The feasibility of bone transport with bone substitute and the factors which are essential for a successful bone transport are unknown. We studied six groups of 12 Japanese white rabbits. Groups A to D received cylindrical autologous bone segments and groups E and F hydroxyapatite prostheses. The periosteum was preserved in group A so that its segments had a blood supply, cells, proteins and scaffold. Group B had no blood supply. Group C had proteins and scaffold and group D had only scaffold. Group E received hydroxyapatite loaded with recombinant human bone morphogenetic protein-2 and group F had hydroxyapatite alone. Distraction osteogenesis occurred in groups A to C and E which had osteo-conductive transport segments loaded with osteo-inductive proteins. We conclude that scaffold and proteins are essential for successful bone transport, and that bone substitute can be used to regenerate bone

Segmental bone transport (SBT) with an external fixator has become a standard method for treatment of large bone defect. However, a long time-application of devices can be very troublesome and complications such as nonunion is sometimes seen at docking site. Although there have been several studies on SBT with large animal models, they were unsuitable for conducting drug application to improve SBT. The purpose of this study was to establish a bone transport model in mice. Six-month-old C57BL/6J mice were divided randomly into bone transport group (group BT) and an immobile control group (group EF). In each group, a 2-mm bone defect was created in the right femur. Group BT was reconstructed by SBT with external fixator (MouseExFix segment transport, RISystem, Switzerland) and group EF was fixed simply with unilateral external fixator (MouseExFix simple). In group BT, a bone segment was transported by 0.2 mm per day. Radiological and histological studies were conducted at 3 and 8 weeks after the surgery. In group BT, radiological data showed regenerative new bone consolidation at 8 weeks after the surgery, whereas high rate of nonunion was observed at the docking site. Histological data showed intramembranous and endochondral ossification. Group EF showed no bone union. In this study, experimental group showed good regenerative new bone formation and was similar ossification pattern to previous large animal models. Thus, the utilization of this bone defect mice model allows to design future studies with standardized mechanical conditions for analyzing mechanisms of bone regeneration induced by SBT

Segmental bone transport (SBT) using an external fixator is currently a standard treatment for large-diameter bone defects at the donor site with low morbidity. However, long-term application of the device is needed for bone healing. In addition, patients who received SBT treatment sometimes fail to show bone repair and union at the docking site, and require secondary surgery. The objective of this study was to investigate whether a single injection of recombinant human bone morphogenetic protein 2 (rhBMP-2)-loaded artificial collagen-like peptide gel (rhBMP-2/ACG) accelerates consolidation and bone union at the docking site in a mouse SBT model. Six-month-old C57BL/6J mice were reconstructed by SBT with external fixator that has transport unit, and a 2.0-mm bone defect was created in the right femur. Mice were divided randomly into four treatment groups with eight mice in each group, Group CONT (immobile control), Group 0.2mm/d, Group 1.0mm/d, and Group BMP-2. Mice in Group 0.2mm/d and 1.0mm/d, bone segment was moved 0.2 mm per day for 10 days and 1.0 mm per day for 2 days, respectively. Mice in Group BMP-2 received an injection of 2.0 μg of rhBMP-2 dissolved in ACG into the bone defect site immediately after the defect-creating surgery and the bone segment was moved 1.0 mm/day for 2 days. All animals were sacrificed at eight weeks after surgery. Consolidation at bone defect site and bone union at docking site were evaluated radiologically and histologically. At the bone defect site, seven of eight mice in Group 0.2mm/d and two of eight mice in Group 1.0mm/d showed bone union. In contrast, all mice in Group CONT showed non-union at the bone defect site. At the docking site, four of eight mice in Group 0.2 mm/d and three of eight mice in Group 1.0 mm/d showed non-union. Meanwhile, all mice in Group BMP-2 showed bone union at the bone defect and docking sites. Bone volume and bone mineral content were significantly higher in Group 0.2mm/d and Group BMP-2 than in Group CONT. HE staining of tissue from Group 0.2mm/d and Group BMP-2 showed large amounts of longitudinal trabecular bone and regenerative new bone at eight weeks after surgery at the bone defect site. Meanwhile, in Group CONT and Group 1.0mm/d, maturation of regenerative bone at the bone defect site was poor. Differences between groups were analyzed using one-way ANOVA and a subsequent Bonferroni's post-hoc comparisons test. P < 0.05 was considered significant. rhBMP-2/ACG combined with SBT may be effective for enhancing bone healing in large bone defects without the need for secondary procedures

Based on Ilizarov's law of tension-stress principle, distraction histogenesis technique has been widely applied in orthopaedic surgery for decades. Derived from this technique, cranial bone transport technique was mainly used for treating cranial deformities and calvarial defects. Recent studies reported that there are dense short vascular connections between skull marrow and meninges for immune cells trafficking, highlighting complex and tight association between skull and brain. Alzheimer's disease (AD) is a progressive neurodegenerative disease and the most common cause of dementia without effective therapy. Meningeal lymphatics have been recognized as an important mediator in neurological diseases. The augmentation of meningeal lymphatic drainage might be a promising therapeutic target for AD. Our proof-of-concept study has indicated that cranial bone transport can promote ischemic stroke recovery via modulating meningeal lymphatic drainage function, providing a rationale for treating AD using cranial bone maneuver (CBM). This study aims to investigate the effects of CBM on AD and to further explore the potential mechanisms. Transgenic 5xFAD mice model was used in this study. After osteotomy, a bone flap was used to perform CBM without damaging the dura. Open filed test, novel object recognition test and Barn's maze test were used to evaluate neurological functions of 5xFAD mice after CBM treatment. Congo red and immunofluorescence staining were used to evaluate amyloid depositions and Aβ plaques in different brain regions. Lymphangiogenesis and the level of VEGF-C were examined after CBM treatment. OVA-A647 was intra-cisterna-magna injected to evaluate meningeal lymphatic drainage function after CBM treatment. CBM significantly improved memory functions and reduced amyloid depositions and Aβ plaques in the hippocampus of 5xFAD mice. A significant increase of meningeal lymphatic vessels in superior sagittal sinus and transverse sinus, and the upregulation of VEGF-C in meninges were observed in 5xFAD mice treated with CBM. Moreover, CBM remarkably enhanced meningeal lymphatic drainage function in 5xFAD mice (n=5-16 mice/group for all studies). CBM may promote meningeal lymphangiogenesis and lymphatic drainage function through VEGF-C-VEGFR3 pathway, and further reduce amyloid depositions and Aβ plaques and alleviate memory deficits in AD

Introduction and Objective. Management of gap non-union of the tibia, the major weight bearing bone of the leg remains controversial. The different internal fixation techniques are often weighed down by relatively high complication rates that include fractures which fail to heal (non-union). Minimally invasive techniques with ring fixators and bone transport (distraction osteogenesis) have come into picture as an alternative allowing alignment and stabilization, avoiding a graduated approach. This study was focused on fractures that result in a gap non-union of > 6 cm. Ilizarov technique was employed for management of such non-unions in this case series. The Ilizarov apparatus consists of rings, rods and kirschner wires that encloses the limb as a cylinder and uses kirschner wires to create tension allowing early weight bearing and stimulating bone growth. Ilizarov technique works on the principle of distraction osteogenesis, that is, pulling apart of bone to stimulate new bone growth. Usually, 4–5 rings are used in the setup depending on fracture site and pattern for stable fixation. In this study, we demonstrate effective bone transport and formation of gap non-union more than 6 cm in 10 patients using only 3 rings construct Ilizarov apparatus. Materials and Methods. This case study was conducted at Dr. D. Y. Patil Medical Hospital, Navi Mumbai, Maharashtra, India. The study involved 10 patients with a non-union or gap > 6 cm after tibial fracture. 3 rings were used in the setup for the treatment of all the patients. Wires were passed percutaneously through the bone using a drill and the projecting ends of the wires were attached to the metal rings and tensioned to increase stability. The outcome of the study was measured using the Oxford Knee scoring system, Functional Mobility Scale, the American Foot and Ankle Score and Visual Analog Scale. Further, follow up of patients was done upto 2 years. Results. All the patients demonstrated good fixation as was assessed clinically and radiologically. 9 patients had a clinical score of > 65 which implied fair to excellent clinical rating. The patients showed good range of motion and were highly satisfied with the treatment as measured by different scoring parameters. Conclusions. In this case study, we demonstrate that the Ilizarov technique using 3 rings is equally effective in treating non-unions > 6 cm as when using 4–5 rings. Obtaining good clinical outcome and low complication rate in all 10 patients shows that this modified technique can be employed for patients with such difficulties in the future

Introduction and Objective. Digital infra-red thermography may have the capability of identifying local inflammations. Nevertheless, the role of thermography in diagnosing pin site infection has not been explored yet and the reliability and validity of this method for pin site surveillance is in question. The purpose of this study was to explore the capability and intra-rater reliability of thermography in detecting pin site infection. Materials and Methods. This explorative proof of concept study follows GRRAS -guidelines for reporting reliability and agreement studies. After clinical assessment of pin sites by one examiner using Modified Gordon Pin Infection Classification (Grade 0 – 6), thermographic images of the pin sites were captured with a FLIR C3 camera and analyzed by the FLIR tools software package. The maximum skin temperature around the pin site and the maximum temperature for the whole thermographic picture was measured. Intra-rater agreement was established and test-retests were performed with different camera angles. Results. Thirteen (4 females) patients (age 9–72 years) were included. Indications for frames: 4 fracture, 2 deformity correction, 1 lengthening, 6 bone transport. Days from surgery to thermography ranged from 27 to 385 days. Overall, 231 pin sites were included. Eleven pin sites were diagnosed with early signs of infection: five grade 1, five grade 2, one grade 3. Mean pin site temperature was 33.9 °C (29.0–35.4). With 34 °C as cut-off value for infection, sensitivity was 73%, specificity 67%, positive predictive value 10% and negative predictive value 98%. Intra-rater reliability for thermography was ICC 0.85 (0.77–0.92). The temperature measured was influenced by the camera postioning in relation to pin site with a variance of 0.2. Conclusions. Measurements of pin sites using the handheld FLIR C3 infrared camera was a reliable method and the temperature was related to infection grading. This study demonstrates that digital thermography with a handheld camera might be used for monitoring the pin sites after operations to detect early infection, however, future larger prospective studies are necessary

Harnessing the potential of mesenchymal stem cell (MSC) mediated endochondral ossification for the repair of large bone defects represents a promising avenue of investigation as an alternative option to autologous bone transplantation. To date, it has been shown that undifferentiated MSCs are somewhat immune-privileged. In order to induce bone formation from MSCs by endochondral ossification it is usually necessary to first differentiate these cells chondrogenically. However, the status of differentiated cells is less clear than that of undifferentiated MSCs. Furthermore, the fate of implanted bone forming constructs in an allogeneic setting is not known. The potential to use allogeneic MSCs for large bone defect repair would offer opportunities to researchers to develop new therapies using more potent MSC sources and in a more readily available manner with regard to the patient. I will present our research investigating the interactions between chondrogenically primed MSCs and immune cell subsets, namely T cells and dendritic cells. Furthermore, I will discuss the ability of human paediatric MSCs to form bone in the in vivo allogeneic setting

Bone has a remarkable capacity to heal. However, in some instances the amount of bone which is needed to heal exceeds its healing capacity. Due to reported issues with current treatments there is continued research into alternative approaches with a view to producing an off the shelf alternative to the gold standard autologous bone transplants. The current investigated the use of a chitosan/hydroxyapatite scaffold, which was used to covalently bone morphogenetic protein and vascular endothelial growth factor using a UV crosslinking process. Results indicate that the incorporation of hydroxyapatite increased the mechanical properties of the scaffold compared to chitosan alone. Furthermore, crosslinking was confirmed using swelling studies and FTIR analysis. Elisa indicated that physiological doses of BMP were released after 10 days while in vitro testing did not indicate a cytotoxic response to the scaffold. In vivo testing in a rat femoral defect model indicated the efficacy of the treatment with scaffolds containing BMP and VEGF in combination resulting in more bone in the defect compared to the scaffold alone 8 weeks post-surgery

Summary Statement. A coupled finite element - analytical model is presented to predict and to elucidate a clinical healing scenario where bone regenerates in a critical-sized femoral defect, bounded by periosteum or a periosteum substitute implant and stabilised via an intramedullary nail. Introduction. Bone regeneration and maintenance processes are intrinsically linked to mechanical environment. However, the cellular and subcellular mechanisms of mechanically-modulated bone (re-) generation are not fully understood. Recent studies with periosteum osteoprogenitor cells exhibit their mechanosensitivity in vitro and in situ. In addtion, while a variety of growth factors are implicated in bone healing processes, bone morphogenetic protein-2 (BMP-2) is recognised to be involved in all stages of bone regeneration. Furthermore, periosteal injuries heal predominantly via endochondral ossification mechanisms. With this background in mind, the current study aims to understand the role of mechanical environment on BMP-2 production and periosteally-mediated bone regeneration. The one-stage bone transport model [1] provides a clinically relevant experimental platform on which to model the mechanobiological process of periosteum-mediated bone regeneration in a critical-sized defect. Here we develop a model framework to study the cellular-, extracellular- and mechanically-modulated process of defect infilling, governed by the mechanically-modulated production of BMP-2 by osteoprogenitor cells located in the periosteum. Methods. Material properties of the healing callus and periosteum contribute to the strain stimulus sensed by osteoprogenitor cells therein. Using a mechanical finite element model, periosteal surface strains are first predicted as a function of callus properties. Strains are then input to a mechanistic mathematical model, where mechanical regulation of BMP-2 production mediates rates of cellular proliferation, differentiation and extracellular matrix (ECM) production, to predict healing outcomes. A parametric approach enables the spatial and temporal prediction of tissue regeneration via endochondral ossification. Predictions are compared with experimental, micro-computed tomographic and histologic, measures of cartilage and mineralised bone tissue regenerates. Model Predictions in Light of Experimental Case Studies: A validated baseline model predicts defect healing via cellular egression, extracellular matrix production and endochondral ossification, using parameters optimised to mimic experimental outcome measures at initial and final stages of healing. To elucidate which predictive model paramenters result in the intrinsic differences in experimental outcomes between defects bounded by either periosteum in situ or a periosteum substitute implant, model parameters are then varied by orders of magnitude to determine which factors exert dominant influence on achievement of experimentally relevant ECM area outcomes. Considering the complete set of parameters relevant to healing, the rate of osteoprogenitor to osteoblast differentiation, as well as rates of chondrocyte and osteoblast proliferation must be reduced and ECM production by chondrocytes must be increased from baseline, to achieve healing outcomes analogous to those observed in experiments. Discussion/Conclusion. The novel model framework presented here integrates a mechanistic feedback system, based on the mechanosensitivity of periosteal osteoprogenitor cells, which allows for modeling and prediction of tissue regeneration on multiple length and time scales

Introduction. The annual incidence of fractures in the UK is almost 4%. Bone grafting procedures and segmental bone transport have been employed for bone tissue regeneration. However, their limited availability, donor site morbidity and increased cost mean that there is still a large requirement for alternative methods and there is considerable research into regeneration using bone morphogenetic proteins (BMPs). The aims of this study are to synthesise and combine BMP-2 with a novel nanocomposite and study its release. Materials and Methods. BMP-2 was synthesised using an E. coli expression system and purified. C2C12 cells were used to test its bioactivity using an alkaline phosphatase (ALP) assay. The modified solution evaporation method was used to fabricate 30% a-TCP/PLGA nanocomposite and it was characterized using SEM, TEM, TGA, XRD, EDX and particle size analysis. The release pattern of adsorbed BMP-2 was studied using an ELISA assay. Results. SEM suggests that there was a homogeneous distribution of a-TCP nanoparticles within the PLGA matrix. The concentration of BMP-2 adsorbed onto a-TCP/PLGA nanocomposites directly correlated with the incubation concentration of BMP-2. Approximately 10-15% of BMP-2 was adsorbed on to the discs, up to an incubation concentration of 25 μg/ml. At a higher incubation concentration (50 μg/ml), however, only 4% of the BMP-2 appears to have been adsorbed. The ALP activity shows that the BMP-2 was bioactive and successfully adsorbed onto the surface of the a-TCP/PLGA nanocomposite. A burst release pattern of BMP-2 was observed over 24h, being maximal at 2 h. Discussion. Increasing incubation concentrations of BMP-2 resulted in an increase of detected adsorbed BMP-2 on the discs, however this was not observed at the highest incubation concentration (50 μg/ml). As adsorption of BMP-2 onto the ground surface of the a-TCP/PLGA nanocomposite occurs primarily through electrostatic interactions between cationic BMP-2 and anionic a-TCP, this might reflect saturation in adsorption secondary to saturation of surface anionic a-TCP by BMP-2, or heterogeneity of the discs' content and/or surface area. Adsorbed BMP-2 was shown to have bioactivity which significantly increased with increasing incubation concentrations of BMP-2 and suggests this nanocomposite could have osteoinductive potential in-vivo. The burst pattern of BMP-2 release has been shown previously from BMP adsorbed onto mPCL/collagen/HA composite and this significantly increased the bone formation of critical-sized defects. Whilst a more sustained release profile of BMP-2 is generally considered desirable, this nanocomposite of a-TCP/PLGA has been shown to possess some osteoconductive and weak osteoinductive properties itself (unpublished). The addition of BMP-2 to the nanocomposite by adsorption results in an early burst release, which can promote the differentiation of mesenchymal cells into osteoblasts. The proliferation of these might then be sustained by the nanocomposite itself, without the need for sustained delivery of BMP-2. Conclusions. Bioactive BMP-2 was synthesised and combined with a-TCP/PLGA nanocomposite, producing a biodegradable and osteoinductive material which has potential for use in bone regeneration

Objectives

Osteophytes are products of active endochondral and intramembranous ossification, and therefore could theoretically provide significant efficacy as bone grafts. In this study, we compared the bone mineralisation effectiveness of osteophytes and cancellous bone, including their effects on secretion of growth factors and anabolic effects on osteoblasts.

Objectives

This study aimed to investigate the functional effects of microRNA (miR)-214-5p on osteoblastic cells, which might provide a potential role of miR-214-5p in bone fracture healing.

The treatment of chronic osteomyelitis often includes surgical debridement and filling the resultant void with antibiotic-loaded polymethylmethacrylate cement, bone grafts or bone substitutes. Recently, the use of bioactive glass to treat bone defects in infections has been reported in a limited series of patients. However, no direct comparison between this biomaterial and antibiotic-loaded bone substitute has been performed.

In this retrospective study, we compared the safety and efficacy of surgical debridement and local application of the bioactive glass S53P4 in a series of 27 patients affected by chronic osteomyelitis of the long bones (Group A) with two other series, treated respectively with an antibiotic-loaded hydroxyapatite and calcium sulphate compound (Group B; n = 27) or a mixture of tricalcium phosphate and an antibiotic-loaded demineralised bone matrix (Group C; n = 22). Systemic antibiotics were also used in all groups.

After comparable periods of follow-up, the control of infection was similar in the three groups. In particular, 25 out of 27 (92.6%) patients of Group A, 24 out of 27 (88.9%) in Group B and 19 out of 22 (86.3%) in Group C showed no infection recurrence at means of 21.8 (12 to 36), 22.1 (12 to 36) and 21.5 (12 to 36) months follow-up, respectively, while Group A showed a reduced wound complication rate.

Our results show that patients treated with a bioactive glass without local antibiotics achieved similar eradication of infection and less drainage than those treated with two different antibiotic-loaded calcium-based bone substitutes.

Cite this article: Bone Joint J 2014; 96-B:845–50.

Objectives

There is increasing application of bone morphogenetic proteins (BMPs) owing to their role in promoting fracture healing and bone fusion. However, an optimal delivery system has yet to be identified. The aims of this study were to synthesise bioactive BMP-2, combine it with a novel α-tricalcium phosphate/poly(D,L-lactide-co-glycolide) (α-TCP/PLGA) nanocomposite and study its release from the composite.

Objectives

To review the systemic impact of smoking on bone healing as evidenced within the orthopaedic literature.