Becerra José, Guerado Enrique, Claros Silvia, Alonso Mônica, Bertrand María L, González Carlos, Andrades José A
University of Málaga, Department of Cell Biology, Genetics and Physiology, Faculty of Sciences, Malaga, Spain.
Regen Med. 2006 Mar;1(2):267-78. doi: 10.2217/17460751.1.2.267.
We report the first clinical case of transplantation of autologous bone marrow-derived cells in vitro exposed to a novel recombinant human transforming growth factor (rhTGF)-beta1 fusion protein bearing a collagen-binding domain (rhTGF-beta(1)-F2), dexamethasone (DEX) and beta-glycerophosphate (beta-GP). When such culture-expanded cells were loaded into porous ceramic scaffolds and transplanted into the bone defect of a 69-year-old man, they differentiated into bone tissue. Marrow cells were obtained from the iliac crest and cultured in collagen gels impregnated with rhTGF-beta1-F2. Cells were selected under serum-restricted conditions in rhTGF-beta(1)-F2-containing medium for 10 days, expanded in 20% serum for 22 days and osteoinduced for 3 additional days in DEX/beta-GP-supplemented medium. We found that the cell number harvested from rhTGF-beta(1)-F2-treated cultures was significantly higher (2.3- to 3-fold) than that from untreated cultures. rhTGF-beta(1)-F2 treatment also significantly increased alkaline phosphatase activity (2.2- to 5-fold) and osteocalcin synthesis, while calcium was only detected in rhTGF-beta(1)-F2-treated cells. Eight weeks after transplantation, most of the scaffold pores were filled with bone and marrow tissue. When we tested the same human cells treated in vitro in a rat model using diffusion chambers, there was subsequent development of cartilage and bone following the subcutaneous transplantation of rhTGF-beta(1)-F2-treated cells. This supports the suggestion that such cells were marrow-derived cells, with chondrogenic and osteogenic potential, whereas the untreated cells were not under the same conditions. The ability for differentiation into cartilage and bone tissues, combined with an extensive proliferation capacity, makes such a marrow-derived stem cell population valuable to induce bone regeneration at skeletal defect sites.
我们报告了首例自体骨髓来源细胞移植的临床病例，这些细胞在体外暴露于一种新型的带有胶原结合域的重组人转化生长因子（rhTGF）-β1融合蛋白（rhTGF-β(1)-F2）、地塞米松（DEX）和β-甘油磷酸（β-GP）。当将这种体外培养扩增的细胞加载到多孔陶瓷支架中，并移植到一名69岁男性的骨缺损处时，它们分化形成了骨组织。骨髓细胞取自髂嵴，在浸渍有rhTGF-β1-F2的胶原凝胶中培养。细胞在含rhTGF-β(1)-F2的培养基中血清限制条件下筛选10天，在20%血清中扩增22天，并在添加了DEX/β-GP的培养基中再进行3天的成骨诱导。我们发现，从rhTGF-β(1)-F2处理的培养物中收获的细胞数量显著高于未处理的培养物（2.3至3倍）。rhTGF-β(1)-F2处理还显著提高了碱性磷酸酶活性（2.2至5倍）和骨钙素合成，而仅在rhTGF-β(1)-F2处理的细胞中检测到钙。移植八周后，大多数支架孔隙被骨和骨髓组织填充。当我们在大鼠模型中使用扩散室对体外处理的相同人类细胞进行测试时，rhTGF-β(1)-F2处理的细胞皮下移植后随后形成了软骨和骨。这支持了这样的观点，即此类细胞是具有软骨生成和成骨潜力的骨髓来源细胞，而未处理的细胞在相同条件下则不具备。分化为软骨和骨组织的能力，以及广泛的增殖能力，使得这样一群骨髓来源的干细胞对于诱导骨骼缺损部位的骨再生具有重要价值。
