CAJ | 학술논문

背景：大量研究证实釉基质蛋白可促进成骨细胞和成牙骨质细胞的再生，将其运用于牙周缺损治疗可达到接近生理性的牙周再生。 　　目的：观察不同质量浓度釉基质蛋白对人牙周膜细胞增生、分化和迁移的影响。 　　方法：取第3代人牙周膜细胞，以含不同质量浓度釉基质蛋白(0，12.5，25，50，100，250 mg/L)的无血清DMEM培养基培养。培养24 h后，采用3H-胸腺嘧啶核苷掺入法检测细胞增殖，MTT法检测细胞活性；培养48 h后，检测细胞碱性磷酸酶活性及骨钙素分泌；待细胞融合为单层，去除细胞培养液，以移液管头将单层细胞制备出1 mm宽的细胞切口，持续24 h观察细胞融合情况。 　　结果与结论：当釉基质蛋白质量浓度在0-100 mg/L范围内，随着其质量浓度的升高，细胞增殖、活性、碱性磷酸酶活性、骨钙素分泌均逐渐升高，以100 mg/L升高最明显；当釉基质蛋白质量浓度增至250 mg/L时，细胞增殖、活性、碱性磷酸酶活性、骨钙素分泌均有所下降，但仍高于0 mg/L组。100 mg/L组在初始观察6 h时，创缘周围的细胞开始向中心生长，待培养12 h时，创缘两侧细胞开始融合，培养20 h后创缘两侧细胞融合完全创缘完全关闭完全，创面愈合优于其他质量浓度组。结果表明釉基质蛋白具有促进牙周膜细胞增殖、分化与迁移的能力。
배경：대량연구증실유기질단백가촉진성골세포화성아골질세포적재생，장기운용우아주결손치료가체도접근생이성적아주재생。 　　목적：관찰불동질량농도유기질단백대인아주막세포증생、분화화천이적영향。 　　방법：취제3대인아주막세포，이함불동질량농도유기질단백(0，12.5，25，50，100，250 mg/L)적무혈청DMEM배양기배양。배양24 h후，채용3H-흉선밀정핵감참입법검측세포증식，MTT법검측세포활성；배양48 h후，검측세포감성린산매활성급골개소분비；대세포융합위단층，거제세포배양액，이이액관두장단층세포제비출1 mm관적세포절구，지속24 h관찰세포융합정황。 　　결과여결론：당유기질단백질량농도재0-100 mg/L범위내，수착기질량농도적승고，세포증식、활성、감성린산매활성、골개소분비균축점승고，이100 mg/L승고최명현；당유기질단백질량농도증지250 mg/L시，세포증식、활성、감성린산매활성、골개소분비균유소하강，단잉고우0 mg/L조。100 mg/L조재초시관찰6 h시，창연주위적세포개시향중심생장，대배양12 h시，창연량측세포개시융합，배양20 h후창연량측세포융합완전창연완전관폐완전，창면유합우우기타질량농도조。결과표명유기질단백구유촉진아주막세포증식、분화여천이적능력。
BACKGROUND:Numerous studies have confirmed that enamel matrix proteins can promote the regeneration of osteoblasts and cementoblast, and then it can achieve approaching physiological periodontal regeneration in the treatment of periodontal defects. OBJECTIVE: To observe the effects of different concentrations of enamel matrix proteins on proliferation, viability, differentiation and migration of human periodontal ligament cels. METHODS: The human periodontal ligament cels at the third generation were gained, and then cultured in serum-free DMEM containing different concentrations of enamel matrix proteins (0, 12.5, 25, 50, 100, 250 mg/L). After 24 hours of culture, proliferation and viability of periodontal ligament cels were measured using [(3)H]-thymidine uptake and MTT assay. After 48 hours of culture, alkaline phosphatase activity and osteocalcin production were detected with commercial available test kits. When the cels grew as a monolayer, the cel culture fluid was removed, and then with a pipette head, a cel incision, 1 mm wideness, was prepared in a monolayer of cels to further observe the cel fusion continuously within 24 hours. RESULTS AND CONCLUSION:The proliferation, viability and differentiation of periodontal ligament cels were gradualy increased with the concentration increase of enamel matrix proteins (0-100 mg/L). When the concentration of enamel matrix proteins was 250 mg/L, these parameters began to decrease, but the levels were stil higher than those in the 0 mg/L enamel matrix protein group. In the 100 mg/L group, the cels in the wound edge began to grow towards the center in the initial 6 hours, then became confluent at 12 hours, and until the 20th hour of culture, the cels in the two sides of the wound edge were completely fused to fuly close the wound edge, indicating a better wound healing in the 100 mg/L group than the other groups. These findings suggest that enamel matrix proteins can stimulate proliferation, viability, differentiation and migration of periodontal ligament cels.