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Comparative Study
. 2015 Feb;48(1):47-58.
doi: 10.1111/cpr.12156. Epub 2014 Dec 11.

Comparison of osteogenic differentiation potential of human adult stem cells loaded on bioceramic-coated electrospun poly (L-lactide) nanofibres

A Ardeshirylajimi  1 , M Mossahebi-Mohammadi, S Vakilian, L Langroudi, E Seyedjafari, A Atashi, M Soleimani
Affiliations
Comparative Study

Comparison of osteogenic differentiation potential of human adult stem cells loaded on bioceramic-coated electrospun poly (L-lactide) nanofibres

A Ardeshirylajimi et al. Cell Prolif. 2015 Feb.

Abstract

Objectives: To compare potential of four types of stem cell in tissue engineering and regenerative medicine applications, osteogenic capacity of newly introduced mesenchymal stem cells (MSCs) derived from buccal fat pads (BFP) (an adipose-encapsulated mass of the oral cavity), was compared to those isolated from bone marrow (BM-MSCs), adipose tissue (AT-MSCs) and unrestricted somatic stem cells (USSCs). Cells were cultured on poly (L-lactide) (PLLA) nanofibres, Bio-Oss(®)-coated PLLA (PLLA-Bio), and culture plates (TCPS) as control.

Materials and methods: Capacity of proliferation and osteogenic differentiation of the stem cells was investigated by MTT assay and common osteogenic markers, alkaline phosphatase activity, calcium mineral deposition and bone-related genes.

Results: Highest proliferation level was observed in cells cultured on PLLA-Bio, but with no significant difference between proliferation levels of the four types of stem cell. Over the period of study, BM-MSCs cultured on PLLA-Bio scaffolds exhibited greatest alkaline phosphatase (ALP) activity and mineralization with BFP-MSCs having the next closest results. However, AT-MSC had the lowest capacity for ALP activity and mineralization during osteogenic differentiation. Gene expression evaluation revealed that highest expression of three important bone-related genes was observed in stem cells cultured on bioceramic-coated nanofibrous scaffolds.

Conclusions: Results indicated Bio-Oss-coated PLLA to compose most appropriate substrates to support proliferation and osteogenic differentiation of stem cells in vitro. BFP-MSCs demonstrated the same osteogenic differentiation capacity as other stem cells tested and thus hold very promising potential for applications in bone tissue engineering and regenerative medicine.

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Figures

Figure 1
Figure 1
Morphology of fabricated scaffolds. Poly (l‐lactide) (PLLA) at two magnifications (×1000, ×2500; a, b). PLLA after being coated with Bio‐Oss® at two magnifications (×1000, ×5000; c, d).
Figure 2
Figure 2
Micrograph of scaffolds after 21 days stem cells culture at two magnifications, unrestricted somatic stem cells (a, b), adipose tissue‐mesenchymal stem cells (c, d), bone marrow‐mesenchymal stem cells (e, f) and buccal fat pads‐mesenchymal stem cells (g, h).
Figure 3
Figure 3
Proliferation of unrestricted somatic stem cells (USSC), adipose tissue‐mesenchymal stem cells (AT‐MSCs), bone marrow‐mesenchymal stem cells (BM‐MSCs) and buccal fat pads‐mesenchymal stem cells (BFP‐MSCs) on scaffolds (PLLA and Bio‐Oss®‐coated PLLA) and tissue culture polystyrene (TCPS) over a 24, 48, 72 and 96‐h culture period (asterisks indicate significant difference between the groups at P < 0.05).
Figure 4
Figure 4
Optical micrographs of stem cells at two magnifications ×10 and ×40, adipose tissue‐mesenchymal stem cells (a, e), buccal fat pads‐mesenchymal stem cells (b, f), bone marrow‐mesenchymal stem cells (c, g) and unrestricted somatic stem cells (d, h) in basal medium.
Figure 5
Figure 5
Flow cytometric analysis of passage 2 stem cells for surface marker profiles CD44, CD90, CD105, CD34, CD45 and HLA‐DR.
Figure 6
Figure 6
Photograph and quantified morphology of stem cells after alizarin red S staining at 14 days adipose tissue‐mesenchymal stem cells (AT‐MSCs), unrestricted somatic stem cells (USSC) (a), bone marrow‐mesenchymal stem cells (BM‐MSCs) (b) and buccal fat pads‐mesenchymal stem cells cultured under induction medium (a). Magnification ×40. Quantified morphology of alizarin red S staining in all groups (asterisks indicate significant difference between the groups at P < 0.05) (b).
Figure 7
Figure 7
Alkaline phosphatase ( ALP ) activity in stem cells on scaffolds (PLLA and Bio‐Oss®‐coated PLLA) and tissue culture polystyrene (TCPS) at 7 and 14 days, during osteogenic differentiation (asterisks indicate significant difference between the groups at P < 0.05).
Figure 8
Figure 8
Calcium content of stem cells on scaffolds (PLLA and Bio‐Oss®‐coated PLLA) and tissue culture polystyrene (TCPS) at 7 and 14 days during osteogenic differentiation (asterisks indicate significant difference between the groups at P < 0.05).
Figure 9
Figure 9
Relative expression of Runx2 , osteonectin and osteocalcin (BGLAP) days 7 and 14, in stem cells on both scaffolds during osteogenesis (asterisks indicate significant difference between the groups at P < 0.05).
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