Soares Diana Gabriela, Rosseto Hebert Luís, Basso Fernanda Gonçalves, Scheffel Débora Salles, Hebling Josimeri, Costa Carlos Alberto de Souza
Department of Physiology and Pathology, Araraquara School of Dentistry, Universidade Estadual Paulista, Araraquara, SP, Brazil.
Ribeirão Preto School of Medicine, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
Braz Oral Res. 2016;30(1):e54. doi: 10.1590/1807-3107BOR-2016.vol30.0054. Epub 2016 Apr 26.
The development of biomaterials capable of driving dental pulp stem cell differentiation into odontoblast-like cells able to secrete reparative dentin is the goal of current conservative dentistry. In the present investigation, a biomembrane (BM) composed of a chitosan/collagen matrix embedded with calcium-aluminate microparticles was tested. The BM was produced by mixing collagen gel with a chitosan solution (2:1), and then adding bioactive calcium-aluminate cement as the mineral phase. An inert material (polystyrene) was used as the negative control. Human dental pulp cells were seeded onto the surface of certain materials, and the cytocompatibility was evaluated by cell proliferation and cell morphology, assessed after 1, 7, 14 and 28 days in culture. The odontoblastic differentiation was evaluated by measuring alkaline phosphatase (ALP) activity, total protein production, gene expression of DMP-1/DSPP and mineralized nodule deposition. The pulp cells were able to attach onto the BM surface and spread, displaying a faster proliferative rate at initial periods than that of the control cells. The BM also acted on the cells to induce more intense ALP activity, protein production at 14 days, and higher gene expression of DSPP and DMP-1 at 28 days, leading to the deposition of about five times more mineralized matrix than the cells in the control group. Therefore, the experimental biomembrane induced the differentiation of pulp cells into odontoblast-like cells featuring a highly secretory phenotype. This innovative bioactive material can drive other protocols for dental pulp exposure treatment by inducing the regeneration of dentin tissue mediated by resident cells.
开发能够驱动牙髓干细胞分化为能分泌修复性牙本质的成牙本质样细胞的生物材料是当前保守牙科的目标。在本研究中，测试了一种由嵌入铝酸钙微粒的壳聚糖/胶原蛋白基质组成的生物膜（BM）。该生物膜是通过将胶原凝胶与壳聚糖溶液（2:1）混合，然后添加生物活性铝酸钙水泥作为矿相制成的。使用惰性材料（聚苯乙烯）作为阴性对照。将人牙髓细胞接种到某些材料的表面，并通过细胞增殖和细胞形态评估细胞相容性，在培养1、7、14和28天后进行评估。通过测量碱性磷酸酶（ALP）活性、总蛋白产量、DMP-1/DSPP的基因表达和矿化结节沉积来评估成牙本质细胞分化。牙髓细胞能够附着在生物膜表面并铺展，在初始阶段显示出比对照细胞更快的增殖速率。该生物膜还作用于细胞，诱导更强烈的ALP活性、14天时的蛋白质产量以及28天时更高的DSPP和DMP-1基因表达，导致矿化基质的沉积比对照组细胞多约五倍。因此，实验性生物膜诱导牙髓细胞分化为具有高度分泌表型的成牙本质样细胞。这种创新的生物活性材料可以通过诱导驻留细胞介导的牙本质组织再生来推动其他牙髓暴露治疗方案。
