J Periodontal Implant Sci.  2011 Apr;41(2):67-72.

Initial adhesion of bone marrow stromal cells to various bone graft substitutes

• Jo YJ
• Kim KH
• Koo KT
• Kim TI
• Seol YJ
• Lee YM
• Ku Y
• Chung CP
• Rhyu IC

Abstract

PURPOSE
The aim of this study is to determine whether certain biomaterials have the potential to support cell attachment. After seeding bone marrow stromal cells onto the biomaterials, we investigated their responses to each material in vitro.
METHODS
Rat bone marrow derived stromal cells were used. The biomaterials were deproteinized bovine bone mineral (DBBM), DBBM coated with fibronectin (FN), synthetic hydroxyapatite (HA), HA coated with FN, HA coated with beta-tricalcium phosphate (TCP), and pure beta-TCP. With confocal laser scanning microscopy, actin filaments and vinculin were observed after 6, 12, and 24 hours of cell seeding. The morphological features of cells on each biomaterial were observed using scanning electron microscopy at day 1 and 7.
RESULTS
The cells on HA/FN and HA spread widely and showed better defined actin cytoskeletons than those on the other biomaterials. At the initial phase, FN seemed to have a favorable effect on cell adhesion. In DBBM, very few cells adhered to the surface.
CONCLUSIONS
Within the limitations of this study, we can conclude that in contrast with DBBM not supporting cell attachment, HA provided a more favorable environment with respect to cell attachment.

Keyword

Bone substitutes; Cell adhesion; Fibronectins; Stem cells

MeSH Terms

Actin Cytoskeleton
Animals
Biocompatible Materials
Bone Marrow
Bone Substitutes
Calcium Phosphates
Cell Adhesion
Durapatite
Fibronectins
Mesenchymal Stromal Cells
Microscopy, Confocal
Microscopy, Electron, Scanning
Rats
Seeds
Stem Cells
Stromal Cells
Transplants
Vinculin
Biocompatible Materials
Bone Substitutes
Calcium Phosphates
Durapatite
Fibronectins
Vinculin

Figure

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  Figure 1 Immunofluorescence images showing cells stained for vinculin (green), actin (red) and the nucleus (blue) at 6 hours (×400). On all the materials but hydroxyapatite/fibronectin (HA/FN), the cells showed round or oval shapes. (A) Deproteinized bovine bone mineral (DBBM). (B) DBBM/FN. (C) HA. (D) HA/FN. (E) HA/tricalcium phosphate (TCP). (F) TCP.

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  Figure 2 Immunofluorescence images showing cells stained for vinculin (green), actin (red) and the nucleus (blue) at 12 hours (×400). In contrast to cells cultured on deproteinized bovine bone mineral (DBBM), cells on the other biomaterials appeared to be widely spread. (A) DBBM. (B) DBBM/fibronectin (FN). (C) Hydroxyapatite (HA). (D) HA/FN. (E) HA/tricalcium phosphate (TCP). (F) TCP.

• 

  Figure 3 Immunofluorescence images showing cells stained for vinculin (green), actin (red) and the nucleus (blue) at 24 hours (×400). On deproteinized bovine bone mineral (DBBM) and DBBM/FN, cells at 24 hours appeared to have round or oval shapes similar to the cells at 6 hours. The cells on the other biomaterials showed more extensive spreading and expressed well-defined actin cytoskeletons. (A) DBBM. (B) DBBM/fibronectin (FN). (C) Hydroxyapatite (HA). (D) HA/FN. (E) HA/tricalcium phosphate (TCP). (F) TCP.

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  Figure 4 Scanning electron microscopy images showing cells on (A) deproteinized bovine bone mineral (DBBM), (B) DBBM/fibronectin (FN), (C) hydroxyapatite (HA), (D) HA/FN, (E) HA/tricalcium phosphate (TCP), and (F) TCP at day 1 (×500). Many of the cells on DBBM and DBBM/FN were less spread out and more round shape. In contrast, the cells on HA and HA/FN appeared to be flattened out throughout the surface.

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  Figure 5 Scanning electron microscopy images showing cells on (A) deproteinized bovine bone mineral (DBBM), (B) DBBM/fibronectin (FN), (C) hydroxyapatite (HA), (D) HA/FN, (E) HA/tricalcium phosphate (TCP), and (F) TCP at day 7 (×500). Only a few, round cells adhered to the surface of DBBM and DBBM/FN. The cells on the other biomaterials adhered close together and showed polygonal shapes.

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