Cryopreservation Effect on Proliferative and Chondrogenic Potential of Human Chondrocytes Isolated from Superficial and Deep Cartilage

[Abstract] Objectives: To compare the proliferative and chondrogenic potential of fresh and frozen chondrocytes isolated from superficial and deep articular cartilage biopsies. Materials and Methodology: The study included 12 samples of fresh and frozen healthy human knee articular cartilage. Cell proliferation was tested at 3, 6 and 9 days. Studies of mRNA quantification, protein expression and immunofluorescence for proliferation and chondrogenic markers were performed. Results: Stimulation of fresh and frozen chondrocytes from both superficial and deep cartilage with fetal bovine serum produced an increase in the proliferative capacity compared to the non-stimulated control group. In the stimulated fresh cells group, the proliferative capacity of cells from the deep biopsy was greater than that from cells from the superficial biopsy (0.046 vs 0.028, respectively, p<0.05). There was also a significant difference between the proliferative capacity of superficial zone fresh (0.028) and frozen (0.051) chondrocytes (p<0.05). CCND1 mRNA and protein expression levels, and immunopositivity for Ki67 revealed a higher proliferative capacity for fresh articular chondrocytes from deep cartilage. Regarding the chondrogenic potential, stimulated fresh cells showed higher SOX9 and Col II expression in chondrocytes from deep than from superficial zone (p<0.05, T student test). Conclusions: The highest rate of cell proliferation and chondrogenic potential of fresh chondrocytes was found in cells obtained from deep cartilage biopsies, whereas there were no statistically significant differences in proliferative and chondrogenic capacity between biopsy origins with frozen chondrocytes. These results indicate that both origin and cryopreservation affect the proliferative and chondrogenic potential of chondrocytes.Servizo Galego de Saúde; PS07/84Instituto de Salud Carlos III; CIBER BBN CB06-01-0040Ministerio Ciencia e Innovacion; PLE2009-0144Ministerio Ciencia e Innovación; PI 08/202

MR imaging of osteochondral grafts and autologous chondrocyte implantation

Surgical articular cartilage repair therapies for cartilage defects such as osteochondral autograft transfer, autologous chondrocyte implantation (ACI) or matrix associated autologous chondrocyte transplantation (MACT) are becoming more common. MRI has become the method of choice for non-invasive follow-up of patients after cartilage repair surgery. It should be performed with cartilage sensitive sequences, including fat-suppressed proton density-weighted T2 fast spin-echo (PD/T2-FSE) and three-dimensional gradient-echo (3D GRE) sequences, which provide good signal-to-noise and contrast-to-noise ratios. A thorough magnetic resonance (MR)-based assessment of cartilage repair tissue includes evaluations of defect filling, the surface and structure of repair tissue, the signal intensity of repair tissue and the subchondral bone status. Furthermore, in osteochondral autografts surface congruity, osseous incorporation and the donor site should be assessed. High spatial resolution is mandatory and can be achieved either by using a surface coil with a 1.5-T scanner or with a knee coil at 3 T; it is particularly important for assessing graft morphology and integration. Moreover, MR imaging facilitates assessment of complications including periosteal hypertrophy, delamination, adhesions, surface incongruence and reactive changes such as effusions and synovitis. Ongoing developments include isotropic 3D sequences, for improved morphological analysis, and in vivo biochemical imaging such as dGEMRIC, T2 mapping and diffusion-weighted imaging, which make functional analysis of cartilage possible

Clinical application of scaffolds for cartilage tissue engineering

The purpose of this paper is to review the basic science and clinical literature on scaffolds clinically available for the treatment of articular cartilage injuries. The use of tissue-engineered grafts based on scaffolds seems to be as effective as conventional ACI clinically. However, there is limited evidence that scaffold techniques result in homogeneous distribution of cells. Similarly, few studies exist on the maintenance of the chondrocyte phenotype in scaffolds. Both of which would be potential advantages over the first generation ACI. The mean clinical score in all of the clinical literature on scaffold techniques significantly improved compared with preoperative values. More than 80% of patients had an excellent or good outcome. None of the short- or mid-term clinical and histological results of these tissue-engineering techniques with scaffolds were reported to be better than conventional ACI. However, some studies suggest that these methods may reduce surgical time, morbidity, and risks of periosteal hypertrophy and post-operative adhesions. Based on the available literature, we were not able to rank the scaffolds available for clinical use. Firm recommendations on which cartilage repair procedure is to be preferred is currently not known on the basis of these studies. Randomized clinical trials and longer follow-up periods are needed for more widespread information regarding the clinical effectiveness of scaffold-based, tissue-engineered cartilage repair

Subacromial impingement syndrome: management challenges

Paolo Consigliere,1 Omar Haddo,2 Ofer Levy,1,3 Giuseppe Sforza1 1The Reading Shoulder Unit, Berkshire Independent Hospital Coley Park, Swallows Croft, Reading, Berkshire, UK; 2The Whittington Hospital NHS FT, London, UK; 3The Reading Shoulder Unit, Royal Berkshire NHS Foundation Trust, Reading, Berkshire, UK Abstract: The painful shoulder is the most common condition seen in specialist shoulder clinics. It is often associated with lack of range of motion and reduced shoulder function. Lack of sleep and difficulties in performing basic daily life activities are common findings. Subacromial Impingement Syndrome (SAIS) has been considered as the most common cause of shoulder pain since it was described in 1852. Charles Neer, in 1972, described the presence of a &ldquo;proliferative spur and ridge&rdquo; on the undersurface of the acromion, which needs to be removed to improve the symptoms (acromioplasty). Neer&rsquo;s &ldquo;impingement&rdquo; hegemony was undisputed for at least 30 years. A more extensive knowledge of the pathogenesis of SAIS, however, has led authors to challenge the role of &ldquo;impingement&rdquo; in the shoulder pain and the role of surgical intervention. The aim of this review was to understand if there is still a role for surgical decompression in patients with SAIS. A literature review was performed in PubMed, PEDro, Embase, and the Cochrane Central Register of Controlled Trials using impingement, subacromial space, rotator cuff tears, tendinopathy, and tendinitis as key words. Randomized clinical trials (RCTs) with long-term follow-up comparing surgical intervention and conservative treatments in SAIS were preferred; however, prospective articles studying the outcome of surgical decompression and physiotherapy were also included. The majority of the studies showed no difference in the outcome between patients randomized to surgical decompression or conservative management. However, some studies reported better results after surgery, especially in the long term. Interpretation of the results is very difficult as most of the studies are of poor quality and have short follow-up. In our opinion, the type of subacromial lesion needs to be considered; this may offer an explanation to the difference in severity of symptoms and to the varying degrees of response to certain treatments, including surgery. Further studies are mandatory to better understand the role of surgery in SAIS. Keywords: impingement, bursitis, supraspinatus tendon, rotator cuff, rotator cuff tears, tendinopathy, tendiniti

Sequential outcome following autologous chondrocyte implantation of the knee: A six-year follow-up

This prospective six-year longitudinal study reviews the clinical outcome of patients undergoing autologous chondrocyte implantation (ACI) and a porcine type I/III collagen membrane cover for deep chondral defects of the knee. We present 57 patients (31 male, 26 female) with a mean age of 31.6 years (range 15–51 years) that have undergone ACI since July 1998. The mean size of the defect was 3.14 cm2 (range 1.0–7.0 cm2). All patients were assessed annually using seven independent validated clinical rating scores with the data analysed using ANOVA. ACI using a porcine type I/III collagen membrane cover produced statistically significant improvements (p < 0.001), maintained for up to six years, in knee symptoms compared to pre-operative levels. This study provides evidence of the medium-term benefit achieved by transplanting autologous chondrocytes to osteochondral defects