space. Conversely, control and AOC-treated defects demonstrated newly formed woven bone rich in cellular activity with no evidence of AOC remaining at the application site.</p> <p>Conclusions</p> <p>Early healing appeared to be impaired by the presence of MFC and impeded by the presence of ORC. In contrast, AOC did not inhibit bone healing and suggest that AOC may be a better bone hemostatic material for procedures where bony fusion is critical and immediate hemostasis required.</p

OPG‐Fc but Not Zoledronic Acid Discontinuation Reverses Osteonecrosis of the Jaws (ONJ) in Mice

Osteonecrosis of the jaws (ONJ) is a significant complication of antiresorptive medications, such as bisphosphonates and denosumab. Antiresorptive discontinuation to promote healing of ONJ lesions remains highly controversial and understudied. Here, we investigated whether antiresorptive discontinuation alters ONJ features in mice, employing the potent bisphosphonate zoledronic acid (ZA) or the receptor activator of NF‐κB ligand (RANKL) inhibitor OPG‐Fc, utilizing previously published ONJ animal models. Mice were treated with vehicle (veh), ZA, or OPG‐Fc for 11 weeks to induce ONJ, and antiresorptives were discontinued for 6 or 10 weeks. Maxillae and mandibles were examined by μCT imaging and histologically. ONJ features in ZA and OPG‐Fc groups included periosteal bone deposition, empty osteocyte lacunae, osteonecrotic areas, and bone exposure, each of which substantially resolved 10 weeks after discontinuing OPG‐Fc but not ZA. Full recovery of tartrate‐resistant acid phosphatase‐positive (TRAP+) osteoclast numbers occurred after discontinuing OPG‐Fc but not ZA. Our data provide the first experimental evidence demonstrating that discontinuation of a RANKL inhibitor, but not a bisphosphonate, reverses features of osteonecrosis in mice. It remains unclear whether antiresorptive discontinuation increases the risk of skeletal‐related events in patients with bone metastases or fracture risk in osteoporosis patients, but these preclinical data may nonetheless help to inform discussions on the rationale for a “drug holiday” in managing the ONJ patient. © 2015 American Society for Bone and Mineral Research.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/113163/1/jbmr2490-sup-0001-SupFigLeg-S1.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/113163/2/jbmr2490.pd

Novel Oxysterols Have Pro-Osteogenic and Anti-Adipogenic Effects In Vitro and Induce Spinal Fusion In Vivo

ABSTRACT Stimulation of bone formation by osteoinductive materials is of great clinical importance in spinal fusion surgery, repair of bone fractures, and in the treatment of osteoporosis. We previously reported that specific naturally occurring oxysterols including 20(S)-hydroxycholesterol (20S) induce the osteogenic differentiation of pluripotent mesenchymal cells, while inhibiting their adipogenic differentiation. Here we report the characterization of two structural analogues of 20S, Oxy34 and Oxy49, which induce the osteogenic and inhibit the adipogenic differentiation of bone marrow stromal cells (MSC) through activation of Hedgehog (Hh) signaling. Treatment of M2-10B4 MSC with Oxy34 or Oxy49 induced the expression of osteogenic differentiation markers Runx2, Osterix (Osx), alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin (OCN), as well as ALP enzymatic activity and robust mineralization. Treatment with oxysterols together with PPARg activator, troglitazone (Tro), inhibited mRNA expression for adipogenic genes PPARg, LPL, and aP2, and inhibited the formation of adipocytes. Efficacy of Oxy34 and Oxy49 in stimulating bone formation in vivo was assessed using the posterolateral intertransverse process rat spinal fusion model. Rats receiving collagen implants with Oxy 34 or Oxy49 showed comparable osteogenic efficacy to BMP2/collagen implants as measured by radiography, MicroCT, and manual inspection. Histological analysis showed trabecular and cortical bone formation by oxysterols and rhBMP2 within the fusion mass, with robust adipogenesis in BMP2-induced bone and significantly less adipocytes in oxysterol-induced bone. These data suggest that Oxy34 and Oxy49 are effective novel osteoinductive molecules and may be suitable candidates for further development and use in orthopedic indications requiring local bone formation

Article Novel Oxysterols Have Pro-Osteogenic and Anti-Adipogenic Effects In Vitro and Induce Spinal Fusion In Vivo

Histological analysis showed trabecular and cortical bone formation by oxysterols and rhBMP2 within the fusion mass, with robust adipogenesis in BMP2-induced bone and significantly less adipocytes in oxysterol-induced bone. These data suggest that Oxy34 and Oxy49 are effective novel osteoinductive molecules and may be suitable candidates for further development and use in orthopaedic indications requiring local bone formation

Regulation of the prostaglandin G/H synthase gene by parathyroid hormone in osteoblastic cells

Parathyroid hormone (PTH), the major regulator of serum calcium levels, has important effects on bone metabolism. Binding of PTH to its receptor initiates protein kinase A (PKA), protein kinase C (PKC) and calcium pathways. PTH induces release of prostaglandins (PGs) by osteoblastic cells. Prostaglandin G/H synthase (PGHS), a central enzyme in the synthesis of PGs, is found in the constitutive PGHS-1 and the inducible PGHS-2 forms. The objective of this thesis was to study the mechanisms by which PTH induces PGHS-2 expression in the osteoblastic MC3T3-E1 cells.^ PTH rapidly and transiently induced PGHS-2 mRNA levels that peaked at one and returned to baseline by six h. PTH increased PGHS-2 transcription suggesting that PTH regulates the PGHS-2 mRNA levels, at least in part, through transcriptional activation. Experiments with inhibitors and mimetics showed that PKA is the main pathway mediating PTH induction of PGHS-2 expression. PTH superinduced PGHS-2 mRNA in the presence of cycloheximide, suggesting that PGHS-2 is a primary response gene. While cycloheximide did not affect PTH induction of PGHS-2 transcription at 0.5 h, cycloheximide did block the subsequent decline of PGHS-2 transcription at 2 h. Therefore, PTH induces the synthesis of a factor(s) that attenuates PGHS-2 transcription.^ The inducible cAMP early repressor (ICER) is a transcriptional inhibitor induced by the cAMP/PKA pathway and transcribed from an intronic promoter of the CREM gene. PTH rapidly induced ICER mRNA levels in MC3T3-E1 cells and calvarial organ cultures suggesting that ICER could mediate the attenuation of the PTH-induced PGHS-2 transcription.^ To identify promoter elements that mediate the PTH regulation, MC3T3-E1 cells were stably transfected with PGHS-2 promoter constructs driving a luciferase reporter. Deletion analysis revealed that elements downstream of $-$150 bp mediate baseline and PTH-mediated promoter activity. Electrophoretic mobility shift assays showed that the strongest binding to the $-$150/ + 70 bp promoter probe was competed by a $-$63/$-$42 bp fragment that contains overlapping CRE and E-box response elements. However, mutations of the CRE and/or E-box sites did not affect PTH stimulation of luciferase activity, suggesting that neither the consensus CRE at $-$57/$-$51 bp, nor the E-box at $-$52/$-$47 bp mediate the PTH and FSK effects.

Imaging of Radiation- and Medication-Related Osteonecrosis

Osteonecrosis is the devitalization of bone and consequent lytic changes. In the jaws, osteonecrosis is a pathologic consequence of prior radiation therapy (osteoradionecrosis) or certain antiresorptive medications. Herein, we review the pathogenesis and clinical manifestations of these lesions, and describe the spectrum of radiologic findings in these conditions, and highlight the similarities and differences between the imaging appearances of these 2 entities