Restor Dent Endod.  2016 May;41(2):98-105. 10.5395/rde.2016.41.2.98.

Biocompatibility of two experimental scaffolds for regenerative endodontics

• Leong DJ
• Setzer FC
• Trope M
• Karabucak

• KMID: 2163291
• DOI: http://doi.org/10.5395/rde.2016.41.2.98

Abstract

OBJECTIVES
The biocompatibility of two experimental scaffolds for potential use in revascularization or pulp regeneration was evaluated.
MATERIALS AND METHODS
One resilient lyophilized collagen scaffold (COLL), releasing metronidazole and clindamycin, was compared to an experimental injectable poly(lactic-co-glycolic) acid scaffold (PLGA), releasing clindamycin. Human dental pulp stem cells (hDPSCs) were seeded at densities of 1.0 × 10(4), 2.5 × 10(4) and 5.0 × 10(4). The cells were investigated by light microscopy (cell morphology), MTT assay (cell proliferation) and a cytokine (IL-8) ELISA test (biocompatibility).
RESULTS
Under microscope, the morphology of cells coincubated for 7 days with the scaffolds appeared healthy with COLL. Cells in contact with PLGA showed signs of degeneration and apoptosis. MTT assay showed that at 5.0 × 10(4) hDPSCs, COLL demonstrated significantly higher cell proliferation rates than cells in media only (control, p < 0.01) or cells co-incubated with PLGA (p < 0.01). In ELISA test, no significant differences were observed between cells with media only and COLL at 1, 3, and 6 days. Cells incubated with PLGA expressed significantly higher IL-8 than the control at all time points (p < 0.01) and compared to COLL after 1 and 3 days (p < 0.01).
CONCLUSIONS
The COLL showed superior biocompatibility and thus may be suitable for endodontic regeneration purposes.

Keyword

Clindamycin; Matrix; Metronidazole; Regenerative endodontics; Revascularization; Scaffold

MeSH Terms

Apoptosis
Cell Proliferation
Clindamycin
Collagen
Dental Pulp
Endodontics*
Enzyme-Linked Immunosorbent Assay
Humans
Interleukin-8
Metronidazole
Microscopy
Regeneration
Stem Cells
Clindamycin
Collagen
Interleukin-8
Metronidazole

Figure

• 

  Figure 1 Comparison of representative cell appearances under the light microscope at ×100 and ×200. 'S' denotes scaffolds. (a - d) Cells grown in proximity to COLL scaffolds demonstrated a generally healthy appearance similar to other areas of the culture wells and controls (arrows 1); (e - f) Co-incubation with the PLGA scaffold often displayed signs of apoptosis and cell degeneration, particularly close to the actual material. Cell debris (arrow 2) and rounded cell morphologies (arrow 3) suggested non-attachment. COLL, lyophilized collagen; PLGA, poly-lactic-co-glycolic acid.

• 

  Figure 2 MTT assay. Absorbance measured using a multiplate reader. hDPSCs were seeded at 3 different densities (1.0 × 104, 2.5 × 104, and 5.0 × 104) with media only (red), PLGA scaffold (blue) or COLL scaffold (green). Mean ± SEM for N = 6 per condition. *p < 0.05, by 2-way ANOVA and Bonferroni post hoc test. hDPSC, human dental pulp stem cells; PLGA, poly-lactic-co-glycolic acid; COLL, lyophilized collagen.

• 

  Figure 3 IL-8 ELISA test. Absorbance measured using a multiplate reader at 3 different time points (Days 1, 3, and 6). hDPSCs were seeded at 5.0 × 104 cell density with media only (red), PLGA scaffold (blue) or COLL scaffold (green). Mean ± SEM for N = 6 per condition. *p < 0.05, by 2-way ANOVA and Bonferroni post hoc test. hDPSC, human dental pulp stem cells; PLGA, poly-lactic-coglycolic acid; COLL, lyophilized collagen.

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