Although many clinical and experimental investigations have shed
light on muscle atrophy and intramuscular accumulation of fat after
rotator cuff disruption, none have reported on their onset in the
absence of muscle retraction. In 30 rabbits, we detached one supraspinatus (SSP) tendon and
repaired it immediately, thus preventing muscle retraction. The
animals were killed in groups of 10 at one, two and six weeks. Both
shoulders of 15 non-operated rabbits served as controls. We measured
the weight and volume of SSP muscles and quantified the cross-sectional
area of intramuscular fat (i-fat) histologically.Objectives
Methods
Short intense electrical pulses transiently increase the permeability of the cell membrane, an effect known as electroporation. This can be combined with antiblastic drugs for ablation of tumours of the skin and subcutaneous tissue. The aim of this study was to test the efficacy of electroporation when applied to bone and to understand whether the presence of mineralised trabeculae would affect the capability of the electric field to porate the membrane of bone cells. Different levels of electrical field were applied to the femoral bone of rabbits. The field distribution and modelling were simulated by computer. Specimens of bone from treated and control rabbits were obtained for histology, histomorphometry and biomechanical testing. After seven days, the area of ablation had increased in line with the number of pulses and/or with the amplitude of the electrical field applied. The osteogenic activity in the ablated area had recovered by 30 days. Biomechanical testing showed structural integrity of the bone at both times. Electroporation using the appropriate combination of voltage and pulses induced ablation of bone cells without affecting the recovery of osteogenic activity. It can be an effective treatment in bone and when used in combination with drugs, an option for the treatment of metastases.
Our understanding of the origin of hip pain in
degenerative disorders of the hip, including primary osteoarthritis, avascular
necrosis and femoroacetabular impingement (FAI), is limited. We
undertook a histological investigation of the nociceptive innervation
of the acetabular labrum, ligamentum teres and capsule of the hip,
in order to prove pain- and proprioceptive-associated marker expression.
These structures were isolated from 57 patients who had undergone
elective hip surgery (44 labral samples, 33 ligamentum teres specimens,
34 capsular samples; in 19 patients all three structures were harvested).
A total of
15 000 histological sections were prepared that were investigated
immunohistochemically for the presence of protein S-100, 68 kDa
neurofilament, neuropeptide Y, nociceptin and substance P. The tissues
were evaluated in six representative areas. Within the labrum, pain-associated free nerve ending expression
was located predominantly at its base, decreasing in the periphery.
In contrast, the distribution within the ligamentum teres showed
a high local concentration in the centre. The hip capsule had an
almost homogeneous marker expression in all investigated areas. This study showed characteristic distribution profiles of nociceptive
and pain-related nerve fibres, which may help in understanding the
origin of hip pain. Cite this article:
The peri-prosthetic tissue response to wear debris
is complex and influenced by various factors including the size, area
and number of particles. We hypothesised that the ‘biologically
active area’ of all metal wear particles may predict the type of
peri-prosthetic tissue response. Peri-prosthetic tissue was sampled from 21 patients undergoing
revision of a small diameter metal-on-metal (MoM) total hip arthroplasty
(THA) for aseptic loosening. An enzymatic protocol was used for
tissue digestion and scanning electron microscope was used to characterise
particles. Equivalent circle diameters and particle areas were calculated.
Histomorphometric analyses were performed on all tissue specimens.
Aspirates of synovial fluid were collected for analysis of the cytokine
profile analysis, and compared with a control group of patients
undergoing primary THA (n = 11) and revision of a failed ceramic-on-polyethylene
arthroplasty (n = 6). The overall distribution of the size and area of the particles
in both lymphocyte and
non-lymphocyte-dominated responses were similar; however, the subgroup
with lymphocyte-dominated peri-prosthetic tissue responses had a
significantly larger total number of particles. 14 cytokines (interleukin (IL)-1ß, IL-2, IL-4, IL-5, IL-6, IL-10,
IL-13, IL-17, interferon (IFN)-γ, and IFN-gamma-inducible protein
10), chemokines (macrophage inflammatory protein (MIP)-1α and MIP-1ß),
and growth factors (granulocyte macrophage colony stimulating factor
(GM-CSF) and platelet derived growth factor) were detected at significantly higher
levels in patients with metal wear debris compared with the control
group. Significantly higher levels for IL-1ß, IL-5, IL-10 and GM-CSF
were found in the subgroup of tissues from failed MoM THAs with
a lymphocyte-dominated peri-prosthetic response compared with those
without this response. These results suggest that the ‘biologically active area’ predicts
the type of
peri-prosthetic tissue response. The cytokines IL-1ß, IL-5, IL-10,
and GM-CSF are associated with lymphocyte-dominated tissue responses
from failed small-diameter MoM THA. Cite this article:
In order to ensure safety of the cell-based therapy for bone
regeneration, we examined BM cells obtained from a total of 13 Sprague-Dawley (SD) green
fluorescent protein transgenic (GFP-Tg) rats were culture-expanded
in an osteogenic differentiation medium for three weeks. Osteoblast-like
cells were then locally transplanted with collagen scaffolds to
the rat model of segmental bone defect. Donor cells were also intravenously infused
to the normal Sprague-Dawley (SD) rats for systemic biodistribution.
The flow cytometric and histological analyses were performed for
cellular tracking after transplantation.Objectives
Methods
Osteoarthritis (OA) is a progressively debilitating disease that
affects mostly cartilage, with associated changes in the bone. The
increasing incidence of OA and an ageing population, coupled with
insufficient therapeutic choices, has led to focus on the potential
of stem cells as a novel strategy for cartilage repair. In this study, we used scaffold-free mesenchymal stem cells (MSCs)
obtained from bone marrow in an experimental animal model of OA
by direct intra-articular injection. MSCs were isolated from 2.8
kg white New Zealand rabbits. There were ten in the study group
and ten in the control group. OA was induced by unilateral transection
of the anterior cruciate ligament of the knee joint. At 12 weeks
post-operatively, a single dose of 1 million cells suspended in 1 ml
of medium was delivered to the injured knee by direct intra-articular
injection. The control group received 1 ml of medium without cells.
The knees were examined at 16 and 20 weeks following surgery. Repair
was investigated radiologically, grossly and histologically using
haematoxylin and eosin, Safranin-O and toluidine blue staining.Introduction
Methods
The nervous system is known to be involved in inflammation and repair. We aimed to determine the effect of physical activity on the healing of a muscle injury and to examine the pattern of innervation. Using a drop-ball technique, a contusion was produced in the gastrocnemius in 20 rats. In ten the limb was immobilised in a plaster cast and the remaining ten had mobilisation on a running wheel. The muscle and the corresponding dorsal-root ganglia were studied by histological and immunohistochemical methods. In the mobilisation group, there was a significant reduction in lymphocytes (p = 0.016), macrophages (p = 0.008) and myotubules (p = 0.008) between three and 21 days. The formation of myotubules and the density of nerve fibres was significantly higher (both p = 0.016) compared with those in the immobilisation group at three days, while the density of CGRP-positive fibres was significantly lower (p = 0.016) after 21 days. Mobilisation after contusional injury to the muscle resulted in early and increased formation of myotubules, early nerve regeneration and progressive reduction in inflammation, suggesting that it promoted a better healing response.
Fracture of a ceramic component in total hip
replacement is a rare but potentially catastrophic complication.
The incidence is likely to increase as the use of ceramics becomes
more widespread. We describe such a case, which illustrates how
inadequate initial management will lead to further morbidity and
require additional surgery. We present the case as a warning that
fracture of a ceramic component should be revised to another ceramic-on-ceramic
articulation in order to minimise the risk of further catastrophic
wear.
The main form of treatment of a chordoma of the
mobile spine is total We report two patients who underwent TES after CIRT for treating
a chordoma in the lumbar spine with good medium-term outcomes. At
operation, there remained histological evidence of viable tumour
cells in both cases. After the combination use of TES following
CIRT, neither patient showed signs of recurrence at the follow-up examination.
These two cases suggest that CIRT should be combined with total
spondylectomy in the treatment of chordoma of the mobile spine. 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.
Platelet-derived growth factor (PDGF) is known
to stimulate osteoblast or osteoprogenitor cell activity. We investigated
the effect of locally applied PDGF from poly- These results indicate that local application of PDGF from biodegradable
PDLLA-coated implants significantly accelerates fracture healing
in experimental animals. Further development may help fracture healing
in the clinical situation.
Although gradual bone transport may permit the
restoration of large-diameter bones, complications are common owing
to the long duration of external fixation. In order to reduce such
complications, a new technique of bone transport involving the use
of an external fixator and a locking plate was devised for segmental
tibial bone defects. A total of ten patients (nine men, one woman) with a mean age
at operation of 40.4 years (16 to 64) underwent distraction osteogenesis
with a locking plate to treat previously infected post-traumatic
segmental tibial defects. The locking plate was fixed percutaneously
to bridge proximal and distal segments, and was followed by external fixation.
After docking, percutaneous screws were fixed at the transported
segment through plate holes. At the same time, bone grafting was
performed at the docking site with the external fixator removed. The mean defect size was 5.9 cm (3.8 to 9.3) and mean external
fixation index was
13.4 days/cm (11.8 to 19.5). In all cases, primary union of the
docking site and distraction callus was achieved, with an excellent
bony result. There was no recurrence of deep infection or osteomyelitis,
and with the exception of one patient with a pre-existing peroneal
nerve injury, all achieved an excellent or good functional result. With short external fixation times and low complication rates,
bone transport with a locking plate could be recommended for patients
with segmental tibial defects. Cite this article:
Surgical marking during tendon surgery is often used for technical
and teaching purposes. This study investigates the effect of a gentian
violet ink marker pen, a common surgical marker, on the viability
of the tissue and cells of tendon.
Objectives
Methods
The aim of this study was to investigate the occurrence of tissue hypoxia and apoptosis at different stages of tendinopathy and tears of the rotator cuff. We studied tissue from 24 patients with eight graded stages of either impingement (mild, moderate and severe) or tears of the rotator cuff (partial, small, medium, large and massive) and three controls. Biopsies were analysed using three immunohistochemical techniques, namely antibodies against HIF-1α (a transcription factor produced in a hypoxic environment), BNip3 (a HIF-1α regulated pro-apoptotic protein) and TUNEL (detecting DNA fragmentation in apoptosis). The HIF-1α expression was greatest in mild impingement and in partial, small, medium and large tears. BNip3 expression increased significantly in partial, small, medium and large tears but was reduced in massive tears. Apoptosis was increased in small, medium, large and massive tears but not in partial tears. These findings reveal evidence of hypoxic damage throughout the spectrum of pathology of the rotator cuff which may contribute to loss of cells by apoptosis. This provides a novel insight into the causes of degeneration of the rotator cuff and highlights possible options for treatment.
Fresh-frozen allograft bone is frequently used
in orthopaedic surgery. We investigated the incidence of allograft-related
infection and analysed the outcomes of recipients of bacterial culture-positive
allografts from our single-institute bone bank during bone transplantation.
The fresh-frozen allografts were harvested in a strict sterile environment
during total joint arthroplasty surgery and immediately stored in
a freezer at -78º to -68º C after packing. Between January 2007
and December 2012, 2024 patients received 2083 allografts with a
minimum of 12 months of follow-up. The overall allograft-associated
infection rate was 1.2% (24/2024). Swab cultures of 2083 allografts
taken before implantation revealed 21 (1.0%) positive findings.
The 21 recipients were given various antibiotics at the individual
orthopaedic surgeon’s discretion. At the latest follow-up, none
of these 21 recipients displayed clinical signs of infection following
treatment. Based on these findings, we conclude that an incidental positive
culture finding for allografts does not correlate with subsequent
surgical site infection. Additional prolonged post-operative antibiotic
therapy may not be necessary for recipients of fresh-frozen bone
allograft with positive culture findings. Cite this article:
The LockDown device (previously called Surgilig)
is a braided polyester mesh which is mostly used to reconstruct the
dislocated acromioclavicular joint. More than 11 000 have been implanted
worldwide. Little is known about the tissue reaction to the device
nor to its wear products when implanted in an extra-articular site
in humans. This is of importance as an adverse immunological reaction
could result in osteolysis or damage to the local tissues, thereby affecting
the longevity of the implant. We analysed the histology of five LockDown implants retrieved
from five patients over the last seven years by one of the senior
authors. Routine analysis was carried out in all five cases and
immunohistochemistry in one. The LockDown device acts as a scaffold for connective tissue
which forms an investing fibrous pseudoligament. The immunological
response at the histological level seems favourable with a limited
histiocytic and giant cell response to micron-sized wear particles.
The connective tissue envelope around the implant is less organised
than a native ligament. Cite this article:
Recent studies have shown that modulating inflammation-related
lipid signalling after a bone fracture can accelerate healing in
animal models. Specifically, decreasing 5-lipoxygenase (5-LO) activity
during fracture healing increases cyclooxygenase-2 (COX-2) expression
in the fracture callus, accelerates chondrogenesis and decreases
healing time. In this study, we test the hypothesis that 5-LO inhibition
will increase direct osteogenesis. Bilateral, unicortical femoral defects were used in rats to measure
the effects of local 5-LO inhibition on direct osteogenesis. The
defect sites were filled with a polycaprolactone (PCL) scaffold
containing 5-LO inhibitor (A-79175) at three dose levels, scaffold
with drug carrier, or scaffold only. Drug release was assessed Objectives
Methods
We attempted to characterise the biological quality
and regenerative potential of chondrocytes in osteochondritis dissecans
(OCD). Dissected fragments from ten patients with OCD of the knee
(mean age 27.8 years (16 to 49)) were harvested at arthroscopy.
A sample of cartilage from the intercondylar notch was taken from
the same joint and from the notch of ten patients with a traumatic
cartilage defect (mean age 31.6 years (19 to 52)). Chondrocytes
were extracted and subsequently cultured. Collagen types 1, 2, and
10 mRNA were quantified by polymerase chain reaction. Compared with
the notch chondrocytes, cells from the dissecate expressed similar
levels of collagen types 1 and 2 mRNA. The level of collagen type
10 message was 50 times lower after cell culture, indicating a loss
of hypertrophic cells or genes. The high viability, retained capacity
to differentiate and metabolic activity of the extracted cells suggests
preservation of the intrinsic repair capability of these dissecates.
Molecular analysis indicated a phenotypic modulation of the expanded
dissecate chondrocytes towards a normal phenotype. Our findings
suggest that cartilage taken from the dissecate can be reasonably
used as a cell source for chondrocyte implantation procedures.
Ligaments which heal spontaneously have a healing process that
is similar to skin wound healing. Menopause impairs skin wound healing
and may likewise impair ligament healing. Our purpose in this study
was to investigate the effect of surgical menopause on ligament
healing in a rabbit medial collateral ligament model. Surgical menopause was induced with ovariohysterectomy surgery
in adult female rabbits. Ligament injury was created by making a
surgical gap in the midsubstance of the medial collateral ligament.
Ligaments were allowed to heal for six or 14 weeks in the presence
or absence of oestrogen before being compared with uninjured ligaments. Molecular
assessment examined the messenger ribonucleic acid levels for collagens,
proteoglycans, proteinases, hormone receptors, growth factors and
inflammatory mediators. Mechanical assessments examined ligament
laxity, total creep strain and failure stress.Objectives
Methods
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