Int J Stem Cells.  2015 May;8(1):90-98. 10.15283/ijsc.2015.8.1.90.

Attachment and Differentiation of Human Umbilical Cord Stem Cells on to the Tooth Root Surface with and without the Use of Fibroblast Growth Factor-An In Vitro Study

• George JP
• Chakravarty P
• Chowdhary
• Purushothama H
• Rao JA

• KMID: 2380802
• DOI: http://doi.org/10.15283/ijsc.2015.8.1.90

Abstract

BACKGROUND AND OBJECTIVES
The purpose of this first of its kind study was to analyse the growth, development and attachment of cultured human umbilical cord stem cells alone or supplemented with basic Fibroblast Growth Factor (bFGF) on both healthy and periodontally diseased tooth surfaces in vitro.
METHODS
Four groups of 12 root surface scaffolds each were classified as Group I- healthy root surfaces; Group II- periodontally diseased; Group III- Healthy with bFGF and Group IV- periodontally diseased root with bFGF. bFGF was applied in the concentration of 8 ng/ml on to the surface followed by incubation of cultured human umbilical cord stem cells (hUCMSCs) on the scaffolds. Scanning electron microscopy observations were made on 14th and 21st days to assess the proliferation and morphology of cells attached on the tooth surface.
RESULTS
Cultured hUCMSCs demonstrated adhesion to tooth root scaffold. All the groups showed a significant increase in the number of cell attachment from 14th day to 21st day. The groups with bFGF showed a significant increase in attachment of cells when compared to the groups without bFGF. The cells showed an increase in number of flat cells from 14th day to 21st day in all the groups indicating an increased maturity of cells. Periodontally diseased groups had less maturity of cells than healthy groups. The groups supplemented with bFGF, had more mature cells than the groups without bFGF.
CONCLUSIONS
hUCMSCs have the propensity to differentiate into cells that have the capacity to bind to root surfaces. hUCMSCs incubated with bFGF showed better proliferation and attachment to tooth root surfaces. The role of hUCMSCs can be further explored for periodontal regeneration.

Keyword

Mesenchymal Stem Cell; Wharton Jelly cells; Multipotent mesenchymal stromal cells; Regeneration; Growth factors

MeSH Terms

Fibroblast Growth Factor 2
Fibroblasts*
Humans
Intercellular Signaling Peptides and Proteins
Mesenchymal Stromal Cells
Microscopy, Electron, Scanning
Regeneration
Stem Cells*
Tooth
Tooth Root*
Umbilical Cord*
Fibroblast Growth Factor 2
Intercellular Signaling Peptides and Proteins

Figure

• 

  Fig. 1 Wharton’s jelly derived human umbilical cord stem cells. (A) Immature hUCMSCs seen as oval/spindle shaped cells in the 3rd passage. (B) Flow cytometry analysis of hUCMSCs after 3 passages of culture showed positive staining for typical mesenchymal markers (CD90, CD105, CD44, CD73).

• 

  Fig. 2 SEM analysis at 500X magnification showing cellular morphology on Day 14. (A) 14th Day scaffold for Group I showing presence of spindle shaped cells (white arrow). (B) 14th Day scaffold for Group II scaffold depicts round cells (white arrow). (C) 14th Day scaffold for Group III shows presence of spindle shaped cells (white arrow). (D) 14th Day scaffold for Group IV shows presence of spindle shaped cells (white arrow).

• 

  Fig. 3 SEM analysis at 500X magnification showing cellular morphology on Day 21. (A) 21st Day scaffold for Group I showing cells with flat morphology (white arrow). (B) 21st Day scaffold for Group II shows cells with flat morphology (white arrow). (C) 21st Day scaffold for Group III show cytoplasmic processes of cells (white arrow). (D) 21st Day scaffold for Group III show cytoplasmic processes of cells (white arrow).

• 

  Fig. 4 Graph showing the difference in cellular counts on Day 14 and Day 21 in all the groups. Student ‘t’ test done to evaluate the number of adherent cells on root specimens in all groups between 14th and 21st day. Day 21 day showed a higher number of cell attachment. Data significant when ‘p’ value<0.05.

• 

  Fig. 5 Graph showing morphologic analysis assessment between all the groups on Day 14 and Day 21. A lesser score with more number of flat cells was seen in day 21.

Reference

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