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. 2017 May 23:16:785-794.
doi: 10.17179/excli2016-741. eCollection 2017.

Osteogenic differentiation of mesenchymal stem cells cultured on PLLA scaffold coated with Wharton's Jelly

Marziehsadat Ahmadi  1 , Ehsan Seyedjafari  2 , Seyed Jalal Zargar  1 , Gebremariam Birhanu  3   4 , Ali Zandi-Karimi  2 , Bahareh Beiki  2 , Kadriye Tuzlakoglu  5
Affiliations

Osteogenic differentiation of mesenchymal stem cells cultured on PLLA scaffold coated with Wharton's Jelly

Marziehsadat Ahmadi et al. EXCLI J. .

Abstract

Poly-L-lactic acid (PLLA) electrospun nanofiber scaffold is one of the most commonly used synthetic polymer scaffolds for bone tissue engineering application. However, PLLA is hydrophobic in nature, hence does not maintain proper cell adhesion and tissue formation, moreover, it cannot provide the osteo-inductive environment due to inappropriate surface characteristic and the lack of surface motives participating in the first cellular events. To modify these shortcomings different approaches have been used, among those the most commonly used one is coating of the surface of the electrospun nanofiber with natural materials. In this work Wharton's jelly (WJ), a tissue which surrounds the umbilical cord vessels, reaches in high amounts of extracellular matrix (ECM) components mainly; collagen, hyaluronic acid and several sulphated glycosaminoglycans (GAGs) were used to cover the surface of electrospun PLLA nanofiber scaffolds. The surface morphology of the nanofiber scaffold was evaluated via scanning electron microscope, and the in vitro osteogenic differentiation potential was determined by MTT assay and common osteogenic marker tests such as alkaline phosphatase (ALP) activity and calcium deposition tests. Coating of WJ could not change the surface morphology and diameter of the nanofibers. However, WJ-PLLA scaffolds showed higher proliferation of human mesenchymal stem cells (MSC) than tissue culture plate (TCP) and pristine PLLA scaffolds, moreover, WJ-PPLA scaffold demonstrated significant alkaline phosphatase activity and calcium mineralization than either TCP or PLLA nanofiber scaffolds.

Keywords: Wharton's Jelly; coating; electrospinning; poly-L-lactic acid.

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Figures

Figure 1
Figure 1. Scanning electron microscope images of electrospun nanofiber scaffolds (A), PLLA (B) WJ- PLLA
Figure 2
Figure 2. Proliferation of MSC on scaffolds (PLLA and WJ coated PLLA) and TCP. The results given are representative of three independent experiments.
Figure 3
Figure 3. Alkaline phosphatase (ALP) activity of stem cells of PLLA scaffolds, WJ coated PLLA scaffolds and tissue culture plate (TCP) on 7th, 14th and 21st day, during osteogenic differentiation. The results given are representative of three independent experiments.
Figure 4
Figure 4. Total calcium content of stem cells on scaffolds (PLLA and WJ coated PLLA) and tissue culture plate (TCP) on 7th, 14th and 21st day of osteogenic differentiation. The results given are representative of three independent experiments.
Figure 5
Figure 5. Alizarin red S staining confirming the differentiation of human MSC into the osteoblast at day 14 (A), control (B) osteogenic media
See this image and copyright information in PMC

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