雌激素缺乏对大鼠根尖牙乳头干细胞增殖及牙向/骨向分化能力的影响  

Effects of estrogen deficiency on the proliferation and odonto/osteogenic differentiation of rat stem cells from the apical papilla

作　　者：李瑶 曹灵[3] 夏涵[1] 周俊波 LI Yao;CAO Ling;XIA Han;ZHOU Junbo(Department of Stomatology,Nanjing Integrated Traditional Chinese and Western Medicine Hospital,Nanjing 210014,China;Jiangsu Province Key Laboratory of Oral Diseases,Nanjing Medical University,Nanjing 210029,China;Department of Pediatric and Preventive Dentistry,The Affiliated Stomatological Hospital of Nanjing Medical Universi-ty,Jiangsu Province Key Laboratory of Oral Diseases,Jiangsu Province Engineering Research Center of Stomatological Translational Medicine,Nanjing 210029,China)

机构地区：[1]南京市中西医结合医院口腔科,江苏南京210014 [2]南京医科大学江苏省口腔疾病研究重点实验室,江苏南京210029 [3]南京医科大学附属口腔医院儿童口腔预防科,江苏省口腔疾病研究重点实验室,江苏省口腔转化医学工程研究中心,江苏南京210029

出　　处：《口腔生物医学》2023年第3期156-161,共6页Oral Biomedicine

基　　金：国家自然科学基金(81873707,82170940)。

摘　　要：目的:探讨雌激素缺乏对大鼠根尖牙乳头干细胞(SCAPs)增殖及牙向/骨向分化能力的影响。方法:选取20只8周龄雌性SD大鼠随机分为假手术(Sham)组和卵巢去势(OVX)组,每组各10只。全麻下OVX组摘除双侧卵巢,Sham组行相同切口保留双侧卵巢,术前、术后1个月分别检测两组大鼠血清雌二醇水平。1个月后每组各处死5只大鼠,分离切牙根尖牙乳头组织,酶消化法分离培养两组SCAPs,采用MTT、碱性磷酸酶(ALP)活性检测、实时荧光定量PCR、Western blot检测雌激素缺乏对大鼠SCAPs增殖及成牙/成骨向分化能力的影响。将两组SCAPs细胞以明胶海绵为载体分别植入对应组别大鼠肾被膜下,8周后取出植入组织,使用数字化X线牙片机对取出团块曝光成像及HE染色观察其矿化能力。结果:OVX组大鼠去势1个月后血清雌二醇水平显著低于术前(P<0.001),Sham组大鼠血清雌二醇水平术前、术后差异无统计学意义(P>0.05)。MTT结果显示,OVX组SCAPs增殖能力显著下降(P<0.001)。ALP活性检测结果显示,OVX组SCAPs的ALP活性低于Sham组(P<0.001)。实时荧光定量PCR和Western blot结果均表明OVX组SCAPs的牙本质涎磷蛋白(DSPP)、成骨细胞特异性转录因子Osterix(OSX)、骨钙素(OCN)等成牙/成骨相关基因和蛋白表达低于Sham组(P<0.01)。体内移植结果显示,Sham组形成密度较高的较规则牙髓-牙本质复合体,而OVX组形成不规则结构。结论:雌激素缺乏减弱大鼠SCAPs的增殖及牙向/骨向分化能力。Objective:To evaluate the effects of estrogen deficiency on the proliferation and odonto/osteogenic differentiation of rat stem cells from the apical papilla(SCAPs).Methods:Twenty 8⁃week⁃old female SD rats were randomly divided into two groups,sham⁃operation(Sham)group and ovariectomized(OVX)group,10 rats in each group.All rats were anesthetized and both ovaries of OVX⁃rats were gently removed.Sham⁃operation rats were treated with the same incisions to expose the ovaries but without any hurt to them.Serum estradiol levels of OVX⁃rats and Sham⁃rats were detected before and 1 month after the operation.One month after the oper⁃ation,5 rats were killed in each group,the apical papillae of rat incisors were gently separated and SCAPs were isolated and incubated.Methyl thiazolyl tetrazolium(MTT)assay,alkaline phosphatase(ALP)activity,real⁃time reverse transcription PCR and Western blot assay were respectively used to determine the proliferation and odonto/osteogenic differentiation potential of rat SCAPs.Cell pellets of OVX⁃SCAPs and Sham⁃SCAPs were respectively transplanted into the renal capsules of immunodeficient female rats.After in vivo cul⁃ture for 8 weeks,the retrieved implants were processed for X⁃ray observation and hematoxylin and eosin(HE)staining.Results:Ser⁃um estradiol levels of OVX⁃rats significantly decreased 1 month after ovariectomy compared with those before the operation(P<0.001),while serum estradiol levels of Sham⁃rats had no obvious changes before and after the operation(P>0.05).MTT results showed that OVX⁃SCAPs exhibited the decreased proliferative activity as compared with Sham⁃SCAPs(P<0.001).ALP activity of OVX⁃SCAPs was lower than that of Sham⁃SCAPs(P<0.001).Real⁃time reverse transcription PCR and Western blot results showed that the expression of odonto/osteogenic markers dentin sialophosphoprotein(DSPP),Osterix(OSX),osteocalcin(OCN)in OVX⁃SCAPs was significantly inhibited as compared with Sham⁃SCAPs(P<0.01).Sham⁃SCAPs implants showed the typi

关 键 词：雌激素 根尖牙乳头干细胞 增殖 牙向/骨向分化 

分 类 号：R781[医药卫生—口腔医学]