本文選題：羊皮 + 脫細(xì)胞真皮基質(zhì)��； 參考：《中南大學(xué)學(xué)報(醫(yī)學(xué)版)》2016年10期

【摘要】：目的:利用溶血反應(yīng)、熱原反應(yīng)和細(xì)胞毒性反應(yīng)實(shí)驗(yàn)了解羊脫細(xì)胞真皮基質(zhì)(acellular dermal matrix,ADM)在動物體內(nèi)外的毒性反應(yīng)強(qiáng)度。方法:溶血反應(yīng):分別制作交聯(lián)型和非交聯(lián)型羊ADM的浸提液,為交聯(lián)組和非交聯(lián)組;并設(shè)置陽性對照組10 m L滅菌注射用水,陰性對照組10 m L 0.9%氯化鈉溶液。將新鮮抗凝稀釋兔血加入實(shí)驗(yàn)組和對照組試管中,離心后取上清液測量吸光度值,計算溶血度。熱原實(shí)驗(yàn):按照條件入選16只新西蘭兔,分為初試A,B兩組各3只和復(fù)試C,D兩組各5只。分別將交聯(lián)與未交聯(lián)的兩種ADM浸提液注射入兩組新西蘭兔體內(nèi),注射后每隔0.5 h測量一次體溫,共測量6次,以6次中最高一次減去正常體溫即為該兔的體溫升高度數(shù)。據(jù)此判斷是否符合生物制品熱原實(shí)驗(yàn)的評價標(biāo)準(zhǔn)。MTT實(shí)驗(yàn):收集對數(shù)期的小鼠成纖維細(xì)胞細(xì)胞,種植于含有不同濃度羊ADM浸提液的培養(yǎng)基,測量吸光度值進(jìn)行細(xì)胞活力測定。結(jié)果:實(shí)驗(yàn)A,B組溶血度均小于5%,熱原反應(yīng)測得新西蘭兔體溫升高之和低于1.8℃。MTT實(shí)驗(yàn)顯示10%~90%羊ADM浸提液培養(yǎng)基對體外細(xì)胞的毒性為1級,100%的羊浸提液對體外細(xì)胞的毒性為2級,交聯(lián)型與非交聯(lián)型組的浸提液對體外細(xì)胞的毒性的無顯著性差異(P0.05)。均對體外細(xì)胞的增殖影響輕微。結(jié)論:羊ADM植入新西蘭兔體內(nèi)引起的溶血反應(yīng)和熱原反應(yīng)在評價標(biāo)準(zhǔn)之內(nèi),在體外對細(xì)胞的增殖和活力影響不明顯。
[Abstract]:Objective: to investigate the intensity of acellular dermal adm in vivo and in vitro by hemolytic reaction pyrogen reaction and cytotoxicity test. Methods: hemolysis reaction: the extract of cross-linked and non-cross-linked sheep ADM was made into cross-linked group and non-cross-linked group respectively, and 10 mL sterilizing water was set up in the positive control group and 0.9% sodium chloride solution in the negative control group. The fresh anticoagulant diluted rabbit blood was added to the test tube of the experimental group and the control group. The absorbance of the supernatant was measured after centrifugation and the hemolysis was calculated. Pyrogen test: sixteen New Zealand rabbits were selected according to the conditions and divided into two groups: group A (n = 3) and group C (n = 5). The cross-linked and uncrosslinked ADM extractions were injected into the two groups of New Zealand rabbits respectively. The body temperature was measured every 0.5 h after injection. According to the results, the mouse fibroblasts in logarithmic phase were collected and planted in the medium containing different concentrations of sheep ADM extract, and the cell viability was measured by measuring the absorbance value. Results: the hemolysis degree of group A and B was less than 5. The sum of the elevated body temperature of New Zealand rabbits was less than 1.8 鈩，

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