Gierloff M, Nitsche T, Adam-Klages S, Liebs K, Hedderich J, Gassling V, Wiltfang J, Kabelitz D, Açil Y
Department of Oral and Maxillofacial Surgery, Christian-Albrechts-University Kiel, Arnold-Heller-Strasse 16, 24105, Kiel, Germany,
Clin Oral Investig. 2014 Jan;18(1):247-59. doi: 10.1007/s00784-013-0956-9. Epub 2013 Mar 6.
Injectable or implantable scaffolds seeded with autologous chondrogenic cells may represent a promising option for treatment of cartilage defects in the future. Current problems with the autologous chondrocyte implantation including dedifferentiation and the development of fibrocartilage suggest the use of alternative chondrogenic cell sources such as mesenchymal stromal cells (MSCs). The aim of this study was to compare the early effects of different scaffolds on the proliferation and metabolic activity of chondrogenic MSCs in vitro.
Multipotent stromal cells were isolated from rat bone marrow, phenotyped by flow cytometry, and differentiated into distinct lineages proved by lineage-specific staining and gene expression (RT-PCR) pattern. Cell proliferation on Tutodent® Membrane, Bio-Gide®, TissuFleece E, and Belotero® Soft was quantified by the MTT and WST-1 assay and direct determination of total cell numbers. Potential cytotoxic effects of eluates obtained from the materials were quantified by lactate dehydrogenase (LDH) and 5-bromo-2-deoxyuridine (BrdU) assay.
TissuFleece E displayed the best results regarding cell proliferation on the biomaterials and metabolic activity (MTT, WST-1) (p < 0.001). Yet, the eluates of TissuFleece E caused an increased LDH release and lower values in the BrdU test. Cell proliferations on Bio-Gide®, Tutodent® Membrane, and Belotero® Soft were similar to the control. The eluates of Belotero® Soft exhibited the highest LDH release and lowest values in the BrdU assay (p < 0.05).
Our results support the use of Tissufleece E as scaffold for chondrogenic rat MSCs. However, it should be prewashed with culture medium before seeding of the cells.
Tissufleece E may serve as a promising carrier material for chondrogenic MSCs for cartilage tissue engineering attempts.
接种自体软骨生成细胞的可注射或可植入支架可能是未来治疗软骨缺损的一个有前景的选择。目前自体软骨细胞植入存在的问题,包括去分化和纤维软骨的形成,提示可使用间充质基质细胞(MSC)等替代软骨生成细胞来源。本研究的目的是比较不同支架在体外对软骨生成MSC增殖和代谢活性的早期影响。
从大鼠骨髓中分离多能基质细胞,通过流式细胞术进行表型分析,并通过谱系特异性染色和基因表达(RT-PCR)模式证明其分化为不同谱系。通过MTT和WST-1测定以及直接测定总细胞数来量化Tutodent®膜、Bio-Gide®、TissuFleece E和Belotero® Soft上的细胞增殖。通过乳酸脱氢酶(LDH)和5-溴-2-脱氧尿苷(BrdU)测定来量化从材料中获得的洗脱液的潜在细胞毒性作用。
TissuFleece E在生物材料上的细胞增殖和代谢活性(MTT、WST-1)方面显示出最佳结果(p < 0.001)。然而,TissuFleece E的洗脱液导致LDH释放增加,并且在BrdU试验中值较低。Bio-Gide®、Tutodent®膜和Belotero® Soft上的细胞增殖与对照组相似。Belotero® Soft的洗脱液在BrdU试验中显示出最高的LDH释放和最低的值(p < 0.05)。
我们的结果支持使用Tissufleece E作为大鼠软骨生成MSC的支架。然而,在接种细胞之前,应用培养基对其进行预洗。
TissuFleece E可能作为软骨生成MSC用于软骨组织工程尝试的一种有前景的载体材料。