Wittenburg Gretel, Flade Viktoria, Garbe Annette I, Lauer Günter, Labudde Dirk
Clinic of Oral and Maxillofacial Surgery, University of Technology Dresden, University Hospital Carl Gustav Carus, Fetscherstr. 74, D-01307 Dresden, Germany; Department of Bioinformatics, University of Applied Sciences Mittweida, Technikumplatz 17, D-09648 Mittweida, Germany; Centre for Translational Bone, Joint and Soft Tissue Research, Fetscherstr. 74, D-01307 Dresden, Germany.
Clinic of Oral and Maxillofacial Surgery, University of Technology Dresden, University Hospital Carl Gustav Carus, Fetscherstr. 74, D-01307 Dresden, Germany.
Br J Oral Maxillofac Surg. 2014 May;52(5):409-14. doi: 10.1016/j.bjoms.2014.02.023. Epub 2014 Mar 27.
We have analysed the growth and differentiation of mesenchymal stromal cells (MSC) from bone marrow, and of adipose derived stem cells (ASC) from murine abdominal fat tissue, of green fluorescent protein (GFP) transgenic animals grown directly on two types of hydroxyapatite ceramic bone substitutes. BONITmatrix® and NanoBone® have specific mechanical and physiochemical properties such as porosity and an inner surface that influence cellular growth. Both MSC and ASC were separately seeded on 200mg of each biomaterial and cultured for 3 weeks under osteogenic differentiation conditions. The degree of mineralisation was assessed by alizarin red dye and the specific alkaline phosphatase activity of the differentiated cells. The morphology of the cells was examined by scanning electron microscopy and confocal microscopy. The osteoblastic phenotype of the cells was confirmed by analysing the expression of bone-specific genes (Runx2, osteocalcin, osteopontin, and osteonectin) by semiquantitative reverse transcriptase polymerase chain reaction (PCR). Comparison of BONITmatrix® and NanoBone® showed cell type-specific preferences in terms of osteogenic differentiation. MSC-derived osteoblast-like cells spread optimally on the surface of NanoBone® but not BONITmatrix® granules. In contrast BONITmatrix® granules conditioned the growth of osteoblast-like cells derived from ASC. The osteoblastic phenotype of the cultured cells on all matrices was confirmed by specific gene expression. Our results show that the in vitro growth and osteogenic differentiation of murine MSC or ASC of GFP transgenic mice are distinctly influenced by the ceramic substratum. While NanoBone® granules support the proliferation and differentiation of murine MSC isolated from bone marrow, the growth of murine ASC is supported by BONITmatrix® granules. NanoBone® is therefore recommended for use as scaffold in tissue engineering that requires MSC, whereas ASC can be combined with BONITmatrix® for in vitro bone engineering.
我们分析了来自骨髓的间充质基质细胞（MSC）以及来自小鼠腹部脂肪组织的脂肪衍生干细胞（ASC）在直接生长于两种羟基磷灰石陶瓷骨替代物上的绿色荧光蛋白（GFP）转基因动物中的生长和分化情况。BONITmatrix®和NanoBone®具有特定的机械和物理化学性质，如孔隙率和内表面，这些性质会影响细胞生长。将MSC和ASC分别接种在每种200mg的生物材料上，并在成骨分化条件下培养3周。通过茜素红染料评估矿化程度以及分化细胞的特异性碱性磷酸酶活性。通过扫描电子显微镜和共聚焦显微镜检查细胞形态。通过半定量逆转录聚合酶链反应（PCR）分析骨特异性基因（Runx2、骨钙素、骨桥蛋白和骨连接蛋白）的表达，以确认细胞的成骨细胞表型。BONITmatrix®和NanoBone®的比较显示，在成骨分化方面存在细胞类型特异性偏好。源自MSC的成骨样细胞在NanoBone®表面而非BONITmatrix®颗粒上最佳地铺展。相反，BONITmatrix®颗粒调节源自ASC的成骨样细胞的生长。通过特异性基因表达证实了所有基质上培养细胞的成骨细胞表型。我们的结果表明，GFP转基因小鼠的鼠MSC或ASC的体外生长和成骨分化受到陶瓷基质的明显影响。虽然NanoBone®颗粒支持从骨髓分离的鼠MSC的增殖和分化，但鼠ASC的生长则由BONITmatrix®颗粒支持。因此，推荐将NanoBone®用作需要MSC的组织工程中的支架，而ASC可与BONITmatrix®结合用于体外骨工程。
