Semipermeable Capsules Wrapping a Multifunctional and Self-regulated Co-culture Microenvironment for Osteogenic Differentiation

Clara R Correia; Rogério P Pirraco; Mariana T Cerqueira; Alexandra P Marques; Rui L Reis; João F Mano

doi:10.1038/srep21883

Semipermeable Capsules Wrapping a Multifunctional and Self-regulated Co-culture Microenvironment for Osteogenic Differentiation

Sci Rep. 2016 Feb 24:6:21883. doi: 10.1038/srep21883.

Authors

Clara R Correia^{1

2}, Rogério P Pirraco^{1

2}, Mariana T Cerqueira^{1

2}, Alexandra P Marques^{1

2}, Rui L Reis^{1

2}, João F Mano^{1

2}

Affiliations

¹ 3B's Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, 4805-017 Barco, Guimarães, Portugal.
² ICVS/3B's - PT Government Associate Laboratory, Braga/Guimarães, Portugal.

Abstract

A new concept of semipermeable reservoirs containing co-cultures of cells and supporting microparticles is presented, inspired by the multi-phenotypic cellular environment of bone. Based on the deconstruction of the "stem cell niche", the developed capsules are designed to drive a self-regulated osteogenesis. PLLA microparticles functionalized with collagen I, and a co-culture of adipose stem (ASCs) and endothelial (ECs) cells are immobilized in spherical liquified capsules. The capsules are coated with multilayers of poly(L-lysine), alginate, and chitosan nano-assembled through layer-by-layer. Capsules encapsulating ASCs alone or in a co-culture with ECs are cultured in endothelial medium with or without osteogenic differentiation factors. Results show that osteogenesis is enhanced by the co-encapsulation, which occurs even in the absence of differentiation factors. These findings are supported by an increased ALP activity and matrix mineralization, osteopontin detection, and the up regulation of BMP-2, RUNX2 and BSP. The liquified co-capsules also act as a VEGF and BMP-2 cytokines release system. The proposed liquified capsules might be a valuable injectable self-regulated system for bone regeneration employing highly translational cell sources.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alginates / chemistry
Bone Morphogenetic Protein 2 / genetics
Bone Morphogenetic Protein 2 / metabolism
Capsules / chemistry*
Cell Differentiation
Cells, Cultured
Chitosan / chemistry
Coculture Techniques
Core Binding Factor Alpha 1 Subunit / genetics
Core Binding Factor Alpha 1 Subunit / metabolism
Cytokines / analysis
Cytokines / metabolism
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Fluorescent Dyes / chemistry
Glucuronic Acid / chemistry
Hexuronic Acids / chemistry
Humans
Integrin-Binding Sialoprotein / genetics
Integrin-Binding Sialoprotein / metabolism
Microscopy, Electron, Scanning
Microscopy, Fluorescence
Nanostructures / chemistry
Osteoblasts / cytology
Osteoblasts / metabolism
Osteogenesis
Osteopontin / metabolism
Platelet Endothelial Cell Adhesion Molecule-1 / genetics
Platelet Endothelial Cell Adhesion Molecule-1 / metabolism
Polylysine / chemistry
Real-Time Polymerase Chain Reaction
Vascular Endothelial Growth Factor A / genetics
Vascular Endothelial Growth Factor A / metabolism

Substances

Alginates
Bone Morphogenetic Protein 2
Capsules
Core Binding Factor Alpha 1 Subunit
Cytokines
Fluorescent Dyes
Hexuronic Acids
Integrin-Binding Sialoprotein
Platelet Endothelial Cell Adhesion Molecule-1
Vascular Endothelial Growth Factor A
Osteopontin
Polylysine
Glucuronic Acid
Chitosan