Lambricht Laure, De Berdt Pauline, Vanacker Julie, Leprince Julian, Diogenes Anibal, Goldansaz Hadi, Bouzin Caroline, Préat Véronique, Dupont-Gillain Christine, des Rieux Anne
Université catholique de Louvain, Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials, 1200 Brussels, Belgium.
Department of Endodontics, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA.
Dent Mater. 2014 Dec;30(12):e349-61. doi: 10.1016/j.dental.2014.08.369. Epub 2014 Aug 31.
The goal of the present work was to evaluate in vitro and in vivo the influence of various types and compositions of natural hydrogels on the viability and metabolic activity of SCAPs.
Two alginate, three hyaluronic-based (Corgel™) hydrogel formulations and Matrigel were characterized for their mechanical, surface and microstructure properties using rheology, X-ray photoelectron spectroscopy and scanning electron microscopy, respectively. A characterized SCAP cell line (RP89 cells) was encapsulated in the different experimental hydrogel formulations. Cells were cultured in vitro, or implanted in cyclosporine treated mice. In vitro cell viability was evaluated using a Live/Dead assay and in vitro cellular metabolic activity was evaluated with a MTS assay. In vivo cell apoptosis was evaluated by a TUNEL test and RP89 cells were identified by human mitochondria immunostaining.
Hydrogel composition influenced their mechanical and surface properties, and their microstructure. In vitro cell viability was above 80% after 2 days but decreased significantly after 7 days (60-40%). Viability at day 7 was the highest in Matrigel (70%) and then in Corgel 1.5 (60%). Metabolic activity increased over time in all the hydrogels, excepted in alginate SLM. SCAPs survived after 1 week in vivo with low apoptosis (<1%). The highest number of RP89 cells was found in Corgel 5.5 (140cells/mm(2)).
Collectively, these data demonstrate that SCAP viability was directly modulated by hydrogel composition and suggest that a commercially available hyaluronic acid-based formulation might be a suitable delivery vehicle for SCAP-based dental pulp regeneration strategies.
本研究旨在体外和体内评估各种类型和组成的天然水凝胶对根尖乳头干细胞(SCAPs)活力和代谢活性的影响。
分别使用流变学、X射线光电子能谱和扫描电子显微镜对两种藻酸盐、三种基于透明质酸的(Corgel™)水凝胶配方以及基质胶的力学、表面和微观结构特性进行表征。将经过表征的SCAP细胞系(RP89细胞)封装在不同的实验水凝胶配方中。细胞在体外培养,或植入经环孢素处理的小鼠体内。使用活/死检测法评估体外细胞活力,使用MTS检测法评估体外细胞代谢活性。通过TUNEL检测评估体内细胞凋亡情况,并通过人线粒体免疫染色鉴定RP89细胞。
水凝胶的组成影响其力学和表面特性以及微观结构。2天后体外细胞活力高于80%,但7天后显著下降(60%-40%)。第7天的活力在基质胶中最高(70%),其次是Corgel 1.5(60%)。除藻酸盐SLM外,所有水凝胶中的代谢活性均随时间增加。SCAPs在体内1周后存活,凋亡率较低(<1%)。在Corgel 5.5中发现的RP89细胞数量最多(140个细胞/mm²)。
总体而言,这些数据表明SCAP的活力直接受水凝胶组成的调节,并表明一种市售的基于透明质酸的配方可能是基于SCAP的牙髓再生策略的合适递送载体。