HUMAN PERIODONTAL LIGAMENT STEM CELLS CULTURED ONTO CORTICO-CANCELLOUS SCAFFOLD DRIVE BONE REGENERATIVE PROCESS
HUMAN PERIODONTAL LIGAMENT STEM CELLS CULTURED ONTO CORTICO-CANCELLOUS SCAFFOLD DRIVE BONE REGENERATIVE PROCESS
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DOI：
10.22203/ecm.v032a12
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发表时间：
2016-07-01
影响因子：
3.1
通讯作者：
Trubiani, O.
中科院分区：
文献类型：
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作者：
Diomede, F.;Zini, N.;Trubiani, O.
本工作的目的是在体外和体内测试由猪皮质-松质骨支架（Osteobiol Dual Block）（DB）和人牙周韧带干细胞（hPDLSC）的无异种离体培养物构成的新的组织工程构建体。在无异种培养基配方中培养的hPDLSC保留了干细胞的形态学特征、干细胞性和多能性标志物的表达以及它们分化成间充质谱系的能力。透射电子显微镜分析表明，经过一周的培养，无论是非诱导和成骨分化诱导细胞加入DB分泌细胞外基质（ECM），在成骨诱导样品中的纤维分层组装和生长。定量RT-PCR（qRT-PCR）显示在用DB培养的分化和未分化的hPDLSC（hPDLSC/DB）中参与骨分化途径的关键基因的上调。功能研究显示，在DB存在下，在用降钙素和甲状旁腺素刺激的未分化和分化细胞中，钙瞬变的响应显著增加，表明生物材料可以驱动hPDLSC的成骨分化过程。这些数据证实了L型电压依赖性Ca 2+（VDCCL），亚基α 1C和α 2D 1在DB的存在下，在未分化的细胞中的基因表达的增加。hPDLSC/DB活构建体在小鼠颅骨中的体内植入证明了早熟的骨整合和血管形成过程。我们的研究结果表明，考虑DB作为一种生物相容性，骨诱导性和骨传导性生物材料，使其成为一个有前途的工具，以调节细胞在生物环境中的活动，并在新的定制组织工程的发展中的潜在用途。
The purpose of this work was to test, in vitro and in vivo, a new tissue-engineered construct constituted by porcine cortico-cancellous scaffold (Osteobiol Dual Block) (DB) and xeno-free ex vivo culture of human Periodontal Ligament Stem Cells (hPDLSCs). hPDLSCs cultured in xeno-free media formulation preserved the stem cells' morphological features, the expression of stemness and pluripotency markers, and their ability to differentiate into mesenchymal lineage. Transmission electron microscopy analysis suggested that after one week of culture, both noninduced and osteogenic differentiation induced cells joined and grew on DB secreting extracellular matrix (ECM) that in osteogenic induced samples was hierarchically assembled in fibrils. Quantitative RT-PCR (qRT-PCR) showed the upregulation of key genes involved in the bone differentiation pathway in both differentiated and undifferentiated hPDLSCs cultured with DB (hPDLSCs/DB). Functional studies revealed a significant increased response of calcium transients in the presence of DB, both in undifferentiated and differentiated cells stimulated with calcitonin and parathormone, suggesting that the biomaterial could drive the osteogenic differentiation process of hPDLSCs. These data were confirmed by the increase of gene expression of L-type voltage-dependent Ca2+ (VDCCL), subunits alpha 1C and alpha 2D1 in undifferentiated cells in the presence of DB. In vivo implantation of the hPDLSCs/DB living construct in the mouse calvaria evidenced a precocious osteointegration and vascularisation process. Our results suggest consideration of DB as a biocompatible, osteoinductive and osteoconductive biomaterial, making it a promising tool to regulate cell activities in biological environments and for a potential use in the development of new custom-made tissue engineering.