Nanoscaled and microscaled parallel topography promotes tenogenic differentiation of ASC and neotendon formation in vitro

Abstract

BackgroundTopography at different scales plays an important role in directing mesenchymal stem cell differentiation including adipose-derived stem cells (ASCs) and the differential effect remains to be investigated.PurposeThis study aimed to investigate the similarity and difference between micro- and nanoscaled aligned topography for inducing tenogenic differentiation of human ASCs (hASCs).MethodsParallel microgrooved PDMS membrane and a parallel aligned electrospun nanofibers of gelatin/poly-ε-caprolactone mixture were employed as the models for the study.ResultsAligned topographies of both microscales and nanoscales could induce an elongated cell shape with parallel alignment, as supported by quantitative cell morphology analysis (cell area, cell body aspect, and cell body major axis angle). qPCR analysis also demonstrated that the aligned topography at both scales could induce the gene expressions of various tenogenic markers at the 7th day of in vitro culture including tenomodulin, collagen I and collagen VI, decorin, tenascin-C and biglycan, but with upregulated expression of scleraxis and tenascin-C only in microscaled topography. Additionally, tenogenic differentiation at the 3rd day was confirmed only at microscale. Furthermore, microscaled topography was confirmed for its tenogenic induction at tissue level as neotendon tissue was formed with the evidence of mature type I collagen fibers only in parallel aligned polyglycolic acid (PGA) microfibers after in vitro culture with mouse ASCs. Instead, only fat tissue was formed in random patterned PGA microfibers.ConclusionBoth microscaled and nanoscaled aligned topographies could induce tenogenic differentiation of hASCs and micro-scaled topography seemed better able to induce elongated cell shape and stable tenogenic marker expression when compared to nanoscaled topography. The microscaled inductive effect was also confirmed at tissue level by neotendon formation in vitro.

Macromechanics and polycaprolactone fiber organization drive macrophage polarization and regulate inflammatory activation of tendon in vitro and in vivo

“…Across this work, our findings were consistent with previous literature on mechanical and surface structure regulation of macrophages and tendon cells, individually [19,21,22,40]. Our work represents a knowledge advance in identifying how these individual responses augment and in some cases differ when these biophysical cues are applied in co-culture systems.…”

“…How biomaterial substrate mechanics and biochemistry interact to guide cell behavior has emerged as an important research topic with broad implications to biomaterial design [15][16][17]. Specifically for tendon repair, an aligned micro-or nanofiber structure is now understood to be a highly instructive biophysical cue that can direct tendon https://doi.org/10.1016/j.biomaterials.2020.120034 Received 30 March 2019; Received in revised form 4 April 2020; Accepted 5 April 2020 specific lineage differentiation [18][19][20][21][22][23][24][25]. Our own work has shown that deviations from aligned biomaterial surface structures may play a potentially important role in predisposing inflammatory response of attached tendon fibroblasts [26].…”

3D Bone Biomimetic Scaffolds for Basic and Translational Studies with Mesenchymal Stem Cells

“…To this end, the authors employed PCL electrospun fibers with three different types of alignment: random (REF), aligned (AEF), and mesh (MEF). In addition to the influence on cell morphology exerted by the way of alignment and likely due to topography-related contact cues directing cell behavior (as reported also by others [ 9 , 70 , 111 , 112 ]), they observed an impact on MSC secretory profile. Namely, the expression and release of anti-inflammatory and wound healing-promoting factors, including COX2, PGE2, iNOS, TSG6, and TGFβ, and of the proangiogenic factors VEGF, HGF, and bFGF, which were increased in 3D-cultured MSCs as compared to cells cultured in a monolayer and, in particular, were higher in cells on AEF and MEF, as compared to those on REF [ 110 ].…”

Nanofibrillar cellulose wound dressing supports the growth and characteristics of human mesenchymal stem/stromal cells without cell adhesion coatings

“…Differing properties of biomaterials, caused by different manufacturing processes, are well known to alter the function of cells [42][43][44][45]. In this study, three different NFC dressings varying in topography and amount of NFC were studied with multiple cell densities in order to identify the optimal culture conditions for hASCs.…”