医药前沿
醫藥前沿
의약전연
YIAYAO QIANYAN
2014年
12期
45-47
,共3页
宿学家%张慧颖%易定华%魏旭峰%张志勇
宿學傢%張慧穎%易定華%魏旭峰%張誌勇
숙학가%장혜영%역정화%위욱봉%장지용
组织工程瓣%去细胞%钙化%环氧氯丙烷
組織工程瓣%去細胞%鈣化%環氧氯丙烷
조직공정판%거세포%개화%배양록병완
tissue engineering valve%aceelular%calcification%epichlorohydrin
目的：观察去细胞过程及环氧氯丙烷预处理（EC）对猪主动脉瓣纤维结构的影响，探讨EC对去细胞猪主动脉瓣钙化的抑制作用。方法实验分为三组：新鲜瓣叶组（F组）：新鲜猪主动脉瓣未作任何处理。去细胞瓣叶（A组）：以胰酶消化，TrintonX100漂洗法去除瓣叶的细胞成分。EC预处理去细胞瓣叶（E组）：以2％的EC /PBS溶液预处理去细胞猪主动脉瓣48h。以电子万能拉力计测定各组瓣叶的断裂强度。傅立叶变换红外光谱仪测定各组瓣叶的游离羧基、酰胺基团变化。原子吸收法测定SD幼年大鼠皮下埋藏8周后钙含量，von kossa染色观察钙沉积。结果1、A组的断裂强度与F组比较没有明显的差异（619±98.9 g/mm2vs632±89.8 g/mm2 p＞0.05），E组较F组稍有增加（799±37.4g/mm2 vs 619±98.9g/mm2 p＜0.05）。2、红外光谱图显示：F组瓣叶在1700~1750cm-1波长范围（游离羧基的特征性吸收光谱）内没有明显的吸收峰，A组吸收峰深大，而E组又有明显的回落。在1600～1650cm-1范围内（酰胺基的特征性光谱） F组吸收峰明显，A组吸收峰回落，E组吸收峰介入两者之间且较前两者均左移。3、体内埋藏2周后各组瓣叶钙含量开始增加，到达第六周时，新鲜瓣膜组钙含量有显著增加。去细胞瓣膜组增加缓慢，而经EC处理去细胞瓣膜组钙含量始终维持较低水平。4、von kossa染色显示：F组瓣叶被破坏吸收，未见有新的细胞生长，有大量钙化斑存在。A组及E组瓣叶均可见大量的新细胞生长，前者仅出现散在的微小钙化灶，后者未见有明显的钙化灶。结论去细胞后瓣叶的机械强度没有明显减低，游离羧基有明显的暴露，酰胺基团减少。经EC预处理可封闭游离羧基，增加酰胺基团含量，使瓣叶结构紧密，增加去细胞瓣叶的机械强度，具有减缓钙化的作用。
目的：觀察去細胞過程及環氧氯丙烷預處理（EC）對豬主動脈瓣纖維結構的影響，探討EC對去細胞豬主動脈瓣鈣化的抑製作用。方法實驗分為三組：新鮮瓣葉組（F組）：新鮮豬主動脈瓣未作任何處理。去細胞瓣葉（A組）：以胰酶消化，TrintonX100漂洗法去除瓣葉的細胞成分。EC預處理去細胞瓣葉（E組）：以2％的EC /PBS溶液預處理去細胞豬主動脈瓣48h。以電子萬能拉力計測定各組瓣葉的斷裂彊度。傅立葉變換紅外光譜儀測定各組瓣葉的遊離羧基、酰胺基糰變化。原子吸收法測定SD幼年大鼠皮下埋藏8週後鈣含量，von kossa染色觀察鈣沉積。結果1、A組的斷裂彊度與F組比較沒有明顯的差異（619±98.9 g/mm2vs632±89.8 g/mm2 p＞0.05），E組較F組稍有增加（799±37.4g/mm2 vs 619±98.9g/mm2 p＜0.05）。2、紅外光譜圖顯示：F組瓣葉在1700~1750cm-1波長範圍（遊離羧基的特徵性吸收光譜）內沒有明顯的吸收峰，A組吸收峰深大，而E組又有明顯的迴落。在1600～1650cm-1範圍內（酰胺基的特徵性光譜） F組吸收峰明顯，A組吸收峰迴落，E組吸收峰介入兩者之間且較前兩者均左移。3、體內埋藏2週後各組瓣葉鈣含量開始增加，到達第六週時，新鮮瓣膜組鈣含量有顯著增加。去細胞瓣膜組增加緩慢，而經EC處理去細胞瓣膜組鈣含量始終維持較低水平。4、von kossa染色顯示：F組瓣葉被破壞吸收，未見有新的細胞生長，有大量鈣化斑存在。A組及E組瓣葉均可見大量的新細胞生長，前者僅齣現散在的微小鈣化竈，後者未見有明顯的鈣化竈。結論去細胞後瓣葉的機械彊度沒有明顯減低，遊離羧基有明顯的暴露，酰胺基糰減少。經EC預處理可封閉遊離羧基，增加酰胺基糰含量，使瓣葉結構緊密，增加去細胞瓣葉的機械彊度，具有減緩鈣化的作用。
목적：관찰거세포과정급배양록병완예처리（EC）대저주동맥판섬유결구적영향，탐토EC대거세포저주동맥판개화적억제작용。방법실험분위삼조：신선판협조（F조）：신선저주동맥판미작임하처리。거세포판협（A조）：이이매소화，TrintonX100표세법거제판협적세포성분。EC예처리거세포판협（E조）：이2％적EC /PBS용액예처리거세포저주동맥판48h。이전자만능랍력계측정각조판협적단렬강도。부립협변환홍외광보의측정각조판협적유리최기、선알기단변화。원자흡수법측정SD유년대서피하매장8주후개함량，von kossa염색관찰개침적。결과1、A조적단렬강도여F조비교몰유명현적차이（619±98.9 g/mm2vs632±89.8 g/mm2 p＞0.05），E조교F조초유증가（799±37.4g/mm2 vs 619±98.9g/mm2 p＜0.05）。2、홍외광보도현시：F조판협재1700~1750cm-1파장범위（유리최기적특정성흡수광보）내몰유명현적흡수봉，A조흡수봉심대，이E조우유명현적회락。재1600～1650cm-1범위내（선알기적특정성광보） F조흡수봉명현，A조흡수봉회락，E조흡수봉개입량자지간차교전량자균좌이。3、체내매장2주후각조판협개함량개시증가，도체제륙주시，신선판막조개함량유현저증가。거세포판막조증가완만，이경EC처리거세포판막조개함량시종유지교저수평。4、von kossa염색현시：F조판협피파배흡수，미견유신적세포생장，유대량개화반존재。A조급E조판협균가견대량적신세포생장，전자부출현산재적미소개화조，후자미견유명현적개화조。결론거세포후판협적궤계강도몰유명현감저，유리최기유명현적폭로，선알기단감소。경EC예처리가봉폐유리최기，증가선알기단함량，사판협결구긴밀，증가거세포판협적궤계강도，구유감완개화적작용。
AIM: To observe the effect of acel ular and pretreated with epichlorohydrin(EC) on structure of canine aortic valve fibrinous scaffold and investigate the anti-cailcification of EC.METHODS:This study was divided three groups:Fresh valve group(F),the valve was not treated.Acel ular valve group(A),the valve were acel ulared by trypsogen and Triton X100. Acel ular valve pretreated with EC group(E),acel ular valve was pretreated with EC/PBS 48h.Fracture tension was mensured by omnipotent pul dynamometer. Free carboxyl and acyl-amine wre tested by Fourier infraed spectrometer.Calcific content of valve after implanted subdermal was mensured by atomic absorbtion pectroscopy.Calcifical deposition was observed by von kossa staining. RESULTS:1.The fracture tension of A group was not different from F group(619±98.9 g/mm2 vs 632±89.8 g/mm2 p>0.05).In E group,the fractrure tension was stronger than A group(799±37.4g/mm2vs 619±98.9g/mm2 p<0.05).2.On infrared spectrum,no absorb peak appeared in wavelength of 1700cm-1 to 1750cm-1 which was eigenvalue of free carboxyl,while one absorb peak appeared significantly in A group.In E group ,the absorb peak in this wavelength was lower than A group.In wavelength of 1600cm-1 to 1650cm-1 which was eigencalue of acyl-amine,the absorb peak of F group was deeper than A group while that of E group was between. A and F groups,and the position of absorb peak in E group was left-moved than A and F group.3. Calcific cintent of every group valves was increased begin 2 weeks after implanted.In F group, it was increased distinctly which was increased slowly.But,in E group the calcifical cintent was maintained lower level.4.In von kossa staining,F group valve was destroyed and absorbed and none of new cel s grew into it.A mass of cel s grew into A and E group valve.Disperse microcalcifical nods were found in A valve bu not in E valve.CONCLUSION: The machinical tension was not decreased after acel ular,but the free carboxyl exposed and acyl-amine decreased.After pretreated by EC,the machinical tension of valve higher than fresh and acel ular valve,free carboxyl was blocked out and acyl-amine increased while structure of E valve was tighter than A and F valve.
