RESUMO
Background/purpose: Periodontal ligament stem cells (PDLSCs) have the potential for regenerating periodontal tissue. The study aims to investigate the impact of demographics (ages, gender, disease) and culture techniques (shipping storage time and culture method) on the success of primary culture. Materials and methods: PDLSCs were collected from 51 teeth of 26 patients and cultured via outgrowth (OG) and enzymatic digestion (ED) methods. Cells characteristics were confirmed by flow cytometry, MTT, and ARS. The primary culture success rate was evaluated with a serial chi-square test to determine the relationship with culture technique (ED/OG and ≤4 h/prolonged culture) and patient demographics (Young/Old, Female/Male, and Health/Periodontitis). Results: The overall success rate of Health group (69.7%) was higher than Periodontitis (38.9%). Culturing within 4 h possessed a higher success rate (71.8%) than prolonged group (16.7%) regardless of patient demographics, and using OG method (81.5%) revealed more promising. Subgroup analysis of 39 cases (culture within 4 h) found that the success rate of OG was higher than ED in the Old group (87.5%-25.0%) and in the Periodontitis group (83.3%-25.0%). Conclusion: Primary culturing of PDLSCs within 4 h and using the outgrowth method led to higher success rates regardless of patient demographics. It can achieve successful PDLSCs culture of older patients or patients with periodontal disease by appropriate culture technique.
RESUMO
Periodontitis, a chronic inflammatory disease caused by microbial communities carrying pathogens, leads to the loss of tooth-supporting tissues and is a significant contributor to tooth loss. This study aims to develop a novel injectable cell-laden hydrogel consisted of collagen (COL), riboflavin, and a dental light-emitting diode (LED) photo-cross-linking process for periodontal regeneration. Utilizing α-SMA and ALP immunofluorescence markers, we confirmed the differentiation of human periodontal ligament fibroblasts (HPLFs) into myofibroblasts and preosteoblasts within collagen scaffolds in vitro. Twenty-four rats with three-wall artificial periodontal defects were divided into four groups, Blank, COL_LED, COL_HPLF, and COL_HPLF_LED, and histomorphometrically assessed after 6 weeks. Notably, the COL_HPLF_LED group showed less relative epithelial downgrowth (p < 0.01 for Blank, p < 0.05 for COL_LED and COL_HPLF), and the relative residual bone defect was significantly reduced in the COL_HPLF_LED group compared to the Blank and the COL_LED group (p < 0.05). The results indicated that LED photo-cross-linking collagen scaffolds possess sufficient strength to withstand the forces of surgical process and biting, providing support for HPLF cells embedded within them. The secretion of cells is suggested to promote the repair of adjacent tissues, including well-oriented periodontal ligament and alveolar bone regeneration. The approach developed in this study demonstrates clinical feasibility and holds promise for achieving both functional and structural regeneration of periodontal defects.
