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. 2022 Sep 6;10(9):167.
doi: 10.3390/dj10090167.

Viability and Adhesion of Periodontal Ligament Fibroblasts on a Hydroxyapatite Scaffold Combined with Collagen, Polylactic Acid-Polyglycolic Acid Copolymer and Platelet-Rich Fibrin: A Preclinical Pilot Study

Leonor C Espitia-Quiroz  1   2 , Andrés L Fernández-Orjuela  1 , Lina M Anaya-Sampayo  3 , Adriana P Acosta-Gómez  4 , Luis Gonzalo Sequeda-Castañeda  3   5 , Sandra Janeth Gutiérrez-Prieto  3   6 , Nelly S Roa-Molina  3   7 , Dabeiba A García-Robayo  3   7
Affiliations

Viability and Adhesion of Periodontal Ligament Fibroblasts on a Hydroxyapatite Scaffold Combined with Collagen, Polylactic Acid-Polyglycolic Acid Copolymer and Platelet-Rich Fibrin: A Preclinical Pilot Study

Leonor C Espitia-Quiroz et al. Dent J (Basel). .

Abstract

Background: Conventional periodontal therapy relies on bone regeneration strategies utilizing scaffolds made of diverse materials, among which collagen, to promote cell adhesion and growth. Objective: To evaluate periodontal ligament fibroblast (HPdLF) cell adhesion and viability for periodontal regeneration purposes on hydroxyapatite scaffolds containing collagen (HAp-egg shell) combined with polylactic acid−polyglycolic acid copolymer (PLGA) and Platelet-Rich Fibrin (PRF). Methods: Four variations of the HAp-egg shell were used to seed HPdLF for 24 h and evaluate cell viability through a live/dead assay: (1) (HAp-egg shell/PLGA), (2) (HAp-egg shell/PLGA + collagen), (3) (HAp-egg shell/PLGA + PRF) and (4) (HAp-egg shell/PLGA + PRF + collagen). Cell adhesion and viability were determined using confocal microscopy and quantified using central tendency and dispersion measurements; significant differences were determined using ANOVA (p < 0.05). Results: Group 1 presented low cell viability and adhesion (3.70−10.17%); groups 2 and 3 presented high cell viability and low cell adhesion (group 2, 59.2−11.1%, group 3, 58−4.6%); group 4 presented the highest cell viability (82.8%) and moderate cell adhesion (45%) (p = 0.474). Conclusions: The effect of collagen on the HAp-egg shell/PLGA scaffold combined with PRF favored HPdLF cell adhesion and viability and could clinically have a positive effect on bone defect resolution and the regeneration of periodontal ligament tissue.

Keywords: collagen; fibroblasts; hydroxyapatite; periodontal ligament; platelet-rich fibrin; polylactic acid–polyglycolic acid copolymer; tissue engineering.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Experimental processes. (A) Human periodontal ligament fibroblasts culture; the cells show their characteristic spindle shape. (B) HAp-egg shell/PLGA scaffold used for all groups in the form of a 5 mm disc with 2 mm height. (C) Platelet-Rich Fibrin (PRF), with gelatinous consistency, a characteristic of this biomaterial. (D) Experimental design in a 96-well plate, containing the HPdLF culture medium. (E) Figure illustrating the four evaluated groups: red (HAp-egg shell/PLGA + HPdLF), blue (HAp-egg shell/PLGA + HPdLF + Collagen), yellow (HAp-egg shell/PLGA + HPdLF + PRF) and green (HAp-egg shell/PLGA + HPdLF + Collagen + PRF).
Figure 2
Figure 2
Images captured under a confocal fluorescence microscope (CFM); n = 3 (i, ii, iii). (A). HAp-egg shell/PLGA + HPdLF. (B). HAp-egg shell/PLGA + HPdLF + Collagen. (C). HAp-egg shell/PLGA + HPdLF + PRF. (D). HAp-egg shell/PLGA + HPdLF + PRF + Collagen. Live cells are green, dead cells are red.
Figure 3
Figure 3
Comparison among the evaluated groups of cell adhesion (A) and viability (B). Mean ± SD.
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