Influence of Culture Period on Osteoblast Differentiation of Tissue-Engineered Bone Constructed by Apatite-Fiber Scaffolds Using Radial-Flow Bioreactor

Abstract

With the limitation of autografts, the development of alternative treatments for bone diseases to alleviate autograft-related complications is highly demanded. In this study, a tissue-engineered bone was formed by culturing rat bone marrow cells (RBMCs) onto porous apatite-fiber scaffolds (AFSs) with three-dimensional (3D) interconnected pores using a radial-flow bioreactor (RFB). Using the optimized flow rate, the effect of different culturing periods on the development of tissue-engineered bone was investigated. The 3D cell culture using RFB was performed for 0, 1 or 2 weeks in a standard medium followed by 0, 1 or 2 weeks in a differentiation medium. Osteoblast differentiation in the tissue-engineered bone was examined by alkaline phosphatase (ALP) and osteocalcin (OC) assays. Furthermore, the tissue-engineered bone was histologically examined by hematoxylin and eosin and alizarin red S stains. We found that the ALP activity and OC content of calcified cells tended to increase with the culture period, and the differentiation of tissue-engineered bone could be controlled by varying the culture period. In addition, the employment of RFB and AFSs provided a favorable 3D environment for cell growth and differentiation. Overall, these results provide valuable insights into the design of tissue-engineered bone for clinical applications.

Nature-Inspired Effects of Naturally Occurring Trace Element-Doped Hydroxyapatite Combined with Surface Interactions of Mineral-Apatite Single Crystals on Human Fibroblast Behavior

“…Although similar investigations were conducted for biomaterials such as titanium [ 49 , 50 ]; texturized hydroxyapatite ceramics [ 14 ]; lithium-doped hydroxyapatites [ 51 ]; fluorapatite/glucan composites [ 52 ]; and rutile surfaces [ 53 ] using osteoblasts, preosteoblasts, and hepatocytes, to the best of our knowledge, the relation between the specific atomic structure of the surface and the normal human fibroblast biological performance is presented for the first time in our study. Despite recent advances in knowledge, few articles concern complex issues of potential HAp application in tissue engineering [ 54 ]. The presented interactions between human cells and apatite-crystal surfaces revealed the aspects of molecular mechanisms that may improve the process of evaluation of apatite bioavailability and advance the designing of biomaterials more suitable for medical interventions in tissue regeneration and controlled drug release [ 55 ].…”

Integrating physicomechanical and biological strategies for BTE: biomaterials-induced osteogenic differentiation of MSCs

“…Oscillatory perfusion helps to distribute the cells more uniformly within the scaffold 147 . According to recent studies, MSCs were cultured in osteogenic induction medium supplemented with dexamethasone, β-glycerophosphate and ascorbic acid 2-phosphate to further enhance osteogenic differentiation 149 . However, there are certain differences in the appropriate perfusion velocity and culture time required by different devices and scaffolds.…”

“…MSCs produced the highest angiogenic markers at 7 days under perfusion culture at a flow rate of 3 mL/min, but the DNA content and osteogenic differentiation markers stabilized at 14 days 168 . Suzuki K et al used a different culture protocol, using standard medium for one week and then osteogenic differentiation medium for one week 149 . The highest ALP activity was observed at this time point, however, the activity of ALP decreased at 2 weeks of osteogenic differentiation medium culture.…”