Aim

This study aimed to assess whether the severity of symptoms (assessed with the Oxford Hip Score (OHS)) can relate to the levels of mRNA expression of markers for muscle inflammation (tumour necrosis factor alpha (TNFα), interleukin 6 (IL-6)) in the proximal vastus lateralis (VL) of patients with severe OA undergoing THR.

Aim

To assess whether the Oxford Hip Score (OHS), is reflective of objectively assessed functional performance (timed up and go (TUG), 30 sec sit to stand (ST), 6 minute walk test (6MWT), stair climb performance (SCP), and gait speed (GS)) in patients undergoing total hip arthroplasty (THA).

Aim

To assess the relationship between mRNA expression of genetic markers of inflammation (tumour necrosis factor-alpha (TNFα)) and interleukin-6 (IL-6) in the vastus lateralis (VL) of the operated leg, and the strength of the operated leg quadriceps, in patients following THR.

Introduction

Late (commenced 6 months to 4 years post-op) home-based progressive resistance training programs are proven to improve muscle strength and function after total hip replacement (THR). This study assessed whether early (commenced < 1 week post-op) HBPRT post-THR improves muscle mass, strength and function relative to routine physiotherapy rehabilitation (RPR) at up to 12 months follow up.

Introduction

Malalignment of some designs of stem is associated with an increased risk of aseptic loosening and revision. We investigated whether the alignment of the cemented polished, double-taper design adversely affected outcome, in a multicentre prospective study.

INTRODUCTION

Bone marrow derived mesenchymal stem cells are a potential source of cells for the repair of articular cartilage defects. Hypoxia has been shown to improve chondrogenesis in adult stem cells. In this study we characterised bone marrow derived stem cells and investigated the effects of hypoxia on gene expression changes and chondrogenesis.

Introduction

Preoperative psychological distress has been reported to predict poor outcome and patient dissatisfaction after total hip replacement (THR). We investigated this relationship in a prospective multi-centre study between January 1999 and January 2002.

Introduction

Mesenchymal stem cells are a potential source of cells for the repair of articular cartilage defects. We have previously demonstrated that the infrapatellar synovial fat pad is a rich source of mesenchymal stem cells and these cells are able to undergo chondrogenic differentiation. Although synovial fat pad derived mesenchymal stem cells may represent a heterogenous population, clonal populations derived from the synovial fat pad have not previously been studied.

INTRODUCTION

Bone marrow derived mesenchymal stem cells are a potential source of cells for the repair of articular cartilage defects. Hypoxia has been shown to improve chondrogenesis in adult stem cells. In this study we characterised bone marrow derived stem cells and investigated the effects of hypoxia on gene expression changes and chondrogenesis.

Introduction

Mesenchymal stem cells are a potential source of cells for the repair of articular cartilage defects. We have previously demonstrated that the infrapatellar synovial fat pad is a rich source of mesenchymal stem cells and these cells are able to undergo chondrogenic differentiation. Although synovial fat pad derived mesenchymal stem cells may represent a heterogenous population, clonal populations derived from the synovial fat pad have not previously been studied.

Introduction: Bone marrow derived mesenchymal stem cells are a potential source of cells for the repair of articular cartilage defects. Hypoxia has been shown to improve chondrogenesis in adult stem cells. In this study we characterised bone marrow derived stem cells and investigated the effects of hypoxia on gene expression changes and chondrogenesis.

Material and Methods: Adherent colony forming cells were isolated and cultured from the stromal component of bone marrow. The cells at passage 2 were characterised for stem cell surface epitopes, and then cultured as cell aggregates in chondrogenic medium under normoxic (20% oxygen) or hypoxic (5% oxygen) conditions for 14 days. Gene expression analysis, glycosoaminoglycan and DNA assays, and immunohistochemical staining were determined to assess chondrogenesis.

Results: Bone marrow derived adherent colony forming cells stained strongly for markers of adult mesenchymal stem cells including CD44, CD90 and CD105, and they were negative for the haematopoietic cell marker CD34 and for the neural and myogenic cell marker CD56. Interestingly, a high number of cells were also positive for the pericyte marker 3G5. Cell aggregates showed a chondrogenic response and in lowered oxygen there was increased matrix accumulation of proteoglycan, but less cell proliferation, which resulted in 3.2-fold more glycosoaminoglycan per DNA after 14 days of culture. In hypoxia there was increased expression of key transcription factor SOX6, and the expression of collagens II and XI, and aggrecan was also increased.

Discussion: Pericytes are a candidate stem cell in many tissue and our results show that bone marrow derived mesenchymal stem cells express the pericyte marker 3G5. The response to chondrogenic culture in these cells was enhanced by lowered oxygen tension, which up-regulated SOX6 and increased the synthesis and assembly of matrix during chondrogenesis. This has important implications for tissue engineering applications of bone marrow derived stem cells.

Introduction: There is an ever-increasing clinical need for the regeneration and replacement of tissue to replace soft tissue lost due to trauma, disease and cosmetic surgery. A potential alternative to the current treatment modalities is the use of tissue engineering applications using mesenchymal stem cells that have been identified in many tissues including the fat pad. In this study, stem cells isolated from the fat pad were characterised and their differentiation potential assessed.

Materials and Methods: The infrapatellar fat pad was obtained from total knee replacement for osteoarthritis. Cells were isolated, expanded and stained for a number of stem cell markers. For adipogenic differentiation, cells were cultured in adipogenic inducing medium (10ug/ml insulin, 1uM dexamthasone, 100uM indomethacin and 500uM 3-isobutyl-1-methyl xanthine). Gene expression analyses and Oil red O staining was performed to assess adipogenesis.

Results: Cells at passage 2 stained strongly for CD13, CD29, CD44, CD90 and CD105 (mesenchymal stem cell markers). The cells stained sparsely for 3G5 (peri-cyte marker). On gene expression analyses, the cells cultured under adipogenic conditions had almost a 1,000 fold increase in expression of peroxisome proliferator-activated receptor gamma-2 (PPAR gamma-2) and 1,000,000 fold increase in expression of lipoprotein lipase (LPL). Oil red O staining revealed triglyceride accumulation within typical adipogenic morphology, confirming the adipogenic nature of the observed vacuoles, and showed failure of staining in control cells.

Discussion: Fat pad derived stem cells expressed a cell surface epitope profile of mesenchymal stem cells, and exhibited the potential to undergo adipogenic differentiation. Our results show that the human fat pad is a viable potential autogeneic source for mesenchymal stem cells capable of adipogenic differentiation as well as previously documented ostegenic and chondrogenic differentiation. This cell source has potential use in tissue engineering applications.

Bone marrow derived mesenchymal stem cells are a potential source of cells for the repair of articular cartilage defects. Hypoxia has been shown to improve chondrogenesis in adult stem cells. In this study we characterised bone marrow derived stem cells and investigated the effects of hypoxia on gene expression changes and chondrogenesis.

Adherent colony forming cells were isolated and cultured from the stromal component of bone marrow. The cells at passage 2 were characterised for stem cell surface epitopes, and then cultured as cell aggregates in chondrogenic medium under normoxic (20% oxygen) or hypoxic (5% oxygen) conditions for 14 days. Gene expression analysis, glycosoaminoglycan and DNA assays, and immunohistochemical staining were determined to assess chondrogenesis.

Bone marrow derived adherent colony forming cells stained strongly for markers of adult mesenchymal stem cells including CD44, CD90 and CD105, and they were negative for the haematopoietic cell marker CD34 and for the neural and myogenic cell marker CD56. Interestingly, a high number of cells were also positive for the pericyte marker 3G5. Cell aggregates showed a chondrogenic response and in lowered oxygen there was increased matrix accumulation of proteoglycan, but less cell proliferation, which resulted in 3.2-fold more glycosoaminoglycan per DNA after 14 days of culture. In hypoxia there was increased expression of key transcription factor SOX6, and the expression of collagens II and XI, and aggrecan was also increased.

Pericytes are a candidate stem cell in many tissue and our results show that bone marrow derived mesenchymal stem cells express the pericyte marker 3G5. The response to chondrogenic culture in these cells was enhanced by lowered oxygen tension, which up-regulated SOX6 and increased the synthesis and assembly of matrix during chondrogenesis. This has important implications for tissue engineering applications of bone marrow derived stem cells.

Introduction: We prospectively followed all hip fracture patients admitted between 2004–2006, identified cases where the intention was to treat conservatively and compared their functional outcome and mortality with a similar cohort treated surgically over the same period.

Methods: We recorded length of hospital stay, place of discharge, pre and post-fracture mobility and residence, 30 day and 1 yr mortality, re-admission and delayed surgery. The group treated surgically was recruited and matched for age, gender, pre and post fracture mobility, mental confusion and independence with the conservatively treated group.

Results: 25 patients were treated conservatively. 22 patients treated surgically over the same period were recruited. The mean hospital stay was 13 days in both groups. There were 4 extracapsular (3 displaced) and 21 intracapsular fractures (5 displaced) in the conservative arm and 11 extracapsular and 9 intracapsular fractures in the surgically treated arm. 4 patients from the conservative treatment group underwent late surgery 20 days – 2 months after the index event. Surgically treated group had 11 dynamic screw fixation, 1 cannulated screw, 1 total hip replacement and 7 hemiarthroplasty. 9/14 of the conservatively treated patients were mobile independently or with aid after treatment compared to 11/16 patients after surgery. 7/16 patients treated conservatively were living independently in their own residence, compared to 10/14 patients in the operatively treated patients. 1 month and 1 year mortality in conservatively treated group was 4/21 and 7/21 respectively compared to 1/20 and 5/20 in the operative fixation group. There was no statistically significant difference in mobility, residence or mortality between the two groups (Fisher exact test, p > 0.05).

Discussion: Conservative management after hip fracture in medically unfit patients does not result in statistically significant difference in functional outcome or mortality compared to patients treated surgically.

Introduction: We investigated the relationship between psychological distress and outcome after total hip replacement (THR) in the Exeter Primary Outcome Study (EPOS).

Materials & Methods: Data were collected from a number of centres across England between January 1999 and January 2002 for patients undergoing primary hip replacement using the cemented Exeter femoral component (Stryker). We recorded the Oxford Hip Score (OHS) for physical function and SF36 questionnaire for both physical and mental domain assessment annually for five years. We dichotomised the patients into the mentally distressed (MHS < 50) and the not mentally distressed (MHS ≥ 50) groups based on their pre-operative Mental Health Score (MHS) from the SF36 score.

Results: Complete data were available for 455 (407 not distressed and 48 distressed) patients. Pre-operative OHS and SF-36 score was significantly worse in the distressed group (both p< 0.001). Mean OHS improved from 43 to 20 at 1 year after surgery and remained the same thereafter in the non distressed group. In the mentally distressed group pre-operative mean OHS of 48 improved to 22 at 1 year after surgery. Maximum improvement in OHS occurred in the 1st yr after surgery. Mean MHS improved from 76 to 81 at 1 year after surgery and remained the same thereafter in the non distressed group. Mean MHS improved from 35 to 62 at 1 year after surgery, reaching 65 at 5 years after surgery in the mentally distressed group. The maximum improvement in MHS occurred in the 1st yr after surgery.

Conclusion: Pre-operative psychological distress did not compromise functional outcome after hip arthroplasty. There was a substantial improvement in mental distress in patients with mental distress prior to surgery. Both groups of patients experienced improvement in Oxford Hip Score, which was maximal by 1 year after surgery and was maintained over the 5 year follow up.

There is an ever-increasing clinical need for the regeneration and replacement of tissue to replace soft tissue lost due to trauma, disease and cosmetic surgery. A potential alternative to the current treatment modalities is the use of tissue engineering applications using mesenchymal stem cells that have been identified in many tissue including the infrapatellar fat pad. In this study, stem cells isolated from the infrapatellar fat pad were characterised to ascertain their origin, and allowed to undergo adipogenic differentiation to confirm multilineage differentiation potential.

The infrapatellar fat pad was obtained from total knee replacement for osteoarthritis. Cells were isolated and expanded in monolayer culture. Cells at passage 2 stained strongly for CD13, CD29, CD44, CD90 and CD105 (mesenchymal stem cell markers). The cells stained poorly for LNGFR and STRO1 (markers for freshly isolated bone marrow derived stem cells), and sparsely for 3G5 (pericyte marker). Staining for CD34 (haematopoetic marker) and CD56 (neural and myogenic lineage marker) was negative. {BR}For adipogenic differentiation, cells were cultured in adipogenic inducing medium consisting of basic medium with 10ug/ml insulin, 1uM dexamthasone, 100uM indomethacin and 500uM 3-isobutyl-1-methyl xanthine. By day 16, many cells had lipid vacuoles occupying most of the cytoplasm. On gene expression analyses, the cells cultured under adipogenic conditions had almost a 1,000 fold increase in expression of peroxisome proliferator-activated receptor gamma-2 (PPAR gamma-2) and 1,000,000 fold increase in expression of lipoprotein lipase (LPL). Oil red O staining confirmed the adipogenic nature of the observed vacuoles and showed failure of staining in control cells.

Our results show that the human infrapatellar fat pad is a viable potential autogeneic source for mesenchymal stem cells capable of adipogenic differentiation as well as previously documented ostegenic and chondrogenic differentiation. This cell source has potential use in tissue engineering applications.

Purpose: To investigate the value of various clinical signs to identify occult hip fracture.

Methodology: MRI register was searched to identify all patients who had MRI scan between July 2000–June 2006 for suspected occult hip fracture. 64 patients were identified. 33 patients had occult proximal femoral fractures. 27 patients had no fracture.

Results: 19 patients with fracture were not independently mobile compared to 6 patients without fracture. 7 patients with fracture and 2 patients without fracture were able to weight bear. 13 patients with fracture and 10 patients without fracture had unrestricted straight leg raise ability. 7 patients with fracture and 16 patients without fracture had no pain on axial loading. Fisher exact and chi square analysis was performed; with Bonferroni correction for multiple comparisons (10 tests) p< 0.005 was deemed significant. Pain on axial loading of limb and pre-fracture patient mobility were associated with fracture (p< 0.005). Both factors had positive predictive value = 0.76 and post-test probability of disease given a negative test = .30. Predictive values remained the same when both factors were considered together.

Discussion: These data indicate that although patients who were independently mobile before the fall and who do not have pain on axial compression of the limb are less likely to have a fracture, these signs alone or in combination will not exclude a fracture. It is essential to perform MRI scanning of patients with severe hip pain but normal x rays after fall as it does not seem possible to clinically exclude fracture.

Introduction: The possibility of occult hip fracture in older patients after a fall is a common problem. The value of various clinical signs to determine which patients require further investigation has not been reported.

Methodology: MRI register was hand searched to identify all patients who had MRI scan between July 2000–June 2006 for suspected occult hip fracture. 64 patients were identified. 33 patients had occult proximal femoral fractures. 27 patients had no fracture.

Results: More patients with fracture were living in their own home (20/26), were independent for daily living (20/26) and were not independently mobile(19/26) compared to patients without a fracture (14/22, 11/22 and 6/22 respectively). 7 patients with fracture and 2 patients without fracture were able to weight bear. 13 patients with fracture and 10 patients without fracture had unrestricted straight leg raise ability. 7 patients with fracture and 16 patients without fracture had no pain on axial loading. The value of individual tests was evaluated using Fisher exact and chi square analysis; with Bonferroni correction for multiple comparisons (10 tests) p< 0.005 was deemed significant. Pain on axial loading of the limb and pre-fracture patient mobility were both associated with the presence of a fracture (p< 0.005).

Discussion: These data indicate that although patients who were independently mobile before the fall and who do not have pain on axial compression of the limb are less likely to have a fracture, these signs alone or in combination will not exclude a fracture. Other widely used signs (eg ability to straight leg raise) appear of little predictive value. On the basis of our data, we believe it is essential to have a policy of MRI scanning of patients with severe hip pain but normal x rays after a fall as it does not seem possible to clinically exclude a fracture.

Introduction: Autologous chondrocytes harvested from articular cartilage are being used for the repair of focal cartilage defects. The procedure involves injury to the cartilage and alternative sources of stem cells for use in repair are being explored. Stem cells have been found in many tissue including bone marrow and the infrapa-tellar fat pad. Infrapatellar fat pad derived stem cells present a viable and easily accessible source of stem cells for the repair of cartilage defects and tissue engineering applications.

Hypoxia has been shown to improve chondrogenesis in stem cells derived from the bone marrow. We explore the hypothesis that this effect would also apply to stem cells derived from the infrapatellar fat pad.

Materials and methods: Cell aggregates from early passage stem cells isolated from the infrapatellar fat pad were placed in chondrogenic media for 14 days either in a normoxic (20% oxygen) or hypoxic (5% oxygen) environment. Gene expression analysis, DNA and glycosoaminoglycan assays and immunohistochemi-cal studies were performed on the aggregates to assess chondrogenesis.

Results: Cells grown under hypoxic conditions showed significantly improved chondrogenesis as determined by relatively higher gene expression of proteoglycans, collagens and SOX genes. The cell aggregates also had a higher glycosoaminoglycan content and glycosoamino-glycan content per DNA. Immunohistochemical studies confirm enhanced production of collagen types I and II and aggrecan.

Discussion: These findings confirm the previously documented effects of hypoxic culture conditions on stem cells and extend the findings to include infrapatellar fat pad derived stem cells. Our findings suggest that oxygen tension has a role in regulating the function of stem cells as they undergo chondrogenesis. In culture these cells appear to function optimally in an atmosphere of reduced oxygen that more closely approximates documented in vivo oxygen tension. This has important implications in future tissue engineering applications of these cells.

Measurement of the rate of fracture healing is a major problem in fracture research. Bone mineral density (BMD) of fracture callus has been used as a measure of healing in diaphyseal fractures. However, metaphyseal fractures (especially in the elderly) are now the commonest type of fracture and are a significant public health problem. This study investigated whether measurement of BMD at the fracture site in the distal radius can be used as a measure of fracture healing.

We recruited 28 patients who had sustained a dorsally displaced distal radial fracture which was deemed to require treatment by intrafocal wire fixation. All patients had acceptable correction of dorsal and radial angle at final x ray (3 months). Wrist function was measured using the Patient Rated Wrist Evaluation (PRWE – a validated outcome measure for use after distal radial fractures), grip strength,and range of motion. All measurements were made at 6, 12 and 26 weeks. BMD was measured at the fracture site (examining the BMD of the medullary bone at the fracture site after removal of wires), in the opposite wrist and the lumbar spine using QCT at 6 weeks after fracture.

There was no correlation between fracture site BMD and BMD at the other wrist or the lumbar spine (r < 0.3). The BMD at the fracture site was higher than the BMD at the other wrist (mean 168 vs 70 HU; p< 0.001 paired T test). There was no relationship between fracture site BMD or the ratio of BMDs fracture site / normal wrist, and any of the functional assessments (proportion grip strength recovered, range of motion or PRWE (r < 0.3)).

15 of these patients underwent a second QCT at 12 weeks after fracture. There was no significant change in fracture site BMD between the first and second scan.

These data indicate that fracture site BMD is unlikely to be a useful method of measuring metaphyseal bone healing. The increase in BMD at the fracture site was unexpected; possible explanations include impaction of bone or high BMD in woven bone (the relationship of which to bone stiffness is uncertain).