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

Abstract

Mimicking the complex intricacies of the extra cellular matrix including 3D configurations and aligned fibrous structures were traditionally perused for producing cartilage tissue from stem cells. This study shows that human adipose derived mesenchymal stem cells (hADMSCs) establishes significant chondrogenic differentiation and may generate quality cartilage when cultured on 2D and randomly oriented fibrinogen/poly‐lactic acid nanofibers compared to 3D sandwich‐like environments. The adhering cells show well‐developed focal adhesion complexes and actin cytoskeleton arrangements confirming the proper cellular interaction with either random or aligned nanofibers. However, quantitative reverse transcription‐polymerase chain reaction analysis for Collagen 2 and Collagen 10 genes expression confirms favorable chondrogenic response of hADMSCs on random nanofibers and shows substantially higher efficacy of their differentiation in 2D configuration versus 3D constructs. These findings introduce a new direction for cartilage tissue engineering through providing a simple platform for the routine generation of transplantable stem cells derived articular cartilage replacement that might improve joint function.

Principles of Fibrinogen Fiber Assembly In Vitro

“…[3,120] Good biocompatibility of electrospun fibrinogen fibers was found with neonatal rat cardiac fibroblasts, [121] human bladder smooth muscle cells, [122] endothelial cells, [123] and human adipose derived mesenchymal stem cells. [124] However, a major disadvantage of electrospinning is the high protein concentration of more than 100 mg mL −1 required to produce nanofibrous fibrinogen scaffolds with reproducible diameters and porosity. [3,117,125] Acidic pH Conditions, Heating, and Protein Binding: Wei and co-workers also observed fiber assembly of fibrinogen on hydrophilic mica under acidic pH conditions.…”

Osteogenic differentiation of mesenchymal stem cells using hybrid nanofibers with different configurations and dimensionality

“…In this respect, our sandwich-like construct provides ADMSCs with opportunity to receive stimuli from the third dimension, which enables comparison of their behavior in 2-D and 3-D environments. 18 It has to be mentioned that we already explored this construct for studies on the chondrogenic differentiation of ADMSCs 38 and surprisingly found that cells produces favorably cartilage in 2-D random samples versus their aligned counterparts, and also more than in 3-D conditions. 38 The same approach, but now applied for the osteogenic response, revealed a generally better ADMSCs differentiation in 3-D environment, while the effect of alignment was found only in respect to OSP gene expression.…”

“…18 It has to be mentioned that we already explored this construct for studies on the chondrogenic differentiation of ADMSCs 38 and surprisingly found that cells produces favorably cartilage in 2-D random samples versus their aligned counterparts, and also more than in 3-D conditions. 38 The same approach, but now applied for the osteogenic response, revealed a generally better ADMSCs differentiation in 3-D environment, while the effect of alignment was found only in respect to OSP gene expression. Similar studies with aim to emulate in vivo tissue structures used layer-by-layer approach to stack cell-seeded nanofiber meshes (aligned or random) into 3-D multilayered constructs.…”

Fabrication and properties of an injectable sodium alginate/PRP composite hydrogel as a potential cell carrier for cartilage repair

“…The ideal scaffold should have a uniform structure, excellent biocompatibility and biodegradability, as well as suitable mechanical strength, thus providing a specific environment for cell and tissue function in vitro and in vivo (Yang, Du, & Chua, ). So far, various types of materials such as biopolymer‐based hydrogels (Sá‐Lima, Caridade, Mano, & Reis, ) or nanofibers (Forget et al, ), synthetic polymer‐derived scaffolds (Izal et al, ) or hydrogels (Grant et al, ), and inorganic particle‐based nanocomposite hydrogels (Asadi et al, ) have been employed for cartilage regeneration. Particularly, synthetic, biodegradable polymers have been considered as useful scaffold materials, since their properties can be readily adjusted through the control of the polymerization reaction (Liu, Holzwarth, & Ma, ), but their biological efficiency as well as safety remain unsettled.…”

“…The ideal scaffold should have a uniform structure, excellent biocompatibility and biodegradability, as well as suitable mechanical strength, thus providing a specific environment for cell and tissue function in vitro and in vivo (Yang, Du, & Chua, 2001). So far, various types of materials such as biopolymer-based hydrogels (Sá-Lima, Caridade, Mano, & Reis, 2010) or nanofibers (Forget et al, 2016), synthetic polymer-derived scaffolds (Izal et al, 2013) or hydrogels (Grant et al, 2008), and inorganic particle-based nanocomposite hydrogels (Asadi et al, 2018) have been employed for cartilage regeneration.…”