Díaz-Arca Anabel, Ros-Tárraga Patricia, Tomé María J Martínez, De Aza Antonio H, Meseguer-Olmo Luis, Mazón Patricia, De Aza Piedad N
Instituto de Bioingeniería, Universidad Miguel Hernández, 03202 Elche, Spain.
Instituto de Investigación, Desarrollo e Innovación en Biotecnología Sanitaria de Elche, 03202 Elche, Spain.
Materials (Basel). 2021 Mar 16;14(6):1439. doi: 10.3390/ma14061439.
Micro-/nano-structured scaffolds with a weight composition of 46.6% α-tricalcium phosphate (α-TCP)-53.4% silicocarnotite (SC) were synthesized by the polymer replica method. The scanning electron microscopy (SEM) analysis of the scaffolds and natural cancellous bone was performed for comparison purposes. Scaffolds were obtained at three cooling rates via the eutectoid temperature (50 °C/h, 16.5 °C/h, 5.5 °C/h), which allowed the surface nanostructure and mechanical strength to be controlled. Surface nanostructures were characterized by transmission electron microscopy (TEM) and Raman analysis. Both phases α-TCP and SC present in the scaffolds were well-identified, looked compact and dense, and had neither porosities nor cracks. The non-cytotoxic effect was evaluated in vitro by the proliferation ability of adult human mesenchymal stem cells (ah-MSCs) seeded on scaffold surfaces. There was no evidence for cytotoxicity and the number of cells increased with culture time. A dense cell-hydroxyapatite layer formed until 28 days. The SEM analysis suggested cell-mediated extracellular matrix formation. Finally, scaffolds were functionalized with the alkaline phosphatase enzyme (ALP) to achieve biological functionalization. The ALP was successfully grafted onto scaffolds, whose enzymatic activity was maintained. Scaffolds mimicked the micro-/nano-structure and chemical composition of natural cancellous bone by considering cell biology and biomolecule functionalization.
采用聚合物复型法合成了重量组成为46.6%α-磷酸三钙（α-TCP）-53.4%硅钙钛矿（SC）的微/纳米结构支架。为了进行比较，对支架和天然松质骨进行了扫描电子显微镜（SEM）分析。通过共析温度（50℃/h、16.5℃/h、5.5℃/h）以三种冷却速率获得支架，这使得表面纳米结构和机械强度得以控制。通过透射电子显微镜（TEM）和拉曼分析对表面纳米结构进行了表征。支架中存在的α-TCP和SC两相均得到了很好的识别，看起来致密且紧实，既无孔隙也无裂纹。通过接种在支架表面的成人骨髓间充质干细胞（ah-MSCs）的增殖能力在体外评估了其无细胞毒性作用。没有细胞毒性的证据，并且细胞数量随培养时间增加。直到28天时形成了致密的细胞-羟基磷灰石层。SEM分析表明存在细胞介导的细胞外基质形成。最后，用碱性磷酸酶（ALP）对支架进行功能化以实现生物功能化。ALP成功接枝到支架上，其酶活性得以维持。通过考虑细胞生物学和生物分子功能化，支架模拟了天然松质骨的微/纳米结构和化学成分。
