Fibrin hydrogels functionalized with cartilage extracellular matrix and incorporating freshly isolated stromal cells as an injectable for cartilage regeneration

Abstract

An alternative to the use of in vitro expanded cells in regenerative medicine is the use of freshly isolated stromal cells, where a bioactive scaffold or hydrogel is used to provide an environment that enhances their proliferation and tissue-specific differentiation in vivo. The objective of this study was to develop an injectable fibrin hydrogel functionalized with cartilage ECM micro-particles and the growth factor TGF-β3 as a therapeutic for articular cartilage regeneration. This study demonstrates that freshly isolated stromal cells generate cartilage tissue in vivo when incorporated into such a fibrin hydrogels functionalized with cartilage ECM particles. These findings open up new possibilities for in-theatre, single-stage, cell-based therapies for joint regeneration.

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

“…cECM independently enhanced chondrogenesis at day 7, but the chondroinductive effect decreased by day 21. Rather, medium supplementation with TGF-β was required for sustained upregulation of chondrogenic markers, with corresponding deposition of cartilage ECM proteins; this finding agrees with related work [27, 28, 65]. Despite the apparent necessity of exogenous TGF-β for robust cartilage formation, cECM within the hydrogel interacted synergistically with the supplemented TGF-β, as demonstrated by the greatest increases in chondrogenic gene expression and sGAG deposition seen in the cECM + TGF-β group.…”

“…The inclusion of cartilage ECM structural proteins such as collagen type VI [62], collagen type II, and proteoglycans [63, 64] within MSC-seeded hydrogels has been found to enhance chondrogenic differentiation. Similarly, Almeida et al [28, 65] reported similar improvements when using decellularized cartilage ECM particles. In this study, cECM was mixed with photocrosslinkable GelMA hydrogels, a biomaterial which we previously found to support robust chondrogenesis [8].…”

Anatomical region-dependent enhancement of 3-dimensional chondrogenic differentiation of human mesenchymal stem cells by soluble meniscus extracellular matrix

“…Additionally, the inclusion of a water-soluble, visible light-responsive photoinitiatior (i.e., LAP) obviates concerns of possible cellular damage caused by UV light exposure, as required by most photoinitiators (e.g., Irgacure) [24]. While the effect of meniscal ECM supplementation on MSC-seeded GelMA hydrogels was previously unexplored, functionalization of GelMA with particulated (hyaline) cartilage ECM has been reported to enhance chondrogenesis of encapsulated MSCs [29, 30]. Although the particulated ECM was presumably chondroinductive due to retention of intrinsic chondrogenic cues, robust chondrogenesis required stimulation with exogenous TGF-β3.…”

Injectable hydrogels for cartilage and bone tissue engineering

“…128,129,130,131 Benavides et al 132 have applied fibrin-based hydrogels, together with PEG and human amniotic fluid-derived stem cells, to develop a novel injectable hydrogel system that is able to induce a fibrin-driven angiogenic host response and promote in situ amniotic fluid-derived stem cell-derived neovascularization. Almeida et al 133 have developed an injectable, cartilaginous ECM microparticle-functionalized fibrin-based hydrogel, which transforms growth factor transforming growth factor-β 3 into a putative therapeutic for articular cartilage regeneration. The capacity of the hydrogel to promote chondrogenes is of freshly isolated stromal cells in vivo suggests that the hydrogel can induce cartilage formation and has the potential for cartilage repair, and thus may have the potential to overcome several current challenges related to cartilage tissue engineering.…”