Synergistic Effect of Polyelectrolyte Multilayers and Osteogenic Growth Medium on Differentiation of Human Mesenchymal Stem Cells

• Published in: Macromolecular bioscience 2010-09, Vol.10 (9), p.1043-1054
• Main Authors: Liu, Zhen-Mei, Gu, Qinyi, Xu, Zhi-Kang, Groth, Thomas
• Format: Article
• Language: English
• Subjects: Cell Adhesion - drug effects; Cell Culture Techniques; Cell Differentiation - drug effects; Cell migration; Cell Proliferation - drug effects; Chitosan; Differentiation; Gelatin; glycosaminoglycan; Heparin; human mesenchymal stem cell; Humans; Hyaluronic Acid; Macromolecules; Male; Mesenchymal Stromal Cells - cytology; Mesenchyme; Osteoblastogenesis; Osteoblasts; Osteogenesis - drug effects; osteogenic differentiation; Polyamines - chemistry; Polyanions; Polycations; Polyelectrolytes; polyesters; Polyesters - chemistry; Polyesters - pharmacology; Polymers - chemistry; Spreading; Static Electricity; Stem cells; surface plasmon resonance
• ISSN: 1616-5187; 1616-5195; 1616-5195
• DOI: 10.1002/mabi.201000086

Layer‐by‐layer assembly of biogenic polyelectrolytes (PEL) was carried out on the surface of poly (L‐lactide) to generate polyelectrolyte multilayers (PEM) that foster osteogenic differentiation of human mesenchymal stem cell (hMSC). Gelatin (GEL), hyaluronic acid (HA) and heparin (HEP) were chosen as polyanions, while chitosan (CHI) was employed as polycation. Multilayer formation was monitored by surface plasmon resonance and water contact angle measurements showing that layer formation process and surface wetting properties depended on the type of polyanions. While HEP as strong PEL led to thicker and more hydrophilic PEM, layer mass was lower for weak polyanions GEL and HA. Short‐term adhesion studies with hMSC showed strong adhesion and spreading of cells on PEM composed of GEL/CHI and low spreading, motile phenotype and aggregation of hMSC on HEP/CHI or HA/CHI. Long term studies over three weeks were carried out to follow growth and differentiation of hMSC on the PEM. Weak osteogenic differentiation of hMSC was observed on GEL/CHI if cells were cultured in normal medium while no osteogenic phenotypes were observed on HEP/CHI or HA/CHI. When cells were cultured in osteogenic differentiation medium, however, PEM composed of HEP/CHI or HA/CHI promoted differentiation of hMSC towards osteoblasts, while PEM composed of GEL/CHI failed to do so. Overall, the composition of PEMs can be used as additional tool to control osteogenic differentiation of hMSC. Formation of multilayers from gelatin (GEL), hyaluronic acid (HA) and heparin (HEP) as polyanions and chitosan as polycation was conducted on poly‐L‐lactide. Long‐term culture (three weeks) revealed monolayer growth on GEL, while predominant growth in nodules on HA or HEP was observed. Staining with alizarin red showed strong calcification of nodules in osteogenic medium (HA), while calcified areas were found on gelatin‐terminated multilayers (GEL) in normal medium.