本文選題：AZ31鎂合金 + 頸內(nèi)動脈支架��； 參考：《中國人民解放軍醫(yī)學(xué)院》2016年碩士論文

【摘要】：研究背景：鎂合金作為性能優(yōu)越的可降解合金被廣泛應(yīng)用于醫(yī)療內(nèi)置物材料,已在骨植入物、心血管支架方面進入臨床觀察階段,而這種鎂合金材料能否作為腦血管支架材料仍是未知數(shù).目前神經(jīng)介入治療中所應(yīng)用的永久性支架存在植入后對血管造成的機械性損傷、支架植入處再狹窄、且后續(xù)無法順利行MRI檢查等缺點,而可降解鎂合金支架的優(yōu)點在于可完全降解,能有效避免了永久性支架帶來的損傷。已知鎂合金在降解過程中會釋放Mg離子、Al離子、以及會造成局部酸堿度的改變,而這些改變對神經(jīng)系統(tǒng)是否會產(chǎn)生影響目前尚未見研究,也是鎂合金支架能否運用于腦血管內(nèi)重建治療的關(guān)鍵。本實驗就可降解鎂合金支架AZ31的降解對神經(jīng)細(xì)胞SY5Y的安全性進行研究.目的：明確鎂合金支架AZ31對神經(jīng)細(xì)胞SYSY生物學(xué)行為的影響。方法：本實驗采用間接法和直接法兩種評估方法。間接法：測定A1及Mg的濃度變化對SY5Y死亡率的影響.配制含不同濃度梯度MgCl2及AlCl3(如下表1、表2所示)的細(xì)胞培養(yǎng)液,用于培養(yǎng)SY5Y細(xì)胞,根據(jù)其MTT實驗結(jié)果,評估A1及Mg的濃度與SY5Y細(xì)胞死亡率間的關(guān)系.收集降解過程中的降解液,每3天檢測其中Al及Mg濃度,根據(jù)估測的安全濃度范圍,評估降解過程中所產(chǎn)生的Mg、Al濃度對神經(jīng)細(xì)胞是否存在細(xì)胞毒性。直接法：將收集所得降解液直接用于SY5Y細(xì)胞的培養(yǎng),為了確定降解液局部引起的pH值變化對細(xì)胞生存率是否會產(chǎn)生影響,將pH調(diào)節(jié)至正常范圍定為pH調(diào)節(jié)組,與未調(diào)節(jié)pH的降解液組對比。對照組：普通DMEM/F12培養(yǎng)液；實驗組1(未調(diào)節(jié)pH值)：每隔三日收集的降解液處理細(xì)胞組---第1d組,第4d組,第7d組...第88d組；實驗組2(調(diào)節(jié)pH值)：每隔三日收集的降解液調(diào)整pH值后處理細(xì)胞組--第1d組,第4d組,第7d組...第88d組；MTT法測定各組OD值,并計算細(xì)胞死亡率。結(jié)果間接法：SY5Y細(xì)胞的死亡率均隨著Mg濃度及A1濃度的增高而上升,并且,SY5Y細(xì)胞的死亡率與Mg離子濃度呈線性相關(guān)(P0.01).Mg：體外降解的過程中,約10%天數(shù)降解產(chǎn)生Mg濃度低于1.8mmol/L,細(xì)胞毒性為1級,其他時間可能產(chǎn)生2級細(xì)胞毒性.Al：測定得出A1在降解液中濃度極低,故可以認(rèn)為,AZ31鎂合金可降解支架在降解過程中產(chǎn)生的Al濃度范圍對SY5Y神經(jīng)細(xì)胞是安全的.直接法：pH未調(diào)節(jié)組細(xì)胞死亡率主要與pH值相關(guān)(P0.01),pH調(diào)節(jié)組死亡率與Mg濃度相關(guān)(P0.01),兩組細(xì)胞死亡率均與Al無關(guān).不考慮pH值的影響,降解過程中約有10天,Mg對SY5Y產(chǎn)生2級的細(xì)胞毒性。結(jié)論1.AZ31可降解鎂合金產(chǎn)生的降解產(chǎn)物Al對神經(jīng)細(xì)胞是安全的,不產(chǎn)生細(xì)胞毒性。2.AZ31可降解鎂合金產(chǎn)生的降解產(chǎn)物中Mg濃度偏高,對于神經(jīng)細(xì)胞存在2級毒性反應(yīng)。3.體外降解引起的pH值變化,對神經(jīng)細(xì)胞存在細(xì)胞毒性.4.實驗結(jié)果可以為鎂合金材料改良提供依據(jù),但不能絕對否定其體內(nèi)的應(yīng)用,仍需要體內(nèi)試驗進一步證實其安全性及可行性。
[Abstract]:Background: magnesium alloys, as degradable alloys with superior properties, have been widely used in medical implants, and have entered the clinical observation stage in bone implants and cardiovascular stents. But whether this magnesium alloy material can be used as cerebral vascular stent material is still unknown. At present, the permanent stent used in nerve interventional therapy has the disadvantages of mechanical injury to blood vessel after implantation, restenosis of stent implantation, and the failure of subsequent MRI examination. The advantage of degradable magnesium alloy scaffold is that it is completely degradable and can effectively avoid the damage caused by permanent scaffold. It is known that magnesium alloys release mg ion and Al ions during the degradation process and cause local pH changes, and whether these changes have any effect on the nervous system has not been studied. It is also the key whether magnesium alloy stents can be used in the treatment of cerebral vascular reconstruction. The safety of AZ31 degradation of degradable magnesium alloy scaffold on nerve cell SY5Y was studied in this experiment. Objective: to investigate the effect of magnesium alloy scaffold AZ31 on the biological behavior of nerve cell SYSY. Methods: indirect method and direct method were used in this experiment. Indirect method: the effects of changes of Al and mg concentrations on SY5Y mortality were determined. The cell culture medium containing different concentration gradient MgCl2 and ALCL 3 (as shown in table 1 and table 2) was used to culture SY5Y cells. The relationship between the concentration of A1 and mg and the death rate of SY5Y cells was evaluated according to the MTT results. The degradation solution was collected and the concentrations of Al and mg were detected every 3 days. According to the estimated safe concentration range, the toxicity of MgN Al concentration to nerve cells was evaluated. Direct method: the biodegradable solution collected was directly used in the culture of SY5Y cells. In order to determine whether the pH change caused by the degradation solution had an effect on cell survival, the pH was adjusted to a normal range. The results were compared with the unadjusted pH group. Control group: normal DMEM/F12 culture medium, experimental group 1 (unadjusted pH value: treated cell group every 3 days-group 1 d, group 4 d, group 7 d). Group 2 (adjusting pH value: treated cell group after adjusting pH value every 3 days-group 1 d, group 4 d, group 7 d). OD value was measured by MTT assay and cell death rate was calculated in 88 d group. Results the mortality of the cell line was increased with the increase of mg concentration and A1 concentration, and there was a linear correlation between the death rate of SY5Y cell and mg ion concentration in vitro. About 10% of the days of degradation resulted in mg concentration below 1.8 mmol / L, cytotoxicity of grade 1, other time may produce grade 2 cytotoxicity. Al: the concentration of A1 in the degradation solution was very low. It can be concluded that the concentration range of Al produced in the degradable scaffold of AZ31 magnesium alloy is safe for SY5Y nerve cells. The cell death rate in the unadjusted group was mainly related to the pH value (P 0.01) and mg concentration (P 0.01). The cell death rate was not related to Al in both groups. In the process of degradation, mg produced second-order cytotoxicity to SY5Y for about 10 days without taking into account the effect of pH value. Conclusion the degradation product Al produced by 1.AZ31 degradable magnesium alloy is safe to nerve cells. In vitro degradation caused by pH changes, there is cytotoxicity to nerve cells. 4. The experimental results can provide the basis for the improvement of magnesium alloy materials, but can not absolutely deny its application in vivo. It is still necessary to further verify the safety and feasibility of the in vivo test.
【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R318.08;R651.12

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