Despite the increasing availability of bone grafting materials, the regeneration of large bone defects remains a challenge. Especially infection prevention while fostering regeneration is a crucial issue. Therefore, loading of grafting material with antibiotics for direct delivery to the site of need is desired. This study evaluates the concept of local delivery using in vitro and in vivo investigations. We aim at verifying safety and reliability of a perioperative enrichment procedure of demineralized bone matrix (DBM) with gentamicin. DBM (DBMputty, DIZG, Germany) was mixed with antibiotic using a syringe with an integrated mixing propeller (Medmix Systems, Switzerland). Gentamicin, as powder or solution, was mixed with DBM at different concentrations (25 −100 mg/g DBM), release and cytotoxicity was analyzed. For in vivo analysis, sterile drill hole defects (diameter: 6 mm, depth: 15 mm) were created in diaphyseal and metaphyseal bones of sheep (Pobloth et al. 2016). Defects (6 – 8 per group and time point) were filled with DBM or DBM enriched with gentamicin (50 mg/g DBM) or left untreated. After three and nine weeks, defect regeneration was analyzed by µCT and histology. The release experiments revealed a burst release of gentamicin from DBM independent of the used amount, the sampling strategy, or the formulation (powder or solution). Gentamicin was almost completely released after three days in all set-ups. Eluates showed an antimicrobial activity against S. aureus over at least three days. Eluates had no negative effect on viability and alkaline phosphatase activity of osteoblast-like cells (partially published Bormann et al. 2014). µCT and histology of the drill hole defects revealed a reduced bone formation with gentamicin loaded DBM. After nine weeks significantly less mineralized tissue was detectable in metaphyseal defects of the gentamicin group. Histological evaluation revealed new bone formation starting at the edges of the drill holes and growing into the center over time. The amount of DBM decreased over time due to the active removal by osteoclasts while osteoblasts formed new bone. Using this mixing procedure, loading of DBM was fast, reliable and possible during surgical setting. In vitro experiments revealed a burst and almost complete release after three days, antimicrobial activity and good biocompatibility of the eluates. Gentamicin/DBM concentration was in the range of clinically used antibiotic-loaded-cement for prophylaxis and treatment in joint replacement (Jiranek et al. 2006). The delayed healing seen in vivo was unexpected due to the good biocompatibility found in vitro. A reduced healing was also seen in spinal fusion where DBM was mixed with vancomycin (Shields et al. 2017), whereas DBM with gentamicin or DBM/bioactive glass with tobramycin had no negative effect on osteoinductivity or femur defect healing, respectively (Lewis et al. 2010, Shields et al. 2016). In conclusion, loading of DBM with gentamicin showed a proper antibiotic delivery over several days, covering the critical phase shortly after surgery. Due to the faster and complete release of the antibiotic compared to antibiotic loaded cement, the amount of antibiotic should be much lower in the DBM compared to cement

Autologous bone graft has been used in the treatment of complex bone defects for more than a century. Morbidity associated with the harvest of this bone graft has led orthopaedic surgeons to seek alternative therapies in the treatment of long bone non-unions. The aim of this study was to determine whether the use of demineralised bone matrix as a bone healing adjunct improves clinical outcomes in adult patients with long bone non-union. A systematic search was carried out of the peer-reviewed English language literature to identify all relevant studies. The search strategy returned a total of 47 studies. Five of these studies were relevant to the research question. The studies were critically assessed and where appropriate combined in a meta-analysis. 4 non-comparative studies and one comparative study were reviewed. An overall estimate of the rate of union for the five studies was 86% (95%CI: 71–94%). The one comparative study demonstrated the relative risk (RR) of healing was not significantly better than in patients treated with autologous bone graft; RR=1.03 (95%CI 0.96–1.12). There are limited data to support the use of demineralised bone matrix in the treatment of long bone non-union. Demineralised bone matrix is likely to be similarly effective to other treatments in the management of non-union. This study confirms the clinical and ethical requirements to proceed with a randomised controlled trial to test the effectiveness of this intervention

The purpose of this study was to understand the effects of terminal sterilisation and residual calcium on human demineralised bone matrix (DBM) in ectopic bone formation in nude rat. The intramuscular implantation of human DBM prepared by the Queensland Bone Bank (QBB) from four donors into eight male athymic rats was used to assess osteoinductivity. The DBM contained different levels of residual calcium and treated with or without gamma-irradiation at 11kGy. At 6 weeks post-implantation, calcium deposition was assessed by manual palpitation and radiological imaging. Tissue morphology and cellular interactions was analysed using various histological staining methods whilst protein expression of anabolic and catabolic biomarkers were examined through immunohistochemistry. All results were then analysed in qualitative, semi-quantitative and quantitative manners and tested for statistical significance. Bone formation was observed in all specimens at the gross level. This was confirmed by histology which revealed bony capsules surrounded by soft tissue in the muscle pockets and differences in tissue components. On a cellular level, variations in osteoclast expression were found between the two groups as well as amongst individual donors through statistical analysis which resulted in an imbalance of the expression of anabolic and catabolic markers. Furthermore, a positive relationship between residual calcium and new bone formation in gamma irradiated DBM samples was found. To date, no studies have compared the effect of calcium in gamma irradiated DBM. Our results suggest that gamma irradiation even at low doses and residual calcium may affect new bone formation. Taken together, this study stresses the importance of selecting ideal conditions for graft processing and the need to identify an optimal level of irradiation and remaining calcium levels that confers a balance between osteoinductivity and sterility

Introduction. Tendon injuries remain challenging, secondary healing and prolonged immobilisation result in suboptimal outcome. Previous study by our group showed that demineralised bone matrix (DBM) can result in faster healing of a tendon enthesis. The aim of this study is to test different ways augmenting tendon with DBM to enhance tendon repair and regeneration. Methods. DBM strips were prepared from tibias of mature ewes. Patella, patellar tendon and tibias were dissected and the distal 1 cm of the patellar tendon was excised. 4 models were designed;. Model-1, DBM strip was used to bridge the gap between the tendon and the tibial tuberosity. The DBM strip was stitched to the tendon using one bone anchor. Model-2, similar to model 1 with the use of 2 anchors. Model-3, similar to model 2, construct was off loaded by continuous thread looped twice through bony tunnels sited in the patella and in the tibial tuberosity. Model-4, similar to model 3 with 3 threads as off loading loop. All models were tested for pullout force and mode of failure. Results. The median failure force for model-1 (N=5) was 250N while for model-2 (N=5) was 290N. In model-3 and model-4 failure of the off loading loop was used as end point, 6 samples were tested in each model. Median failure force of model-3 was 767N and for model-4 was 934N. There was no statistical significance between model-1 and model-2 (p=0.249), however statistical significance was found between other models (p=< 0.006). Discussion. A study published in 1996 proved that cortical DBM can be used as ACL graft with evidence of ligamentisation. DBM provides a biologic scaffold with potential for use as ligament and tendon replacement. Our study shows that a tendon rupture can be augmented with DBM giving intial appropriate mechanical strength suitable for in-vivo use

Introduction. Demineralised Bone Matrix (DBM) is widely used in Orthopaedics and dentistry as a bone graft substitute and may be used to augment bone formation in load bearing applications. In this study we examine the effect of gamma irradiation and freeze drying on the tensile strength of Demineralised Cortical Bone (DCB). Methods. Tibias were harvested from mature ewes and cut into bony strips. Demineralisation was done using 0.6M HCL and confirmed by X-ray. Specimens were washed until a pH of 7.0 +/_ 0.2 was achieved in the washing solutions. Specimens were allocated into 4 groups; group (A) non freeze dried non gamma irradiated, group (B) freeze dried non gamma irradiated, group (C) non freeze dried gamma irradiated mention the level of gamma irradiation and group (D) freeze dried and gamma irradiated. The maximum tensile force and stress were measured. Statistical analysis using the Mann-Whitney U test was carried out. Results. The Median of maximum tensile force for group (A) was 218N, group (B) was 306N, group (C) was 263N and for group (D) was 676N. Group (D) results were statistically higher (p=< 0.05) compared to group (A) and (C), while there was no statistical significance compared to group (B). Conclusion. Previously published studies suggested the possibility of using DCB as ACL graft substitute. We examined the effect of gamma radiation as the most common sterilisation technique in medical field and the freeze drying as a possible technique for long term storage on the tensile strength of the DCB. Freeze drying significantly increases the tensile strength of the DCB while gamma irradiation has no significant effect. Our results indicate that freeze dried gamma irradiated DCB can be used as a ligament substitute

Aim. infected segmental bone defect (ISBD) is frequent in developing countries. The aim of this study was to assess the efficacy of the Masquelet technique in the treatment of ISBD in a low-resource setting. Patients and Method. We performed a prospective cohort study during the period from 2018 to 2022. Patients with infected bone defect of long bones were included. Management protocol consisted of two stages in all patients. The first stage consisted in debridement, tissues biopsy for microbiological culture, stabilization with external fixator and defect filling with gentamicin cement spacer. The second stage consisted of reconstruction using a cancellous bone autograft alone, or a mixture of autograft with allograft (demineralized bone matrix + tricalcium phosphate) and 1 gram of vancomycin powder. All patients were followed-up for at least one year. The results were assessed based on both objective (clinical and radiographic evaluation) and subjective (limb function and patient satisfaction) criteria. Main outcomes were bone union, reoperation and failure rates, union time, and limb function. Results. We included 31 patients in this study (80.6% men), with a median age of 35 [9 – 80] years. The tibia was affected in 12 cases and the femur in 15 cases. The median size of bone defect was 4 [1.5 – 12] cm. The most prevalent microorganisms were Klebsiella pneumoniae and Staphylococcus aureus. The mean interval between both stages was 14 (8 – 36) weeks and the median follow-up period after the second stage was 20 [12-62] months. External fixation was used in both stages in 25(80%) cases. Bone union was achieved in 26 (83.8%) patients of whom 24 without recurrence of infection, over a median time of 9 [6 – 16] months. All patients with a mixed graft (allograft and autograft) impregnated with local antibiotics achieved bone union. Two patients needed reoperation for relapse of infection between both stages, and subsequently achieved bone union without recurrence of infection. There were three cases of failure related to persistent infection or insufficient fixation stability in the second stage. Conclusions. Masquelet technique is a reliable procedure that can be safely performed in limited resources settings with satisfactory results. The mixture of autograft and allograft when available, all mixed with vancomycin seems to give promising results

Previously, we reported impaired biomechanical bone properties and inferior bone matrix quality in tachykinin1 (Tac1)-deficient mice lacking the sensory neuropeptide substance P (SP). Additionally, fracture callus development is affected by the absence of SP indicating a critical effect of sensory nerve fibers on bone health and regeneration. For α-calcitonin gene-related peptide (α-CGRP)-deficient mice, a profound distortion of bone microarchitecture has also been described. We hypothesize that SP and α-CGRP modulate inflammatory as well as pain-related processes and positively affect bone regeneration during impaired fracture healing under osteoporotic conditions. Therefore, this study investigates the effects of SP and α-CGRP on fracture healing and fracture-related pain processes under conditions of experimental osteoporosis using SP- and α-CGRP-deficient mice and WT controls. We ovariectomized female WT, Tac1−/− and α-CGRP−/− mice (age 10 weeks, all strains on C57Bl/6J background) and set intramedullary fixed femoral fractures in the left femora 28 days later. We analyzed pain threshold (Dynamic Plantar Aesthesiometer Test) and locomotion (recorded at day and night, each for 1 hour, EthoVision®XT, Noldus) at 5, 9, 13, 16 and 21 days after fracture. At each time point, fractured femora were prepared for histochemical analysis of callus tissue composition (alcian blue/sirius red staining). Pain threshold is significantly higher in Tac1−/− mice 13 days after fracture and tends to be higher after 21 days compared to WT controls. In contrast, touch sensibility was similar in α-CGRP−/− mice and WT controls but compared to Tac1−/− mice pain threshold was significantly lower in α-CGRP−/− mice 13 and 16 days and tends to be lower 21 days after fracture. Locomotion of Tac1−/− mice during daylight was by trend higher 9 days after fracture and significantly higher 16 days after fracture whereas nightly locomotion is reduced compared to WT mice. Analysis of locomotion during daylight or night revealed no differences between α-CGRP−/− and WT mice. During early fracture healing phase, 5 and 9 days after fracture, transition of mesenchymal to cartilaginous callus tissue tends to be faster in Tac1−/− mice compared to WT controls whereas no difference was observed during late stage of fracture healing, 13, 16 and 21 days after fracture. In contrast, callus tissue maturation seems to be similar in α-CGRP−/− and WT mice. Our data indicate different effects of SP and α-CGRP on fracture healing under conditions of experimental osteoporosis as a model for impaired bone tissue. Lack of α-CGRP seems to have no effects, but loss of SP affects locomotion throughout osteoporotic fracture healing and fracture-related pain processes during late phases of osteoporotic fracture healing. This indicates a modified role of SP during fracture healing under impaired versus healthy conditions, where SP changed early fracture-related pain processes and had no influence on callus tissue composition

Aims

Disorders of bone integrity carry a high global disease burden, frequently requiring intervention, but there is a paucity of methods capable of noninvasive real-time assessment. Here we show that miniaturized handheld near-infrared spectroscopy (NIRS) scans, operated via a smartphone, can assess structural human bone properties in under three seconds.

Objectives. Despite promising results have shown by osteogenic cell-based demineralized bone matrix composites, they need to be optimized for grafts that act as structural frameworks in load-bearing defects. The aims of this study is attempt to assess the effects of laser perforations on osteoinduction in cortical bone allografts. Methods. Sixteen wistar rats were divided into two groups according to the type of structural bone allograft; the first: partially demineralized only (Donly) and the second: partially demineralized laser-perforated (DLP). Trans-cortical holes were achieved by Er:YAG laser at a wave length of 2.94 µm in four rows of three holes approximated cylindrical holes 0.5 mm in diameter, with centres 2.5 mm apart. Histologic and histomorphometric analysis were performed at 12 weeks. Results. Statistical analysis showed significant differences between the 2 groups at 3 months. Results showed that partially demineralized laser-perforated grafts had substantially higher incorporation by woven bone than partially demineralized grafts; yet this difference at the interface gap remained insignificant. In DLP allografts healing at the junction was more complete and a wider area was in contact with host and graft surfaces. Conclusions. Based on the results of this study, it may be concluded that surface changes induced by Er:YAG laser, accelerated bone healing with good osteoinduction results and the process might have improved, if they could have been supplemented with the proper stipulations

Background. Despite promising results have shown by osteogenic cell-based demineralized bone matrix composites, they need to be optimized for grafts that act as structural frameworks in load-bearing defects. The purpose of this experiment is to determine the effect of bone marrow mesenchymal stem cells seeding on partially demineralized laser-perforated structural allografts that have been implanted in critical femoral defects. Materials and Methods. Thirty-two wistar rats were divided into four groups according to the type of structural bone allograft; the first: partially demineralized only (Donly), the second: partially demineralized stem cell seeded (DST), the third: partially demineralized laser-perforated (DLP), and the fourth: partially demineralized laser-perforated and stem cell seeded (DLPST). Trans-cortical holes were achieved in four rows of three holes approximated cylindrical holes 0.5 mm in diameter, with centres 2.5 mm apart. P3 MSCs were used for graft seeding. Histologic and histomorphometric analysis were performed at 12 weeks. Results. DLP grafts had the highest woven bone formation, where most parts of laser pores were completely healed by woven bone. DST and DLPST grafts surfaces had extra vessel-ingrowth-like porosities. Furthermore, in the DLPST grafts, a distinct bone formation at the interfaces was noted. Conclusion. This study indicated that surface changes induced by laser perforation, accelerated angiogenesis induction by MSCs, which resulted in endochondral bone formation at the interface. Despite non-optimal results, stem cells showed a tendency to improve osteochondrogenesis, and the process might have improved, if they could have been supplemented with the proper stipulations

Wear and osteolysis are the major problems limiting the longevity of total hip arthroplasty. There is general agreement that if left untreated osteolysis will eventually lead to loosening of the acetabular component. In many cases polyethylene liner exchange may be preferable to revision of a well-fixed acetabular component. If there is osteolysis present the question is when should the polyethylene liner exchange be performed? The answer to that question has not been definitively defined at the present time. There are few studies available that evaluate the timing of surgical intervention when acetabular osteolysis is present. Indications for surgical intervention include prevention of complete wear of the head through the polyethylene liner (liner thickness < 1.5 mm) and when the osteolysis involves 50% or more of the shell circumference on AP or lateral x-rays. Of course persistent pain with wear or osteolysis is another indication for surgery. Contraindications to cup retention and liner exchange include: 1) Malpositioned component; 2) Non-modular component; 3) Unable to obtain hip stability; 4) Thin polyethylene liner (relative); 5) Severe damage to acetabular shell; and 6) Poor track record of the acetabular component. If one decides to retain the component the following steps are generally involved in operative management. Remove the liner and assess component stability. Assess the locking mechanism for the polyethylene. If the locking mechanism is not intact one can consider cementing the liner in place. In general, it is recommended to debride and bone graft the osteolytic lesion. The author prefers to use an access hole at the periphery of the component or at times a trapdoor can be made in the ilium. It is essential not to de-stabilise the acetabular component. At the present time there is no optimal graft material to use. Potential graft options include demineralised bone matrix or cancellous bone chips. Since dislocation is the number one complication after polyethylene liner exchange, it is a good idea to use a larger femoral head whenever possible. In some cases it is also worthwhile to consider bracing the patient after the surgery. It is essential to be ready to perform a complete revision. Therefore, when planning to perform a polyethylene liner exchange one needs to have the appropriate components available to completely revise the acetabular component

Aim. This study describes and correlates the radiographic and histologic changes which develop in a Gentamicin-eluting synthetic bone graft substitute. *. in the management of bone defects after resection of chronic osteomyelitis (COM). Method. 100 patients with COM were treated with a single stage procedure, including management of the dead space with insertion of a Gentamicin-eluting synthetic bone graft substitute. *. Radiographs of 73 patients with a follow-up of at least 12 months (range 12–33 months) were available for review. Bone defects were diaphyseal in 32, metaphyseal in 34 and combined in 7 patients. In 3 patients, radiographs were not of sufficient quality to allow analysis. Five patients had subsequent surgery, not related to recurrence of infection, which allowed biopsy of the implanted material. These biopsies were harvested between 12 days and 9 months after implantation. Tissue was fixed in formalin and stained with haematoxylin-eosin and immunohistochemically for bone matrix markers. Results. Radiographic: 31 of 34 diaphyseal implantations (91%) demonstrated remodelling of the biocomposite, gradually over many months, producing new bone and resulting in a “normal post-osteomyelitic” appearance. In metaphyseal implantations, new bone filled two-thirds or more of the defect in 55% of cases, one to two-thirds was filled in 31% and one third or less was filled in 14%. 22% of patients exhibited radiographic signs of dissolution and remodelling which are specific to this material. The ‘Halo’ sign of peripheral zone remodelling, the ‘Marble’ sign of dissolution and the ‘Puddle’ sign of distal migration can be described. Histologic: Histological assessment revealed early active remodelling of the biocomposite. The material was osteoconductive with accumulation of osteoblasts and osteoid and woven bone formation on the surface of the Gentamicin-eluting synthetic bone graft substitute. *. separated by fibrous tissue at the edge of the defect beneath reactive viable host bone. Fibrous tissue contained a heavy macrophage infiltrate and the newly formed matrix contained the specific bone proteins, dentine matrix protein-1 and podoplanin. There was limited evidence of remodelling into lamellar bone at 20 weeks after implantation. Conclusions. The Gentamicin-eluting synthetic bone graft substitute. *. exhibits a specific pattern of radiographic change over many months after implantation. The resolution of the bone defect would appear to be due to bone formation, as seen in the histologic and immunohistochemical analysis

Evaluation of the effectiveness of biodegradable bone substitute with high doses of antibiotics in cavitary osteomyelitis and infected nonunions. The authors evaluated 8 cases, 5 of them related to osteomyelitis with bone sequestration and other 3 regarding infected nonunions. All of them had in common the persistence of infection after antibiotic therapy. All infections were confirmed by microbiological studies. In all cases the surgeons conducted a thorough surgical debridement and filling of bone defects with Herafill®. Later a tight clinical, analytical and imagiological control was performed. Five of the cases were a success with simultaneous healing of the bone loss and treatment of the infection. These corresponded to the cases of cavitary osteomyelitis. In the remaining 3 cases, despite infection eradication, union was not achieved and additional surgical procedures were required for definitive treatment of nonunion. In the treatment of bone infection, use of high doses of antibiotics at the site is a consensus as it allows eradication of the infection with lower systemic effects. With the emergence of biodegradable bone substitutes, the need for a new surgical intervention for their removal can be avoided. Properties of calcium sulfate and calcium carbonate stimulate osteogenesis at the site, allowing their absorption and replacement by bone matrix. These properties make them ideal to usage in cases of cavitary bone defects. Our experience supports the idea that the use of high doses of antibiotics locally permits remission of the infection. However, when this is implemented through a bone substitute, it is possible to achieve osteogenesis in bony cavities. Nevertheless, when applied to infected nonunions, their role seems to be limited to the eradication of the infection

Background. In 2012, the National Joint Registry recorded 86,488 primary total hip replacements (THR) and 9,678 revisions (1). To date aseptic loosening remains the most common cause of revision in hip and knee arthroplasty, accounting for 40% and 32% of all cases respectively and emphasising the need to optimise osseointegration in order to reduce revisions. Clinically, osseointegration results in asymptomatic stable durable fixation of orthopaedic implants. Osseointegration is a complex process involving a number of distinct mechanisms affected by the implant surface topography, which is defined by surface orientation and surface roughness. Micro- and nano-topography levels have discrete effects on implant osseointegration and yet the role on cell function and subsequent bone implant function is unknown. Nanotopography such as collagen banding is a critical component influencing the SSC niche in vivo and has been shown to influence a range of cell behaviours in vitro (2,3). We have used unique fabricated nanotopographical pillar substrates to examine the function of human bone stem cells on titanium surfaces. Aim. To investigate the effect of nanotopographical cues on adult skeletal stem cell (SSC) fate, phenotype and function within in-vitro environments. Materials and methods. Adult human skeleltal stem cells (SSCs) were immunoselected and enriched using STRO-1 antibody and cultured on tissue culture plastic (TCP) and titanium-coated nanotopgraphical substrates (illustrated in Figure 1). Following culture, metabolic activity of SSCs on TCP and Ti substrates was compared. Subsequently, osteoinductive potential was analysed under basal and osteogenic conditions (four groups: TCP in basal media, TCP in osteogenic media, Ti planar substrates basal and Ti pillar substrates basal). Results. At 7 days, cell metabolic activity was significantly enhanced on Ti substrates, specifically on Ti pillars of defined height in comparison to TCP (Figure 2). Following culture on defined topographies for 21 days, expression of the bone matrix protein, osteopontin, on Ti pillars was significantly enhanced when compared to TCP or Ti planar (Figure 3). Conclusion. We demonstrate the ability of discrete raised nanopillars to modualte adult SSC populations in the absence of any chemical cues. These results indicate the potential of discrete and defined nanopillar constructs to stimulate SSC function, an effect not observed on planar Ti constructs. These findings herald exciting opportunities to improve the bioactivity of implant design and, ultimately, osseointegration with clinical implications therein

Long-term biological fixation and stability of uncemented acetabular implant are influenced by peri-prosthetic bone ingrowth which is known to follow the principle of mechanoregulatory tissue differentiation algorithm. A tissue differentiation is a complex set of cellular events which are largely influenced by various mechanical stimuli. Over the last decade, a number of cell-phenotype specific algorithms have been developed in order to simulate these complex cellular events during bone ingrowth. Higher bone ingrowth results in better implant fixation. It is hypothesized that these cellular events might influence the peri-prosthetic bone ingrowth and thereby implant fixation. Using a three-dimensional (3D) microscale FE model representing an implant-bone interface and a cell-phenotype specific algorithm, the objective of the study is to evaluate the influences of various cellular activities on peri-prosthetic tissue differentiation. Consequently the study aims at identifying those cellular activities that may enhance implant fixation. The 3D microscale implant-bone interface model, comprising of Porocast Bead of BHR implant, granulation tissue and bone, was developed and meshed in ANSYS (Fig. 1b). Frictional contact (µ=0.5) was simulated at all interfaces. The displacement fields were transferred and prescribed at the top and bottom boundaries of the microscale model from a previously investigated macroscale implanted pelvis model (Fig. 1a) [4]. Periodic boundary conditions were imposed on the lateral surfaces. Linear elastic, isotropic material properties were assumed for all materials. Young's modulus and Poisson's ratios of bone and implant were mapped from the macroscale implanted pelvis [4]. A cell-phenotype specific mechanoregulatory algorithm was developed where various cellular activities and tissue formation were modeled with seven coupled differential equations [1, 2]. In order to evaluate the influence of various cellular activities, a Plackett-Burman DOE scheme was adopted. In the present study each of the cellular activity was assumed to be an independent factor. A total of 20 independent two-level factors were considered in this study which resulted in altogether 24 different combinations to be investigated. All these cellular activities were in turn assumed to be regulated by local mechanical stimulus [3]. The mechano-biological simulation was run until a convergence in tissue formation was attained. The cell-phenotype specific algorithm predicted a progressive transformation of granulation tissue into bone, cartilage and fibrous tissue (Fig. 1c). Various cellular activities were found to influence the time to reach equilibrium in tissue differentiation and, thereby, attainment of sufficient implant fixation (Fig. 2, Table 1). Negative regression coefficients were predicted for the significant factors, differentiation rate of MSCs and bone matrix formation rate, indicating that these cellular activities favor peri-prosthetic bone ingrowth by facilitating rapid peri-prosthetic bone ingrowth. Osteoblast differentiation rate, on the contrary, was found to have the highest positive regression coefficient among the other cellular activities, indicating that an increase in this cellular activity delays the attainment of equilibrium in bone ingrowth prohibiting rapid implant fixation. To view tables/figures, please contact authors directly

Peri-prosthetic osteolysis and subsequent aseptic loosening is the most common reason for revising total hip replacements. Wear particles originating from the prosthetic components interact with multiple cell types in the peri-prosthetic region resulting in an inflammatory process that ultimately leads to peri-prosthetic bone loss. These cells include macrophages, osteoclasts, osteoblasts and fibroblasts. The majority of research in peri-prosthetic osteolysis has concentrated on the role played by osteoclasts and macrophages. The purpose of this review is to assess the role of the osteoblast in peri-prosthetic osteolysis. In peri-prosthetic osteolysis, wear particles may affect osteoblasts and contribute to the osteolytic process by two mechanisms. First, particles and metallic ions have been shown to inhibit the osteoblast in terms of its ability to secrete mineralised bone matrix, by reducing calcium deposition, alkaline phosphatase activity and its ability to proliferate. Secondly, particles and metallic ions have been shown to stimulate osteoblasts to produce pro inflammatory mediators in vitro. In vivo, these mediators have the potential to attract pro-inflammatory cells to the peri-prosthetic area and stimulate osteoclasts to absorb bone. Further research is needed to fully define the role of the osteoblast in peri-prosthetic osteolysis and to explore its potential role as a therapeutic target in this condition. Cite this article: Bone Joint J 2013;95-B:1021–5

Osteocytes (OCY) are the end stage differentiation cells of the osteoblast lineage, and are incorporated in the bone matrix during bone formation. In doing so, OCY control the mineralisation of osteoid. OCY form a dense inter-connected network of cell bodies and cell processes throughout the mineralised matrix of bone. OCY viability depends on interstitial fluid flow along the OCY canaliculi, driven by pulsatile blood flow and loading of the skeleton. Maintenance of the density and viability of OCY are essential for bone health because OCY perform many important functions in bone. Firstly, OCY appear to initiate bone repair of bone microdamage. Secondly, OCY are almost certainly the cells, which initiate new bone formation in response to increased loading of bone. Thirdly, OCY are able to regulate the amount of new bone formation in bone remodelling cycles, at least in part by the production of a molecule called sclerostin (SCL). Mutations in the SCL gene, or deletion of the SCL gene in transgenic mice, are associated with particularly dense, fracture resistant bones. This information has led to development of anti-SCL antibodies as a potential anabolic therapy for bones. Bone loss in ovariectomised aged rats was shown recently to be reversed by treatment with neutralising SCL antibodies. There is also some data to suggest that these antibodies may promote fracture healing. Reduced OCY viability and/or density have been reported in association with osteoporotic fracture. OCY viability seems to be dependent on skeletal loading, adequate skeletal blood flow and estrogen in females. OCY viability is adversely affected by hypoxia, unloading of the skeleton and pharmacobiology, such as chronic exposure to glucocorticoids. Both micro and macro-fractures result in disruption of the OCY network, as do procedures such as drilling and cutting of bone. Because of the important roles of OCY in bone, new approaches to bone health may require the identification of agents to protect these cells from harmful influences in disease and ageing

Aim. To compare a variety of commercially available bone graft substitutes (BGS) in terms of promoting adherence, proliferation and differentiation of osteoprogenitor cells. Materials and methods. A fixed number of porcine mononuclear cells obtained from cancellous bone of the proximal femur was mixed with a standard volume of BGS and then cultured for one week in media followed by two weeks in osteogenic media. BGS included commercially available β-Tricalcium Phosphate (□-TCP), highly porous β-TCP, Hydroxyapatite/Tricalcium phosphate composite, calcium sulphate (CS), Hydroxyapatite (HA), Demineralised bone matrix (DBM), polygraft, and polymers (PGA, PLGA). Staining for live/dead cells as well as scanning electron microscopy (SEM) were carried out on all samples to determine viability and cellular binding. Further outcome measures included alkaline phosphatase assays with normalisation for DNA content to quantify osteogenic potential. Negative (BGS without cells) and positive (culture expanded osteoprogenitors) control experiments were carried out in parallel to validate the results. Results. Live/dead and SEM imaging showed higher cellular viability and attachment with β-TCP than with other BGS. In the experimental setup the average alkaline phosphatase activity in nmol/ml (normalised value for DNA content in nmol/μg DNA) per sample was 657.58 (132.03) for β-TCP, 36.22 (unable to normalise) for calcium sulphate, 19.93 (11.39) for the HA/ TCP composite, 14.79 (18.53) for polygraft, 13.98 (8.15) for the highly porous β-TCP, 5.56 (10.0) for PLGA, 3.82 (3.8) and for HA. It was not possible to analyse data for either DBM or PGA. Conclusion. Under theses experimental conditions, β-TCP has apparent favourable characteristics in terms of maintaining viability of osteoprogenitor cells and allowing proliferation and differentiation. Further work will be carried out to characterise the effect that BGS have on osteoprogenitor cells

Purpose. The focus of current management of soft tissue sarcoma on limb preservation often necessitates that patients undergo multimodal treatment, including both surgery and external beam radiotherapy. Pathologic fracture is a serious, late complication of radiotherapy. In patients who have also undergone wide excision of soft tissue sarcoma, nonunion rates of 80–90% persist despite optimal internal fixationMany sequelae of the treatments for soft tissue sarcoma exhibit the potential to perpetuate failure of bony union. Limb salvage surgery is associated with extensive periosteal excision, disruption of vascular supply and eradication of local osteoprogenitor cells. External beam radiotherapy leads to obliterative endarteritis, decreased osteoblast proliferation and reduction in bone matrix production. We hypothesize that the combination of radiotherapy and surgical periosteal stripping leads to greater impairment in the fracture repair process than either intervention alone will produce. Method. We developed a method for creating a reproducible, low energy, simple femoral fracture in an animal model designed to proceed to nonunion. Female Wistar, retired breeder rats were separated into four treatment groups of 18 animals each: control, radiotherapy, surgery and combination radiotherapy and surgery. Animals were then further randomized to temporal end-points of 21, 28 and 35 days post-fracture. Designated animals first underwent external beam radiotherapy, followed by surgical stripping of the periosteum three weeks later and femoral fracture with fixation after another three weeks. Animals were sacrificed at their randomly assigned end-points. Results. The fracture device was shown to produce simple, transverse or short oblique femoral fractures using x-rays obtained immediately following fracture, validating the reliability of the model. No significant differences were observed in the force required for fracture between treatment groups. Mineralized callus was observed in control animals and those undergoing periosteal stripping alone, but was absent in all animals receiving radiotherapy. Reactive bone formation was observed in animals undergoing periosteal stripping alone, but was absent when preceded by radiotherapyMicroCT analysis confirmed the results visualized on plain x-ray. No callus formation was observed in animals undergoing radiotherapy and significantly less mineralized callus was produced in animals undergoing periosteal stripping when compared to control. Preliminary studies have shown an absence of cellular activity in animals undergoing radiotherapy, suggesting that fracture in these animals will proceed to nonunion. Conclusion. Early results suggest that this pre-clinical model of combined radiation and surgical periosteal stripping prior to controlled fracture reliably results in nonunion. We expect to utilize this model to examine interventions designed to improve fracture healing in this difficult clinical situation

The most frequent cause of failure after total hip replacement in all reported arthroplasty registries is peri-prosthetic osteolysis. Osteolysis is an active biological process initiated in response to wear debris. The eventual response to this process is the activation of macrophages and loss of bone.

Activation of macrophages initiates a complex biological cascade resulting in the final common pathway of an increase in osteolytic activity. The biological initiators, mechanisms for and regulation of this process are beginning to be understood. This article explores current concepts in the causes of, and underlying biological mechanism resulting in peri-prosthetic osteolysis, reviewing the current basic science and clinical literature surrounding the topic.