IntroductionMesenchymal stromal cells (MSCs) are multipotent cells that can differentiate to adipogenic, osteogenic, and chondrogenic lineages (Pittenger et al., 1999). MSCs play critical roles in coordinating tissue regeneration through their versatile differentiation capacity and immunomodulatory effects. Human MSCs can readily be isolated from different body sites (i.e. bone marrow, adipose tissue) and easily be cultured in vitro (Pittenger et al., 1999; Zuket al., 2002), making them an attractive cell source for biomedical research. Identifying the lineage commitment potential of MSCs is necessary to establish a deeper understanding of their therapeutic potential. Depending on culture conditions and their tissue of origin, MSCs have variable potency and molecular properties, including a global gene expression profile (Lee et al., 2004;Picchi et al., 2013;Wang et al., 2016).Because of its validity and sensitivity, quantitative polymerase chain reaction (qPCR) has been a technique of choice in assessing gene expression of MSCs derived from distinct body sites (Picchi et al., 2013;Wang et al., 2016). The reliability of qPCR results (i.e. target gene expression) can only be judged when optimum conditions are used. Using an accurate reference gene (RG) with a stable expression level is compulsory for normalizing target gene expression. Mostly genes that function in cellular integrity (e.g., beta-actin. ACTB) or metabolism (e.g., glyceraldehyde 3-phosphate dehydrogenase, GAPDH) are used as RGs. However, these frequently used genes have been reported to have variable expression levels under different experimental procedures that make them unsuitable to be used for normalization. Several studies showed that ACTB and GAPDH are improper RGs in MSC expansion and differentiation (
Polyhydroxybutyrate (PHB) is a polymer used to restore tissues or regenerate bones. However, its compatibility with human bone marrow-derived mesenchymal stem cells (MSCs) has not been examined. PHB membranes of random (r-PHB) or aligned (a-PHB) electrospun nanofibers that generate bone and spinal axon scaffolds were combined with human MSCs. The adhesion and proliferation of cells on these membranes were examined. The orientation of cells on PHB membranes was analyzed by confocal microscopy and scanning electron microscopy (SEM). The MSCs maintained their characteristic properties on the membranes, adhered to the membranes, and preserved their viability. The cell morphology was different when they were grown on differentially designed matrices. Cells expanded on the a-PHB membrane and showed fibroid-like morphology. Conversely, cells interacting with the r-PHB membrane were located homogeneously and demonstrated polygonal morphology. The adhesion and proliferation of human MSCs was higher on the a-PHB membrane than on the randomly oriented one. Fiber orientation influenced the phenotype and biological behavior of human MSCs. This property may be useful in the selection of specifically designed scaffolds for the desired tasks. The results of this preliminary study indicated that PHB membranes designed for bone or nerve tissue engineering are compatible with human MSCs.
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