临床和实验医学杂志
臨床和實驗醫學雜誌
림상화실험의학잡지
JOURNAL OF CLINICAL AND EXPERIMENTAL MEDICINE
2014年
24期
2009-2011
,共3页
余锦强%柯峰%李儒华%凌佼佼%华远峰%邵杰
餘錦彊%柯峰%李儒華%凌佼佼%華遠峰%邵傑
여금강%가봉%리유화%릉교교%화원봉%소걸
兔%骨髓间充质干细胞%移植%角膜缘干细胞
兔%骨髓間充質榦細胞%移植%角膜緣榦細胞
토%골수간충질간세포%이식%각막연간세포
Rabbits%Bone marrow mesenchymal stem cells%Transplantation%Corneal limbal stem cells
目的：探讨兔自体骨髓间充质干细胞（ BMSCs）移植对角膜缘干细胞（ LSCs）损伤的初步疗效。方法36只大白兔分为三组：BMSCs组、LSCs组与羊膜对照组，每组12只。BMSCs组采用附着有BMSCs的羊膜治疗角膜缘干细胞损伤，LSCs组将附着有角膜内皮细胞的羊膜治疗角膜缘干细胞损伤，羊膜对照组则仅将羊膜用缝合线固定于浅层巩膜上治疗角膜缘干细胞损伤。于术前、术后1周、术后2周、3周和4周对眼表的形态进行评分；并于术前、术后1周、4周、8周利用聚焦显微镜观察细胞密度和表面积。结果在移植手术实施之前三组兔的眼表形态综合评分之间的差异无统计学意义（ P ＞0.05）；手术之后4周BMSCs组和LSCs组兔的眼表形态评分得以显著改善，差异有统计学意义（ P＜0.05）；手术之后4周羊膜对照组兔的眼表形态评分与手术前相比，差异无统计学意义（ P ＞0.05）；手术之后4周BMSCs组和LSCs组之间的眼表形态评分之间差异无统计学意义（ P ＞0.05）。与羊膜对照组相比，其它两组手术4周之后的眼表形态评分改善更显著，差异有统计学意义（ P ＜0.05）。经过共焦显微镜观察，BMSCs组兔移植1周之后的细胞形态为圆形，而非初始的梭边形，细胞较小，密度较高，而随着时间推移，个数逐渐减少，体积变大。随着时间的推移BMSCs组与LSCs组兔移植后的平均细胞密度出现下降。手术前BMSCs组与LSCs组细胞表面积差异无统计学意义（ P ＞0.05）；手术后1周、4周BMSCs组的细胞表面积显著低于LSCs组，差异有统计学意义（ P ＜0.05），手术之后8周和12周时BMSCs组与LSCs组细胞表面积差异无统计学意义（ P ＞0.05）。结论与角膜缘干细胞相似，兔自体骨髓间充质干细胞移植治疗角膜缘干细胞可以重建受损的眼表，减轻水肿，抑制血管生成。
目的：探討兔自體骨髓間充質榦細胞（ BMSCs）移植對角膜緣榦細胞（ LSCs）損傷的初步療效。方法36隻大白兔分為三組：BMSCs組、LSCs組與羊膜對照組，每組12隻。BMSCs組採用附著有BMSCs的羊膜治療角膜緣榦細胞損傷，LSCs組將附著有角膜內皮細胞的羊膜治療角膜緣榦細胞損傷，羊膜對照組則僅將羊膜用縫閤線固定于淺層鞏膜上治療角膜緣榦細胞損傷。于術前、術後1週、術後2週、3週和4週對眼錶的形態進行評分；併于術前、術後1週、4週、8週利用聚焦顯微鏡觀察細胞密度和錶麵積。結果在移植手術實施之前三組兔的眼錶形態綜閤評分之間的差異無統計學意義（ P ＞0.05）；手術之後4週BMSCs組和LSCs組兔的眼錶形態評分得以顯著改善，差異有統計學意義（ P＜0.05）；手術之後4週羊膜對照組兔的眼錶形態評分與手術前相比，差異無統計學意義（ P ＞0.05）；手術之後4週BMSCs組和LSCs組之間的眼錶形態評分之間差異無統計學意義（ P ＞0.05）。與羊膜對照組相比，其它兩組手術4週之後的眼錶形態評分改善更顯著，差異有統計學意義（ P ＜0.05）。經過共焦顯微鏡觀察，BMSCs組兔移植1週之後的細胞形態為圓形，而非初始的梭邊形，細胞較小，密度較高，而隨著時間推移，箇數逐漸減少，體積變大。隨著時間的推移BMSCs組與LSCs組兔移植後的平均細胞密度齣現下降。手術前BMSCs組與LSCs組細胞錶麵積差異無統計學意義（ P ＞0.05）；手術後1週、4週BMSCs組的細胞錶麵積顯著低于LSCs組，差異有統計學意義（ P ＜0.05），手術之後8週和12週時BMSCs組與LSCs組細胞錶麵積差異無統計學意義（ P ＞0.05）。結論與角膜緣榦細胞相似，兔自體骨髓間充質榦細胞移植治療角膜緣榦細胞可以重建受損的眼錶，減輕水腫，抑製血管生成。
목적：탐토토자체골수간충질간세포（ BMSCs）이식대각막연간세포（ LSCs）손상적초보료효。방법36지대백토분위삼조：BMSCs조、LSCs조여양막대조조，매조12지。BMSCs조채용부착유BMSCs적양막치료각막연간세포손상，LSCs조장부착유각막내피세포적양막치료각막연간세포손상，양막대조조칙부장양막용봉합선고정우천층공막상치료각막연간세포손상。우술전、술후1주、술후2주、3주화4주대안표적형태진행평분；병우술전、술후1주、4주、8주이용취초현미경관찰세포밀도화표면적。결과재이식수술실시지전삼조토적안표형태종합평분지간적차이무통계학의의（ P ＞0.05）；수술지후4주BMSCs조화LSCs조토적안표형태평분득이현저개선，차이유통계학의의（ P＜0.05）；수술지후4주양막대조조토적안표형태평분여수술전상비，차이무통계학의의（ P ＞0.05）；수술지후4주BMSCs조화LSCs조지간적안표형태평분지간차이무통계학의의（ P ＞0.05）。여양막대조조상비，기타량조수술4주지후적안표형태평분개선경현저，차이유통계학의의（ P ＜0.05）。경과공초현미경관찰，BMSCs조토이식1주지후적세포형태위원형，이비초시적사변형，세포교소，밀도교고，이수착시간추이，개수축점감소，체적변대。수착시간적추이BMSCs조여LSCs조토이식후적평균세포밀도출현하강。수술전BMSCs조여LSCs조세포표면적차이무통계학의의（ P ＞0.05）；수술후1주、4주BMSCs조적세포표면적현저저우LSCs조，차이유통계학의의（ P ＜0.05），수술지후8주화12주시BMSCs조여LSCs조세포표면적차이무통계학의의（ P ＞0.05）。결론여각막연간세포상사，토자체골수간충질간세포이식치료각막연간세포가이중건수손적안표，감경수종，억제혈관생성。
Objective To explore the efficacy of transplantation in rabbits with autologous bone marrow mesenchymal stem cells( BMSCs) for treatment of injury of limbal stem cells( LSCs). Methods A total of 36 rabbits were divided into three groups:BMSCs group,LSCs group and amniotic membrane control group,12 rabbits in each group. Rabbits in BMSCs group were applied with BMSCs attached to amniotic membrane for treatment of corneal limbal stem cell damage,rabbits in LSCs group were applied with corneal endothelial cells attached to amniotic membrane for treatment of corneal limbal stem cell injury,and rabbits in amniotic membrane control group were only applied with amniotic membrane fixed on superficial sclera for treatment of corneal limbal stem cell injury with suture. Morphological scores on surface of eyes were recorded before opera-tion,1 week,2 weeks,3 weeks and 4 weeks after surgery,and ocular surface area and cell density were observed before operation,and 1 week, 4 weeks,8 weeks after surgery by using confocal microscopy. Results There was no statistically significant difference in ocular surface shape comprehensive scores between 3 groups of rabbits in results before implementation of transplantation operation( P >0. 05);rabbit ocular surface morphology scoring could be obviously improved in BMSCs group and LSCs group at 4 weeks after operation,there was statistically significant difference( P <0. 05);there was no significant difference in rabbit scores of control group with operation at 4 weeks after the amnion eye pheno-typic state compared with pre operation( P >0. 05);the difference in ocular surface morphology scores at 4 weeks after operation between BMSCs group and LSCs group had no statistical significance( P >0. 05). In comparison with amniotic membrane control group,the ocular surface mor-phology scores in other 2 groups had been improved more significantly,there was statistically significant difference,there was statistically signifi-cant difference( P <0. 05)at 4 weeks after operation. Through confocal microscopy,cell shape in rabbits of BMSCs group at 1 week after trans-plantation was round,rather than the initial shuttle edge shape,these cells were smaller,with higher density,but with the passage of time,their number had been gradually reduced. With the average cell density,time of BMSCs group and LSCs group rabbits had been decreased after trans-plantation. Surface area of cells in BMSCs group and LSCs group had no significant difference between before operation( P >0. 05);cell surface area at 1 week,4 weeks after operation in BMSCs group was lower than that of LSCs group( P <0. 05),there was no significant difference in cell surface area in BMSCs group and LSCs group in 8 and 12 weeks after operation( P >0. 05). Conclusion In comparison with limbal stem cells, autologous bone marrow mesenchymal stem cells transplantation in rabbits for treatment of corneal limbus stem cells can rebuild the ocular surface damage,alleviate edema and inhibit angiogenesis.
