Lima Eric G, Grace Chao Pen-Hsiu, Ateshian Gerard A, Bal B Sonny, Cook James L, Vunjak-Novakovic Gordana, Hung Clark T
Department of Biomedical Engineering, Columbia University, 351 Engineering Terrace MC8904, 1210 Amsterdam Avenue, New York, NY 10027, United States.
Biomaterials. 2008 Nov;29(32):4292-9. doi: 10.1016/j.biomaterials.2008.07.018. Epub 2008 Aug 20.
In the current study, evidence is presented demonstrating that devitalized trabecular bone has an inhibitory effect on in vitro chondral tissue development when used as a base material for the tissue-engineering of osteochondral constructs for cartilage repair. Chondrocyte-seeded agarose hydrogel constructs were cultured alone or attached to an underlying bony base in a chemically defined medium formulation that has been shown to yield engineered cartilaginous tissue with native Young's modulus (E(Y)) and glycosaminoglycan (GAG) content. By day 42 in culture the incorporation of a bony base significantly reduced these properties (E(Y)=87+/-12 kPa, GAG=1.9+/-0.8%ww) compared to the gel-alone group (E(Y)=642+/-97 kPa, GAG=4.6+/-1.4%ww). Similarly, the mechanical and biochemical properties of chondrocyte-seeded agarose constructs were inhibited when co-cultured adjacent to bone (unattached), suggesting that soluble factors rather than direct cell-bone interactions mediate the chondro-inhibitory bone effects. Altering the method of bone preparation, including demineralization, or the timing of bone introduction in co-culture did not ameliorate the effects. In contrast, osteochondral constructs with native cartilage properties (E(Y)=730+/-65 kPa, GAG=5.2+/-0.9%ww) were achieved when a porous tantalum metal base material was adopted instead of bone. This work suggests that devitalized bone may not be a suitable substrate for long-term cultivation of osteochondral grafts.
在当前研究中,有证据表明,当失活的小梁骨用作软骨修复的骨软骨构建体组织工程的基础材料时,其对体外软骨组织发育具有抑制作用。接种软骨细胞的琼脂糖水凝胶构建体单独培养,或附着于化学定义培养基配方中的下层骨基上,该培养基已被证明能产生具有天然杨氏模量(E(Y))和糖胺聚糖(GAG)含量的工程软骨组织。培养至第42天时,与仅凝胶组(E(Y)=642±97 kPa,GAG=4.6±1.4%ww)相比,加入骨基显著降低了这些特性(E(Y)=87±12 kPa,GAG=1.9±0.8%ww)。同样,接种软骨细胞的琼脂糖构建体与骨(未附着)共培养时,其力学和生化特性受到抑制,这表明可溶性因子而非直接的细胞-骨相互作用介导了软骨抑制性骨效应。改变骨制备方法,包括脱矿,或共培养中骨引入的时间,均未改善这种效应。相比之下,采用多孔钽金属基础材料而非骨时,可获得具有天然软骨特性(E(Y)=730±65 kPa,GAG=5.2±0.9%ww)的骨软骨构建体。这项工作表明,失活骨可能不是骨软骨移植物长期培养的合适基质。