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Updated: Dec 21, 2025
Formation of Human Periodontal Ligament Cell Spheroids on Chitosan Films
Published on: June 19, 2019
Periodontal Ligament Cell Sheets and RGD-Modified Chitosan Improved Regeneration in the Horizontal Periodontal Defect
Lisa R Amir1, Yuniarti Soeroso2, Dewi Fatma1
1Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia.
Objective:
The aim of this study was to examine the potential of periodontal ligament (PDL) cells sheet and arginine-glycyl-aspartic acid (RGD)-modified chitosan scaffold for periodontal tissue regeneration in horizontal periodontal defect model.
Materials And Methods:
PDL cell cytotoxicity was tested with 3-[4,5- dimethylthiazol-2yl]-2,5-diphenyl-2H-tetrazolium bromide assay. Cell migration toward the chitosan-based materials was analyzed with trans-well migration assay. Horizontal periodontal defect model was created in four maxillary and mandibular lateral incisors of Macaque nemestrina. Following periodontal therapy, the sites were transplanted with various regenerative materials: (1) chitosan, (2) RGD-modified chitosan, (3) PDL cell sheet with chitosan, (4) PDL cell sheet with RGD-modified chitosan. The periodontal tissue regeneration was evaluated clinically and radiographically. Gingival crevicular fluids were collected each week to evaluate cementum protein-1 (CEMP-1) expression with enzyme-linked immunosorbent assay, while the biopsies were retrieved after 4 weeks for histological and microcomputed tomography evaluation.
Statistical Analysis:
Data was statistically analyzed using GraphPad Prism 6 for MacOS X. Normality was tested using the Shapiro-Wilk normality test. The Kruskal-Wallis test was used to compare the groups. Significance was accepted when p < 0.05.
Results:
Clinical examination revealed more epithelial attachment was formed in the group with PDL cell sheet with RGD-modified chitosan. Similarly, digital subtraction radiography analysis showed higher gray scale, an indication of higher alveolar bone density surrounded the transplanted area, as well as higher CEMP-1 protein expression in this group. The incorporation of RGD peptide to chitosan scaffold in the group with or without PDL cells sheet reduced the distance of cement-enamel junction to the alveolar bone crest; hence, more periodontal tissue formed.
Conclusions:
Horizontal periodontal defect model could be successfully created in M. nemestrina model. Combination of PDL cell sheet and RGD-modified chitosan resulted in the higher potential for periodontal tissue regeneration. The results of this study highlight the PDL cell sheet and RGD-modified chitosan as a promising approach for future clinical use in periodontal regeneration.
