Freeze-dry processing of three-dimensional cell constructs for bone graft materials.
Freeze-dry processing of three-dimensional cell constructs for bone graft materials.
复制标题
用于骨移植材料的三维细胞结构的冷冻干燥处理。
DOI：
10.1002/jbm.b.34448
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Imazato S.
中科院分区：
文献类型：
--
作者：
Sasaki JI;Yoshimoto I;Katata C;Tsuboi R;Imazato S.
冻干加工提高了细胞生物材料的可操作性和稳定性，并有利于临床应用的灭菌。然而，目前还没有针对工程组织的既定冷冻干燥方案。最近，我们报道了可以使用无支架三维（3D）细胞结构来制造仿生骨组织，具有作为骨移植材料的潜在应用。本研究的目的是评估冷冻干燥对 3D 细胞结构的形态和成分的影响。在磷酸盐缓冲盐水 (PBS) 中冷冻干燥的细胞构建体保留了有机和无机成分；而柠檬酸钠缓冲液（SCB）处理的构建体的钙和骨相关蛋白含量显着降低。通过在含 10% 蔗糖的 PBS 中冷冻干燥，细胞构建体中的碱性磷酸酶 (ALP) 活性得到维持，而用不含蔗糖的 PBS 或含蔗糖的 SCB 处理的细胞构建体显示 ALP 活性显着降低。在这项研究中，我们发现含蔗糖的磷酸盐缓冲液适合冷冻干燥，以维持 3D 细胞结构内的矿物质和蛋白质功能，而柠檬酸盐缓冲液则不合适。这项研究获得的见解可能有助于开发通过组织工程方法和骨移植生物材料制造的新型细胞基生物材料。
Freeze‐dry processing improves the operability and stability of cell‐based biomaterials and facilitates sterilization for clinical application. However, there is no established freeze‐drying protocol for engineered tissues. Recently, we reported that biomimetic bone tissues can be fabricated using scaffold‐free three‐dimensional (3D) cell constructs with potential applications as bone graft materials. The purpose of this study was to assess the influence of freeze drying on the morphology and components of 3D cell constructs. Cell constructs freeze dried in phosphate buffered saline (PBS) maintained organic and inorganic components; whereas sodium citrate buffer (SCB)‐treated constructs had significantly lower amounts of calcium and bone‐related proteins. Alkaline phosphatase (ALP) activity in cell constructs was maintained by freeze drying in 10% sucrose‐containing PBS, whereas cell constructs treated with PBS without sucrose or with sucrose‐containing SCB showed significant reductions of ALP activity. In this study, we found that sucrose‐containing phosphate buffer was suitable for freeze drying to maintain minerals and protein functions within 3D cell constructs, whereas citrate buffer was inappropriate. The insights gained by this study may facilitate the development of novel cell‐based biomaterials fabricated by tissue engineering approaches and bone graft biomaterials.