J Dent Anesth Pain Med.  2017 Sep;17(3):191-198. 10.17245/jdapm.2017.17.3.191.

Effects of nerve cells and adhesion molecules on nerve conduit for peripheral nerve regeneration

• Chung JR
• Choi JW
• Fiorellini JP
• Hwang KG
• Park CJ

• KMID: 2390803
• DOI: http://doi.org/10.17245/jdapm.2017.17.3.191

Abstract

BACKGROUND
For peripheral nerve regeneration, recent attentions have been paid to the nerve conduits made by tissue-engineering technique. Three major elements of tissue-engineering are cells, molecules, and scaffolds.
METHODS
In this study, the attachments of nerve cells, including Schwann cells, on the nerve conduit and the effects of both growth factor and adhesion molecule on these attachments were investigated.
RESULTS
The attachment of rapidly-proliferating cells, C6 cells and HS683 cells, on nerve conduit was better than that of slowly-proliferating cells, PC12 cells and Schwann cells, however, the treatment of nerve growth factor improved the attachment of slowly-proliferating cells. In addition, the attachment of Schwann cells on nerve conduit coated with fibronectin was as good as that of Schwann cells treated with glial cell line-derived neurotrophic factor (GDNF).
CONCLUSIONS
Growth factor changes nerve cell morphology and affects cell cycle time. And nerve growth factor or fibronectin treatment is indispensable for Schwann cell to be used for implantation in artificial nerve conduits.

Keyword

Cellular Attachment; Fibronectins; Nerve Growth Factor; Nerve Cell; Tenascin

MeSH Terms

Animals
Attention
Cell Cycle
Fibronectins
Glial Cell Line-Derived Neurotrophic Factor
Nerve Growth Factor
Neurons*
PC12 Cells
Peripheral Nerves*
Regeneration*
Schwann Cells
Tenascin
Fibronectins
Glial Cell Line-Derived Neurotrophic Factor
Nerve Growth Factor
Tenascin

Figure

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  Fig. 1 Morphological appearance of tissue-engineered nerve conduit. Grossly, it has hollow and round structure with 1-mm inner diameter and 0.2-mm thickness and prepared in 5-mm length (A). The SEM view shows the multi-porous outer surface (B, × 1000, left; × 3000, right).

• 

  Fig. 2 Cellular attachment on tissue-engineered nerve conduit by MTT assay in control, Group I and Group II after 48 hr incubation (*P < 0.05, compared with control; SC = Schwann cell).

• 

  Fig. 3 Cellular attachment on tissue-engineered nerve conduit by MTT assay in control, Group II and Group III after 48 hr incubation. (*P < 0.05, compared with control, #P < 0.05, compared with Group II; SC = Schwann cell).

• 

  Fig. 4 SEM views of cell-nerve conduit complexes in Group I and Group II. C6 cells (A; ×1000), HS683 cells (B; ×1000) and Schwann cells (D; ×1000) show polygonal shape but PC12 cells (C; ×1000) show small and round shape.

• 

  Fig. 5 SEM views of cell-nerve conduit complexes in Group III. The cells in Group III, PC 12 cells treated with NGF (A; ×1000) and Schwann cells with GDNF (B; ×1000), were morphologically changed from small and round shape to spindle shape with elongated cytoplasmic extension and reached confluency (C; ×1000) at the same 48 hr incubation.

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  Fig. 6 Cellular attachment on tenascin-coated nerve conduit by MTT assay in C6 cells and Schwann cells after 48 hr incubation (TN = Tenascin, SC = Schwann cell).

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  Fig. 7 Cellular attachment on fibronectin-coated nerve conduit by MTT assay in C6 cells and Schwann cells after 48 hr incubation (*P < 0.05, compared with none-coated nerve conduit; FN = Fibronectin, SC = Schwann cell).

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