Jones Gemma L, Motta Antonella, Marshall Mike J, El Haj Alicia J, Cartmell Sarah H
Guy Hilton Research Centre, Institute of Science and Technology in Medicine, University of Keele, Thornburrow Drive, Hartshill, Stoke-on-Trent, Keele ST4 7QB, UK.
Biomaterials. 2009 Oct;30(29):5376-84. doi: 10.1016/j.biomaterials.2009.07.028. Epub 2009 Aug 3.
This study investigates the growth of a co-culture of osteoblasts and osteoclasts on four different types of degradable biomaterials with bone tissue engineering potential. Single or co-cultures of osteoblasts and osteoclasts (used at a ratio of 1:100 osteoblast:osteoclasts) were cultured on vapour stabilised silk fibroin, methanol stabilised silk fibroin, chitosan and poly (l lactic acid) (PLLA) films for 10 days. Osteoclast differentiation was determined by tartrate resistant acid phosphatase (TRAP) staining, total cell number by a picogreen DNA assay, cell morphology by scanning electron microscopy (SEM) and the material topography by atomic force microscopy (AFM). Samples were also monitored for degradation by differential scanning calorimetry (DSC) and fourier transform infrared (FTIR). Results demonstrated that vapour stabilised silk fibroin, methanol stabilised silk fibroin and chitosan all support the growth of osteoblasts and osteoclasts in both single and co-cultures. PLLA showed poor osteoclast differentiation in both single and co-cultures but supported osteoblast attachment and proliferation. Both silk fibroin materials showed sign of early degradation in the ten-day period, but very little change was seen in chitosan and PLLA samples. This study indicates that this novel co-culture approach for bone tissue engineering may be possible if scaffolds are created from silk fibroin or chitosan.
本研究调查了成骨细胞和破骨细胞在四种具有骨组织工程潜力的不同类型可降解生物材料上的共培养生长情况。将成骨细胞和破骨细胞的单培养或共培养（成骨细胞与破骨细胞的比例为1:100）在气相稳定的丝素蛋白、甲醇稳定的丝素蛋白、壳聚糖和聚（L-乳酸）（PLLA）薄膜上培养10天。通过抗酒石酸酸性磷酸酶（TRAP）染色测定破骨细胞分化，通过皮考绿DNA测定法测定总细胞数，通过扫描电子显微镜（SEM）观察细胞形态，通过原子力显微镜（AFM）观察材料表面形貌。还通过差示扫描量热法（DSC）和傅里叶变换红外光谱（FTIR）监测样品的降解情况。结果表明，气相稳定的丝素蛋白、甲醇稳定的丝素蛋白和壳聚糖在单培养和共培养中均支持成骨细胞和破骨细胞的生长。PLLA在单培养和共培养中破骨细胞分化均较差，但支持成骨细胞附着和增殖。两种丝素蛋白材料在10天内均显示出早期降解迹象，但壳聚糖和PLLA样品变化很小。本研究表明，如果用丝素蛋白或壳聚糖制作支架，这种用于骨组织工程的新型共培养方法可能可行。
