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Periodontal healing with preameloblast-conditioned medium concentrated mixed protein

Authors
 유상준 
Issue Date
2015
Description
Dept. of Dentistry/석사
Abstract
Background and Objective: The predictability of conventional periodontal treatments to damaged periodontal ligament tissue are limited, especially on the regeneration of new cementum and attachment. And as signaling molecules, various growth factors have been used for promoting periodontal regeneration on periodontal ligament and cementum defects. Preameloblast conditioned medium (PA-CM) was prepared from cultured murine apical bud cells (ABCs) which can differentiate into ameloblasts. The aim of this study was to investigate the effects of PA-CM on cementoblasts and periodontal ligament cells in vitro and evaluate histologically the possibility of PA-CM on a regeneration for experimentally induced periodontal ligament defects in vivo.Materials and Methods : In vitro, the effect of cell migration by PA-CM on periodontal ligament(PDL) fibroblasts was studied using scratch wound healing assay and transwell assay. And the mineralization potential of the treated murine cementoblast cells (OCCM-30) was investigated by Alizarin red S staining and realtime PCR analysis. In vivo, 6 mongrel dogs(12 to 16 kg; 6 to 8 months old) were used. Total thirty-six roots were replanted with 1) no material and no periodontal ligament defect (n=12 ; positive control group); 2) no material and periodontal ligament defect (n=12 ; negative control group); 3) PA-CM soaking and periodontal ligament defect (n=12 ; experimental group). At experimental group, periodontal ligament and cementum between notchs was removed with Gracey curette and soaked in 0.08 ml water containing 80 μg of preameloblast-conditioned medium (PA-CM) for 2 minutes. At 4 and 8 weeks post-surgery, dogs were sacrificed.Results: In vitro results demonstrated that PA-CM stimulate migration of PDL cells and promote differentiation and mineralization of cementoblasts. PA-CM significantly increased cell migration (five

folds) at transwell assay. And mineralized nodules were observed in PA-CM treated OCCM-30 from the 4th day. RT-PCR analysis revealed that stronger expression of Runx2, Osx, OC, Bsp, and Cap in PA-CM treated OCCM-30 cells than those in control cells. In vivo, the newly formed periodontal ligament-like tissue and cementum-like tissue was observed partially between the root surfaces and the new bone at experimental group. The regenerated periodontal ligament-like tissue in the experimental group was significantly higher than that in the negative control group at 8 weeks (p<0.05). And the replacement resorption on the experimental group was significantly less than that in the negative control group at 8weeks (p<0.05). And the newly formed cementum-like tissue in the experimental group was significantly higher than that in the negative control group at 4 and 8weeks (p<0.05).Conclusion: The results suggest that PA-CM have the potential to regenerate periodontal tissues in the damaged periodontal ligament of replanted teeth.
Files in This Item:
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Appears in Collections:
2. College of Dentistry (치과대학) > Others (기타) > 2. Thesis
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/146130
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