Chondrogenic Differentiation of Human Mesenchymal Stem Cells on Oriented Nanofibrous Scaffolds: Engineering the Superficial Zone of Articular Cartilage

Abstract

Cell differentiation, adhesion, and orientation are known to influence the functionality of both natural and engineered tissues, such as articular cartilage. Several attempts have been devised to regulate these important cellular behaviors, including application of inexpensive but efficient electrospinning that can produce patterned extracellular matrix (ECM) features. Electrospun and oriented polycaprolactone (PCL) scaffolds (500 or 3000 nm fiber diameter) were created, and human mesenchymal stem cells (hMSCs) were cultured on these scaffolds. Cell viability, morphology, and orientation on the fibrous scaffolds were quantitatively determined as a function of time. While the fiber-guided initial cell orientation was maintained even after 5 weeks, cells cultured in the chondrogenic media proliferated and differentiated into the chondrogenic lineage, suggesting that cell orientation is controlled by the physical cues and minimally influenced by the soluble factors. Based on assessment by the chondrogenic markers, use of the nanofibrous scaffold (500 nm) appears to enhance the chondrogenic differentiation. These findings indicate that hMSCs seeded on a controllable PCL scaffold may lead to an alternate methodology to mimic the cell and ECM organization that is found, for example, in the superficial zone of articular cartilage.

Differentiation of Human Mesenchymal Stem Cells Toward Quality Cartilage Using Fibrinogen‐Based Nanofibers

“…Hence, our results are not in agreement with what the traditionally thought of in terms of having the aligned nanofi bers to be better suited than random nanofi bers for producing cartilage tissue in vitro. [ 13,19,39 ] Regarding the role of nanofi bers dimensionality, our PCR data unexpectedly show a signifi cantly higher cartilage specifi c gene activity in 2D environment versus the 3D environment, in agreement with the visual morphological inspection above for favored Collagen II and glycosaminoglycans deposition. We eliminated possible differences in medium or nutrition exposure of the two environments by measuring the activity of only middle part of construct.…”

“…[ 13,43 ] Typically, organized fi brils within a scaffold are thought to promote cell migration and thus probably contribute to improving cartilage regeneration. [ 13,39,44 ] Our previous studies with endothelial cells also support this view [ 27 ] contrary to typical observations. We generally found a better organized cartilage matrix on random nanofi bers representing more homogenous structure in both, 2D or 3D environment as can be seen on Figures 7 and 8 .…”

Polymeric Nanofibers in Tissue Engineering

“…Additionally, the nanofibrous scaffolds were found to be easily fixed to the surrounding tissue with sutures and did not require a periosteal covering, which reduced the morbidity associated with the procedure. 109 While researchers have found higher levels of chondrogenesis with nanofiber scaffolds compared to microfiber scaffolds, 110 some groups have found higher levels of chondrogenic gene expression in progenitor cells grown on microfiber scaffolds compared to nanofiber scaffolds. 61 This occurrence could be due to the larger pore sizes in microfiber constructs.…”

Nanotechnology Advances in Drug Delivery

“…Given the properties of biocompatibility and biodegradability of the nanoparticles, many studies now exist on the use of nanomaterials to prolong the expression of proteins for improved angiogenesis and recruitment of stem cells for tissue and organ regeneration (i.e., cardiac and vascular tissues [57,58]). Among the others, carbon nanotubes (CNTs), PEGylated multi walled carbon nanotubes (MWCNTs) [59], chitosan nanoparticles (CSNPs) [60], poly(lactic-co-glycolic acid) (PLGA) scaffolds [61], polycaprolactones (PCL) scaffolds [62], poly-L-lactic acid (PLLA) [63], polyethyleneimine (PEI) have been already explored [64].…”