中华烧伤杂志
中華燒傷雜誌
중화소상잡지
16
2008年
2期
114-117
,共4页
顾华%何黎%刘玲%金以超
顧華%何黎%劉玲%金以超
고화%하려%류령%금이초
胶原%戊二醛%紫外线%成纤维细胞%共同培养技术
膠原%戊二醛%紫外線%成纖維細胞%共同培養技術
효원%무이철%자외선%성섬유세포%공동배양기술
Collagen%Glutaral%Ultraviolet rays%Fibroblast%Coculture techniques
目的 比较不同预冻温度及交联方法对胶原膜内部超微结构及成纤维细胞增殖的影响. 方法 将牛Ⅰ型胶原溶液(10 g/L)分别在-20℃和-80℃预冻12 h后,放入-70℃冻干机内冷冻干燥48 h.利用扫描电镜测量2种预冻温度下胶原膜的内部孔径,比较戊二醛交联法及紫外线+戊二醛双重交联法对胶原孔径的影响,通过噻唑蓝(MTT)法检测人成纤维细胞在不同交联法制备的胶原膜中的增殖情况. 结果 -20℃预冻胶原膜的孔径为(172±37)μm,-80℃预冻胶原膜孔径为(99±24)μm,选择后者进行后续实验.经戊二醛交联后胶原膜孔径缩小,种植后第8天,成纤维细胞的吸光度值为1.534±0.013;紫外线联合戊二醛双重交联后胶原膜原有孔径不变,种植后第8天成纤维细胞的吸光度值为3.778±0.010,与前者比较,差异有统计学意义(P<0.05). 结论 -80℃预冻后经紫外线+戊二醛双重交联法构建的胶原膜,可作为体外真皮支架替代物.
目的 比較不同預凍溫度及交聯方法對膠原膜內部超微結構及成纖維細胞增殖的影響. 方法 將牛Ⅰ型膠原溶液(10 g/L)分彆在-20℃和-80℃預凍12 h後,放入-70℃凍榦機內冷凍榦燥48 h.利用掃描電鏡測量2種預凍溫度下膠原膜的內部孔徑,比較戊二醛交聯法及紫外線+戊二醛雙重交聯法對膠原孔徑的影響,通過噻唑藍(MTT)法檢測人成纖維細胞在不同交聯法製備的膠原膜中的增殖情況. 結果 -20℃預凍膠原膜的孔徑為(172±37)μm,-80℃預凍膠原膜孔徑為(99±24)μm,選擇後者進行後續實驗.經戊二醛交聯後膠原膜孔徑縮小,種植後第8天,成纖維細胞的吸光度值為1.534±0.013;紫外線聯閤戊二醛雙重交聯後膠原膜原有孔徑不變,種植後第8天成纖維細胞的吸光度值為3.778±0.010,與前者比較,差異有統計學意義(P<0.05). 結論 -80℃預凍後經紫外線+戊二醛雙重交聯法構建的膠原膜,可作為體外真皮支架替代物.
목적 비교불동예동온도급교련방법대효원막내부초미결구급성섬유세포증식적영향. 방법 장우Ⅰ형효원용액(10 g/L)분별재-20℃화-80℃예동12 h후,방입-70℃동간궤내냉동간조48 h.이용소묘전경측량2충예동온도하효원막적내부공경,비교무이철교련법급자외선+무이철쌍중교련법대효원공경적영향,통과새서람(MTT)법검측인성섬유세포재불동교련법제비적효원막중적증식정황. 결과 -20℃예동효원막적공경위(172±37)μm,-80℃예동효원막공경위(99±24)μm,선택후자진행후속실험.경무이철교련후효원막공경축소,충식후제8천,성섬유세포적흡광도치위1.534±0.013;자외선연합무이철쌍중교련후효원막원유공경불변,충식후제8천성섬유세포적흡광도치위3.778±0.010,여전자비교,차이유통계학의의(P<0.05). 결론 -80℃예동후경자외선+무이철쌍중교련법구건적효원막,가작위체외진피지가체대물.
Objeetive To investigate the effects of preemptive freezing with different temDerature and cross-linking methods on the ultrastructure of collagen membrane and its influence on human fibroblast Droliferation. Methods Bovine collagen type Ⅰ solution in concentration of 10 g/L was preliminarilv frozen at -20℃or-80℃for 12 hours,and lyophilized at-70℃fnr 48 hours.The diameter of apertures in collagen membranes prepared with two different preliminary temperatures were observed by scanning electron microscope(SEM)and compared.The preliminary freezing temperature of-80℃was used tor the following study.The apertures of collagen membrane performed with cross-linking glutaraldehyde and uhraviolet (UV)radiation cross-linking with glutaraldehyde(double cross-linking)after preliminary freezing were also compared.The proliferation of human fibroblasts inoculated in above cross-linking collagens were assessed by MTT assay,in terms of absorption value. Results The mean diameter of apertures of collagen membrane pre-frozen at-20℃was(172±37)μm,while that at-80℃was(99±24)μm.The apertures of collagen membrane were reduced in size after glutaraldehyde cross-linking,while those of double cross-linking showed no change in size.There was obvious difierence in absorption value of fibroblasts 8 days after seeding between above two cross-linking methods(1.534±0.01 3 for glutaraldehyde cross-linking,3.778±0.010 for double cross-linking,P<0.05). Conclusion The collagen membrane after preliminary freezing at-80℃and double cross-linking with UV radiation and glutaraldehyde may be used as a dermal skeleton substitute.
