We hypothesised that cells obtained via a Reamer–Irrigator–Aspirator
(RIA) system retain substantial osteogenic potential and are at
least equivalent to graft harvested from the iliac crest. Graft
was harvested using the RIA in 25 patients (mean age 37.6 years
(18 to 68)) and from the iliac crest in 21 patients (mean age 44.6
years (24 to 78)), after which ≥ 1 g of bony particulate graft material
was processed from each. Initial cell viability was assessed using Trypan
blue exclusion, and initial fluorescence-activated cell sorting
(FACS) analysis for cell lineage was performed. After culturing
the cells, repeat FACS analysis for cell lineage was performed and
enzyme-linked immunosorbent assay (ELISA) for osteocalcin, and Alizarin
red
Objectives. 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. Methods. 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 t-test (two tailed; p < 0.05 was considered significant). Results. Average PMMA spacer in vivo time was 11.9 weeks (six to 18). Trabecular bone was present in 33.3% of the biomembrane specimens; bone presence did not correlate with spacer duration. Biomembrane morphology showed high vascularity and collagen content and positive
Objectives. Nonunion is one of the most troublesome complications to treat
in orthopaedics. Former authors believed that atrophic nonunion
occurred as a result of lack of mesenchymal stem cells (MSCs). We
evaluated the number and viability of MSCs in site of atrophic nonunion compared
with those in iliac crest. Methods. We enrolled five patients with neglected atrophic nonunions of
long bones confirmed by clinical examinations and plain radiographs
into this study. As much as 10 ml bone marrow aspirate was obtained
from both the nonunion site and the iliac crest and cultured for
three weeks. Cell numbers were counted using a haemocytometer and
vitality of the cells was determined by trypan blue
This study aimed to investigate the role of quantitative histological
analysis in the diagnosis of fracture-related infection (FRI). The clinical features, microbiology culture results, and histological
analysis in 156 surgically treated nonunions were used to stratify
the likelihood of associated infection. There were 64 confirmed
infected nonunions (one or more confirmatory criteria: pus, sinus,
and bacterial growth in two or more samples), 66 aseptic nonunions
(no confirmatory criteria), and 26 possibly infected nonunions (pathogen
identified from a single specimen and no confirmatory criteria).
The histological inflammatory response was assessed by average neutrophil
polymorph (NPs) counts per high-power field (HPF) and compared with
the established diagnosis.Aims
Patients and Methods
In this retrospective observational cohort study,
we describe 17 patients out of 1775 treated for various fractures who
developed mycobacterium tuberculosis (MTB) infection after surgery.
The cohort comprised 15 men and two women with a mean age of 40
years (24 to 70). A total of ten fractures were open and seven were
closed. Of these, seven patients underwent intramedullary nailing
of a fracture of the long bone, seven had fractures fixed with plates,
two with Kirschner-wires and screws, and one had a hemiarthroplasty
of the hip with an Austin Moore prosthesis. All patients were followed-up
for two years. In all patients, the infection resolved, and in 14
the fractures united. Nonunion was seen in two patients one of whom
underwent two-stage total hip arthroplasty (THA) and the other patient
was treated using excision arthoplasty. Another patient was treated
using two-stage THA. With only sporadic case reports in the literature,
MTB infection is rarely clinically suspected, even in underdeveloped
and developing countries, where pulmonary and other forms of TB
are endemic. In developed countries there is also an increased incidence
among immunocompromised patients. In this paper we discuss the pathogenesis
and incidence of MTB infection after surgical management of fractures
and suggest protocols for early diagnosis and management. Cite this article:
Compartment syndrome, a devastating consequence
of limb trauma, is characterised by severe tissue injury and microvascular
perfusion deficits. We hypothesised that leucopenia might provide
significant protection against microvascular dysfunction and preserve
tissue viability. Using our clinically relevant rat model of compartment syndrome,
microvascular perfusion and tissue injury were directly visualised
by intravital video microscopy in leucopenic animals. We found that
while the tissue perfusion was similar in both groups (38.8% (standard
error of the mean ( Cite this article:
Compartment syndrome results from increased intra-compartmental
pressure (ICP) causing local tissue ischaemia and cell death, but
the systemic effects are not well described. We hypothesised that
compartment syndrome would have a profound effect not only on the affected
limb, but also on remote organs. Using a rat model of compartment syndrome, its systemic effects
on the viability of hepatocytes and on inflammation and circulation
were directly visualised using intravital video microscopy.Aims
Methods
Demineralised bone matrix (DBM) is rarely used for the local
delivery of prophylactic antibiotics. Our aim, in this study, was
to show that a graft with a bioactive glass and DBM combination,
which is currently available for clinical use, can be loaded with
tobramycin and release levels of antibiotic greater than the minimum
inhibitory concentration for Antibiotic was loaded into a graft and subsequently evaluated
for drug elution kinetics and the inhibition of bacterial growth.
A rat femoral condylar plug model was used to determine the effect
of the graft, loaded with antibiotic, on bone healing.Aims
Materials and Methods
MicroRNAs (miRNAs ) are small non-coding RNAs
that regulate gene expression. We hypothesised that the functions
of certain miRNAs and changes to their patterns of expression may
be crucial in the pathogenesis of nonunion. Healing fractures and
atrophic nonunions produced by periosteal cauterisation were created
in the femora of 94 rats, with 1:1 group allocation. At post-fracture
days three, seven, ten, 14, 21 and 28, miRNAs were extracted from
the newly generated tissue at the fracture site. Microarray and
real-time polymerase chain reaction (PCR) analyses of day 14 samples
revealed that five miRNAs, miR-31a-3p, miR-31a-5p, miR-146a-5p,
miR-146b-5p and miR-223-3p, were highly upregulated in nonunion.
Real-time PCR analysis further revealed that, in nonunion, the expression
levels of all five of these miRNAs peaked on day 14 and declined
thereafter. Our results suggest that miR-31a-3p, miR-31a-5p, miR-146a-5p,
miR-146b-5p and miR-223-3p may play an important role in the development
of nonunion. These findings add to the understanding of the molecular mechanism
for nonunion formation and may lead to the development of novel
therapeutic strategies for its treatment. Cite this article:
The scarcity of mesenchymal stem cells (MSCs) in iliac crest bone marrow aspirate (ICBMA), and the expense and time in culturing cells, has led to the search for alternative harvest sites. The reamer-irrigation-aspirator (RIA) provides continuous irrigation and suction during reaming of long bones. The aspirated contents pass via a filter, trapping bony fragments, before moving into a ‘waste’ bag from which MSCs have been previously isolated. We examined the liquid and solid phases, performed a novel digestion of the solid phase, and made a comparative assessment in terms of number, phenotype and differentiation capacity with matched ICBMA. The solid fraction from the filtrate was digested for 60 minutes at 37°C with collagenase. Enumeration was performed via the colony-forming unit fibroblast (CFU-F) assay. Passage (P2) cells were differentiated towards osteogenic, adipogenic and chondrogenic lineages, and their phenotypes assessed using flow cytometry (CD33, CD34, CD45, CD73, CD90, and CD105). MSCs from the RIA phases were able to differentiate at least as well as those from ICBMA, and all fractions had phenotypes consistent with other established sources. The median number of colonies for the three groups was: ICBMA = 8.5 (2 to 86), RIA-liquid = 19.5 (4 to 90), RIA-solid = 109 (67 to 200) per 200 μl. The mean total yield of cells for the three groups was: ICBMA = 920 (0 to 4275), RIA-liquid = 114 983 (16 500 to 477 750), RIA-solid = 12 785 (7210 to 28 475). The RIA filtrate contains large numbers of MSCs that could potentially be extracted without enzymatic digestion and used for bone repair without prior cell expansion.
Small animal models of fracture repair primarily investigate
indirect fracture healing via external callus formation. We present
the first described rat model of direct fracture healing. A rat tibial osteotomy was created and fixed with compression
plating similar to that used in patients. The procedure was evaluated
in 15 cadaver rats and then Objectives
Methods
Gunshot injuries to the shoulder are rare and
difficult to manage. We present a case series of seven patients
who sustained a severe shoulder injury to the non-dominant side
as a result of a self-inflicted gunshot wound. We describe the injury
as ‘suicide shoulder’ caused by upward and outward movement of the
gun barrel as the trigger is pulled. All patients were male, with
a mean age of 32 years (21 to 48). All were treated at the time
of injury with initial repeated debridement, and within four weeks
either by hemiarthroplasty (four patients) or arthrodesis (three patients).
The hemiarthroplasty failed in one patient after 20 years due to
infection and an arthrodesis was attempted, which also failed due
to infection. Overall follow-up was for a mean of 26 months (12
to 44). All four hemiarthroplasty implants were removed with no
feasible reconstruction ultimately possible, resulting in a poor functional
outcome and no return to work. In contrast, all three primary arthrodeses
eventually united, with two patients requiring revision plating
and grafting. These patients returned to work with a good functional
outcome. We recommend arthrodesis rather than replacement as the
treatment of choice for this challenging injury. Cite this article:
Several bisphosphonates are now available for the treatment of osteoporosis. Porous hydroxyapatite/collagen (HA/Col) composite is an osteoconductive bone substitute which is resorbed by osteoclasts. The effects of the bisphosphonate alendronate on the formation of bone in porous HA/Col and its resorption by osteoclasts were evaluated using a rabbit model. Porous HA/Col cylinders measuring 6 mm in diameter and 8 mm in length, with a pore size of 100 μm to 500 μm and 95% porosity, were inserted into a defect produced in the lateral femoral condyles of 72 rabbits. The rabbits were divided into four groups based on the protocol of alendronate administration: the control group did not receive any alendronate, the pre group had alendronate treatment for three weeks prior to the implantation of the HA/Col, the post group had alendronate treatment following implantation until euthanasia, and the pre+post group had continuous alendronate treatment from three weeks prior to surgery until euthanasia. All rabbits were injected intravenously with either saline or alendronate (7.5 μg/kg) once a week. Each group had 18 rabbits, six in each group being killed at three, six and 12 weeks post-operatively. Alendronate administration suppressed the resorption of the implants. Additionally, the mineral densities of newly formed bone in the alendronate-treated groups were lower than those in the control group at 12 weeks post-operatively. Interestingly, the number of osteoclasts attached to the implant correlated with the extent of bone formation at three weeks. In conclusion, the systemic administration of alendronate in our rabbit model at a dose-for-weight equivalent to the clinical dose used in the treatment of osteoporosis in Japan affected the mineral density and remodelling of bone tissue in implanted porous HA/Col composites.
Fracture repair occurs by two broad mechanisms:
direct healing, and indirect healing with callus formation. The effects
of bisphosphonates on fracture repair have been assessed only in
models of indirect fracture healing. A rodent model of rigid compression plate fixation of a standardised
tibial osteotomy was used. Ten skeletally mature Sprague–Dawley
rats received daily subcutaneous injections of 1 µg/kg ibandronate
(IBAN) and ten control rats received saline (control). Three weeks
later a tibial osteotomy was rigidly fixed with compression plating.
Six weeks later the animals were killed. Fracture repair was assessed
with mechanical testing, radiographs and histology. The mean stress at failure in a four-point bending test was significantly
lower in the IBAN group compared with controls (8.69 Nmm-2 ( Bisphosphonate treatment in a therapeutic dose, as used for risk
reduction in fragility fractures, had an inhibitory effect on direct
fracture healing. We propose that bisphosphonate therapy not be
commenced until after the fracture has united if the fracture has
been rigidly fixed and is undergoing direct osteonal healing. Cite this article:
Osteochondral injuries, if not treated adequately, often lead
to severe osteoarthritis. Possible treatment options include refixation
of the fragment or replacement therapies such as Pridie drilling,
microfracture or osteochondral grafts, all of which have certain
disadvantages. Only refixation of the fragment can produce a smooth
and resilient joint surface. The aim of this study was the evaluation
of an ultrasound-activated bioresorbable pin for the refixation of
osteochondral fragments under physiological conditions. In 16 Merino sheep, specific osteochondral fragments of the medial
femoral condyle were produced and refixed with one of conventional
bioresorbable pins, titanium screws or ultrasound-activated pins.
Macro- and microscopic scoring was undertaken after three months. Objectives
Methods
In this retrospective study we evaluated the method of acute shortening and distraction osteogenesis for the treatment of tibial nonunion with bone loss in 17 patients with a mean age of 36 years (10 to 58). The mean bone loss was 5.6 cm (3 to 10). In infected cases, we performed the treatment in two stages. The mean follow-up time was 43.5 months (24 to 96). The mean time in external fixation was 8.0 months (4 to 13) and the mean external fixator index was 1.4 months/cm (1.1 to 1.8). There was no recurrence of infection. The bone evaluation results were excellent in 16 patients and good in one, while functional results were excellent in 15 and good in two. The complication rate was 1.2 per patient. We conclude that acute shortening and distraction osteogenesis is a safe, reliable and successful method for the treatment of tibial nonunion with bone loss, with a shorter period of treatment and lower rate of complication.
We reviewed 13 patients with infected nonunion of the distal femur and bone loss, who had been treated by radical surgical debridement and the application of an Ilizarov external fixator. All had severely restricted movement of the knee and a mean of 3.1 previous operations. The mean length of the bony defect was 8.3 cm and no patient was able to bear weight. The mean external fixation time was 309.8 days. According to Paley’s grading system, eight patients had an excellent clinical and radiological result and seven excellent and good functional results. Bony union, the ability to bear weight fully, and resolution of the infection were achieved in all the patients. The external fixation time was increased when the definitive treatment started six months or more after the initial trauma, the patient had been subjected to more than four previous operations and the initial operation had been open reduction and internal fixation.
