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

Kitaru Suzuki,Michiyo Honda,Mamoru Aizawa,Jun Fukasawa,Maiko Miura,Poon Nian Lim,Tomokazu Matsuura

doi:10.3390/ijms222313080

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.

Highlights

In orthopedic surgery, autologous bone graft is the gold standard treatment for repairing bone tissue damage [1]
The X-ray diffraction (XRD) patterns indicated that apatite fibers (AFs) were single-phase HAp (Figure 1a)
The XRD patterns of AFs showed a strong 300 reflection peak compared to those of HAp (HAp listed in ICDD card #9-432)

Summary

Introduction

Autologous bone graft is the gold standard treatment for repairing bone tissue damage [1]. This approach involves osteoinduction, which is important for bone defect treatment [2]. Autografts are the preferred technique in clinical sites, two major autografting-associated problems remain to be addressed: (i) Limited amount of the grafted bone and (ii) secondary invasion of the healthy bone tissue [1]. Allografts can solve both of these problems. These limitations lead to a strong impetus for developing alternative treatments for bone regeneration

Objectives

Methods

Results