在灌注生物反应器中紧密多层纳米羟基磷灰石-聚己内酯电纺支架上的成骨分化与矿化
Osteogenic Differentiation and Mineralization on Compact Multilayer nHA-PCL Electrospun Scaffolds in a Perfusion Bioreactor.
作者信息
Yaghoobi Maliheh, Hashemi-Najafabadi Sameereh, Soleimani Masoud, Vasheghani-Farahani Ebrahim, Mousavi Seyyed Mohammad
机构信息
Biomedical Engineering Group, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran.
Hematology Group, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
出版信息
Iran J Biotechnol. 2016 Jun;14(2):41-49. doi: 10.15171/ijb.1382.
BACKGROUND
Monolayer electrospun scaffolds have already been used in bone tissue engineering due to their high surface-tovolume ratio, interconnectivity, similarity to natural bone extracellular matrix (ECM), and simple production.
OBJECTIVES
The aim of this study was to evaluate the dynamic culture effect on osteogenic differentiation and mineralizationi into a compact cellular multilayer nHA-PCL electrospun construct. The dynamic culture was compared with static culture.
MATERIALS AND METHODS
The calcium content, alkaline phosphatase (ALP) activity and cell viability were investigated on days 3 and 7.
RESULTS
When the dynamic culture compared to static culture, the mineralization and ALP activity were increased in dynamic culture. After 7 days, calcium contents were 41.24 and 20.44 μg.(cm), and also normalized ALP activity were 0.32 and 0.19 U.mg in dynamic and static culture, respectively. Despite decreasing the cell viability until day 7, the scanning electron microscopy (SEM) results showed that, due to higher mineralization, a larger area of the construct was covered with calcium deposition in dynamic culture.
CONCLUSIONS
The dynamic flow could improve ALP activity and mineralization into the compact cellular multilayer construct cultured in the perfusion bioreactor after 7 days. Fluid flow of media helped to facilitate the nutrients transportation into the construct and created uniform cellular construct with high mineralization. This construct can be applied for bone tissue engineering.
背景
单层电纺支架因其高比表面积、连通性、与天然骨细胞外基质(ECM)的相似性以及简单的生产工艺,已被用于骨组织工程。
目的
本研究旨在评估动态培养对致密细胞多层nHA-PCL电纺构建体中成骨分化和矿化的影响,并将动态培养与静态培养进行比较。
材料与方法
在第3天和第7天研究钙含量、碱性磷酸酶(ALP)活性和细胞活力。
结果
与静态培养相比,动态培养时矿化和ALP活性增加。7天后,动态培养和静态培养中的钙含量分别为41.24和20.44μg·(cm),归一化后的ALP活性分别为0.32和0.19U·mg。尽管到第7天时细胞活力有所下降,但扫描电子显微镜(SEM)结果显示,由于矿化程度更高,动态培养中构建体的更大面积被钙沉积覆盖。
结论
动态流动可在7天后提高灌注生物反应器中培养的致密细胞多层构建体的ALP活性和矿化程度。培养基的流体流动有助于促进营养物质向构建体的运输,并形成具有高矿化程度的均匀细胞构建体。这种构建体可应用于骨组织工程。