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The Bone & Joint Journal
Vol. 105-B, Issue 6 | Pages 657 - 662
1 Jun 2023
Meaike JJ Meaike JD Collins MS Bishop AT Shin AY

Aims. The benefit of MRI in the preoperative assessment of scaphoid proximal fragment vascularity remains controversial. The purpose of this study is to compare preoperative MRI findings to intraoperative bleeding of the proximal scaphoid. Methods. A retrospective review of 102 patients who underwent surgery for scaphoid nonunion between January 2000 and December 2020 at a single institution were identified. Inclusion criteria were: isolated scaphoid nonunion; preoperative MRI assessing the proximal fragment vascularity; and operative details of the vascularity of the proximal fragment with the tourniquet deflated. MRI results and intraoperative findings were dichotomized as either ‘yes’ or ‘no’ for the presence of vascularity. A four-fold contingency table was used to analyze the utility of preoperative MRI with 95% confidence intervals. Relative risk was calculated for subgroups to analyze the association between variables and MRI accuracy. Results. Preoperative MRI identified 55 proximal scaphoid fragments with ischaemia and 47 with vascularized proximal fragments. After the proximal fragment was prepared, the tourniquet was deflated and assessed for bleeding; 63 proximal fragments had no bleeding and 39 demonstrated bleeding. MRI was not reliable or accurate in the assessment of proximal fragment vascularity when compared with intraoperative assessment of bleeding. No patient or MRI factors were identified to have a statistical impact on MRI accuracy. Conclusion. Current preoperative MRI protocols and diagnostic criteria do not provide a high degree of correlation with observed intraoperative assessment of proximal fragment bleeding. While preoperative MRI may assist in surgical planning, intraoperative assessment remains the best means for assessing proximal fragment vascularity in scaphoid nonunion. Future efforts should focus on the development of objective measures of osseous blood flow that may be performed intraoperatively. Cite this article: Bone Joint J 2023;105-B(6):657–662


The Bone & Joint Journal
Vol. 106-B, Issue 8 | Pages 842 - 848
1 Aug 2024
Kriechling P Whitefield R Makaram NS Brown IDM Mackenzie SP Robinson CM

Aims. Vascular compromise due to arterial injury is a rare but serious complication of a proximal humeral fracture. The aims of this study were to report its incidence in a large urban population, and to identify clinical and radiological factors which are associated with this complication. We also evaluated the results of the use of our protocol for the management of these injuries. Methods. A total of 3,497 adult patients with a proximal humeral fracture were managed between January 2015 and December 2022 in a single tertiary trauma centre. Their mean age was 66.7 years (18 to 103) and 2,510 (72%) were female. We compared the demographic data, clinical features, and configuration of those whose fracture was complicated by vascular compromise with those of the remaining patients. The incidence of vascular compromise was calculated from national population data, and predictive factors for its occurrence were investigated using univariate analysis. Results. A total of 18 patients (0.5%) had a proximal humeral fracture and clinical evidence of vascular compromise, giving an annual incidence of 0.29 per 100,000 of the population. Their mean age was 68.7 years (45 to 92) and ten (56%) were female. Evidence of a mixed pattern neurological deficit (brachial plexus palsy) (odds ratio (OR) 380.6 (95% CI 85.9 to 1,685.8); p < 0.001), complete separation of the proximal shaft from the humeral head with medial displacement (OR 39.5 (95% CI 14.0 to 111.8); p < 0.001), and a fracture-dislocation (OR 5.0 (95% CI 1.6 to 15.3); p = 0.015) were all associated with an increased risk of associated vascular compromise. A policy of reduction and fixation of the fracture prior to vascular surgical intervention had favourable outcomes without vascular sequelae. Conclusion. The classic signs of distal ischaemia are often absent in patients with proximal injuries to major vessels. We were able to identify specific clinical and radiological ‘red flags’ which, particularly when present in combination, should increase the suspicion of a fracture with an associated vascular injury, and facilitate early diagnosis and appropriate combined orthopaedic and vascular intervention. Cite this article: Bone Joint J 2024;106-B(8):842–848


Bone & Joint Research
Vol. 12, Issue 3 | Pages 212 - 218
9 Mar 2023
Buchalter DB Kirby DJ Anil U Konda SR Leucht P

Aims. Glucose-insulin-potassium (GIK) is protective following cardiac myocyte ischaemia-reperfusion (IR) injury, however the role of GIK in protecting skeletal muscle from IR injury has not been evaluated. Given the similar mechanisms by which cardiac and skeletal muscle sustain an IR injury, we hypothesized that GIK would similarly protect skeletal muscle viability. Methods. A total of 20 C57BL/6 male mice (10 control, 10 GIK) sustained a hindlimb IR injury using a 2.5-hour rubber band tourniquet. Immediately prior to tourniquet placement, a subcutaneous osmotic pump was placed which infused control mice with saline (0.9% sodium chloride) and treated mice with GIK (40% glucose, 50 U/l insulin, 80 mEq/L KCl, pH 4.5) at a rate of 16 µl/hr for 26.5 hours. At 24 hours following tourniquet removal, bilateral (tourniqueted and non-tourniqueted) gastrocnemius muscles were triphenyltetrazolium chloride (TTC)-stained to quantify percentage muscle viability. Bilateral peroneal muscles were used for gene expression analysis, serum creatinine and creatine kinase activity were measured, and a validated murine ethogram was used to quantify pain before euthanasia. Results. GIK treatment resulted in a significant protection of skeletal muscle with increased viability (GIK 22.07% (SD 15.48%)) compared to saline control (control 3.14% (SD 3.29%)) (p = 0.005). Additionally, GIK led to a statistically significant reduction in gene expression markers of cell death (CASP3, p < 0.001) and inflammation (NOS2, p < 0.001; IGF1, p = 0.007; IL-1β, p = 0.002; TNFα, p = 0.012), and a significant reduction in serum creatine kinase (p = 0.004) and creatinine (p < 0.001). GIK led to a significant reduction in IR-related pain (p = 0.030). Conclusion. Systemic GIK infusion during and after limb ischaemia protects murine skeletal muscle from cell death, kidneys from reperfusion metabolites, and reduces pain by reducing post-ischaemic inflammation. Cite this article: Bone Joint Res 2023;12(3):212–218


The Journal of Bone & Joint Surgery British Volume
Vol. 50-B, Issue 3 | Pages 482 - 492
1 Aug 1968
McQuillan WM Nolan B

1. The types of ischaemia that accompany injuries to a limb are described. 2. The ischaemia may be total or local. 3. Thirty-seven cases of total or local ischaemia of a limb are analysed and conclusions are drawn therefrom


The Journal of Bone & Joint Surgery British Volume
Vol. 68-B, Issue 4 | Pages 620 - 624
1 Aug 1986
James J Steijn-Myagkaya G

Ischaemia kills osteocytes, but opinions differ as to how long they can survive. These differences are due to the varying methods of inducing ischaemia, and to the different criteria for diagnosing cell death. Using rabbit bone and a technique of in vitro ischaemia at 37 degrees C, we have shown by electron microscopy that, after up to two hours, the changes which occur are probably reversible; after four hours, the cells were irreversibly damaged. This difference could not be detected by light microscopy. After 24 hours of ischaemia, most lacunae were empty or contained only osteocyte debris. We conclude that osteocytes suffer irreversible damage after in vitro ischaemia of about two hours, which is much the same response as that of most other mammalian cells


The Journal of Bone & Joint Surgery British Volume
Vol. 48-B, Issue 1 | Pages 150 - 152
1 Feb 1966
Monk CJE

1. A patient with complete ischaemia of the muscles of the calf and anterior compartment of the leg is described. 2. In diagnosis, tenseness of the calf, equinus and pain on attempted dorsiflexion of the ankle are most important. 3. Peripheral pulses may be present and do not contra-indicate decompression by operation to prevent ischaemia


The Journal of Bone & Joint Surgery British Volume
Vol. 49-B, Issue 2 | Pages 268 - 275
1 May 1967
Owen R Tsimboukis B

1. The frequency of variable degrees of ischaemia of soft tissue in closed tibial and fibular shaft fractures is emphasised. 2. Two cases with ischaemia of calf musculature are described. 3. A follow-up study of 100 cases of closed tibial shaft fractures is analysed; ten patients were found to be affected. 4. The anatomical and pathological changes are discussed. 5. A plea is made for early surgical exploration in cases manifesting signs of ischaemia


The Journal of Bone & Joint Surgery British Volume
Vol. 66-B, Issue 3 | Pages 434 - 440
1 May 1984
Newman R

A model of tourniquet ischaemia was developed in the hind limb of the rat, and the metabolic changes that occurred in the calf muscles were monitored by the non-invasive technique of phosphorus-31 nuclear magnetic resonance spectroscopy. During ischaemia the intramyocellular pH became acidic as the level of phosphocreatine declined and that of inorganic phosphate rose. Phosphocreatine was no longer detectable after approximately 2 hours and ATP was depleted after approximately 3.5 hours. Metabolic recovery was rapid (1 hour) if ATP was present when the tourniquet was released but was prolonged (3 or more hours) if ATP was depleted. Hourly release of the tourniquet for 10 minutes ensured the maintenance of ATP and rapid metabolic recovery. Release for intervals of only 5 minutes did not have the same protective effect and in fact worsened tissue pH during the period of tourniquet ischaemia. Heparin and corticosteroids were without effect during and after periods of tourniquet ischaemia


The Journal of Bone & Joint Surgery British Volume
Vol. 78-B, Issue 6 | Pages 930 - 933
1 Nov 1996
Farrar MJ Bennet GC Wilson NIL Azmy A

Peripheral limb ischaemia is rare in children. We have treated only 12 infants and children with this condition in the past 15 years at the Royal Hospital for Sick Children in Glasgow. There were nine neonates and three older children. Most were suffering from life-threatening illnesses or severe infection. Two were born with ischaemic arms with no apparent cause. We have analysed the factors leading to ischaemia, the outcome of the initial treatment and the later orthopaedic problems. Two required amputation of both legs, one of an arm, two of feet and one of toes. Two had skin grafts. All surgery was performed after demarcation was well established and delayed closure was used after amputation. Five children developed limb-length discrepancy or an angular deformity. To date two have required additional corrective surgery


The Journal of Bone & Joint Surgery British Volume
Vol. 91-B, Issue 11 | Pages 1487 - 1492
1 Nov 2009
Blakey CM Biant LC Birch R

A series of 26 children was referred to our specialist unit with a ‘pink pulseless hand’ following a supracondylar fracture of the distal humerus after a mean period of three months (4 days to 12 months) except for one referred after almost three years. They were followed up for a mean of 15.5 years (4 to 26). The neurovascular injuries and resulting impairment in function and salvage procedures were recorded. The mean age at presentation was 8.6 years (2 to 12). There were eight girls and 18 boys. Only four of the 26 patients had undergone immediate surgical exploration before referral and three of these four had a satisfactory outcome. In one child the brachial artery had been explored unsuccessfully at 48 hours. As a result 23 of the 26 children presented with established ischaemic contracture of the forearm and hand. Two responded to conservative stretching. In the remaining 21 the antecubital fossa was explored. The aim of surgery was to try to improve the function of the hand and forearm, to assess nerve, vessel and muscle damage, to relieve entrapment and to minimise future disturbance of growth. Based on our results we recommend urgent exploration of the vessels and nerves in a child with a ‘pink pulseless hand’, not relieved by reduction of a supracondylar fracture of the distal humerus and presenting with persistent and increasing pain suggestive of a deepening nerve lesion and critical ischaemia


The Journal of Bone & Joint Surgery British Volume
Vol. 53-B, Issue 1 | Pages 113 - 117
1 Feb 1971
Robins RHC Murrell JS

A case is reported of fracture of the tibia in a haemophiliac, complicated by severe ischaemia. Extensive decompression of the limb followed later by skin grafting was required. The haematological management which made such surgery practicable is described. It involved the use of cryoprecipitate for a period of seven weeks


The Journal of Bone & Joint Surgery British Volume
Vol. 48-B, Issue 4 | Pages 627 - 636
1 Nov 1966
Seddon HJ

1. Volkmann's ischaemia of the lower limb is more common in adults than in children and occurs with sufficient frequency after injuries of the femur, knee and leg to warrant a more determined effort to prevent it. 2. The first and most essential step is to recognise the early signs of ischaemic damage. Incision of the deep fascia may then save the threatened underlying muscle, though it may also be necessary to seek for and evacuate a haematoma beneath the muscle. When the femoral or popliteal artery is injured, exploration and repair may be imperative. 3. The treatment of established ischaemic contracture is by whatever measures are necessary to correct the deformity. These are lengthening of shortened tendons, or excision of them if they are involved in dense fibrosis at the periphery of the ischaemic mass; and excision of all totally destroyed muscle. Amputation may be necessary


The Journal of Bone & Joint Surgery British Volume
Vol. 52-B, Issue 4 | Pages 738 - 740
1 Nov 1970
Kirby NG

1. A bilateral case of chronic intermittent ischaemia of the soleus muscle within its fascial compartment is described in a dental surgeon aged twenty-seven. 2. Symptoms were promptly relieved by decompression. 3. The mechanism of the condition is compared with that of the anterior tibial syndrome


The Journal of Bone & Joint Surgery British Volume
Vol. 82-B, Issue 3 | Pages 383 - 386
1 Apr 2000
Davies MS Nadel S Habibi P Levin M Hunt DM

Between March 1993 and February 1999, 14 children aged from eight months to 14.75 years were admitted to the paediatric intensive-care unit with meningococcal septicaemia in association with severe peripheral ischaemia. Of these, 13 were operated upon, eight of whom had early fasciotomies. Five children died. Of the nine survivors, one had no amputations while in the other eight 14 limb segments were amputated. We review the case histories and propose a protocol for the early management of these children


The Bone & Joint Journal
Vol. 105-B, Issue 3 | Pages 231 - 238
1 Mar 2023
Holme TJ Crate G Trompeter AJ Monsell FP Bridgens A Gelfer Y

Aims

The ‘pink, pulseless hand’ is often used to describe the clinical situation in which a child with a supracondylar fracture of the humerus has normal distal perfusion in the absence of a palpable peripheral pulse. The management guidelines are based on the assessment of perfusion, which is difficult to undertake and poorly evaluated objectively. The aim of this study was to review the available literature in order to explore the techniques available for the preoperative clinical assessment of perfusion in these patients and to evaluate the clinical implications.

Methods

A systematic literature review was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines and registered prospectively with the International Prospective Register of Systematic Reviews. Databases were explored in June 2022 with the search terms (pulseless OR dysvascular OR ischaemic OR perfused OR vascular injury) AND supracondylar AND (fracture OR fractures).


The Journal of Bone & Joint Surgery British Volume
Vol. 67-B, Issue 2 | Pages 297 - 301
1 Mar 1985
Irving G Noakes T

The protective effect of local hypothermia was studied in pig's limbs made ischaemic by long, repeated application of a pneumatic tourniquet. Twenty-one Landrace pigs were anaesthetised on two separate occasions six days apart and a pneumatic tourniquet at 500 mmHg pressure was applied to the same forelimb for three and two hours respectively. Ten of the pigs had local hypothermia from cold gel packs placed around the limb during the first tourniquet application; the other 11 had the ischaemic limb exposed to room temperature. In comparison with the normothermic limbs, the hypothermic ischaemic limbs had significant slowing of metabolism. The hypothermic limbs also showed less inflammatory response and a faster rate of recovery, both immediately after removal of the tourniquet, and by the end of the experiment, 10 days after the first tourniquet. Local hypothermia produced by this technique was shown to be safe and effective, while appearing to protect muscles exposed to prolonged tourniquet-induced ischaemia


The Journal of Bone & Joint Surgery British Volume
Vol. 70-B, Issue 3 | Pages 465 - 467
1 May 1988
Bongard O Krahenbuhl B

The predictive value of the pedal transcutaneous oxygen tension (tcPO2) and of the distal systolic blood pressure (SBP) in forecasting the necessity for later amputation has been studied in 26 patients suffering from severe chronic ischaemia of the lower limbs. In all these patients vascular surgery had failed or not been possible, and they were threatened by amputation; they suffered from trophic lesions, or pain at rest, or both. The great toe SBP averaged 10 mmHg (range 0 to 60 mmHg) and the pedal tcPO2 10 mmHg (range 2 to 45 mmHg). After six minutes of oxygen inhalation there was an increase in pedal tcPO2 of 9 mmHg (0 to 50 mmHg). After a follow-up period averaging 7 months (range 10 days to 13 months), 13 patients underwent an amputation and nine (five of whom had been amputated) died. The great toe SBP in the patients who required amputation was initially lower than in those who did not. The pedal tcPO2 also was lower in amputated than in non-amputated patients. There was no amputation in the group showing an increase of at least 10 mmHg after six minutes of oxygen inhalation; and conversely, all patients in whom the pedal tcPO2 increased less than 10 mmHg were amputated. Thus increase in the pedal tcPO2 after oxygen inhalation appears the best criterion for estimating the prognosis of severely ischaemic limbs


The Journal of Bone & Joint Surgery British Volume
Vol. 61-B, Issue 3 | Pages 265 - 266
1 Aug 1979
Eadie D


The Journal of Bone & Joint Surgery British Volume
Vol. 50-B, Issue 3 | Pages 472 - 481
1 Aug 1968
MacGowan W

1. Experience with twelve cases of arterial trauma are presented.

2. Six limbs were amputated and six limbs saved.

3. Delay in effective treatment was the major cause of failure to save the limb.

4. Five types of limb injury involving vessels are described.


The Journal of Bone & Joint Surgery British Volume
Vol. 34-B, Issue 1 | Pages 68 - 69
1 Feb 1952
Wilson JN


The Journal of Bone & Joint Surgery British Volume
Vol. 44-B, Issue 4 | Pages 760 - 761
1 Nov 1962
Barnes R


The Journal of Bone & Joint Surgery British Volume
Vol. 83-B, Issue 3 | Pages 461 - 463
1 Apr 2001
FARRAR MJ BENNETT GC WILSON NIL AZMY A


The Journal of Bone & Joint Surgery British Volume
Vol. 42-B, Issue 1 | Pages 110 - 125
1 Feb 1960
Brookes M

1. Twenty-five lower limbs, amputated above the knee for senile atherosclerosis with peripheral gangrene, have been investigated radiologically and histologically to determine the vascular patterns in ischaemic bone with particular reference to the tibia. These have been contrasted with the patterns found in non-atherosclerotic tubular bone.

2. The principal changes are the development of a diffuse vascularisation of compact and spongy bone; a widening of Haversian spaces which come to contain a variable number of sinusoidal blood vessels; and an increasing periosteal participation in cortical nutrition which is related to the severity and chronicity of the ischaemic process.

3. Views on the normal blood supply of long bones are discussed, and evidence is presented for regarding this as discrete and end-arterial in nature; in particular it is suggested that the normal cortex has a wholly medullary, centrifugal, arterial supply.


The Journal of Bone & Joint Surgery British Volume
Vol. 71-B, Issue 1 | Pages 102 - 104
1 Jan 1989
Falstie-Jensen N Christensen K Brochner-Mortensen J

We analysed the complication rate in 140 below-knee amputations in relation to surgical technique and the presence of diabetes. In all cases, the skin perfusion pressure was measured below the knee before operation to provide an objective evaluation of the microcirculation. In diabetic patients we found a significantly higher complication rate after using a long posterior flap than after equal sagittal flaps. No such difference could be demonstrated in non-diabetic patients. We suggest that the higher incidence of atherosclerotic lesions in the three major arteries below the knee in diabetic patients may account for the difference. We recommend the use of the sagittal technique for below-knee amputation in diabetic patients.


Bone & Joint Research
Vol. 10, Issue 6 | Pages 363 - 369
1 Jun 2021
MacDonald DRW Neilly DW Elliott KE Johnstone AJ

Aims. Tourniquets have potential adverse effects including postoperative thigh pain, likely caused by their ischaemic and possible compressive effects. The aims of this preliminary study were to determine if it is possible to directly measure intramuscular pH in human subjects over time, and to measure the intramuscular pH changes resulting from tourniquet ischaemia in patients undergoing knee arthroscopy. Methods. For patients undergoing short knee arthroscopic procedures, a sterile calibrated pH probe was inserted into the anterior fascial compartment of the leg after skin preparation, but before tourniquet inflation. The limb was elevated for three minutes prior to tourniquet inflation to 250 mmHg or 300 mmHg. Intramuscular pH was recorded at one-second intervals throughout the procedure and for 20 minutes following tourniquet deflation. Probe-related adverse events were recorded. Results. A total of 27 patients were recruited to the study. Mean tourniquet time was 21 minutes (10 to 56). Tourniquet pressure was 300 mmHg for 21 patients and 250 mmHg for six patients. Mean muscle pH prior to tourniquet inflation was 6.80. Muscle pH decreased upon tourniquet inflation, with a steeper fall in the first ten minutes than for the rest of the procedure. Change in muscle pH was significant after five minutes of tourniquet ischaemia (p < 0.001). Mean muscle pH prior to tourniquet release was 6.58 and recovered to 6.75 within 20 minutes following release. No probe related adverse events were recorded. Conclusion. It is possible to directly measure skeletal muscle pH in human subjects over time. Tourniquet ischaemia results in a decrease in human skeletal muscle pH over time during short procedures. Cite this article: Bone Joint Res 2021;10(6):363–369


Bone & Joint Research
Vol. 11, Issue 4 | Pages 189 - 199
13 Apr 2022
Yang Y Li Y Pan Q Bai S Wang H Pan X Ling K Li G

Aims. Treatment for delayed wound healing resulting from peripheral vascular diseases and diabetic foot ulcers remains a challenge. A novel surgical technique named ‘tibial cortex transverse transport’ (TTT) has been developed for treating peripheral ischaemia, with encouraging clinical effects. However, its underlying mechanisms remain unclear. In the present study, we explored the potential biological mechanisms of TTT surgery using various techniques in a rat TTT animal model. Methods. A novel rat model of TTT was established with a designed external fixator, and effects on wound healing were investigated. Laser speckle perfusion imaging, vessel perfusion, histology, and immunohistochemistry were used to evaluate the wound healing processes. Results. Gross and histological examinations showed that TTT technique accelerated wound closure and enhanced the quality of the newly formed skin tissues. In the TTT group, haematoxylin and eosin (H&E) staining demonstrated a better epidermis and dermis recovery, while immunohistochemical staining showed that TTT technique promoted local collagen deposition. The TTT technique also benefited to angiogenesis and immunomodulation. In the TTT group, blood flow in the wound area was higher than that of other groups according to laser speckle imaging with more blood vessels observed. Enhanced neovascularization was seen in the TTT group with double immune-labelling of CD31 and α-Smooth Muscle Actin (α-SMA). The number of M2 macrophages at the wound site in the TTT group was also increased. Conclusion. The TTT technique accelerated wound healing through enhanced angiogenesis and immunomodulation. Cite this article: Bone Joint Res 2022;11(4):189–199



The Journal of Bone & Joint Surgery British Volume
Vol. 61-B, Issue 3 | Pages 296 - 300
1 Aug 1979
Holden C

Ischaemia is a rare complication of injury to a limb but must be excluded in every case. Two distinct types occur: Type I, where a proximal arterial injury gives rise to ischaemia distally; and Type II, where a direct injury gives rise to ischaemia at the site of the injury. Whatever the nature of the insult, an ischaemic contracture only develops as a result of swelling of the soft tissues where these soft tissues are contained in un unyielding osteofascial compartment. This secondary ischaemia can only be relieved by a timely fasciotomy. The diagnosis of ischaemia in an injured limb and the indications to operate on it can usually be made on clinical grounds alone


The Journal of Bone & Joint Surgery British Volume
Vol. 84-B, Issue 8 | Pages 1189 - 1193
1 Nov 2002
Bushell AJ Klenerman L Davies H Grierson I McArdle A Jackson MJ

We have previously shown that prior exposure of rat hind limbs to ischaemia for five minutes and reperfusion for five minutes reduced the structural damage to skeletal muscle which followed a subsequent period of ischaemia for four hours and reperfusion for one hour. We have now examined the potential mechanisms by which this ischaemic preconditioning protocol may be effective in reducing damage to skeletal muscle induced by prolonged ischaemia and reperfusion. Prior exposure of the hindlimb to ischaemia for five minutes and reperfusion for five minutes did not prevent the fall in the ATP content of tibialis anterior which occurred after a subsequent period of ischaemia for four hours and reperfusion for one hour. Similarly, no effect of the preconditioning protocol was seen on the elevated muscle myeloperoxidase, indicative of an elevated neutrophil content, or abnormal muscle cation content. Reperfused ischaemic muscle was also found to have an increased content of heat-shock protein (HSP) 72, but the preconditioning protocol did not further increase the content of this or other HSPs indicating that it was not acting by increasing the expression of these cytoprotective proteins. The protective effects of preconditioning appeared to be mimicked by the infusion of adenosine to animals immediately before exposure to the four-hour period, indicating a potential mechanism by which skeletal muscle may be preconditioned to maintain structural viability


The Journal of Bone & Joint Surgery British Volume
Vol. 51-B, Issue 1 | Pages 165 - 174
1 Feb 1969
Rösingh GE James J

1. An investigation was made of the tolerance of the cells in the femoral head in rabbits for ischaemia brought about by transecting the ligament of the femoral head and applying a ligature around the femoral neck. The animals were killed two, six, twelve, twenty-four and seventy-two hours after operation. 2. In the cells of the bone marrow and in the osteoblasts distinct histological signs of disintegration were present six hours after operation. Pyknosis of the osteocyte nuclei was found after twenty-four hours' ischaemia; sometimes vacuolar clarifications could be observed in these pyknotic nuclei. After three days of ischaemia the staining affinity for Feulgen and haematoxylin of a number of osteocyte nuclei had visibly decreased. 3. The Feulgen-DNA content of the osteocyte nuclei-as measured in individual nuclei by means of an integrated microdensitometer-was significantly reduced as compared with similar nuclei from the control side as early as after six hours of ischaemia. This DNA loss was progressive with the period of ischaemia. From these facts, the conclusion was reached that in the femoral head of the rabbit the period of reversible damage for osteocytes must have ended within six hours


The Journal of Bone & Joint Surgery British Volume
Vol. 84-B, Issue 8 | Pages 1184 - 1188
1 Nov 2002
Bushell AJ Klenerman L Taylor S Davies H Grierson I Helliwell TR Jackson MJ

Ischaemic preconditioning is a process by which exposure of a tissue to a short period of non-damaging ischaemic stress leads to resistance to the deleterious effects of a subsequent prolonged ischaemic stress. It has been extensively described in the heart, but few studies have examined the possibility that it can occur in skeletal muscle. We have used a rat model of ischaemia of one limb to examine this possibility. Exposure of the hind limb to a period of ischaemia of five minutes and reperfusion for five minutes significantly protected the tibialis anterior muscle against the structural damage induced by a subsequent period of limb ischaemia for four hours and reperfusion for one hour. This protection was evident on examination of the muscle by both light and electron microscopy. Longer or shorter times of prior ischaemia had no effect


The Bone & Joint Journal
Vol. 98-B, Issue 8 | Pages 1132 - 1137
1 Aug 2016
Lawendy A Bihari A Sanders DW Badhwar A Cepinskas G

Aims. 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. Methods. 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. Results. We found that hepatocellular injury was significantly higher in the compartment syndrome group (192 PI-labelled cells/10. -1 . mm. 3. , standard error of the mean (. sem. ) 51) compared with controls (30 PI-labelled cells/10. -1 . mm. 3. , . sem . 12, p < 0.01). The number of adherent venular white blood cells was significantly higher for the compartment syndrome group (5 leukocytes/30s/10 000 μm. 2. , . sem 1. ) than controls (0.2 leukocytes/30 s/10 000 μm. 2. , . sem . 0.2, p < 0.01). Volumetric blood flow was not significantly different between the two groups, although there was an increase in the heterogeneity of perfusion. Conclusions. Compartment syndrome can be accompanied by severe systemic inflammation and end organ damage. This study provides evidence of the relationship between compartment syndrome in a limb and systemic inflammation and dysfunction in a remote organ. Cite this article: Bone Joint J 2016; 98-B:1132–7


The Bone & Joint Journal
Vol. 97-B, Issue 11 | Pages 1447 - 1455
1 Nov 2015
Alshameeri Z Bajekal R Varty K Khanduja V

Vascular injuries during total hip arthroplasty (THA) are rare but when they occur, have serious consequences. These have traditionally been managed with open exploration and repair, but more recently there has been a trend towards percutaneous endovascular management. We performed a systematic review of the literature to assess if this change in trend has led to an improvement in the overall reported rates of morbidity and mortality during the last 22 years in comparison with the reviews of the literature published previously. . We found a total of 61 articles describing 138 vascular injuries in 124 patients. Injuries because of a laceration were the most prevalent (n = 51, 44%) and the most common presenting feature, when recorded, was bleeding (n = 41, 53.3%). Delay in diagnosis was associated with the type of vascular lesion (p < 0.001) and the clinical presentation (p = 0.002). Open exploration and repair was the most common form of management, however percutaneous endovascular intervention was used in one third of the injuries and more constantly during the last 13 years. The main overall reported complications included death (n = 9, 7.3%), amputation (n = 2, 1.6%), and persistent ischaemia (n = 9, 7.3%). When compared with previous reviews there was a similar rate of mortality but lower rates of amputation and permanent disability, especially in patients managed by endovascular strategies. Cite this article: Bone Joint J 2015;97-B:1447–55


The Journal of Bone & Joint Surgery British Volume
Vol. 38-B, Issue 1 | Pages 152 - 174
1 Feb 1956
Seddon HJ

1 . In the common type of Volkmann's ischaemic contracture affecting the forearm flexors, the infarct takes the form of an ellipsoid with its axis in the line of the anterior interosseous artery and with its central point a little above the middle of the forearm. The greatest damage is at the centre and usually falls most heavily on flexor digitorum profundus and flexor pollicis longus, which are often necrotic. Those muscles more superficially placed, and sometimes the deep extensors, are more likely to exhibit fibrosis. 2. The median nerve runs near the centre of the ellipsoid and may exhibit profound ischaemia. The ulnar nerve, lying at the edge of the ischaemic zone, tends to be less severely affected. 3. The treatment for this condition is excision of all tissues irreparably damaged by ischaemia. If this operation is performed within twelve months from the time of injury, correction of the contracture should be almost complete. The tendons of shortened but active muscles are lengthened or transplanted. 4. After such excision it is possible to carry out reconstructive procedures commonly used in the surgery of lower motor neurone disorders and of trauma. A wide variety of tendon transplantations is available. The median nerve may be repaired either by a free graft or, in cases where both nerves have been extensively damaged by ischaemia, by an ulnar to median nerve-pedicle graft


The Bone & Joint Journal
Vol. 105-B, Issue 6 | Pages 649 - 656
1 Jun 2023
Dagneaux L Amundson AW Larson DR Pagnano MW Berry DJ Abdel MP

Aims

Nonagenarians (aged 90 to 99 years) have experienced the fastest percent decile population growth in the USA recently, with a consequent increase in the prevalence of nonagenarians living with joint arthroplasties. As such, the number of revision total hip arthroplasties (THAs) and total knee arthroplasties (TKAs) in nonagenarians is expected to increase. We aimed to determine the mortality rate, implant survivorship, and complications of nonagenarians undergoing aseptic revision THAs and revision TKAs.

Methods

Our institutional total joint registry was used to identify 96 nonagenarians who underwent 97 aseptic revisions (78 hips and 19 knees) between 1997 and 2018. The most common indications were aseptic loosening and periprosthetic fracture for both revision THAs and revision TKAs. Mean age at revision was 92 years (90 to 98), mean BMI was 27 kg/m2 (16 to 47), and 67% (n = 65) were female. Mean time between primary and revision was 18 years (SD 9). Kaplan-Meier survival was used for patient mortality, and compared to age- and sex-matched control populations. Reoperation risk was assessed using cumulative incidence with death as a competing risk. Mean follow-up was five years.


Bone & Joint Research
Vol. 3, Issue 4 | Pages 130 - 138
1 Apr 2014
Shapiro F Connolly S Zurakowski D Flynn E Jaramillo D

Objectives. An experimental piglet model induces avascular necrosis (AVN) and deformation of the femoral head but its secondary effects on the developing acetabulum have not been studied. The aim of this study was to assess the development of secondary acetabular deformation following femoral head ischemia. Methods. Intracapsular circumferential ligation at the base of the femoral neck and sectioning of the ligamentum teres were performed in three week old piglets. MRI was then used for qualitative and quantitative studies of the acetabula in operated and non-operated hips in eight piglets from 48 hours to eight weeks post-surgery. Specimen photographs and histological sections of the acetabula were done at the end of the study. . Results. The operated-side acetabula were wider, shallower and misshapen, with flattened labral edges. At eight weeks, increased acetabular cartilage thickness characterised the operated sides compared with non-operated sides (p < 0.001, ANOVA). The mean acetabular width on the operated side was increased (p = 0.015) while acetabular depth was decreased anteriorly (p = 0.007) and posteriorly (p = 0.44). The cartilage was thicker, with delayed acetabular bone formation, and showed increased vascularisation with fibrosis laterally and focal degenerative changes involving chondrocyte hypocellularity, chondrocyte cloning, peripheral pannus formation and surface fibrillation. . Conclusions. We demonstrate that femoral head AVN in the young growing piglet also induced, and was coupled with, secondary malformation in acetabular shape affecting both articular and adjacent pelvic cartilage structure, and acetabular bone. The femoral head model inducing AVN can also be applied to studies of acetabular maldevelopment, which is less well understood in terms of developing hip malformation. Cite this article: Bone Joint Res 2014;3:130–8


The Journal of Bone & Joint Surgery British Volume
Vol. 87-B, Issue 8 | Pages 1102 - 1106
1 Aug 2005
Stenning M Drew S Birch R

We describe 20 patients, aged between 43 and 88 years, with delayed nerve palsy or deepening of an initial palsy caused by arterial injury from low-energy injuries to the shoulder. The onset of palsy ranged from immediately after the injury to four months later. There was progression in all the patients with an initial partial nerve palsy. Pain was severe in 18 patients, in 16 of whom it presented as neurostenalgia and in two as causalgia. Dislocation of the shoulder or fracture of the proximal humerus occurred in 16 patients. There was soft-tissue crushing in two and prolonged unconsciousness from alcoholic intoxication in another two. Decompression of the plexus and repair of the arterial injury brought swift relief from pain in all the patients. Nerve recovery was generally good, but less so in neglected cases. The interval from injury to the repair of the vessels ranged from immediately afterwards to 120 days. Delayed onset of nerve palsy or deepening of a nerve lesion is caused by bleeding and/or impending critical ischaemia and is an overwhelming indication for urgent surgery. There is almost always severe neuropathic pain


The Bone & Joint Journal
Vol. 105-B, Issue 4 | Pages 347 - 355
15 Mar 2023
Birch NC Cheung JPY Takenaka S El Masri WS

Initial treatment of traumatic spinal cord injury remains as controversial in 2023 as it was in the early 19th century, when Sir Astley Cooper and Sir Charles Bell debated the merits or otherwise of surgery to relieve cord compression. There has been a lack of high-class evidence for early surgery, despite which expeditious intervention has become the surgical norm. This evidence deficit has been progressively addressed in the last decade and more modern statistical methods have been used to clarify some of the issues, which is demonstrated by the results of the SCI-POEM trial. However, there has never been a properly conducted trial of surgery versus active conservative care. As a result, it is still not known whether early surgery or active physiological management of the unstable injured spinal cord offers the better chance for recovery. Surgeons who care for patients with traumatic spinal cord injuries in the acute setting should be aware of the arguments on all sides of the debate, a summary of which this annotation presents.

Cite this article: Bone Joint J 2023;105-B(4):347–355.


The Bone & Joint Journal
Vol. 95-B, Issue 12 | Pages 1645 - 1649
1 Dec 2013
Bernhoff K Rudström H Gedeborg R Björck M

Popliteal artery injury (PAI) is a feared complication during knee replacement. Our aim was to investigate those injuries that occurred in association with knee replacement in terms of the type of injury, treatment and outcomes. From our national vascular registry (Swedvasc) and the Swedish Patient Insurance databases a total of 32 cases were identified. Prospective data from the registries was supplemented with case-records, including long-term follow-up. We estimated the incidence during 1998 to 2010 to be 0.017%. In our series of 32 patients with PAI occurring between 1987 and 2011, 25 (78%) were due to penetrating trauma and seven were caused by blunt trauma. The patients presented in three ways: bleeding (14), ischaemia (7) and false aneurysm formation (11), and five occurred during revision surgery. A total of 12 injuries were detected intra-operatively, eight within 24 hours (3 to 24) and 12 at more than 24 hours post-operatively (2 to 90 days). Treatment comprised open surgery in 28 patients. Patency of the vascular repair at 30 days was 97% (31 of 32, one amputation). At the time of follow-up (median 546 days, mean 677 days (24 to 1251)), 25 patients had residual symptoms. Of seven patients with a complete recovery, six had had an early diagnosis of the PAI during the procedure, and were treated by a vascular surgeon in the same hospital. PAI is a rare adverse event during knee replacement surgery. The outcome following such events is often adversely affected by diagnostic and therapeutic delay. Bleeding and false aneurysm were the most common clinical presentations. Cite this article: Bone Joint J 2013;95-B:1645–9


The Journal of Bone & Joint Surgery British Volume
Vol. 87-B, Issue 8 | Pages 1134 - 1139
1 Aug 2005
Schäfer M Elke R Young JR Gancs P Kindler CH

Using a computer-based quality assurance program, we analysed peri-operative data on 160 patients undergoing one-stage bilateral hip or knee arthroplasties under regional anaesthesia with routine anaesthetic monitoring and only using peripheral intravenous access for peri-operative safety. We monitored defined intra-operative adverse events such as hypotension, myocardial ischaemia, arrhythmias, hypovolaemia, hypertension and early post-operative complications. We also determined post-operative hip and knee function, and patient satisfaction with different aspects of the anaesthetic management. Those patients undergoing one-stage bilateral arthroplasties were matched according to a cross-stratification which used three variables (American Society of Anesthesiologists’ physical status scoring system, age and joint replaced) to patients undergoing unilateral hip or knee arthroplasties. Serious intra-operative adverse events were, with the exception of intra-operative hypotension, very infrequent in patients undergoing bilateral (nine adverse events) as well as unilateral arthroplasties (five adverse events). Early post-operative complications were also infrequent in both groups. However, the risks of receiving a heterologous blood transfusion (odds ratio 2.5; 95% confidence interval (CI) 1.3 to 5.0, estimated by exact conditional logistic regression) or vasoactive drugs (odds ratio 3.9; 95% CI 2.0 to 7.8) were significantly greater for patients undergoing bilateral operations. Patient satisfaction with anaesthesia was high; all patients who underwent the one-stage bilateral operation would choose the same anaesthetic technique again


The Bone & Joint Journal
Vol. 105-B, Issue 4 | Pages 449 - 454
15 Mar 2023
Zhang C Wang C Duan N Zhou D Ma T

Aims

The aim of this study was to assess the safety and clinical outcome of patients with a femoral shaft fracture and a previous complex post-traumatic femoral malunion who were treated with a clamshell osteotomy and fixation with an intramedullary nail (IMN).

Methods

The study involved a retrospective analysis of 23 patients. All had a previous, operatively managed, femoral shaft fracture with malunion due to hardware failure. They were treated with a clamshell osteotomy between May 2015 and March 2020. The mean age was 42.6 years (26 to 62) and 15 (65.2%) were male. The mean follow-up was 2.3 years (1 to 5). Details from their medical records were analyzed. Clinical outcomes were assessed using the quality of correction of the deformity, functional recovery, the healing time of the fracture, and complications.


The Bone & Joint Journal
Vol. 106-B, Issue 9 | Pages 994 - 999
1 Sep 2024
El-Khaldi I Gude MH Gundtoft PH Viberg B

Aims

Pneumatic tourniquets are often used during the surgical treatment of unstable traumatic ankle fractures. The aim of this study was to assess the risk of reoperation after open reduction and internal fixation of ankle fractures with and without the use of pneumatic tourniquets.

Methods

This was a population-based cohort study using data from the Danish Fracture Database with a follow-up period of 24 months. Data were linked to the Danish National Patient Registry to ensure complete information regarding reoperations due to complications, which were divided into major and minor. The relative risk of reoperations for the tourniquet group compared with the non-tourniquet group was estimated using Cox proportional hazards modelling.


Bone & Joint Open
Vol. 5, Issue 7 | Pages 560 - 564
7 Jul 2024
Meißner N Strahl A Rolvien T Halder AM Schrednitzki D

Aims

Transfusion after primary total hip arthroplasty (THA) has become rare, and identification of causative factors allows preventive measures. The aim of this study was to determine patient-specific factors that increase the risk of needing a blood transfusion.

Methods

All patients who underwent elective THA were analyzed retrospectively in this single-centre study from 2020 to 2021. A total of 2,892 patients were included. Transfusion-related parameters were evaluated. A multiple logistic regression was performed to determine whether age, BMI, American Society of Anesthesiologists (ASA) grade, sex, or preoperative haemoglobin (Hb) could predict the need for transfusion within the examined patient population.


The Journal of Bone & Joint Surgery British Volume
Vol. 79-B, Issue 4 | Pages 621 - 623
1 Jul 1997
Ballaro A Gibbons CLM Murray DM Kettlewell MGW Benson MK

Acute colonic pseudo-obstruction is a poorly recognised and potentially fatal complication of hip surgery. Between 1991 and 1994 six patients were observed who required laparotomy after failure of medical management. In three the indication was signs of peritonism, while in the other three exploration was required to exclude segmental ischaemia and to decompress the bowel. In all, there was no evidence of mechanical obstruction. Patients having total hip replacement are at risk of developing pseudo-obstruction due to their age, comorbidity, high doses of analgesics and the nature of the operation. If postoperative ileus persists for more than 48 hours acute colonic pseudo-obstruction should be suspected and confirmed by plain radiography. Prompt recognition and treatment with early referral to a colorectal unit are indicated. Laparotomy appears to carry less risk than that for patients with idiopathic pseudo-obstruction, but should be performed only if colonic ischaemia is suspected


The Journal of Bone & Joint Surgery British Volume
Vol. 83-B, Issue 5 | Pages 751 - 759
1 Jul 2001
Sato M Sugano N Ohzono K Nomura S Kitamura Y Tsukamoto Y Ogawa S

Using in situ hybridisation and the terminal deoxynucleotidyl transferase-mediated biotin-dUTP nick end-labelling (TUNEL) reaction in rats with osteonecrosis of the femoral head we have studied the effect of ischaemia on the gene expression of the stress proteins oxygen-regulated protein 150 (ORP150) and haemoxygenase 1 (HO1) and the death mechanism of the cells involved in osteonecrosis. Both ORP150 and HO1 have been reported to have important roles in the successful adaptation to oxygen deprivation. ORP150 and HO1 mRNA expression was induced by ischaemia in osteoblasts and osteocytes. In proliferative chondrocytes, these signals were detected constitutively. During the development of ischaemic osteonecrosis, the mechanism of cell death was apoptosis as indicated by DNA fragmentation and the presence of apoptotic bodies in osteocytes, chondrocytes and bone-marrow cells. After the initial ischaemic event, expression of ORP150 and HO1 mRNA, the TUNEL-positive reaction and empty lacunae were found sequentially. These findings were exclusive and may be considered to be markers for each stage in the development of osteonecrosis


The Bone & Joint Journal
Vol. 106-B, Issue 1 | Pages 53 - 61
1 Jan 2024
Buckland AJ Huynh NV Menezes CM Cheng I Kwon B Protopsaltis T Braly BA Thomas JA

Aims

The aim of this study was to reassess the rate of neurological, psoas-related, and abdominal complications associated with L4-L5 lateral lumbar interbody fusion (LLIF) undertaken using a standardized preoperative assessment and surgical technique.

Methods

This was a multicentre retrospective study involving consecutively enrolled patients who underwent L4-L5 LLIF by seven surgeons at seven institutions in three countries over a five-year period. The demographic details of the patients and the details of the surgery, reoperations and complications, including femoral and non-femoral neuropraxia, thigh pain, weakness of hip flexion, and abdominal complications, were analyzed. Neurological and psoas-related complications attributed to LLIF or posterior instrumentation and persistent symptoms were recorded at one year postoperatively.


Bone & Joint Open
Vol. 5, Issue 4 | Pages 367 - 373
26 Apr 2024
Reinhard J Lang S Walter N Schindler M Bärtl S Szymski D Alt V Rupp M

Aims

Periprosthetic joint infection (PJI) demonstrates the most feared complication after total joint replacement (TJR). The current work analyzes the demographic, comorbidity, and complication profiles of all patients who had in-hospital treatment due to PJI. Furthermore, it aims to evaluate the in-hospital mortality of patients with PJI and analyze possible risk factors in terms of secondary diagnosis, diagnostic procedures, and complications.

Methods

In a retrospective, cross-sectional study design, we gathered all patients with PJI (International Classification of Diseases (ICD)-10 code: T84.5) and resulting in-hospital treatment in Germany between 1 January 2019 and 31 December 2022. Data were provided by the Institute for the Hospital Remuneration System in Germany. Demographic data, in-hospital deaths, need for intensive care therapy, secondary diagnosis, complications, and use of diagnostic instruments were assessed. Odds ratios (ORs) with 95% confidence intervals (CIs) for in-hospital mortality were calculated.


Aims

In this investigation, we administered oxidative stress to nucleus pulposus cells (NPCs), recognized DNA-damage-inducible transcript 4 (DDIT4) as a component in intervertebral disc degeneration (IVDD), and devised a hydrogel capable of conveying small interfering RNA (siRNA) to IVDD.

Methods

An in vitro model for oxidative stress-induced injury in NPCs was developed to elucidate the mechanisms underlying the upregulation of DDIT4 expression, activation of the reactive oxygen species (ROS)-thioredoxin-interacting protein (TXNIP)-NLRP3 signalling pathway, and nucleus pulposus pyroptosis. Furthermore, the mechanism of action of small interfering DDIT4 (siDDIT4) on NPCs in vitro was validated. A triplex hydrogel named siDDIT4@G5-P-HA was created by adsorbing siDDIT4 onto fifth-generation polyamidoamine (PAMAM) dendrimer using van der Waals interactions, and then coating it with hyaluronic acid (HA). In addition, we established a rat puncture IVDD model to decipher the hydrogel’s mechanism in IVDD.


The Journal of Bone & Joint Surgery British Volume
Vol. 30-B, Issue 2 | Pages 309 - 321
1 May 1948
Darmady EM

Traumatic uraemia is of particular significance to orthopaedic surgeons in so far as this complication is responsible for high mortality in cases of severe injury, crushing injury, traumatic shock, gun-shot wounds, incompatible blood transfusions, and the misuse of tourniquets. In this contribution the association between muscle injury and renal failure is reviewed. The syndrome of the "crush kidney," which at first was thought to be the result of deposition of myohaemoglobin in the renal tubules, is almost certainly due to the association of many factors, all of which lead to alteration of the renal circulation and renal ischaemia. Certainly the combination of ischaemia of the kidney with deposition of pigment and haemolysis of blood causes a high mortality in animals, and it is believed that this may account for the serious prognosis in human cases where there is both shock and haemolysis. Disturbance of water balance, allergic phenomena, and chemical nephrotoxic action are also discussed. The pathology is considered together with significant changes in the blood chemistry. A plea is made for early recognition of this clinical syndrome with its characteristic features. Important aspects of treatment are discussed. The dangers of excessive intravenous infusion are emphasised. Closer investigation and further research promise to throw light on the more accurate localisation of function in the nephron, to add to our knowledge of traumatic arterial spasm, and to explain many orthopaedic problems which hitherto appeared insoluble


The Journal of Bone & Joint Surgery British Volume
Vol. 58-B, Issue 3 | Pages 332 - 338
1 Aug 1976
Jensen O Lauritzen J

Specimens of femoral heads were studied at necropsy in two cases of Legg-Calve-Perthes' disease. One was that of a boy aged four years ten months who died from appendicitis; the other was from a boy aged six years who died from a malignant glioma. Both had been treated for one and a half years for Legg-Calve-Perthes' disease which was in a stage of repair at the time of death. The diseased femoral heads were moderately flattened but the surface cartilage was intact. Epiphysial bone and bone marrow were partly replaced by cartilage, fibrous tissue and granulation tissue, and new bone was being formed. Inflammatory reaction was inconspicuous. Enchondral bone formation was only slightly decreased, and the structure of the growth plate was undisturbed. There was no sign of systemic bone disease. In the first case the changes indicated that more than one episode of ischaemia had occurred, and an occlusion--probably from an old thrombus--was demonstrated in the posterior inferior retinacular artery of the femoral head. The last episode of ischaemia, furthermore, had caused infarction of part of the metaphysial bone. In both cases, the central area of the metaphysial bone of the affected femur contained fat, but there were few haemopoietic cells and it therefore looked pale. The findings are discussed in relation to previous work on the pathology in Legg-Calve-Perthes' disease, recent information on the vascularisation of the femoral head in children, and experimental and comparative animal studies