The Institute of Oncology "Prof. Dr. I. Chiricuta" Cluj-Napoca, Cluj-Napoca, Romania ; University of Medicine and Pharmacy "Iuliu Hatieganu" Cluj-Napoca, Cluj-Napoca, Romania.
The Institute of Oncology "Prof. Dr. I. Chiricuta" Cluj-Napoca, Cluj-Napoca, Romania.
J Biol Eng. 2014 Jun 19;8:14. doi: 10.1186/1754-1611-8-14. eCollection 2014.
The development of novel biomaterials able to control cell activities and direct their fate is warranted for engineering functional bone tissues. Adding bioactive materials can improve new bone formation and better osseointegration. Three types of titanium (Ti) implants were tested for in vitro biocompatibility in this comparative study: Ti6Al7Nb implants with 25% total porosity used as controls, implants infiltrated using a sol-gel method with hydroxyapatite (Ti HA) and silicatitanate (Ti SiO2). The behavior of human osteoblasts was observed in terms of adhesion, cell growth and differentiation.
The two coating methods have provided different morphological and chemical properties (SEM and EDX analysis). Cell attachment in the first hour was slower on the Ti HA scaffolds when compared to Ti SiO2 and porous uncoated Ti implants. The Alamar blue test and the assessment of total protein content uncovered a peak of metabolic activity at day 8-9 with an advantage for Ti SiO2 implants. Osteoblast differentiation and de novo mineralization, evaluated by osteopontin (OP) expression (ELISA and immnocytochemistry), alkaline phosphatase (ALP) activity, calcium deposition (alizarin red), collagen synthesis (SIRCOL test and immnocytochemical staining) and osteocalcin (OC) expression, highlighted the higher osteoconductive ability of Ti HA implants. Higher soluble collagen levels were found for cells cultured in simple osteogenic differentiation medium on control Ti and Ti SiO2 implants. Osteocalcin (OC), a marker of terminal osteoblastic differentiation, was most strongly expressed in osteoblasts cultivated on Ti SiO2 implants.
The behavior of osteoblasts depends on the type of implant and culture conditions. Ti SiO2 scaffolds sustain osteoblast adhesion and promote differentiation with increased collagen and non-collagenic proteins (OP and OC) production. Ti HA implants have a lower ability to induce cell adhesion and proliferation but an increased capacity to induce early mineralization. Addition of growth factors BMP-2 and TGFβ1 in differentiation medium did not improve the mineralization process. Both types of infiltrates have their advantages and limitations, which can be exploited depending on local conditions of bone lesions that have to be repaired. These limitations can also be offset through methods of functionalization with biomolecules involved in osteogenesis.
为了构建功能性骨组织,需要开发能够控制细胞活性并指导其命运的新型生物材料。添加生物活性材料可以促进新骨形成和更好的骨整合。在这项比较研究中,测试了三种类型的钛(Ti)植入物的体外生物相容性:作为对照的总孔隙率为 25%的 Ti6Al7Nb 植入物、使用溶胶-凝胶法渗透羟基磷灰石(TiHA)和硅钛酸盐(TiSiO2)的植入物。观察了人类成骨细胞的粘附、细胞生长和分化行为。
两种涂层方法提供了不同的形态和化学性质(SEM 和 EDX 分析)。与 TiSiO2 和多孔未涂层 Ti 植入物相比,TiHA 支架上的细胞在最初 1 小时的附着较慢。Alamar 蓝试验和总蛋白含量评估显示,代谢活性在第 8-9 天达到峰值,TiSiO2 植入物具有优势。通过骨桥蛋白(OP)表达(ELISA 和免疫细胞化学)、碱性磷酸酶(ALP)活性、钙沉积(茜素红)、胶原蛋白合成(SIRCOL 试验和免疫细胞化学染色)和骨钙素(OC)表达评估成骨细胞分化和新矿化,突出了 TiHA 植入物更高的骨诱导能力。在简单成骨分化培养基中培养的细胞中发现了更高水平的可溶性胶原蛋白,用于对照 Ti 和 TiSiO2 植入物。成骨细胞分化的终末标志物骨钙素(OC)在 TiSiO2 植入物上培养的成骨细胞中表达最强。
成骨细胞的行为取决于植入物的类型和培养条件。TiSiO2 支架支持成骨细胞的粘附,并通过增加胶原蛋白和非胶原蛋白(OP 和 OC)的产生促进分化。TiHA 植入物具有较低的诱导细胞粘附和增殖的能力,但具有增加诱导早期矿化的能力。在分化培养基中添加生长因子 BMP-2 和 TGFβ1 并没有改善矿化过程。两种渗透物都有其优点和局限性,可以根据要修复的骨损伤的局部情况加以利用。这些局限性也可以通过功能化与成骨相关的生物分子来抵消。