Can OP-1 stimulate union in a rat model of pathological fracture post treatment for soft tissue sarcoma?

Authors

Fred Nicholls, Adeline H. Ng, Sally Hu, Katarina Janic, Cara Fallis, Thomas Willett, Marc Grynpas, and Peter Ferguson

Abstract

The goal of soft tissue sarcoma management in the extremities is limb preservation, often combining surgery and external beam radiation. In patients who have undergone this therapy in the thigh, pathologic fracture is a serious, late complication. Non-union rates of 80–90% persist. No reliable biologic solution exists. A rat model combining one 18 Gy dose of radiation and diaphyseal periosteal excision reliably generates atrophic non-union of femoral fractures. We hypothesized that augmentation with OP-1 would increase union rate. Female Sprague-Dawley retired breeder rats were randomized to Control, Disease (external beam radiotherapy and periosteal stripping), Control + OP-1 (80 µg) and Disease + OP-1 groups. Animals underwent prophylactic fixation and controlled left femur fracture. Twenty-eight, 35, and 42 days post-fracture were end-points. Femora were analyzed using MicroCT, Back Scattered Electron Microscopy, and Histomorphometry. We observed a 2% union rate in the Disease groups (±OP-1 treatment). The union rate in Control groups was 97%. MicroCT demonstrated a lack of callus volume in Disease groups. Heterotopic ossification was observed in some OP-1 treated animals. The ineffectiveness of OP-1 in stimulating fracture union in this model suggests the endogenous repair mechanism has been compromised beyond the capabilities of osteoinductive biologics.

Link To Article

http://dx.doi.org/10.1002/jor.22661

A novel role for interferon regulatory factor 1 (IRF1) in regulation of bone metabolism

Authors

Sandra Salem, Chan Gao, Ailian Li, Huifen Wang, Loan Nguyen-Yamamoto, David Goltzman, Janet E. Henderson, and Philippe Gros

Abstract

Increased risk of bone fractures is observed in patients with chronic inflammatory conditions, such as inflammatory bowel disease and rheumatoid arthritis. Members of the Interferon Response Factor family of transcriptional regulators, IRF1 and IRF8, have been identified as genetic risk factors for several chronic inflammatory and autoimmune diseases. We have investigated a potential role for the Irf1 gene in bone metabolism. Here, we report that Irf1−/−mutant mice show altered bone morphology in association with altered trabecular bone architecture and increased cortical thickness and cellularity. Ex vivo studies on cells derived from bone marrow stimulated with Rank ligand revealed an increase in size and resorptive activity of tartrate-resistant acid-positive cells from Irf1−/− mutant mice compared with wild-type control mice. Irf1 deficiency was also associated with decreased proliferation of bone marrow-derived osteoblast precursors ex vivo, concomitant with increased mineralization activity compared with control cells. We show that Irf1 plays a role in bone metabolism and suggest that Irf1 regulates the maturation and activity of osteoclasts and osteoblasts. The altered bone phenotype of Irf1−/− mutants is strikingly similar to that of Stat1−/− mice, suggesting that the two interacting proteins play a critical enabling role in the common regulation of these two cell lineages.

Link To Article

http://dx.doi.org/10.1111/jcmm.12327

Increased fracture callus mineralization and strength in cathepsin K knockout mice

Authors

Michael A. Gentile, Do Y. Soung, Carlyle Horrell, Rana Samadfam, Hicham Drissi, Le T. Duong

Abstract

Cathepsin K (CatK) is a cysteine protease, expressed predominantly in osteoclasts (OC) which degrades demineralized bone matrix. Novel selective inhibitors of CatK are currently being developed for the treatment of postmenopausal osteoporosis. Pharmacological inhibition of CatK reduces OC resorption activity while preserving bone formation in preclinical models. Disruption of the CatK gene in mice also results in high bone mass due to impaired bone resorption and elevated formation. Here, we assessed mid-shaft femoral fracture healing in 8–10 week old CatK knock-out (KO) versus wild type (WT) mice. Fracture healing and callus formation were determined in vivo weekly via X-ray, and ex vivo at days 14, 18, 28 and 42 post-fracture by radiographic scoring, micro-computed tomography (μCT), histomorphometry and terminal mechanical four point bend strength testing. Radiological evaluation indicated accelerated bone healing and remodeling for CatK KO animals based on increased total radiographic scores that included callus opacity and bridging at days 28 and 42 post-fracture. Micro-CT based total callus volume was similar in CatK KO and WT mice at day 14. Callus size in CatK KO mice was 25% smaller than that in WT mice at day 18, statistically significant by day 28 and exhibited significantly higher mineralized tissue volume and volumetric BMD as compared to WT by day 18 onward. Osteoclast surface and osteoid surface trended higher in CatK KO calluses at all time-points and osteoblast number was also significantly increased at day 28. Increased CatK KO callus mineral density was reflected in significant increases in peak load and stiffness over WT at day 42 post-fracture. Regression analysis indicated a positive correlation (r = 0.8671; p < 0.001) between callus BMC and peak load indicating normal mineral properties in CatK KO calluses. Taken together, gene deletion of cathepsin K in mice accelerated callus size resolution, significantly increased callus mineralized mass, and improved mechanical strength as compared to wild type mice.

Link To Article

http://dx.doi.org/10.1016/j.bone.2014.04.032

Tissue-engineered bone constructed in a bioreactor for repairing critical-sized bone defects in sheep

Authors

Deqiang Li, Ming Li, Peilai Liu, Yuankai Zhang, Jianxi Lu, Jianmin Li

Abstract

Purpose Repair of bone defects, particularly critical-sized bone defects, is a considerable challenge in orthopaedics. Tissue-engineered bones provide an effective approach. However, previous studies mainly focused on the repair of bone defects in small animals. For better clinical application, repairing critical-sized bone defects in large animals must be studied. This study investigated the effect of a tissue-engineered bone for repairing critical-sized bone defect in sheep.

Methods A tissue-engineered bone was constructed by culturing bone marrow mesenchymal-stem-cell-derived osteoblast cells seeded in a porous β-tricalcium phosphate ceramic (β-TCP) scaffold in a perfusion bioreactor. A critical-sized bone defect in sheep was repaired with the tissue-engineered bone. At the eighth and 16th week after the implantation of the tissue-engineered bone, X-ray examination and histological analysis were performed to evaluate the defect. The bone defect with only the β-TCP scaffold served as the control.

Result X-ray showed that the bone defect was successfully repaired 16 weeks after implantation of the tissue-engineered bone; histological sections showed that a sufficient volume of new bones formed in β-TCP 16 weeks after implantation. Eight and 16 weeks after implantation, the volume of new bones that formed in the tissue-engineered bone group was more than that in the β-TCP scaffold group (P < 0.05).

Conclusion Tissue-engineered bone improved osteogenesis in vivo and enhanced the ability to repair critical-sized bone defects in large animals.

Link To Article

http://dx.doi.org/10.1007/s00264-014-2389-8

Immunohistological expression of human ß-defensin-1 and human ß-defensin-2 in exacerbation of acute and secondary chronic osteomyelitis of the mandible

Authors

Benedicta E. Beck-Broichsitter, Heino Dau, Tobias Moest, Arne Jochens, Philipp Stockmann, Jörg Wiltfang and Stephan T. Becker

Abstract

Background The majority of patients diagnosed with osteomyelitis of the jaw have severe complaints. Unfortunately, the pathogenesis still remains unclear. Human ß-defensins expressed in epithelial and bone tissues as a part of the innate immunity may be involved in disease development. In this study, we hypothesize that expression levels of human ß-defensin-1 and -2 in the acute and secondary chronic osteomyelitis may be altered in comparison with healthy bone and with bisphosphonate-associated necrosis as well as irradiation from a previous study.

Method Bone samples were collected during surgical debridement in a total of eight patients suffering from acute or secondary chronic osteomyelitis of the jaw. Expression levels of hBD-1 and -2 were quantified and related to non-stained cells. Ratios were compared by one-way ANOVA and multiple tests by Holm–Bonferroni.

Results Multiple testing revealed no significant differences for expression levels of human ß-defensin-1 between all groups, whereas labeling index of human ß-defensin-2 was significantly different between specimens of bisphosphonate-associated osteonecrosis of the jaws and all other groups. No significant difference occurred between samples of floride osteomyelitis and healthy bone for expression of hBD-1 and -2.

Conclusions Although the affected patients showed all clinical signs of acute inflammation, expression levels in acute and secondary chronic osteomyelitis in the jaws did not reveal statistically significant differences compared with healthy bone samples. The weak immunological host response in terms of a putative genetically predisposition should be further discussed as pathogenesis factor for osteomyelitis in the future.

Link To Article

http://dx.doi.org/10.1111/jop.12202

Effects of sodium acetate buffer on chitosan sponge properties and in vivo degradation in a rat intramuscular model

Authors

Ashley Cox Parker, James Keaton Smith, Benjamin Reves, Jessica Amber Jennings, Joel D. Bumgardner and Warren O. Haggard

Abstract

Chitosan sponges were developed for adjunctive local antibiotic delivery to reduce bacteria in wounds. There is a need to increase sponge degradation for rapid clearance from the wound site during initial wound care. This work examined the effect of using 0.25 M sodium acetate buffers, at pH 4.6 or 5.6, to fabricate sponges with an amorphous chitosan polymer structure. Sponges were evaluated for their crystallinity, thermal, spectroscopic, and morphological properties, in addition to in vitro degradation, and cytocompatibility analysis using normal human dermal fibroblasts. In vivo degradation and biocompatibility were also examined after 4 and 10 days in rat intramuscular tissues. Both buffered chitosan sponge variations exhibited decreases in crystallinity and thermal decomposition temperatures, and increases in surface roughness, which resulted in over 40% increases in degradation over 10 days in vitro compared to the neutral sponges. There were no significant differences between sponges during in vivo degradation over 10 days with respect to histomorphometric analysis of the recovered sponges. These results demonstrated that the acetate buffer did change characteristic chitosan sponge material properties, and increasing the in vivo sponge degradation rate will require balancing material characteristics and processing.

Link To Article

http://dx.doi.org/10.1002/jbm.b.33204