Bone and cartilage regeneration by using fat tissue without cell culture

成骨细胞的分化从吸脂吸出的人脂肪来源的成体干细胞与或不与bFGF培养。对于成骨细胞分化，用地塞米松或BMP-2处理细胞。测定这些细胞的碱性磷酸酶活性，并通过实时RT-PCR分析成骨细胞标志物。碱性磷酸酶阳性细胞存在于地塞米松或BMP-2处理的细胞中。成骨细胞标志物的基因表达增加。将体外培养的细胞或新鲜细胞接种于胶原支架材料中，植入裸鼠皮下，观察成骨细胞和软骨细胞的生长情况。在8周时通过组织学评估骨形成。组织学上，检测到少量骨形成。免疫组化显示一些细胞来源于人体组织。将新鲜处理的脂肪细胞与胶原凝胶混合并植入大鼠膝关节钻孔的关节软骨缺损中。在4周时通过组织学评估软骨修复。组织学检查可见软骨修复，大部分为纤维软骨。

Differentiation for osteoblastHuman adipose-derived adult stem cells from liposuction aspirates were cultured with or without bFGF. For osteoblastic differentiation, cells were treated with dexamethasone or BMP-2. These cells were determined for alkaline phosphatase activity and analyzed by real-time RT-PCR for osteoblast markers. Alkaline phosphatase-positive cells were present in cells treated with dexamethasone or BMP-2. Gene expression for osteoblast markers were increased. Osteoblastic differentiation was enhanced in cell cultured with bFGF.Bone and cartilage formation in vivoCultuted cells or fresh cells were seeded into Collagraft scaffolds and implanted under the skin of nude mice. Bone formation was assessed by histology at 8 weeks. Histologically, bone formation was detected in small amounts. Immunohistochemistry revealed some cells were derived from human tissue. Fresh processed adipocytes were mixed with collagen gel and implanted into articular cartilage defects drilled into rat knees. Cartilage repair was assessed by histology at 4 weeks. Histologically, cartilage repair was detected and most of area was fibrocartilage.