發(fā)布時間：2018-04-14 17:07

  本文選題：柚皮苷 + MC3T3-E1　； 參考：《重慶醫(yī)科大學》2017年碩士論文

【摘要】：骨組織工程中運用各種生物因子及支架研究骨代謝過程中的骨形成及骨吸收成為當下科學研究及臨床關(guān)注的重點。鑒于現(xiàn)有臨床藥物在骨改建過程中促骨形成時的副作用,植物源性促骨形成藥物成為當下科研焦點,然而研究藥物對單一離體細胞分化成骨作用并不能為藥物的臨床使用提供有力的證據(jù)。本實驗擬研究骨碎補主要活性物質(zhì)-柚皮苷對成骨細胞系MC3T3-E1的促骨形成作用,同時選擇藥物與顱頂骨器官建立共培養(yǎng)模型,運用Micro-CT掃描技術(shù)分析藥物對系列骨細胞(骨細胞、成骨細胞及破骨細胞)、骨基質(zhì)等共存下的顱頂骨器官骨形成的作用。目的:1.在分析區(qū)域包含整個顱頂骨的前提下,改變Micro-CT掃描后二維圖像的重建層數(shù)或者位置,比較進行重建后的骨量參數(shù)(骨表面積、骨體積)是否出現(xiàn)改變。2.分析柚皮苷對成骨細胞系MC3T3-E1和顱頂骨骨形成的作用。方法:1.Micro-CT掃描4日齡CD-1小鼠顱頂骨,三維重建時改變空白分析區(qū)域?qū)訑?shù)或位置來分別分析:(1)996~22層組與800~50層組,分析層數(shù)不同;(2)996~22層組與隨機選擇層組,分析層數(shù)不同;(3)近邊界上緣組與近邊界下緣組,分析層數(shù)相同的骨量參數(shù)變化。2.不同濃度柚皮苷作用于MC3T3-E1后通過CCK-8法檢測細胞生長、定量分析細胞分泌蛋白ALP、RT-PCR法檢測MC3T3-E1細胞RUNX2、OCN和OPN基因m RNA表達量、茜素紅染色柚皮苷作用下細胞礦化結(jié)節(jié)形成。3.不同濃度柚皮苷作用于顱頂骨后通過Micro-CT掃描分析顱頂骨骨量參數(shù)(骨表面積、骨體積)的變化。結(jié)果:1.在分析區(qū)域包含整個顱頂骨的前提下:(1)增加或者減少空白分析層數(shù)會導致顱頂骨骨表面積變化,而骨體積無變化。(2)隨機選擇層數(shù)組與996~22層數(shù)組分析比較時顱頂骨骨表面積、骨體積均有變化。(3)重建層數(shù)相同的前提下,改變分析區(qū)域位置時顱頂骨骨表面積、骨體積無變化。2.不同濃度柚皮苷(0mg/L、1mg/L、10mg/L)均可促進MC3T3-E1的RUNX2、OCN、OPN表達上調(diào)及細胞礦化結(jié)節(jié)形成,但1mg/L濃度柚皮苷促成骨作用較10 mg/L組顯著。3.不同濃度柚皮苷作用的離體培養(yǎng)顱頂骨骨表面積和骨體積無明顯變化。結(jié)論:1.柚皮苷可促體外培養(yǎng)MC3T3-E1細胞成骨向分化,而對體外培養(yǎng)顱頂骨骨形成無明顯作用。2.Micro-CT三維重建時,改變重建層數(shù)可影響骨量參數(shù),而保證分析層數(shù)相同僅改變空白分析區(qū)域的位置不會影響骨量參數(shù)。
[Abstract]:The use of various biological factors and scaffolds in bone tissue engineering to study bone formation and bone resorption during bone metabolism has become the focus of current scientific and clinical research.In view of the side effects of existing clinical drugs in promoting bone formation in the process of bone remodeling, plant-derived osteogenic drugs have become the focus of current scientific research.However, the study of the effect of drugs on the differentiation and osteogenesis of in vitro cells does not provide strong evidence for the clinical use of drugs.The aim of this study was to study the effect of naringin, the main active substance of osteoclastic, on osteogenesis of osteoblast MC3T3-E1. At the same time, the drug was selected to establish coculture model with cranio-parietal organs. Micro-CT scanning technique was used to analyze the effects of naringin on osteoblasts (bone cells).The role of osteoblasts and osteoclasts and bone matrix in the formation of bone in parietal organs.Purpose 1.Under the condition that the region contains the whole cranio-parietal bone, the number or location of the reconstructed layers in the two-dimensional image after Micro-CT scanning is changed, and the bone mass parameters (bone surface area, bone volume) after reconstruction are compared whether the bone mass parameters (bone surface area, bone volume) change. 2.To investigate the effects of naringin on osteoblast MC3T3-E1 and parietal bone formation.Methods: 1. Micro-CT scanning of the parietal bone of 4-day-old CD-1 mice was performed. The number or position of the blank analysis area was changed during 3D reconstruction to analyze the two groups, the two groups were divided into two groups, one was divided into two groups, one was divided into two groups, the other was divided into two groups, one was divided into two groups, the other was divided into two groups, one was divided into two groups, the other was divided into two groups.The changes of bone mass parameters of the upper and lower boundary groups with the same layer number were analyzed.After different concentrations of naringin was treated with MC3T3-E1, cell growth was detected by CCK-8 assay, and the expression of RUNX2OCN and OPN gene m RNA in MC3T3-E1 cells was detected by RT-PCR. The mineralized nodule was formed by alizarin red staining with naringin.The changes of bone mass parameters (bone surface area and bone volume) of parietal bone were analyzed by Micro-CT scanning after different concentrations of naringin were applied to parietal bone.The result is 1: 1.(1) increasing or decreasing the number of blank analytical layers will result in a change in the surface area of the parietal bone, while the bone volume does not change. 2) the surface area of the parietal bone is compared with that of the random array of selected layers compared with the array of 996 ~ 22 layers.On the premise of the same number of reconstructed layers, the surface area of cranio-parietal bone was changed and the volume of bone did not change. 2.Different concentrations of naringin (0 mg / L ~ (-1) mg / L ~ (10) mg / L) could promote the up-regulation of RUNX2 ~ (2) MC3T3-E1 and the formation of mineralized nodule in MC3T3-E1. However, the effect of naringin at 1mg/L concentration on bone formation was significantly higher than that of 10 mg/L group.There was no significant change in surface area and bone volume of in vitro cultured parietal bone with different concentrations of naringin.Conclusion 1.Naringin could promote the osteogenic differentiation of cultured MC3T3-E1 cells, but had no obvious effect on the formation of cranio-parietal bone in vitro .2.When the three-dimensional reconstruction of Micro-CT, changing the number of reconstructed layers could influence the bone mass parameters.But ensuring that the same number of analysis layers only changes the position of blank analysis area will not affect the bone mass parameters.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R782

【參考文獻】

相關(guān)期刊論文 前9條

1 王法琪;嚴亞波;溫鑫鑫;徐超;吳子祥;雷偉;;顯微CT分辨率對松質(zhì)骨微觀結(jié)構(gòu)及生物力學測量的影響[J];現(xiàn)代生物醫(yī)學進展;2015年10期

2 李念虎;徐展望;;柚皮苷促進成骨細胞分化并有效改善卵巢切除所致的骨質(zhì)疏松[J];中國骨質(zhì)疏松雜志;2013年08期

3 陶劍鋒;趙丹慧;吳成愛;綦慧;王宏剛;王娜;;顯微CT旋轉(zhuǎn)步長對牛腰椎松質(zhì)骨重建圖像及數(shù)據(jù)的影響[J];中國脊柱脊髓雜志;2012年09期

4 劉廣鵬;曹誼林;;顯微CT在口腔醫(yī)學研究中的應用[J];上�？谇会t(yī)學;2007年03期

5 郭惠蘭,張曉,揭新明;IL-6/IL-6受體mRNA反義寡核苷酸抑制骨吸收的實驗研究[J];中國老年學雜志;2003年12期

6 李強,史俊南,王鑫源,吳織芬,孫葉芳;白細胞介素1β對培養(yǎng)骨器官鈣代謝的作用[J];牙體牙髓牙周病學雜志;2001年03期

7 劉開泰,王國荃,汪師貞,吳繼堯,肖碧玉;氟對體外骨器官培養(yǎng)長骨細胞周期和細胞凋亡的影響[J];地方病通報;1999年04期

8 鄧廉夫,徐榮輝,柴本甫,齊進;應用趨骨熒光素標記體外培養(yǎng)骨組織[J];臨床與實驗病理學雜志;1999年01期

9 張煒真,于世鳳,鄭麟蕃;用作骨吸收研究的小鼠顱蓋骨器官培養(yǎng)方法[J];中華口腔醫(yī)學雜志;1996年01期

，

本文編號：1750243