Differentiation of Bone Marrow Stromal Cells into Osteoblasts in a Self-assembling Peptide Hydrogel: In Vitro and In Vivo Studies

Abstract

A prerequisite of tissue engineering approaches with regard to autograft is a suitable scaffold that can harbor cells and signals. Conventionally, such scaffolds have been prepared as 3D scaffolds prefabricated from synthetic or natural biomaterials. RAD16 has been introduced as a new biomaterial, where synthetic peptides self-assemble to form a hydrogel. In this study, RAD16 was examined in terms of osteogenic efficacy and feasibility of ectopic mineralization. Two hundred and seventy-one RAD16 was cocultured with 1 × 10(6) bone marrow cells from the femurs of 6-week-old Wistar male rats in alpha minimum essential medium supplemented with or without dexamethasone. Second, the same volume of the RAD16 construct hosting the cells with or without hydroxyapatite (HA) particles was treated in the dexamethasone medium as well, prepared in a Teflon tube, and implanted subcutaneously. Cell proliferation was prominent in the RAD16 coculture with dexamethasone at 1 week and significantly decreased by 2 weeks, whereas the other combinations remained or inclined, and their osteogenic differentiation was accelerated up to 2 weeks, as seen in increasing alkaline phosphatase (ALP) activity and mRNAs of ALP, OPN, and OCN. The RAD16 implant prepared with HA particles allowed more osteoblast-like cells and blood cells to grow inside, which was accompanied by elevating OPN gene expression and the stronger peak of VEGF gene expression at 2 weeks. Furthermore, more OPN mRNA signal was detected around the RAD16 containing HA particles by 4 weeks. On the other hand, the RAD16 alone represented lower expression of OPN gene. During the experiment, however, no ectopic mineralization was observed in both groups. Conclusively, it was suggested that the RAD16 showed feasibility of serving as a matrix for osteogenic differentiation of cocultured bone marrow cells in vitro and in vivo. Proceeding of exploration and modification of RAD16 are continuously required for cell-based tissue engineering.

Self-assembling peptide-enriched electrospun polycaprolactone scaffolds promote the h-osteoblast adhesion and modulate differentiation-associated gene expression

“…Interestingly, scaffolds composed of SApeptides are able to induce adhesion and proliferation of differentiated cells and also to guide cell differentiation of stem cells. Ozeki M et al suggested that RAD16, a self-assembling ionic complementary peptide, serves as a matrix for osteogenic differentiation of cocultured bone marrow cells in vitro and in vivo [26]. On the negative side, hydrogels of SA-peptides have relatively modest moduli of up to about 10 kPa, and thus find application as injectable scaffolds for the treatment of small defects rather than as patches for large ones.…”

Electrospun Scaffolds for Osteoblast Cells: Peptide-Induced Concentration-Dependent Improvements of Polycaprolactone

“…Interestingly, scaffolds made of SAPs support adhesion and proliferation of differentiated cells and also stimulate cell differentiation of immature stem cells [ 11 ]. However, since SAP hydrogels are characterized by a relatively low amount of moduli (up to about 10 kPa), they have mainly been used as injectable scaffolds to treat small defects rather than patches for large ones.…”

The Osteogenetic Efficacy of Goat Bone Marrow-Enriched Self-Assembly Peptide/Demineralized Bone MatrixIn VitroandIn Vivo

“…RADA16 can induce cell differentiation itself (30,31). In this study, we evaluated the osteogenetic induction of gMSCs in the post-enriched SAP/DBM composite.…”