Suzuki Kitaru, Nagata Kohei, Yokota Tomohiro, Honda Michiyo, Aizawa Mamoru
Department of Applied Chemistry, School of Science and Technology, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan.
Biomed Mater Eng. 2017;28(1):57-64. doi: 10.3233/BME-171656.
There is a strong impetus for the development of alternative treatments for bone disease that avoid the complications associated with autografts and allografts. To address this, we previously developed porous apatite-fiber scaffolds (AFSs) which have three-dimensional interconnected pores, and constructed tissue-engineered bone by culturing rat bone marrow cells (RBMCs) using AFSs in a radial-flow bioreactor (RFB).
To generate additional baseline data for the development of tissue-engineered bone constructed for clinical application using a RFB, we cultured RBMCs using AFSs under static conditions (hereafter, RBMC AFS culture), and monitored RBMC growth and differentiation characteristics in vitro, and two weeks after subcutaneous inoculation into recipient rats.
RBMCs were seeded to AFSs and growth, differentiation and calcification were monitored in vitro and in vivo by histological evaluation using hematoxylin eosin, alkaline phosphatase and alizarin red S stains.
RBMCs in/on AFSs proliferated and differentiated normally in vitro and in vivo, and calcification was evident two weeks after subcutaneous AFS culture implantation. Histological assays revealed that AFSs and RBMC AFS cultures were biocompatible, and did not induce inflammation or immunological rejection in vivo.
These findings suggest that AFSs are a conducive microenvironment for bone regeneration and are well tolerated in vivo. The results provide valuable baseline data for the design of implant studies using tissue-engineered bone constructed by RFB.
开发用于骨疾病的替代治疗方法的需求迫切,这类方法需避免自体移植和异体移植相关的并发症。为解决这一问题,我们之前开发了具有三维相互连通孔隙的多孔磷灰石纤维支架(AFS),并通过在径向流生物反应器(RFB)中使用AFS培养大鼠骨髓细胞(RBMC)构建了组织工程骨。
为使用RFB构建用于临床应用的组织工程骨的开发生成更多基线数据,我们在静态条件下使用AFS培养RBMC(以下简称RBMC AFS培养),并在体外以及皮下接种到受体大鼠两周后监测RBMC的生长和分化特征。
将RBMC接种到AFS上,通过苏木精伊红、碱性磷酸酶和茜素红S染色进行组织学评估,在体外和体内监测其生长、分化和钙化情况。
AFS内/上的RBMC在体外和体内均正常增殖和分化,皮下植入AFS培养物两周后钙化明显。组织学分析显示AFS和RBMC AFS培养物具有生物相容性,在体内不会引起炎症或免疫排斥。
这些发现表明AFS是骨再生的有利微环境,在体内耐受性良好。这些结果为使用RFB构建的组织工程骨进行植入研究的设计提供了有价值的基线数据。