Osteoarthritis (OA) is the most common form of joint disease leading to disability and dependence. In severe cases of knee OA, the joint is deemed irrecoverable and total knee replacements are indicated. Tissue engineering is a possible solution for this pathology and previous work from our laboratory has demonstrated that it is possible to isolate and expand chondroprogenitor cells in vitro from healthy knee-joint articular cartilage. Work presented here describes the detection and isolation of chondroprogenitor cells derived from osteoarthritic cartilage following total knee replacement in patients with severe OA, suggesting a pool of viable cells from this degenerate region which has been previously deemed non-recoverable.

Human articular cartilage was excised from tibial plateaux (TP's) obtained from total knee replacements following the diagnoses of severe OA. Cells were isolated by a sequential pronase and collagenase digestion and subject to a fibronectin adhesion assay. Cells were expanded in monolayer in supplemented growth medium. Clonal 3D pellet cultures were established in chondrogenic and osteogenic differentiation media. Adipogenic cultures were also established in monolayer cultures. Histological procedures, immunohistochemistry and molecular biology were undertaken in order to determine the extent of differentiation. In addition, osteochondral plugs were excised from the TP's and wax embedded for further histological and immunohistochemical analysis.

Clonal cell lines obtained from osteoarthritic knee-joint cartilage using the fibronectin adhesion assay were isolated and successfully cultured to a maximum of 60 population doublings whilst still demonstrating a chondrogenic capacity. Three-D pellet cultures after 21 days of chondrogenic induction produced smooth and iridescent pellets which stained positively for toluidine blue and safranin O. Positive labelling for collagen type II and aggrecan were also observed. Following osteogenic induction; evidence of mineralisation was indicated by the von Kossa stain. Adipogenic induction revealed a positive result. Osteochondral plugs demonstrated sporadic positive labelling in the surface region for putative stem cell marker Stro-1.

Chondroprogenitor cells isolated from osteoarthritic display a strong chondrogenic phenotype, and have the ability to be induced into different lineages. These findings suggest the presence of a pool of viable chondroprogenitors from osteoarthritic tissue which was otherwise deemed irrecoverable.

Hyaline cartilage defects are a significant clinical problem for which a plethora of cartilage repair techniques are used. One such technique is cartilage replacement therapy using autologous chondrocyte or mesenchymal stem cell (MSC) implantation (ACI). Mesenchymal stem cells are increasingly being used for these types of repair technique because they are relatively easy to obtain and can be expanded to generate millions of cells. However, implanted MSCs can terminally differentiate and produce osteogenic tissue which is highly undesirable, also, MSCs generally only produce fibrocartilage which does not make biomechanically resilient repair tissue, an attribute that is crucial in high weight-bearing areas. Tissue-specific adult stem cells would be ideal candidates to fill the void, and as we have shown previously in animal model systems [Dowthwaite et al, 2004, J Cell Sci 117;889], they can be expanded to generate hundreds of millions of cells, produce hyaline cartilage and they have a restricted differential potential. Articular chondroprogenitors do not readily terminally differentiate down the osteogenic lineage.

At present, research focused on isolating tissue-specific stem cells from articular cartilage has met with modest success. Our results demonstrate that using differential adhesion it is possible to easily isolate articular cartilage progenitor populations from human hyaline cartilage and that these cells can be subsequently expanded in vitro to a high population doubling whilst maintaining a normal karyotype. Articular cartilage progenitors maintain telomerase activity and telomere length that are a characteristic of progenitor/stem cells and differentiate to produce hyaline cartilage.

In conclusion, we propose the identification and characterisation of a novel articular cartilage progenitor population, resident in human cartilage, which will greatly benefit future cell-based cartilage repair therapies.

A novel scoring system for the grading of osteoarthritis has been developed.

Scoring systems for the measurement of Osteoarthritis (OA) are essential for the understanding of the osteoarthritic process. OA is a mutifactorial degenerative joint disease affecting not only hyaline cartilage but also the surrounding tissues and particularly the subchondral bone. It as questionable as to why the articular cartilage remains the sole component used for histopathological assessment. The intimate relationship between the subchondral bone and overlying cartilage provide major difficulty in their independent measurement.

A new scoring system has been developed to incorporate the subchondral bone into the assessment process and relating it to the structure of the overlying hyaline cartilage, which together permit a more accurate description of the degree of degenerate change.

The new scoring system was developed from the analysis of 26 operative specimens from tibial plateau (TP) from patients who underwent total knee replacement (TKR). Multiple osteochondral plugs were taken from weight-bearing regions of the whole TP. The specimens were fixed and decalcified before being sectioned and stained with Masson's trichrome.

Using a standard imaging system (Photoshop) the areas of bone and hyaline cartilage were identified and measured. Further parameters 1) cartilage thickness 2) tidemark integrity, 3) surface integrity 4) cartilage morphology were measured using a numeric measurement scale.

The scoring system indicated a relationship between the area of subchondral bone and the hyaline cartilage degeneration. The overall sum of scores was also successful in distinguishing between the milder and more severe samples of OA. More comprehensive inter and intra observer variability needs to be tested in order validate the system. Quantifying changes to the subchondral bone may also serve beneficial to clinicians, as it is possible that monitoring these changes clinically could lead to early identification of OA.

Hyaline cartilage defects are a significant clinical problem for which a plethora of cartilage repair techniques are used. One such technique is cartilage replacement therapy using autologous chondrocyte or mesenchymal stem cell (MSC) implantation (ACI). Mesenchymal stem cells are increasingly being used for these types of repair technique because they are relatively easy to obtain and can be expanded to generate millions of cells. However, implanted MSCs can terminally differentiate and produce osteogenic tissue which is highly undesirable, also, MSCs generally only produce fibrocartilage which does not make biomechanically resilient repair tissue, an attribute that is crucial in high weight-bearing areas. Tissue-specific adult stem cells would be ideal candidates to fill the void, and as we have shown previously in animal model systems [Dowthwaite et al, 2004, J Cell Sci 117;889], they can be expanded to generate hundreds of millions of cells, produce hyaline cartilage and they have a restricted differential potential. Articular chondroprogenitors do not readily terminally differentiate down the osteogenic lineage.

At present, research focused on isolating tissue-specific stem cells from articular cartilage has met with modest success. Our results demonstrate that using differential adhesion it is possible to easily isolate articular cartilage progenitor populations from human hyaline cartilage and that these cells can be subsequently expanded in vitro to a high population doubling whilst maintaining a normal karyotype. Articular cartilage progenitors maintain telomerase activity and telomere length that are a characteristic of progenitor/stem cells and differentiate to produce hyaline cartilage.

In conclusion, we propose the identification and characterisation of a novel articular cartilage progenitor population, resident in human cartilage, which will greatly benefit future cell-based cartilage repair therapies.

Introduction

Several disorders have been associated with genetic variants. Copy number variations (CNVs) are documented micro DNA insertions and deletions that may be ten times more frequent than point mutations. We undertook a genome-wide scan to find CNVs associated with adolescent idiopathic scoliosis (AIS).

Background: Arthroscopic femoral osteoplasties might cause prolonged operative times, restricted intraop-erative overview or insufficient localisation of surgical tools. Computer assisted techniques should improve the precision with an overall accuracy is within 1mm/1°. An automated navigated registration process matching preoperative CT data and intraoperative fluoroscopy, should allow for non-invasive registration for FAI surgery. We evaluated the general precision (I) of the CT and fluoroscopic matching process and (II) the precision of identifying the defined osseous lesions in various anatomical areas.

Material and Methods: Three cadavers (6 hip joints) utilizing a conventional navigation system were used. Before preoperative CT scans, defined osseous lesion (0.5x0.5mm) in the femoral neck, head neck junction, head region were created under fluoroscopic control. Following reference marker fixation, two fluoroscopic images (12 inch c-arm) with 30° angle differences of the hip joint were taken. Automated segmentation including CT-fluoro image fusion by the navigation system enabled a noninvasive registration process Precision of registration process was tested with a straight navigated pointer (1mm tip) trough a lateral arthroscopic portal, during virtual contact to the bone, without arthroscopic control After arthroscopic view was enabled the in vivo distance of pointer tip to bone was measured (I). In vivo real distances between inserted navigated shaver and osseous lesions was done over an anterior hip arthrotomy. Under navigated control, blinded to the situ, placement in the lesions should be done. Distances between shaver tip and osseous lesions were measured with a caliper (II).

Results: The precision for registration (I) was within 0.9mm within the femoral neck (SD 0.24mm; 0.6–1.3mm); 1.2 mm (SD 0.33mm; 0.8–2.0mm) (p> 0.05) for the head neck junction; 2.9 mm (SD 0.57mm; 1.8–3.7mm) for the femoral head (p< 0.001 respectively p< 0.001) Mean offset of the navigated shaver to the lesions (II) was 0.93 mm (SD 0.65mm; 0–2mm). Within the femoral neck a mean accuracy of 0.6mm (SD 0.59mm; 0–1.4mm), the head neck junction 0.8 mm (SD 0.78mm; 0.1–1.5mm), the femoral head 1.3 mm (SD 0.50mm; 0.6–1.7mm) was found (p> 0.05; p> 0.05; p> 0.05).

Conclusion: A combined CT-fluoroscopy matching procedure allows for a reproducible noninvasive registration process for navigated FAI surgery. Precision of the registration process itself is more accurate at the femoral neck and head-neck junction than at the femoral head area. However a navigated identification of osseous lesions was possible within 1mm deviations in all regions.

Purpose: Adolescent idiopathic scoliosis (AIS) is know to occur in families and research has shown that in populations or predominantly Northern European origin, 97% of AIS patients are related to families with AIS. It affects 1–2% of the population and results in deformities treated by bracing and surgery. Brace prescription is empirical and surgery is reserved for late cases and brace failures. Identifying the genetic markers for AIS would allow creation of a diagnostic gene-based test that may also have prognostic value for differentiating progressive and non-progressive curves.

Methods: A 21 million name data base of the original European pioneers in Utah was assembled including 3 million descendents and 18 million ancestors. 500 DNA samples from affected and first degree unaffected relatives were collected and genotypes determined with capillary electrophoresis using 763 autosomal markers and gene chip scanning for 116 000 SNPs. Disease haplotypes were also scanned with a 500K SNP chip to further narrow the position of each loci.

Results: Two markers were identified with LOD scores of 7.0 and 7.3. p-values from SNP scanning were highly significant. More detailed descriptions of these genotypes will be presented.

Conclusion: Two genetic markers were identified, one of which was present in 95% of patients with AIS greater than 40°. In our population, no one with AIS less than 40° had these markers. A genotype test for AIS may be possible that would offer both diagnostic and prognostic value. Further characterization of the genes and their mutations could give information concerning the molecular pathway that lead to disease expression.