Characterization of bursa subacromialis-derived mesenchymal stem cells.

INTRODUCTION: The bursa subacromialis (BS) provides the gliding mechanism of the shoulder and regenerates itself after surgical removal. Therefore, we explored the presence of mesenchymal stem cells (MSCs) within the human adult BS tissue and characterized the BS cells compared to MSCs from bone marrow (BMSCs) on a molecular level. METHODS: BS cells were isolated by collagenase digest from BS tissues derived from patients with degenerative rotator cuff tears, and BMSCs were recovered by adherent culture from bone-marrow of patients with osteoarthritis of the hip. BS cells and BMSCs were compared upon their potential to proliferate and differentiate along chondrogenic, osteogenic and adipogenic lineages under specific culture conditions. Expression profiles of markers associated with mesenchymal phenotypes were comparatively evaluated by flow cytometry, immunohistochemistry, and whole genome array analyses. RESULTS: BS cells and BMSCs appeared mainly fibroblastic and revealed almost similar surface antigen expression profiles, which was CD44(+), CD73(+), CD90(+), CD105(+), CD106(+), STRO-1(+), CD14(-), CD31(-), CD34(-), CD45(-), CD144(-). Array analyses revealed 1969 genes upregulated and 1184 genes downregulated in BS cells vs. BMSCs, indicating a high level of transcriptome similarity. After 3 weeks of differentiation culture, BS cells and BMSCs showed a similar strong chondrogenic, adipogenic and osteogenic potential, as shown by histological, immunohistochemical and RT-PCR analyses in contrast to the respective negative controls. CONCLUSIONS: Our in vitro characterizations show that BS cells fulfill all characteristics of mesenchymal stem cells, and therefore merit further attention for the development of improved therapies for various shoulder pathologies.

Indian hedgehog gene transfer is a chondrogenic inducer of human mesenchymal stem cells.

INTRODUCTION: To date, no single most-appropriate factor or delivery method has been identified for the purpose of mesenchymal stem cell (MSC)-based treatment of cartilage injury. Therefore, in this study we tested whether gene delivery of the growth factor Indian hedgehog (IHH) was able to induce chondrogenesis in human primary MSCs, and whether it was possible by such an approach to modulate the appearance of chondrogenic hypertrophy in pellet cultures in vitro. METHODS: First-generation adenoviral vectors encoding the cDNA of the human IHH gene were created by cre-lox recombination and used alone or in combination with adenoviral vectors, bone morphogenetic protein-2 (Ad.BMP-2), or transforming growth factor beta-1 (Ad.TGF-ß1) to transduce human bone-marrow derived MSCs at 5 × 10² infectious particles/cell. Thereafter, 3 × 105 cells were seeded into aggregates and cultured for 3 weeks in serum-free medium, with untransduced or marker gene transduced cultures as controls. Transgene expressions were determined by ELISA, and aggregates were analysed histologically, immunohistochemically, biochemically and by RT-PCR for chondrogenesis and hypertrophy. RESULTS: IHH, TGF-ß1 and BMP-2 genes were equipotent inducers of chondrogenesis in primary MSCs, as evidenced by strong staining for proteoglycans, collagen type II, increased levels of glycosaminoglycan synthesis, and expression of mRNAs associated with chondrogenesis. IHH-modified aggregates, alone or in combination, also showed a tendency to progress towards hypertrophy, as judged by the expression of alkaline phosphatase and stainings for collagen type X and Annexin 5. CONCLUSION: As this study provides evidence for chondrogenic induction of MSC aggregates in vitro via IHH gene delivery, this technology may be efficiently employed for generating cartilaginous repair tissues in vivo.

No increased stem subsidence after arthroplasty in young patients with femoral head osteonecrosis: 41 patients followed for 1-9 years.

BACKGROUND: Poor bone stock in patients with osteonecrosis of the femoral head may be a reason for poor outcome after hip replacement. One way of studying bone quality is to measure implant migration. We thus investigated the clinical and radiographic results of cementless THR in younger patients with femoral head osteonecrosis. PATIENTS AND METHODS: We studied hips in 41 patients (mean age 48 (25-63) years) with a cementless hip arthroplasty after late stage osteonecrosis. Clinical evaluation was by the Harris hip score, the WOMAC score and the SF-36 score. Stem subsidence was measured with the Ein Bild Roentgen Analyse femoral component analysis (EBRA-FCA) at 3, 12, 24, 60, and 72 months after operation. The average duration of follow-up was 7(1-9) years, with less than 2 years for 2 patients. RESULTS: There was no revision of any hip. No radiographic or clinical stem loosening was seen. After 60 months, the cementless stems showed a median subsidence of -0.7 mm (95% CI: -0.9 to -0.2). No femoral osteolysis occurred. Femoral radiolucent lines, all < 1 mm, were seen in 10 hips. At the latest follow-up the Harris hip score was 83 (23-100) points. INTERPRETATION: Our findings for porous-coated stems in patients with femoral osteonecrosis indicate no greater risk of stem subsidence and rate of osteolysis after an average of 7 years follow-up. Thus, we continue to use uncemented stems in younger patients with femoral osteonecrosis. However, continued follow-up will be necessary to evaluate the long-term outcome.