Department of Biomedical Materials Science, School of Dentistry, University of Mississippi Medical Center, Jackson, MS 39216, United States.
Dent Mater. 2013 Feb;29(2):211-20. doi: 10.1016/j.dental.2012.10.003. Epub 2012 Nov 3.
Collagen and elastin are two key structural proteins found in the extra-cellular matrices (ECMs) of most tissues, yet very little is known about the response of bone cells to elastin or its derivatives. Recently, we have designed and characterized a novel class of ECM-based composite scaffolds with collagen and a genetically engineered polymer, elastin-like polypeptide (ELP) and subsequently showed their superior mechanical properties and drug release characteristics compared to collagen scaffolds. The objective of this research was to evaluate osteoblast growth and expression on these composite scaffolds.
A thorough biochemical and morphological characterization was performed on MC3T3-E1 pre-osteoblast cells cultured on collagen and ELP-collagen scaffolds. Cell viability was assessed using a live/dead assay. Total DNA content of all cells present on various surfaces was quantified. Pre-osteoblast differentiation was assessed by measuring the alkaline phosphatase and osteocalcin production. Mineral deposition by the cultured cells was visualized using the Von Kossa stain.
Our results showed that the ELP-collagen scaffolds were suitable substrates for cell culture that allowed MC3T3-E1 pre-osteoblast cell attachment, differentiation, and subsequent mineralization over a period of 3 weeks. The ELP-collagen scaffolds displayed equivalent biocompatibility and cell-interacting properties to those of the neat collagen scaffolds.
The novel ELP-collagen composite material may have future implications as a scaffold material for bone tissue engineering applications, for example, the treatment of alveolar bone loss.
胶原蛋白和弹性蛋白是存在于大多数组织细胞外基质(ECM)中的两种关键结构蛋白,但人们对骨细胞对弹性蛋白或其衍生物的反应知之甚少。最近,我们设计并表征了一类新型的基于细胞外基质的复合支架,其中包含胶原蛋白和一种基因工程聚合物,即弹性蛋白样多肽(ELP),并随后显示出它们与胶原蛋白支架相比具有更好的机械性能和药物释放特性。本研究的目的是评估成骨细胞在这些复合支架上的生长和表达。
对在胶原蛋白和 ELP-胶原蛋白支架上培养的 MC3T3-E1 前成骨细胞进行了全面的生化和形态学表征。使用活/死检测评估细胞活力。通过定量分析各种表面上所有细胞的总 DNA 含量来评估前成骨细胞的分化。通过测量碱性磷酸酶和骨钙素的产生来评估矿化细胞的分化。使用 Von Kossa 染色可视化培养细胞的矿化沉积。
我们的结果表明,ELP-胶原蛋白支架是适合细胞培养的基质,允许 MC3T3-E1 前成骨细胞附着、分化,并在 3 周的时间内随后矿化。ELP-胶原蛋白支架表现出与纯胶原蛋白支架相当的生物相容性和细胞相互作用特性。
新型 ELP-胶原蛋白复合材料可能在骨组织工程应用中作为支架材料具有未来意义,例如治疗牙槽骨丢失。