Extracellular Matrix/Amorphous Magnesium Phosphate Bioink for 3D Bioprinting of Craniomaxillofacial Bone Tissue
Extracellular Matrix/Amorphous Magnesium Phosphate Bioink for 3D Bioprinting of Craniomaxillofacial Bone Tissue
复制标题
DOI：
10.1021/acsami.0c05311
复制
发表时间：
2020-05-27
影响因子：
9.5
通讯作者：
Bottino, Marco C.
中科院分区：
文献类型：
--
作者：
Dubey, Nileshkumar;Ferreira, Jessica A.;Bottino, Marco C.
生物打印是再生医学中一个很有前途的领域，它具有巨大的潜力来创造三维的、特定于缺损区的血管化骨，为解决尚未满足的颅颌面重建挑战提供了巨大的机会。细胞相容的生物墨水是成功再生功能性骨组织的关键前提。人工合成的自组装肽具有类似于天然细胞外基质(ECM)的纳米纤维结构，使其成为优秀的生物墨水成分。与磷酸钙相比，无定形磷酸镁(AMPs)表现出更高的吸收水平，同时保持了高的生物相容性、骨诱导活性和低的炎症反应。在这里，我们建立了一种新的生物墨水配方(ECM/AMP)，它将含有2%八肽FEFEFKFK和98%水的ECM水凝胶与AMP颗粒结合在一起，实现了高细胞功能和理想的生物可印性。我们分析了包裹在生物墨水中的牙髓干细胞(DPSCs)的成骨分化以及体内的骨再生，以确定配方生物墨水作为一种无生长因子成骨策略的潜力。载细胞的AMP修饰的生物印迹构建体显示出改善的细胞形态，但与不含AMP的构建体相比，细胞存活率相似(接近90%)。在功能分析中，AMP修饰的细胞生物印迹构建物在不使用生长因子的情况下表现出高水平的矿化和成骨基因表达，因此提示AMP的存在触发了DPSCs的成骨分化。以无细胞ECM为基础的生物打印构建物被植入体内。与ECM组相比，ECM/1.0AMP组在4周和8周时单位总体积的骨体积分别增加了1.7倍和1.4倍。此外，ECM/1.0AMP组的骨密度从4周到8周显著增加。这些结果表明，AMP在生物墨水中的存在显著增加了骨形成，从而显示了原位生物打印策略的前景。我们预见这种创新的生物墨水在转化为再生牙科患者特有的骨组织方面具有巨大的潜力。
Bioprinting, a promising field in regenerative medicine, holds great potential to create three-dimensional, defect-specific vascularized bones with tremendous opportunities to address unmet craniomaxillofacial reconstructive challenges. A cytocompatible bioink is a critical prerequisite to successfully regenerate functional bone tissue. Synthetic self-assembling peptides have a nanofibrous structure resembling the native extracellular matrix (ECM), making them an excellent bioink component. Amorphous magnesium phosphates (AMPs) have shown greater levels of resorption while maintaining high biocompatibility, osteoinductivity, and low inflammatory response, as compared to their calcium phosphate counterparts. Here, we have established a novel bioink formulation (ECM/AMP) that combines an ECM-based hydrogel containing 2% octapeptide FEFEFKFK and 98% water with AMP particles to realize high cell function with desirable bioprintability. We analyzed the osteogenic differentiation of dental pulp stem cells (DPSCs) encapsulated in the bioink, as well as in vivo bone regeneration, to define the potential of the formulated bioink as a growth factor-free bone-forming strategy. Cell-laden AMP-modified bioprinted constructs showed an improved cell morphology but similar cell viability (similar to 90%) compared to their AMP-free counterpart. In functional assays, the cell-laden bioprinted constructs modified with AMP exhibited a high level of mineralization and osteogenic gene expression without the use of growth factors, thus suggesting that the presence of AMP-triggered DPSCs' osteogenic differentiation. Cell-free ECM-based bioprinted constructs were implanted in vivo. In comparison with the ECM group, bone volume per total volume for ECM/1.0AMP was approximately 1.7- and 1.4-fold higher at 4 and 8 weeks, respectively. Further, a significant increase in the bone density was observed in ECM/1.0AMP from 4 to 8 weeks. These results demonstrate that the presence of AMP in the bioink significantly increased bone formation, thus showing promise for in situ bioprinting strategies. We foresee significant potential in translating this innovative bioink toward the regeneration of patient-specific bone tissue for regenerative dentistry.