CAJ | 학술논문

背景：以往研究用来诱导成牙本质细胞分化的培养基普遍借鉴成骨细胞的诱导浓度，其他浓度的诱导情况尚无相关研究。目的：观察在低浓度β-甘油磷酸钠条件培养下，牙髓干细胞向成牙本质细胞分化过程中，牙本质基质蛋白1、牙本质涎蛋白和细胞外基质磷酸糖蛋白的表达情况。方法：分离培养人牙髓干细胞，用不同浓度的诱导液诱导牙髓干细胞分别向脂肪细胞，成骨细胞分化，证实其多向分化能力。用5 mmol/Lβ-甘油磷酸钠诱导培养牙髓干细胞向牙本质细胞分化，分别在培养第7，14，21，28天提取各组细胞RNA，反转录PCR检测牙本质基质蛋白1、牙本质涎蛋白和细胞外基质磷酸糖蛋白的表达情况；诱导牙髓干细胞向成牙本质细胞分化形成的矿化结节用Alizarin Red S法检测。结果与结论：人牙髓干细胞经过诱导可证实其成功分化为脂肪细胞和成骨细胞；反转录 PCR 结果显示，5 mmol/Lβ-甘油磷酸钠培养牙髓干细胞在培养7，14，21 d，各组细胞牙本质基质蛋白1、牙本质涎蛋白的mRNA表达增加，伴随细胞外基质磷酸糖蛋白表达下调；培养至28 d，行矿化结节检测发现牙髓干细胞向成牙本质细胞成功矿化，可见红染的矿化结节。结果证实，5 mmol/Lβ-甘油磷酸钠诱导培养的牙髓干细胞可成功分化为成牙本质细胞，并下调细胞外基质磷酸糖蛋白mRNA的表达，同时上调牙本质基质蛋白1及牙本质涎蛋白mRNA的表达。
배경：이왕연구용래유도성아본질세포분화적배양기보편차감성골세포적유도농도，기타농도적유도정황상무상관연구。목적：관찰재저농도β-감유린산납조건배양하，아수간세포향성아본질세포분화과정중，아본질기질단백1、아본질연단백화세포외기질린산당단백적표체정황。방법：분리배양인아수간세포，용불동농도적유도액유도아수간세포분별향지방세포，성골세포분화，증실기다향분화능력。용5 mmol/Lβ-감유린산납유도배양아수간세포향아본질세포분화，분별재배양제7，14，21，28천제취각조세포RNA，반전록PCR검측아본질기질단백1、아본질연단백화세포외기질린산당단백적표체정황；유도아수간세포향성아본질세포분화형성적광화결절용Alizarin Red S법검측。결과여결론：인아수간세포경과유도가증실기성공분화위지방세포화성골세포；반전록 PCR 결과현시，5 mmol/Lβ-감유린산납배양아수간세포재배양7，14，21 d，각조세포아본질기질단백1、아본질연단백적mRNA표체증가，반수세포외기질린산당단백표체하조；배양지28 d，행광화결절검측발현아수간세포향성아본질세포성공광화，가견홍염적광화결절。결과증실，5 mmol/Lβ-감유린산납유도배양적아수간세포가성공분화위성아본질세포，병하조세포외기질린산당단백mRNA적표체，동시상조아본질기질단백1급아본질연단백mRNA적표체。
BACKGROUND:The induced concentration for osteoblasts is often introduced as reference to induce odontoblast differentiation. However, there are no reports on other concentrations. OBJECTIVE: To observe the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein during low-dose β-glycerophosphate-induced differentiation of dental pulp stem cels into odontoblasts. METHODS:Human dental pulp stem cels were isolated and cultured, and then induced by different concentrations of inducing solution to differentiate into adipocytes and osteoblasts, which could verify the multi-directional differentiation ability of human dental pulp stem cels. Under 5 mmol/L β-glycerophosphate, dental pulp stem cels differentiated into odontoblasts. At 7, 14, 21, 28 days of culture, RNA samples were extracted from dental pulp stem cels in each group, and reverse-transcription PCR was used to detect the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein. Mineralized nodules were detected by alizarin red S staining to show the successfuly osteogenesis induction. RESULTS AND CONCLUSION: Human dental pulp stem cels could be induced to adipocytes and osteoblasts. The results of reverse-transcription PCR showed that the dental pulp stem cels under 5 mmol/L β-glycerophosphate could increase the expression of dentin matrix protein-1 and dentin sialoprotein, but downregulate the expression of matrix extracelular phosphoglycoprotein at 7, 14, 21 days. At 28 days of culture, dental pulp stem cels were al successfuly mineralized detected by alizarin red S. There were some red mineralized nodules. These findings indicate that the 5 mmol/L β-glycerophosphate can induce the differentiation of dental pulp stem cels into odontoblasts successfuly, up-regulate the mRNA expression of dentin sialoprotein and dentin matrix protein-1, and meanwhile down-regulate the mRNA expression of matrix extracelular phosphoglycoprotein.