In-situ forming injectable GFOGER-conjugated BMSCs-laden hydrogels for osteochondral regeneration

Abstract

The collagen-mimetic peptide GFOGER possesses the chondrogenic potential and has been used as a cell adhesion peptide or chondrogenic inducer. Here, we prepared an injectable in situ forming composite hydrogel system comprising methoxy polyethylene glycol-b-polycaprolactone (MPEG-PCL) and GFOGER-conjugated PEG-PCL (GFOGER-PEG-PCL) with various GFOGER concentrations based on our recently patented technology. The conjugation of GFOGER to PEG-PCL was confirmed by 1H NMR, and the particle size distribution and rheological properties for the sol-gel transition behavior of the samples with respect to the GFOGER content were evaluated systemically. In vitro experiments using rat bone marrow-derived mesenchymal stem cells (BMSCs) revealed that the GFOGER-PEG-PCL hydrogel significantly enhanced expression of integrins (β1, α2, and α11), increased expression of FAK, and induced downstream signaling of ERK and p38. Overexpression of chondrogenic markers suggested that BMSCs have the potential to differentiate into chondrogenic lineages within GFOGER-PEG-PCL samples. In vivo studies using a rat osteochondral defect model revealed that transplanted BMSCs with GFOGER0.8-PEG-PCL survived at the defect with strong chondrogenic expression after 4 weeks. The stem cell-laden GFOGER0.8-PEG-PCL hydrogel produced remarkable osteochondral regeneration at 8 weeks of transplantation, as determined by histological findings and micro-CT analysis. The histomorphological score in the GFOGER0.8-PEG-PCL + BMSCs group was ~1.7-, 2.6-, and 5.3-fold higher than that in the GFOGER0.8-PEG-PCL, MPEG-PCL, and defect groups, respectively. Taken together, these results provide an important platform for further advanced GFOGER-based stem cell research for osteochondral repair.

Three‐dimensional printed biomimetic multilayer scaffolds coordinated with sleep‐related small extracellular vesicles: A strategy for extracellular matrix homeostasis and macrophage polarization to enhance osteochondral regeneration

“…MSCs are capable of differentiation into osteoblasts, adipocytes, and chondrocytes and chondrogenic differentiation is regulated by various factors. 66 Previous study has demonstrated that sEV isolated from MSCs increase the expression of cartilage ECM components and promote cartilage repair. 67 Our experiments have demonstrated that MT-sEV have a stronger ability to promote chondrogenic differentiation of BMSCs and to enhance matrix synthesis of chondrocytes compared with sEV.…”

“…MSCs migrate to the damaged site and differentiate into chondrocytes after cartilage injury, which is beneficial for cartilage regeneration. MSCs are capable of differentiation into osteoblasts, adipocytes, and chondrocytes and chondrogenic differentiation is regulated by various factors 66 . Previous study has demonstrated that sEV isolated from MSCs increase the expression of cartilage ECM components and promote cartilage repair 67 .…”

Injectable smart stimuli-responsive hydrogels: pioneering advancements in biomedical applications

“…175 Moreover, Ha et al developed an in situ -gelation injectable hydrogel consisting of methoxy polyethylene glycol- b -polycaprolactone (MPEG-PCL) and collagen-mimetic peptide GFOGER-conjugated PEG-PCL (GFOGER-PEG-PCL). 176 Owing to the added GFOGER, it significantly increased the expression of integrins and FAK, and induced downstream signaling of p38 and ERK, which finally exhibited remarkable promotion of osteochondral regeneration.…”

Peptide Hydrogels and Nanostructures Controlling Biological Machinery

“…For example, QHREDGS (Q) peptide hydrogel has been shown to improve wound healing by increasing keratinocyte migration and the stimulation of neovascularization . Recently, the collagen-mimetic peptide GFOGER has been used as a chondrogenic inducer . Incorporation of this peptide into hydrogel resulted in the retention of bone marrow-derived mesenchymal stem cells, providing a platform for further research into osteochondral regeneration .…”