Multilayer Nanoscale Encapsulation of Biofunctional Peptides to Enhance Bone Tissue Regeneration In Vivo

Abstract

Bone tissue healing is a dynamic process that is initiated by the recruitment of osteoprogenitor cells followed by their migration, proliferation, differentiation, and development of a mineralizing extracellular matrix. The work aims to manufacture a functionalized porous membrane that stimulates early events in bone healing for initiating a regenerative cascade. Layer‐by‐layer (LbL) assembly is proposed to modify the surface of osteoconductive electrospun meshes, based on poly(lactic‐co‐glycolic acid) and nanohydroxyapatite, by using poly(allylamine hydrochloride) and poly(sodium 4‐styrenesulfonate) as polyelectrolytes. Molecular cues are incorporated by grafting peptide fragments into the discrete nanolayers. KRSR (lysine‐arginine‐serine‐arginine) sequence is grafted to enhance cell adhesion and proliferation, NSPVNSKIPKACCVPTELSAI to guide bone marrow mesenchymal stem cells differentiation in osteoblasts, and FHRRIKA (phenylalanine‐histidine‐arginine‐arginine‐isoleucine‐lysine‐alanine) to improve mineralization matrix formation. Scanning electron microscopy, infrared spectroscopy, and X‐ray photoelectron spectroscopy demonstrate the successful surface functionalization. Furthermore, the peptide incorporation enhances cellular processes, with good viability and significant increase of alkaline phosphatase activity, osteopontin, and osteocalcin. The functionalized membrane induces a favorable in vivo response after implantation for four weeks in nonhealing rat calvarial defect model. It is concluded that the multilayer nanoencapsulation of biofunctional peptides using LbL approach has significant potential as innovative manufacturing technique to improve bone regeneration in orthopedic and craniofacial medical devices.

Advanced Functional Biomaterials for Stem Cell Delivery in Regenerative Engineering and Medicine

“…It has been used alone or in combination with natural materials such as collagen or with growth factors to repair bone defects . Gentile et al reported that peptide‐functionalized nanolayered PLGA scaffolds provided molecular cues to bone marrow MSCs that enhance in vivo bone regeneration in a calvarial defect model . Pretreatment of stem cells is another strategy to improve cell differentiation into osteogenic lineage before adding the cells to a scaffold.…”

Three‐dimensional electrospun nanofibrous scaffolds for bone tissue engineering

“…Gentile et al deposited several polypeptides onto the electrospun mesh by LbL, and found that the osteogenic differentiation of the seeded mesenchymal stem cells was significantly improved. Implantation of functionalized electrospun mesh in skull defects can promote bone regeneration (Gentile et al, ). In summary, for 3D electrospinning scaffolds with complex nanostructures, LbL is a promising surface modification method.…”

Electrospun Fibers for Recruitment and Differentiation of Stem Cells in Regenerative Medicine

“…A recent study on nanoencapsulation of functional biopeptides in electrospun poly(lactic‐co‐glycolic acid) and nanohydroxyapatite fibers using layer‐by‐layer approach showed increased matrix mineralization for bone tissue regeneration . The functional biopeptides are grafted into discrete nanolayers, which include KRSR (lysine‐arginine‐serine‐arginine) sequence to enhance cell adhesion and proliferation, NSPVNSKIPKACCVPTELSAI to guide bone marrow mesenchymal stem cells differentiation, and FHRRIKA (phenylalanine‐histidine‐arginine‐arginine‐isoleucine‐lysine‐alanine) to improve mineralization matrix formation . Studies show that pDA coating stabilizes MSC adhesion and proliferation and maintains the self‐renewal property of pluripotent stem cells for a long time .…”

“…[20] The functional biopeptides are grafted into discrete nanolayers, which include KRSR (lysinearginine-serine-arginine) sequence to enhance cell adhesion and proliferation, NSPVNSKIPKACCVPTELSAI to guide bone marrow mesenchymal stem cells differentiation, and FHRRIKA (phenylalanine-histidine-arginine-arginine-isoleucine-lysine-alanine) to improve mineralization matrix formation. [21] Studies show that pDA coating stabilizes MSC adhesion and proliferation and maintains the self-renewal property of pluripotent stem cells for a long time. [22] The hydrophilicity of pDA coating increases the adhesion of cells to the substrate and also enhances the growth of the attached cells as compared to pristine scaffolds.…”