de Camargo Reis Mello Daphne, Rodrigues Lais Morandini, D'Antola Mello Fabia Zampieri, Gonçalves Thais Fernanda, Ferreira Bento, Schneider Sandra Giacomin, de Oliveira Luciane Dias, de Vasconcellos Luana Marotta Reis
Department of Bioscience and Oral Diagnosis, São José dos Campos School of Dentistry, Universidade Estadual Paulista (UNESP), Av. Engenheiro Francisco José Longo, 777, São José dos Campos, SP, 12245-000, Brazil.
Oakland University, Mathematics and Science, 318 Meadow Brook Rd, Rochester Hills, USA.
Int J Implant Dent. 2020 Oct 25;6(1):65. doi: 10.1186/s40729-020-00261-3.
An effective biomaterial for bone replacement should have properties to avoid bacterial contamination and promote bone formation while inducing rapid cell differentiation simultaneously. Bone marrow stem cells are currently being investigated because of their known potential for differentiation in osteoblast lineage. This makes these cells a good option for stem cell-based therapy. We have aimed to analyze, in vitro, the potential of pure titanium (Ti), Ti-35Nb-7Zr alloy (A), niobium (Nb), and zirconia (Zr) to avoid the microorganisms S. aureus (S.a) and P. aeruginosa (P.a). Furthermore, our objective was to evaluate if the basic elements of Ti-35Nb-7Zr alloy have any influence on bone marrow stromal cells, the source of stem cells, and observe if these metals have properties to induce cell differentiation into osteoblasts.
Bone marrow stromal cells (BMSC) were obtained from mice femurs and cultured in osteogenic media without dexamethasone as an external source of cell differentiation. The samples were divided into Ti-35Nb-7Zr alloy (A), pure titanium (Ti), Nb (niobium), and Zr (zirconia) and were characterized by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS). After predetermined periods, cell interaction, cytotoxicity, proliferation, and cell differentiation tests were performed. For monotypic biofilm formation, standardized suspensions (10 cells/ml) with the microorganisms S. aureus (S.a) and P. aeruginosa (P.a) were cultured for 24 h on the samples and submitted to an MTT test.
All samples presented cell proliferation, growth, and spreading. All groups presented cell viability above 70%, but the alloy (A) showed better results, with statistical differences from Nb and Zr samples. Zr expressed higher ALP activity and was statistically different from the other groups (p < 0.05). In contrast, no statistical difference was observed between the samples as regards mineralization nodules. Lower biofilm formation of S.a and P.a. was observed on the Nb samples, with statistical differences from the other samples.
Our results suggest that the basic elements present in the alloy have osteoinductive characteristics, and Zr has a good influence on bone marrow stromal cell differentiation. We also believe that Nb has the best potential for reducing the formation of microbial biofilms.
一种有效的骨替代生物材料应具备避免细菌污染、促进骨形成并同时诱导细胞快速分化的特性。由于骨髓干细胞已知具有向成骨细胞谱系分化的潜力，目前正在对其进行研究。这使得这些细胞成为基于干细胞治疗的一个良好选择。我们旨在体外分析纯钛（Ti）、Ti-35Nb-7Zr合金（A）、铌（Nb）和氧化锆（Zr）对金黄色葡萄球菌（S.a）和铜绿假单胞菌（P.a）等微生物的抗性。此外，我们的目标是评估Ti-35Nb-7Zr合金的基本元素是否对骨髓基质细胞（干细胞来源）有任何影响，并观察这些金属是否具有诱导细胞分化为成骨细胞的特性。
从小鼠股骨中获取骨髓基质细胞（BMSC），并在不含地塞米松的成骨培养基中培养，作为细胞分化的外部来源。将样本分为Ti-35Nb-7Zr合金（A）、纯钛（Ti）、Nb（铌）和Zr（氧化锆），并通过扫描电子显微镜（SEM）和能量色散X射线光谱（EDS）进行表征。在预定时间段后，进行细胞相互作用、细胞毒性、增殖和细胞分化测试。对于单型生物膜形成，将含有金黄色葡萄球菌（S.a）和铜绿假单胞菌（P.a）的标准化悬浮液（10个细胞/ml）在样本上培养24小时，并进行MTT测试。
所有样本均呈现细胞增殖、生长和铺展。所有组的细胞活力均高于70%，但合金（A）表现出更好的结果，与Nb和Zr样本存在统计学差异。Zr表现出较高的碱性磷酸酶（ALP）活性，与其他组存在统计学差异（p < 0.05）。相比之下，样本之间在矿化结节方面未观察到统计学差异。在Nb样本上观察到S.a和P.a的生物膜形成较少，与其他样本存在统计学差异。
我们的结果表明，合金中存在的基本元素具有骨诱导特性，并且Zr对骨髓基质细胞分化有良好影响。我们还认为Nb具有减少微生物生物膜形成的最佳潜力。
