解剖学报
解剖學報
해부학보
ACTA ANATOMICA SINICA
2014年
4期
561-568
,共8页
邵营宽%严夏霖%饶志恒%黄高建%李嘉威%黄俊杰%梅劲%林刻智
邵營寬%嚴夏霖%饒誌恆%黃高建%李嘉威%黃俊傑%梅勁%林刻智
소영관%엄하림%요지항%황고건%리가위%황준걸%매경%림각지
去细胞%胰腺%细胞外基质%生物相容性%组织工程%大鼠
去細胞%胰腺%細胞外基質%生物相容性%組織工程%大鼠
거세포%이선%세포외기질%생물상용성%조직공정%대서
Decellularization%Pancreas%Extracellular matrix%Biocompatibility%Tissue engineering%Rat
目的:通过离体灌注化学去垢剂的方法制备大鼠胰腺去细胞天然生物支架,并对支架的完整性、生物相容性进行检验。方法健康成年SD大鼠30只,分别经胆管与血管两条途径灌注十二烷基硫酸钠和曲亚通X-100等药品洗脱,获取胰腺去细胞生物支架。通过HE染色、扫描电子显微镜和透射电子显微镜等观察细胞残留于去细胞支架的生物学特性,分光光度计法鉴定残留去垢剂量,酶联免疫吸附剂测定法测定残留支架蛋白含量及生长因子,并用腹壁与皮下包埋实验检验其炎症反应,MTT法测定支架细胞毒性,内皮细胞共培养测定支架生物相容性。结果 HE染色、扫描电子显微镜和透射电子显微镜观察均表明去细胞支架无细胞残留,细胞外支架连续性完好,脉管支架保存完整;药物残留小于规定标准;细胞外支架生长因子及支架蛋白存留量等指标上血管组均优于胆管组;腹壁与皮下包埋实验显示,去细胞支架炎性反应较对照组明显减轻;MTT法显示无细胞毒性;内皮细胞共培养有黏附趋势。结论使用两种灌注法制备胰腺去细胞生物支架,均可将细胞去除彻底,支架生物相容性好。但就血管组细胞外支架保留完整性,生长因子保留更多,则血管灌注途径优于胆管灌注途径。
目的:通過離體灌註化學去垢劑的方法製備大鼠胰腺去細胞天然生物支架,併對支架的完整性、生物相容性進行檢驗。方法健康成年SD大鼠30隻,分彆經膽管與血管兩條途徑灌註十二烷基硫痠鈉和麯亞通X-100等藥品洗脫,穫取胰腺去細胞生物支架。通過HE染色、掃描電子顯微鏡和透射電子顯微鏡等觀察細胞殘留于去細胞支架的生物學特性,分光光度計法鑒定殘留去垢劑量,酶聯免疫吸附劑測定法測定殘留支架蛋白含量及生長因子,併用腹壁與皮下包埋實驗檢驗其炎癥反應,MTT法測定支架細胞毒性,內皮細胞共培養測定支架生物相容性。結果 HE染色、掃描電子顯微鏡和透射電子顯微鏡觀察均錶明去細胞支架無細胞殘留,細胞外支架連續性完好,脈管支架保存完整;藥物殘留小于規定標準;細胞外支架生長因子及支架蛋白存留量等指標上血管組均優于膽管組;腹壁與皮下包埋實驗顯示,去細胞支架炎性反應較對照組明顯減輕;MTT法顯示無細胞毒性;內皮細胞共培養有黏附趨勢。結論使用兩種灌註法製備胰腺去細胞生物支架,均可將細胞去除徹底,支架生物相容性好。但就血管組細胞外支架保留完整性,生長因子保留更多,則血管灌註途徑優于膽管灌註途徑。
목적:통과리체관주화학거구제적방법제비대서이선거세포천연생물지가,병대지가적완정성、생물상용성진행검험。방법건강성년SD대서30지,분별경담관여혈관량조도경관주십이완기류산납화곡아통X-100등약품세탈,획취이선거세포생물지가。통과HE염색、소묘전자현미경화투사전자현미경등관찰세포잔류우거세포지가적생물학특성,분광광도계법감정잔류거구제량,매련면역흡부제측정법측정잔류지가단백함량급생장인자,병용복벽여피하포매실험검험기염증반응,MTT법측정지가세포독성,내피세포공배양측정지가생물상용성。결과 HE염색、소묘전자현미경화투사전자현미경관찰균표명거세포지가무세포잔류,세포외지가련속성완호,맥관지가보존완정;약물잔류소우규정표준;세포외지가생장인자급지가단백존류량등지표상혈관조균우우담관조;복벽여피하포매실험현시,거세포지가염성반응교대조조명현감경;MTT법현시무세포독성;내피세포공배양유점부추세。결론사용량충관주법제비이선거세포생물지가,균가장세포거제철저,지가생물상용성호。단취혈관조세포외지가보류완정성,생장인자보류경다,칙혈관관주도경우우담관관주도경。
Objective To harvest pancreatic tissues from rats , prepare decellularized bio-derived pancreatic scaffolds ( DBPS) , and to examine the integrity and biocompatibility of the scaffolds .Methods Normal pancreases were harvested from healthy adult SD rats .DBPS was prepared by perfusing SDS and Triton X-100 through bile duct and the portal vein, respectively.After decellularization, normal pancreatic tissue and DBPS were compared via HE staining , and transmission electron microscopy ( TEM ) . Abdominal wall and subcutaneous implantations were used to compare biocompatibility , and the remain quantity of residual protein and growth factors were determined via enzyme linked immunosorbent assay(ELISA).MTT assay was used to test the scaffolds’ cytotoxicity.The scaffolds were co-cultured with endotheliocyte .Results HE staining and TEM study indicated no residual cells in the DBPS as well as preservation of the complete extracellular matrix .The remain quantity of residual protein and growth factors in ECM was high .The abdominal wall and subcutaneous implantation revealed that DBPS triggered a lower immune response as compared to the control group.MTT assay showed little cytotoxicity .Endotheliocyte assembled and growed with the scaffolds together .Conclusion DBPS are completely decellularized , and exhibit a higher level of biocompatibility in vivo.Using the way of vessels can make the integrity of extracellular matrix to be fully preserves and contain more growth factors .So using vessels way is better than bile duct .