ABSTRACT
Bone marrow (BM) has been long established as the main source of pluripotential mesenchymal stem cells (MSCs), and has been so far the main recognized source of osteoprogenitor cells that contribute to the turnover of the skeletal scaffold. The existence of an osteoprogenitor cell in other connective tissues such as skeletal muscle has been reported. In light of its availability and because of the relative ease of muscle cell isolation, skeletal muscle is an attractive source of cells for use in tissue engineering applications. The aim of this study was to explore the potential to differentiate into the chondro--osteoblastic lineage of a plastic adhering cell population, referred t as skeletal muscle-derived cells (SMDCs), obtained from biopsies of rat skeletal muscle. SMDCs displayed a fibroblast-like morphology. Our study revealed that the isolated cell population had a mesenchymal origin as indicated by abundant expression of STRO-1 and CD166. Osteogenic markers like osteocalcin (OC), bone sialoprotein (BSP) and osteopontin (OP) gene expressions were detected by RT-PCR. When these cells were cultured in the presence of an osteo-inductive culture medium, positive staining for alkaline phosphatase (ALP) and formation of mineralized matrix were increased. Furthermore SMDCs formed bone and cartilage tissues in vivo when placed inside of diffusion chambers and in demineralized bone matrix (DBM) cylinders, implanted subdermically into the backs of rat for 28 days. In conclusion, this experimental procedure is capable of selecting a cell population obtained from the skeletal muscle that is able to complete the differentiation pathway leading to the formation of cartilage and bone. In this respect SMDCs resemble BM stromal cells (BMSCs) and have demonstrated a potential application for cartilage and bone tissue engineering.
Subject(s)
Cell Differentiation/physiology , Mesenchymal Stem Cells/physiology , Muscle, Skeletal/cytology , Animals , Biomarkers/metabolism , Cell Shape , Cells, Cultured , Chondrocytes/cytology , Chondrocytes/physiology , Mesenchymal Stem Cells/cytology , Osteoblasts/cytology , Osteoblasts/physiology , Osteogenesis/physiology , Rats , Tissue EngineeringABSTRACT
BACKGROUND: Recent studies have demonstrated that adipose-derived adult stromal cells (ADASCs) offer great promise for cell-based therapies due to their ability to differentiate towards bone, cartilage and fat [corrected] The objective of this study was to investigate whether type I collagen would elicit in vivo bone formation of passaged rat adipose-derived adult stromal cells (ADASC) placed extraskeletally. METHODS: After expansion for 1-4 passages (P), cells were incubated in osteogenic medium containing dexamethasone, ascorbic acid and beta-glycerol phosphate for 2-4 weeks. Undifferentiated cells were maintained in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS). Osteogenic differentiation was evaluated by alkaline phosphatase (ALP) and von Kossa staining as well as by gene expression of ALP, osteopontin (OP), osteonectin (ON), osteocalcin (OC), collagen I (colI), collagen II (colII), bone sialoprotein (BSP), periostin (Postn), runx2, osterix (Osx), sox9, msx1 and msx2. Diffusion chambers were filled with 1x10(6) cells mixed with or without type I collagen gel and implanted subcutaneously into rats. Controls included chambers exposed to (1) undifferentiated cells (with or without collagen, (2) collagen without cells and (3) empty chambers (n=5 per group). RESULTS: Four weeks after implantation, in vivo bone and cartilage formation was demonstrated in implants containing 4-week osteo-induced P1 and P4 cells wrapped in the collagen gel, as confirmed by Goldner's trichrome and Alcian blue staining, respectively. Newly formed bone stained positive for type I collagen. Control implants had no bone or cartilage and were primarily filled with fibrous tissue at that time interval. DISCUSSION: Recent studies have demonstrated that ADASC offer great promise for cell-based therapies because of their ability to differentiate toward bone, cartilage and fat. However, the influence of different matrices on the in vivo osteogenic capability of ADASC is not fully understood. These findings suggest that type I collagen may support the survival and expression of osteogenic and chondrogenic phenotypes in passaged rat ADASC in vivo.
Subject(s)
Adipose Tissue/drug effects , Cell Differentiation/drug effects , Chondrogenesis/drug effects , Collagen Type I/pharmacology , Osteogenesis/drug effects , Adipose Tissue/physiology , Alkaline Phosphatase/analysis , Animals , Cell Differentiation/physiology , Cells, Cultured , Chondrogenesis/physiology , Gene Expression , Male , Osteogenesis/physiology , Rats , Rats, Wistar , Stromal Cells/cytology , Stromal Cells/drug effects , Stromal Cells/metabolismABSTRACT
El puente compuesto de protesis tubular de dacron-autoinjerto arterial realizado en 16 pacientes con el objeto de revascularizar la arteria poplitea infrapatelar fue exitoso en 9 (59,9%), con una evolucion promedio de 40 meses. Es un recurso aplicable ante la carencia de vena autogena viable, circunstancia frecuente en esta cirugia
Subject(s)
Middle Aged , Aged , Humans , Male , Female , Femoral Artery , Popliteal Artery , Vascular Surgical Procedures , Surgical Procedures, OperativeABSTRACT
El puente compuesto de protesis tubular de dacron-autoinjerto arterial realizado en 16 pacientes con el objeto de revascularizar la arteria poplitea infrapatelar fue exitoso en 9 (59,9%), con una evolucion promedio de 40 meses. Es un recurso aplicable ante la carencia de vena autogena viable, circunstancia frecuente en esta cirugia
