Bone impaction grafting of the femur is associated with more complications when segmental defects are present. The effect of segmental defect repair on initial stem stability was studied in an in vitro study with fresh frozen goat femora. A standardized medial segmental defect was reconstructed using a cortical strut or a metal mesh. As controls we used intact femora and femora with a non-reconstructed defect. In all four groups impacted bone grafting was performed in combination with a cemented Exeter stem. Each group contained five femora. Reconstructions were dynamically loaded up to 1500N. Migration was measured with Roentgen Stereo-photogrammetric Analysis. All cases with a non-reconstructed segmental defect failed into excessive varus rotation. None of the femora with a reconstructed defect failed. Cortical struts and metal meshes were equally effective in creating a stable stem construction (varus rotation 2.89±2.27 and 2.27±0.57, respectively). Reconstructions with a metal mesh were more reproducible, although the obtained stability was significantly lower (p<
0.01) when compared to impaction grafting in an intact femur (varus rotation 0.58±0.36). Besides, structural grafts may negatively influence the revascularization of the underlying impacted grafts in contrast to an open wire mesh. So, an in vivo study of 12 goats was done. A standardized medial wall defect was reconstructed with a strut or a mesh in six goats per group. In all femora impaction grafting was performed in combination with a cemented Exeter stem. After six weeks the femora were harvested. A high rate of peri-prosthetic fractures was found (43% and 29% for the strut and mesh groups, respectively). Histological and micro-radiological examination showed different revascularization patterns for both reconstruction techniques. In the strut group revascularized graft was found at the edges of the defect. In the mesh group fibrous tissue and blood vessels penetrated through the mesh and a superficial zone of revascularized grafts was found.
Purpose: One of the most difficult challenges for orthopaedic surgeons is the management of bone loss resulting in a segmental bone
It has been shown that mesenchymal stem cells (MSCs) and BMP are involved in bone formation. The aim of the study was to evaluate the osteogenic potential of human bone marrow (hBM), human expanded MSC (hexp-MSC), BMP-7, and hexp-MSC plus BMP-7, to treat a rat femoral segmental defect. Sprague-Dawley (SD) and athymic rats (Nu) were used. SD rats where used in order to define surgical technique. Nu rats groups consisted of: G1-autoclaved bone and human bone marrow (hMNC); G2-bone and hexp-MSC; G3-bone with BMP-7 only; and G4-bone and hexp-MSC with BMP-7. A plate was attached to the femoral diaphysis with two cerclage wires. Then a 6-mm femoral gap was made and filled with a different graft. At regular intervals, the femoral defect was evaluated with radiographs, using a modified six-grade Cook classification. At 8 weeks G1 showed non-visible new bone formation; G2 minimal new disorganised bone; G3 disorganised new bone bridging the graft to host at both ends; and G4 significant new bone and graft remodelling. Histological analysis confirmed these results. Our results showed that although the osteogenic activity may be improved by hMSC (G2) as well as by BMP-7 (G3), the association hexp-MSC plus BMP-7(G4) produced graft osteointegration at 8 weeks after surgery. This may have a remarkable impact on future orthopaedics surgery strategies.
We sought to establish whether fibroblasts transfected ex vivo could be delivered via gelfoam impregnated with a solution of transfected cells to achieve local transgene expression in a fracture site. A 10 millimeter segmental bone defect was created after 12 mm periosteal excision and plated in the middle one third of each rabbit tibia. Dermal tissues were obtained and fibroblasts were cultured with DMEM. Fibroblasts were labeled with CMTMR and 5x106 labeled fibroblasts in 1ml PBS with 1x1 cm? Impregnated gelfoam was placed into the fracture gap (n=2). Twenty four hours after cell injection, the rabbits were killed and specimens were harvested from the fractured leg. Using SuperFect (Qiagen Inc), the primary fibroblasts were transfected with pcDNA-VEGF which was generated with the full length coding sequence of the human VEGF gene. A convenient reporter gene, Efficiency Green Fluorescent Protein (EGFP), was used for monitoring transfection of VEGF by fluorescence intensity. Experimental rabbits received 5.0 X 106 VEGF transfected cells in 1 ml PBS via gelfoam at the fracture sites. The animals were sacrificed at seven days (n=4), fourteen days (n=4) and twenty-one days (n=4) post surgery and the fracture site specimens were collected for analysis. The fluorescently labeled cells with CMTMR were found at the fracture site and surrounding tissues. It was demonstrated that the labeled cells were delivered into the fracture gap, bone marrow and muscle surrounding a segmental defect in the rabbit. In the VEGF group, visualised VEGF immunostaining (brown) was shown in the fracture site around the Gelfoam; as well VEGF was distributed at sites of endochondral ossification. Visible bone formation was shown: VEGF promoted new bone formation by VonKossa staining (dark) and produced numerous vessels by CD31 positive staining (brownish black). The VEGF protein was detected in and around the fracture by ELISA. This data encourages the further development of genetic approaches using cell based VEGF gene transfer without viral vectors to promote fracture healing.
Besides these groups 15 more DM rats were used for PECAM-1 staining for angiogenesis (7 with 1 loss at a 3 week time point) and mechanical testing (8 at a 9 week time point).
Introduction.
The Masquelet or induced membrane technique (IMT) is a two-stage surgical procedure used for the treatment of segmental bone defects. In this technique, the defect is first filled with a polymethyl methacrylate (PMMA) spacer, which triggers the formation of a membrane that will encapsulate the defect. During the second surgery, the spacer is carefully removed and replaced by autologous bone graft while preserving the membrane. This membrane is vascularized, contains growth factors, and provides mechanical stability to the graft, all of which are assumed to prevent graft resorption and promote bone healing. The technique is gaining in popularity and several variations have been introduced in the clinical practice. For instance, orthopaedic surgeons now often include antibiotics in the spacer to treat or prevent infection. However, the consequences of this approach on the properties of the induce membrane are not fully understood. Accordingly, in a small animal model, this study aimed to determine the impact on the induced membrane of impregnating spacers with antibiotics frequently used in the IMT. We surgically created a five-mm segmental defect in the right femur of 25 adult male Sprague Dawley rats. The bone was stabilized with a plate and screws before filling the defect with a PMMA spacer. Animals were divided into five equal groups according to the type and dose of antibiotics impregnated in the spacer: A) no antibiotic (control), B) low-dose tobramycin (1.2 g/40 g of PMMA), C) low-dose vancomycin (1 g/40 g of PMMA), D) high-dose tobramycin (3.6 g/40 g of PMMA), E) high-dose vancomycin (3 g/40 g of PMMA). The animals were euthanized three weeks after surgery and the induced membranes were collected and divided for analysis. We assessed the expression of selected genes (Alpl, Ctgf, Runx2, Tgfb1, Vegfa) within the membrane by quantitative real-time PCR. Moreover, frozen sections of the specimens were used to quantify vascularity by immunohistochemistry (CD31 antigen), proliferative cells by immunofluorescence (Ki-67 antigen), and membrane thickness. Microscopic images of the entire tissue sections were taken and analyzed using FIJI software. Finally, we measured the concentration of vascular endothelial growth factor (VEGF) in the membranes by ELISA. No significant difference was found among the groups regarding the expression of genes related to osteogenesis (Alpl, Runx2), angiogenesis (Vegfa), or synthesis of extracellular matrix (Ctgf, Tgfb1) (n = four or five). Similarly, the density of proliferative cells and blood vessels within the membrane, as well as the membrane thickness, did not vary substantially between the control, low-dose, or high-dose antibiotic groups (n = four or five). The concentration of VEGF was also not significantly influenced by the treatment received (n = four or five). The addition of tobramycin or vancomycin to the spacer, at the defined low and high doses, does not significantly alter the bioactive characteristics of the membrane. These results suggest that orthopaedic surgeons could use antibiotic-impregnated spacers for the IMT without compromising the induced membrane and potentially bone healing.
The management of a significant bony defect following excision
of a diaphyseal atrophic femoral nonunion remains a challenge. We
present the outcomes using a combined technique of acute femoral
shortening, stabilized with a long retrograde intramedullary nail,
accompanied by bifocal osteotomy compression and distraction osteogenesis with
a temporary monolateral fixator. Eight men and two women underwent the ‘rail and nail’ technique
between 2008 and 2016. Proximal locking of the nail and removal
of the external fixator was undertaken once the length of the femur
had been restored and prior to full consolidation of the regenerate.Aims
Patients and Methods
Angiogenesis and osteogenesis are essential for bone growth, fracture repair, and bone remodeling. VEGF has an important role in bone repair by promoting angiogenesis and osteogenesis. In our previous study, endothelial progenitor cells (EPCs) promoted bone healing in a rat segmental bone defect as confirmed by radiological, histological and microCT evaluations (Atesok, Li, Schemitsch 2010); EPC treatment of fractures resulted in a significantly higher strength by biomechanical examination (Li, Schemitsch 2010). In addition, cell-based VEGF gene transfer has been effective in the treatment of segmental bone defects in a rabbit model (Li, Schemitsch et al 2009); Purpose of this study: Evaluation of VEGF gene expression after EPC local therapy for a rat segmental bone defect. Rat bone marrow-derived EPCs were isolated from the rat bone marrow by the Ficoll-paque gradient centrifuge technique. The EPCs were cultured for 7 to 10 days in endothelial cell growth medium with supplements (EGM-2-MV-SingleQuots, Clonetics). and collected for treatment of the rat segmental bone defect. EPCs were identified by immunocytochemistry staining with primary antibodies for CD34, CD133, FLK-1, and vWF. A total of fifty six rats were studied. A five millimeter segmental bone defect was created in the middle 1/3 of each femur followed by mini plate fixation. The treatment group received 1×106 EPCs locally at the bone defect and control animals received saline only. Seven control and seven EPC treated rats were included in each group at 1, 2, 3 and 10 weeks. Animals were sacrificed at the end of the treatment period, and specimens from the fracture gap area were collected and immediately frozen. Rat VEGF mRNA was measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and quantified by VisionWorksLS. All measurements were performed in triplicate.Purpose
Method
The biomembrane (induced membrane) formed around polymethylmethacrylate (PMMA) spacers has value in clinical applications for bone defect reconstruction. Few studies have evaluated its cellular, molecular or stem cell features. Our objective was to characterise induced membrane morphology, molecular features and osteogenic stem cell characteristics. Following Institutional Review Board approval, biomembrane specimens were obtained from 12 patient surgeries for management of segmental bony defects (mean patient age 40.7 years, standard deviation 14.4). Biomembranes from nine tibias and three femurs were processed for morphologic, molecular or stem cell analyses. Gene expression was determined using the Affymetrix GeneChip Operating Software (GCOS). Molecular analyses compared biomembrane gene expression patterns with a mineralising osteoblast culture, and gene expression in specimens with longer spacer duration (> 12 weeks) with specimens with shorter durations. Statistical analyses used the unpaired student Objectives
Methods
Conclusion: The results of this study demonstrate that EPCs are effective as cell-based therapy for healing critical sized bone defects in a rat model. In this model EPCs demonstrated superiority to MSCs with regard to bone healing. In addition, EPCs demonstrated superior angiogenesis over controls in a rat model of fracture healing. These results strongly suggest that EPCs are effective for therapeutic angiogenesis and osteogenesis in fracture healing. There is a clinical need for effective strategies in the management of traumatic bone defects and nonunions. Investigation into the use of MSCs as an effective alternative to autologous bone grafting has failed to translate into clinical use. It is possible that EPCs are more effective at the regeneration of bone in segmental defects because of their synergistic effect on angiogenesis and osteogenesis. Further research into EPC based therapies for fracture healing is warranted.
Impaction grafting is an excellent option for acetabular revision. It is technique specific and very popular in England and the Netherlands and to some degree in other European centers. The long term published results are excellent. It is, however, technique dependent and the best results are for contained cavitary defects. If the defect is segmental and can be contained by a single mesh and impaction grafting, the results are still quite good. If, however, there is a larger segmental defect of greater than 50% of the acetabulum or a pelvic discontinuity, other options should be considered.
Objectives. Long bone defects often require surgical intervention for functional restoration. The ‘gold standard’ treatment is autologous bone graft (ABG), usually from the patient’s iliac crest. However, autograft is plagued by complications including limited supply, donor site morbidity, and the need for an additional surgery. Thus, alternative therapies are being actively investigated. Autologous bone marrow (BM) is considered as a candidate due to the presence of both endogenous reparative cells and growth factors. We aimed to compare the therapeutic potentials of autologous bone marrow aspirate (BMA) and ABG, which has not previously been done. Methods. We compared the efficacy of coagulated autologous BMA and ABG for the repair of ulnar defects in New Zealand White rabbits.
Background:.
Nonunions and segmental bone defects associated with infection are challenging problems faced by the orthopaedic surgeon. Antibiotic cement-coated (ACC) interlocking nails, prepared in the operating theatre using nails and materials generally available, can be used to treat these conditions. Two different types of moulds can be used (reusable or disposable). Materials and Methods: The infected nonunion/segmental bone defect was treated by débridement followed by ACC nailing in 52 patients (12 female, 40 male, age range 16–86 years). Other procedures for deformity correction, bone defect etc were carried out simultaneously as indicated. Infected nonunion was seen in 34 patients, 1 was an acute fracture after external fixator.
The goals of revision arthroplasty of the hip are to restore the anatomy and achieve stable fixation for new acetabular and femoral components. It is important to restore bone stock, thereby creating an environment for stable fixation for the new components. The bone defects encountered in revision arthroplasty of the hip can be classified either as contained (cavitary) or uncontained (segmental). Contained defects on both the acetabular and femoral sides can be addressed by morselised bone graft that is compacted into the defect. Severe uncontained defects are more of a problem particularly on the acetabular side where bypass fixation such as distal fixation on the femoral side is not really an alternative. Most authors agree that the use of morselised allograft bone for contained defects is the treatment of choice as long as stable fixation of the acetabular component can be achieved and there is a reasonable amount of contact with bleeding host bone for eventual ingrowth and stabilisation of the cup. On the femoral side, contained defects can be addressed with impaction grafting for very young patients or bypass fixation in the diaphysis of the femur using more extensively coated femoral components or taper devices.
Purpose. Traditionally, the gold standard for bone grafting has been either autografts or allografts. Whilst autografts are still widely used, drawbacks such as donor site morbidity are shifting the market rapidly toward the use of orthobiologic bone graft substitutes. This study investigated the in vivo performance of a novel (W02008096334) collagen-hydroxyapatite (CHA) bone graft substitute material as an osteoinductive tissue engineering scaffold. This highly porous CHA scaffold offers significantly increased mechanical strength over collagen-only scaffolds while still exhibiting an extremely high porosity (≈ 99%), and an osteoinductive hydroxyapatite phase [1]. This study assessed the ability of the CHA scaffolds to heal critical-sized (15 mm) long bone segmental defects in vivo, as a viable alternative to autologous bone grafts. Method. Collagen-HA (CHA) composite scaffolds were fabricated based on a previously-described freeze-drying technique [1]. After freeze-drying, these scaffolds were subjected to a dehydrothermal treatment and subsequently chemically crosslinked using EDAC. In vivo performance was assessed using a critical size segmental radial defect (15 mm) introduced into 16 young adult New Zealand White Rabbits under Irish Government license. Animals were divided into three groups; (i) an empty defect group (negative control), (ii) an autogenous bone graft group (positive control) and (iii) a CHA scaffold group (CHA).
This study was designed to characterize the recurrence incidence and risk factors of antibiotic-loaded cement spacer (ALCS) for definitive bone defect treatment in limb osteomyelitis. We included adult patients with limb osteomyelitis who received debridement and ALCS insertion into the bone defect as definitive management between 2013 and 2020 in our clinical centre. The follow-up time was at least two years. Data on patients’ demographics, clinical characteristics, and infection recurrence were retrospectively collected and analyzed.Aims
Methods