J Periodontal Implant Sci.  2019 Aug;49(4):258-267. 10.5051/jpis.2019.49.4.258.

Assessment of stem cell viability in the initial healing period in rabbits with a cranial bone defect according to the type and form of scaffold

• Kang SH
• Park JB
• Kim I
• Lee W
• Kim H

• KMID: 2455803
• DOI: http://doi.org/10.5051/jpis.2019.49.4.258

Abstract

PURPOSE
Increased bone regeneration has been achieved through the use of stem cells in combination with graft material. However, the survival of transplanted stem cells remains a major concern. The purpose of this study was to evaluate the viability of transplanted mesenchymal stem cells (MSCs) at an early time point (24 hours) based on the type and form of the scaffold used, including type I collagen membrane and synthetic bone.
METHODS
The stem cells were obtained from the periosteum of the otherwise healthy dental patients. Four symmetrical circular defects measuring 6 mm in diameter were made in New Zealand white rabbits using a trephine drill. The defects were grafted with 1) synthetic bone (β-tricalcium phosphate/hydroxyapatite [β-TCP/HA]) and 1×105 MSCs, 2) collagen membrane and 1×105 MSCs, 3) β-TCP/HA+collagen membrane and 1×105 MSCs, or 4) β-TCP/HA, a chipped collagen membrane and 1×105 MSCs. Cellular viability and the cell migration rate were analyzed.
RESULTS
Cells were easily separated from the collagen membrane, but not from synthetic bone. The number of stem cells attached to synthetic bone in groups 1, 3, and 4 seemed to be similar. Cellular viability in group 2 was significantly higher than in the other groups (P<0.05). The cell migration rate was highest in group 2, but this difference was not statistically significant (P>0.05).
CONCLUSIONS
This study showed that stem cells can be applied when a membrane is used as a scaffold under no or minimal pressure. When space maintenance is needed, stem cells can be loaded onto synthetic bone with a chipped membrane to enhance the survival rate.

Keyword

Bone transplantation; Cell survival; Membranes; Stem cells; Tissue scaffolds

MeSH Terms

Bone Regeneration
Bone Transplantation
Cell Movement
Cell Survival
Collagen
Collagen Type I
Humans
Membranes
Mesenchymal Stromal Cells
Periosteum
Rabbits*
Space Maintenance, Orthodontic
Stem Cells*
Survival Rate
Tissue Scaffolds
Transplants
Collagen
Collagen Type I

Figure

• 

  Figure 1 Stem cells used for transplantation. (A) Cultured stem cells derived from the periosteum. (B) Periosteum-derived stem cells before transplantation. Fluorescent staining was performed before transplantation to confirm the survival of transplanted stem cells.

• 

  Figure 2 Surgical procedure. (A) Bone defects were made in the rabbit calvarium. A circular defect with a diameter of 6 mm was formed in the skull. (B) The groups were designated as follows; group 1: β-TCP/HA+1×105 MSCs, group 2: type I collagen membrane+1×105 MSCs, group3: β-TCP/HA+type I collagen membrane+1×105 MSCs and group 4: β-TCP/HA+type I collagen chipped membrane+1×105 MSCs. (C) A Teflon sheet was applied and titanium screws were placed before suturing. The periosteum and scalp were replaced with non-absorbable sutures. (D) Harvesting of the grafted tissue was done at 24 hours after surgery. β-TCP/HA: β-tricalcium phosphate/hydroxyapatite, MSCs: mesenchymal stem cells.

• 

  Figure 3 Fluorescent microscopic evaluation of the grafted tissue. (A) Group 1 (β-TCP/HA+MSCs): Viable MSCs were attached to β-TCP/HA. (B) Group 2 (collagen membrane+MSCs): The surviving MSCs that had been transplanted were easily separated from the collagen membrane. (C) Group 3 (β-TCP/HA+collagen membrane+MSCs). The surviving MSCs that had been transplanted were abundantly attached to the β-TCP/HA, and a small amount of MSCs were scattered around the β-TCP/HA. (D) Group 4 (β-TCP/HA+chipped collagen membrane+MSCs). The results were similar to those of group 3. β-TCP/HA: β-tricalcium phosphate/hydroxyapatite, MSCs: mesenchymal stem cells.

• 

  Figure 4 Cell viability in each group was calculated by dividing the number of harvested live cells by the total number of harvested cells on day 1. a)A statistically significant difference was seen between group 2 and group 1 on day 1.

• 

  Figure 5 The cell migration rate was calculated by dividing the total number of harvested cells by the total number of grafted cells on day 1.

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  Figure 6 The survival rate of cells was calculated by dividing the number of harvested live cells by the number of grafted live cells on day 1.

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