CAJ | 학술논문

背景：釉基质衍生物已经在临床上用于治疗严重的牙周炎，发现其可促进牙周组织修复、再生，但其中的机制还未阐明。目的：探讨釉基质衍生物对牙周膜干细胞分化和增殖的作用。方法：分离培养人牙周膜干细胞，检测其克隆形成率、表面抗原表达情况，鉴定其多向分化潜能。将不同质量浓度的釉基质衍生物(20，50或100 mg/L)作用于牙周膜干细胞培养2，4周，用Trichrome和Von Kosa’s染色法检测牙周膜干细胞胶原合成及矿化结节形成情况，Real time RT-PCR方法检测成骨分化相关基因Ⅰ型胶原、骨钙素、RUX2的表达，MTT法和生长率法检测牙周膜干细胞的增殖情况。结果与结论：牙周膜干细胞呈梭形，其克隆形成率较牙周膜细胞高，表面抗原CD105，CD29，CD45，CD44的表达率分别为99.8%，99.7%，1.26%，98.8%，具备多向分化潜能。釉基质衍生物呈现一定的时间剂量效应关系促进牙周膜干细胞胶原合成及矿化结节形成，促进成骨分化相关基因Ⅰ型胶原、骨钙素、RUX2的表达，促进牙周膜干细胞的增殖，可能在牙周组织修复、再生中发挥作用。
배경：유기질연생물이경재림상상용우치료엄중적아주염，발현기가촉진아주조직수복、재생，단기중적궤제환미천명。목적：탐토유기질연생물대아주막간세포분화화증식적작용。방법：분리배양인아주막간세포，검측기극륭형성솔、표면항원표체정황，감정기다향분화잠능。장불동질량농도적유기질연생물(20，50혹100 mg/L)작용우아주막간세포배양2，4주，용Trichrome화Von Kosa’s염색법검측아주막간세포효원합성급광화결절형성정황，Real time RT-PCR방법검측성골분화상관기인Ⅰ형효원、골개소、RUX2적표체，MTT법화생장솔법검측아주막간세포적증식정황。결과여결론：아주막간세포정사형，기극륭형성솔교아주막세포고，표면항원CD105，CD29，CD45，CD44적표체솔분별위99.8%，99.7%，1.26%，98.8%，구비다향분화잠능。유기질연생물정현일정적시간제량효응관계촉진아주막간세포효원합성급광화결절형성，촉진성골분화상관기인Ⅰ형효원、골개소、RUX2적표체，촉진아주막간세포적증식，가능재아주조직수복、재생중발휘작용。
BACKGROUND:The enamel matrix derivative has been used in the clinical treatment of severe periodontitis; however, the mechanism(s) by which enamel matrix derivative promotes periodontal regeneration is stil obscure. OBJECTIVE:To explore the effects of enamel matrix derivatives on the differentiation and proliferation of periodontal ligament stem cels. METHODS:Periodontal ligament stem cels were isolated and identified from human teeth. Cloning forming efficiency, surface antigen expression and pluripotency were detected and identified. Enamel matrix derivatives with different concentrations (20, 50, 100 mg/L) were used to culture periodontal ligament stem cels for 2 and 4 weeks. Colagen synthesis and mineralized nodule formation were detected using Trichrom staining and Von Kosa’s staining, respectively; real-time RT-PCR was employed to detect expressions of colagen type I, osteocalcin, and RUX2; MTT and cel growth rate assay were used to detect the proliferation of periodontal ligament stem cels. RESULTS AND CONCLUSION:Periodontal ligament stem cels were spindle-shaped and showed a higher colony forming efficiency than periodontal ligament cels. The expressions of surface antigens of periodontal ligament stem cels-CD105, CD29, CD45, CD44 were respectively 99.8%, 99.7%, 1.26%, 98.8%, indicating periodontal ligament stem cels have the multilineage differentiation potential. Enamel matrix derivatives improve the colagen synthesis and mineralization nodule formation of periodontal ligament stem cels in a time-dose dependent manner. They also can improve the expression of osteogensis-related genes colagen type I, osteocalcin, RUX2 and proliferation of periodontal ligament stem cels.