A comparative study on in vitro osteogenic priming potential of electron spun scaffold PLLA/HA/Col, PLLA/HA, and PLLA/Col for tissue engineering application

Hanumantha Rao Balaji Raghavendran; Subramaniam Puvaneswary; Sepehr Talebian; Malliga R Murali; Sangeetha V Naveen; G Krishnamurithy; Robert McKean; Tunku Kamarul

doi:10.1371/journal.pone.0104389

A comparative study on in vitro osteogenic priming potential of electron spun scaffold PLLA/HA/Col, PLLA/HA, and PLLA/Col for tissue engineering application

PLoS One. 2014 Aug 20;9(8):e104389. doi: 10.1371/journal.pone.0104389. eCollection 2014.

Authors

Hanumantha Rao Balaji Raghavendran¹, Subramaniam Puvaneswary¹, Sepehr Talebian², Malliga R Murali, Sangeetha V Naveen¹, G Krishnamurithy¹, Robert McKean³, Tunku Kamarul⁴

Affiliations

¹ Tissue Engineering Group (TEG), National Orthopaedic Centre of Excellence in Research and Learning (NOCERAL), Department of Orthopaedic Surgery, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
² Department of Mechanical Engineering, Engineering Faculty, University of Malaya, Kuala Lumpur, Malaysia.
³ The Electrospinning Company Ltd, Rutherford Appleton Laboratory, Harwell Oxford, Didcot, Oxfordshire, United Kingdom.
⁴ Tissue Engineering Group (TEG), National Orthopaedic Centre of Excellence in Research and Learning (NOCERAL), Department of Orthopaedic Surgery, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia; Clinical Investigative Centre, Faculty of Medicine, University Malaya Medical Center, Kuala Lumpur, Malaysia.

Abstract

A comparative study on the in vitro osteogenic potential of electrospun poly-L-lactide/hydroxyapatite/collagen (PLLA/HA/Col, PLLA/HA, and PLLA/Col) scaffolds was conducted. The morphology, chemical composition, and surface roughness of the fibrous scaffolds were examined. Furthermore, cell attachment, distribution, morphology, mineralization, extracellular matrix protein localization, and gene expression of human mesenchymal stromal cells (hMSCs) differentiated on the fibrous scaffolds PLLA/Col/HA, PLLA/Col, and PLLA/HA were also analyzed. The electrospun scaffolds with a diameter of 200-950 nm demonstrated well-formed interconnected fibrous network structure, which supported the growth of hMSCs. When compared with PLLA/H%A and PLLA/Col scaffolds, PLLA/Col/HA scaffolds presented a higher density of viable cells and significant upregulation of genes associated with osteogenic lineage, which were achieved without the use of specific medium or growth factors. These results were supported by the elevated levels of calcium, osteocalcin, and mineralization (P<0.05) observed at different time points (0, 7, 14, and 21 days). Furthermore, electron microscopic observations and fibronectin localization revealed that PLLA/Col/HA scaffolds exhibited superior osteoinductivity, when compared with PLLA/Col or PLLA/HA scaffolds. These findings indicated that the fibrous structure and synergistic action of Col and nano-HA with high-molecular-weight PLLA played a vital role in inducing osteogenic differentiation of hMSCs. The data obtained in this study demonstrated that the developed fibrous PLLA/Col/HA biocomposite scaffold may be supportive for stem cell based therapies for bone repair, when compared with the other two scaffolds.

Publication types

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

MeSH terms

Cells, Cultured
Collagen
Durapatite
Humans
Mesenchymal Stem Cells / cytology*
Osteogenesis / physiology*
Polyesters
Tissue Engineering / methods*
Tissue Scaffolds*

Substances

Polyesters
hydroxyapatite-polylactide
Collagen
Durapatite

Grants and funding

This work was supported in part by UMRG from the University of Malaya and major grant support (Reference number -UM.C/625/1/HIR/MOHE/CHAN/03, account number - A000003-50001), University of Malaya. The Electrospinning Company Ltd provided support in the form of salaries for author RM. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.