Ligament-Derived Matrix Stimulates a Ligamentous Phenotype in Human Adipose-Derived Stem Cells

Abstract

Human adipose stem cells (hASCs) can differentiate into a variety of phenotypes. Native extracellular matrix (e.g., demineralized bone matrix or small intestinal submucosa) can influence the growth and differentiation of stem cells. The hypothesis of this study was that a novel ligament-derived matrix (LDM) would enhance expression of a ligamentous phenotype in hASCs compared to collagen gel alone. LDM prepared using phosphate-buffered saline or 0.1% peracetic acid was mixed with collagen gel (COL) and was evaluated for its ability to induce proliferation, differentiation, and extracellular matrix synthesis in hASCs over 28 days in culture at different seeding densities (0, 0.25Â10 6 , 1Â106 , or 2Â10 6 hASC/mL). Biochemical and gene expression data were analyzed using analysis of variance. Fisher's least significant difference test was used to determine differences between treatments following analysis of variance. hASCs in either LDM or COL demonstrated changes in gene expression consistent with ligament development. hASCs cultured with LDM demonstrated more dsDNA content, sulfated-glycosaminoglycan accumulation, and type I and III collagen synthesis, and released more sulfated-glycosaminoglycan and collagen into the medium compared to hASCs in COL ( p 0.05). Increased seeding density increased DNA content incrementally over 28 days in culture for LDM but not COL constructs ( p 0.05). These findings suggest that LDM can stimulate a ligament phenotype by hASCs, and may provide a novel scaffold material for ligament engineering applications.

Tendon-Derived Extracellular Matrix Enhances Transforming Growth Factor-β3-Induced Tenogenic Differentiation of Human Adipose-Derived Stem Cells

“…Our findings indicate that the regulatory effects of tECM on tendon cell behavior also apply to ASCs. Consistent with our results, Little et al 42 found increased proliferation and partial adoption of tendon phenotype by ASCs seeded on acellular tendon/ligament matrix. The tECM contains not only collagen but also a number of Most fibers in the aligned scaffold were oriented at between 80°and 100°with respect to cross axis.…”

2010 Nicolas Andry Award: Multipotent Adult Stem Cells from Adipose Tissue for Musculoskeletal Tissue Engineering

“…Our findings and those of others have shown that ASCs exhibit multipotent differentiation capabilities in the mesenchymal lineage, similar to other adult stem cells such as MSCs, with evidence of adipogenic, chondrogenic, myogenic, neurogenic, osteogenic, and tenogenic differentiation [36,53,86]. Similarly, the culture conditions used to induce lineage-specific differentiation of ASCs generally are similar to those used for MSCs; although increasing evidence indicates that different adult stem cells possess unique characteristics and properties depending on their site of origin.…”

Genipin-Crosslinked Cartilage-Derived Matrix as a Scaffold for Human Adipose-Derived Stem Cell Chondrogenesis

“…48 ECM proteins are generally well conserved among different species and, as such, may provide the opportunity for use as functional scaffolds for cell attachment, proliferation, and differentiation. 26,27,29,36,49 We previously showed that CDM, a porous scaffold derived from articular cartilage, is capable of inducing chondrogenic differentiation of ASCs without exogenous growth factors. 28 By crosslinking the CDM in a genipin solution (0.5%, 0.05% and 0.005%) at 37°C for 3 days, constructs were produced that were microscopically unique relative to noncrosslinked CDM, though similar to the unmodified matrix, genipin-crosslinked materials were highly porous (94.7%-95.9%).…”