Acellular mineral deposition in collagen-based biomaterials incubated in cell culture media
Acellular mineral deposition in collagen-based biomaterials incubated in cell culture media
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DOI：
10.1007/s002230010041
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发表时间：
2000-03-01
影响因子：
4.2
通讯作者：
Herbage, D
中科院分区：
文献类型：
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作者：
Andre-Frei, V;Chevallay, B;Herbage, D
组织工程的快速发展重新引起了人们对可生物降解三维结构的兴趣，例如胶原基生物材料。在体外种植成纤维细胞、成骨细胞和软骨细胞的胶原基质可以形成类似皮肤、骨和软骨的组织，可以作为受损组织的功能替代品。胶原蛋白与胶原种植体的营养不良钙化和骨矿化有关。我们在这里报告了胶原海绵在无细胞培养液中孵育的钙化特性。通过X射线、电子衍射、傅里叶变换红外光谱、钙磷摩尔比(Ca：P)等方法对沉积在海绵中的矿物进行了鉴定，结果表明，沉积在海绵中的磷灰石是一种结晶较差的类骨磷灰石。钙化程度随培养时间的延长和培养液中钙磷含量的增加而增加，在含1.6-3 mM钙和2-3 mm(2)CaxP摩尔积的培养液中培养21天后，钙化程度达到10-15%，而在含1.33 mM钙和1.7 mm(2)CaxP摩尔积的培养基中培养21d，钙化程度仅为2%。在经过广泛洗涤且初始P含量低于0.01%的海绵中，矿物沉积被完全抑制。在这些海绵中重新添加磷酸盐以及随后的冷冻干燥和灭菌恢复了它们的矿化能力，这表明胶原本身不能启动钙化，与胶原制备过程相关的无机磷含量处于固态时是潜在的成核剂。在海绵中加入4-硫酸软骨素可部分或全部抑制矿物沉积，即使80%-90%的化合物在24小时内释放。这些结果表明，在目前组织工程中使用的培养条件下，胶原基生物材料可以发生脱细胞钙化。
Rapid developments in tissue engineering have renewed interest in biodegradable three-dimensional structures such as collagen-based biomaterials. Collagen matrices seeded in vitro with fibroblasts, osteoblasts, and chondrocytes can form tissues resembling skin, bone, and cartilage that could be used as functional substitutes for damaged tissues. Collagen is associated with both dystrophic calcification of collagenous implants and bone mineralization. We report here the calcification properties of collagen sponges incubated in cell-free media. Mineral deposited in sponges was identified by X-ray and electron diffraction, Fourier transform infrared spectroscopy, and the molar ratio of calcium:phosphorus (Ca:P) as a poorly crystalline apatite similar to bone. The degree of calcification increased with length of incubation and the Ca and P content of the media, with 10-15% Ca (dry weight) after 21 days' incubation in media containing 1.6-3 mM Ca and a Ca x P molar product of 2-3 mM(2), but only 2% Ca after incubation in medium with 1.33 mM Ca and a 1.7 mM(2) Ca x P molar product. Mineral deposition was completely inhibited in sponges that were washed extensively and initially contained less than 0.01% P. Readdition of phosphate in these sponges and subsequent freeze drying and sterilization restore their mineralization capacity, suggesting that collagen per se cannot initiate calcification and that the inorganic phosphate content associated with the collagen preparation process is in the solid state a potential nucleator. Addition of chondroitin 4-sulfate to the sponges partially or totally inhibited mineral deposition, even though 80-90% of the compound was released within 24 hours. These results indicate that acellular calcification of collagen-based biomaterials can occur under the culture conditions currently used in tissue engineering.