Tissue-specific bioactivity of soluble tendon-derived and cartilage-derived extracellular matrices on adult mesenchymal stem cells

Abstract

BackgroundBiological scaffolds composed of tissue-derived extracellular matrix (ECM) can promote homologous (i.e., tissue-specific) cell differentiation through preservation of biophysical and biochemical motifs found in native tissues. Solubilized ECMs derived from decellularized tendon and cartilage have recently been promoted as tissue-specific biomaterials, but whether tissue-specific bioactivity is preserved following solubilization is unknown. This study explored the tissue-specific bioactivity of soluble decellularized tendon and cartilage ECMs on human bone marrow-derived mesenchymal stem cells (MSCs) presented across different culture microenvironments, including two-dimensional (2D) tissue culture plastic, aligned electrospun nanofibers, cell pellets, and cell-seeded photocrosslinkable hydrogels.MethodsTendon and cartilage ECMs were decellularized using established methods and solubilized either via pepsin digestion or urea extraction. The effect of soluble ECMs on cell proliferation and differentiation was initially explored by supplementing basal medium of human MSCs cultured on 2D tissue culture plastic. In subsequent experiments, MSCs were cultured on aligned electrospun nanofibers, ascell pellets, or encapsulated within photocrosslinkable methacrylated gelatin (GelMA) hydrogels. Urea-extracted tendon and cartilage ECMs were added as supplements.ResultsPepsin-digested ECMs did not promote homologous differentiation in human MSCs, whether provided as a medium supplement or three-dimensional (3D) hydrogels. In contrast, urea-extracted ECMs tended to promote tissue-specific differentiation of MSCs cultured in 2D and 3D microenvironments. The application of the small molecule TGF-β signaling inhibitor SB-431542 largely negated the tissue-specific gene expression patterns mediated by tendon and cartilage ECMs. This suggests that the action of endogenous TGF-β was required, but was not sufficient, to impart tissue-specific bioactivity of urea-extracted ECMs. When urea-extracted cartilage ECM was incorporated within a photocurable GelMA hydrogel it independently enhanced chondrogenesis in encapsulated MSCs, and showed additive prochondrogenesis upon TGF-β supplementation in the medium.ConclusionsUrea-extracted ECM fractions of decellularized tendon and cartilage are soluble supplements capable of enhancing tissue-specific differentiation of adult stem cells.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0580-8) contains supplementary material, which is available to authorized users.

Meniscus ECM‐functionalised hydrogels containing infrapatellar fat pad‐derived stem cells for bioprinting of regionally defined meniscal tissue

“…Together, these results demonstrate that different ECMs can be combined with specific GFs to differentially regulate the phenotype of FPSCs for region‐specific meniscus tissue engineering. This is in agreement with recent studies that demonstrated cartilage ECM incorporated into a GelMA hydrogel‐enhanced chondrogenesis of encapsulated MSCs and showed additive pro‐chondrogenesis upon additional TGF‐β supplementation (Rothrauff et al, ). This may facilitate the development of biological implants with a superior capacity to promote meniscal regeneration than constructs that rely on GF stimulation alone (Lee et al, ).…”

“…It has been demonstrated that the inner meniscus ECM also contains higher level of proteoglycans and GFs such as TGF2 and TGF3, whereas the outer meniscus ECM is rich in bFGF and insulin (Rothrauff et al, ). Endogenous GFs within ECMs are believed to play a key role in determining their bioactivity (Rothrauff, Yang, & Tuan, ). Further work is required to completely characterise the composition of different regions of the meniscus ECM.…”

Do There Exist Innovative Perspectives in Tissue Engineering-Based Ligament Reconstruction?

“…6,10,[12][13][14][15] Aligned, interwoven and multilayered composite structures most likely reflect the natural aligned ligament ultrastructure and biomechanics. 10 Subsequently, an extensive biomaterial testing is necessary starting with the pure material to evaluate crucial parameters such as hydrophobicity/hydrophilicity, degradation, permeability, porosity, interconnectivity, surface texture and biomechanics. Later, in combination with a model cell line and then, with the respective ligamentogenic primary cell types, cellular parameters should be assessed such as cytocompatibility, cell adherence, distribution, morphology, cytoskeletal architecture, gene and protein expression profiles including established ligament and fibroblast markers such as scleraxis, tenomodulin and tenascin C. 16 Unfortunately, many synthetic polymers such as PLA or natural polymers such as silk with promising mechanical properties allow only limited cell adherence and growth.…”

“…8,9 In addition, the ECMs promote differentiation of precursor cells. 10,11 Scaffolds for ligament tissue engineering can be produced by various techniques such as braiding, embroidering, 3D printing, electrospinning, electro hydrodynamic jet printing and combinations of them. 6,10,[12][13][14][15] Aligned, interwoven and multilayered composite structures most likely reflect the natural aligned ligament ultrastructure and biomechanics.…”

“…10,11 Scaffolds for ligament tissue engineering can be produced by various techniques such as braiding, embroidering, 3D printing, electrospinning, electro hydrodynamic jet printing and combinations of them. 6,10,[12][13][14][15] Aligned, interwoven and multilayered composite structures most likely reflect the natural aligned ligament ultrastructure and biomechanics. 10 Subsequently, an extensive biomaterial testing is necessary starting with the pure material to evaluate crucial parameters such as hydrophobicity/hydrophilicity, degradation, permeability, porosity, interconnectivity, surface texture and biomechanics.…”