CAJ | 학술논문

目的观察单抗CD44-生物素(Biotin) -亲和素(Avidin)绑定系统在构建组织工程软骨中能否提高软骨细胞与支架的黏附能力.方法分别制备软骨细胞二维和三维培养体系,分3组:A:壳聚糖+软骨细胞；B:生物素+亲和素化壳聚糖+软骨细胞；C:生物素化单抗CD44+亲和素化壳聚糖+软骨细胞.分别测定细胞展平面积、细胞接种脱落率、细胞增殖率,逆转录-聚合酶链反应(RT-PCR)法检测Ⅱ型胶原(ColⅡ)、聚集蛋白聚糖(aggrecan )、sox9的mRNA表达,组织学切片观察壳聚糖支架内细胞黏附生长情况.结果细胞展平面积、细胞增殖率及ColⅡ、aggrecan、sox9的mRNA表达量皆为:C组＞B组＞A组(P＜0.05).细胞接种脱落率:A组为C组的3.71倍、为B组的2.17倍,而B组为C组的1.71倍,A组＞B组＞C组(P＜0.05).组织切片显示细胞在支架内C组增殖旺盛、胞外基质表达较丰富,A组增殖较弱、胞外基质表达较弱,B组介于C与A组间.结论单抗CD44-Biotin-Avidin绑定系统能比Biotin-Avidin绑定系统更显著的提高软骨细胞与支架的黏附能力,促进组织工程软骨种子细胞的增殖和软骨细胞表型的表达.
목적 관찰단항CD44-생물소(Biotin) -친화소(Avidin)방정계통재구건조직공정연골중능부제고연골세포여지가적점부능력.방법 분별제비연골세포이유화삼유배양체계,분3조:A:각취당+연골세포；B:생물소+친화소화각취당+연골세포；C:생물소화단항CD44+친화소화각취당+연골세포.분별측정세포전평면적、세포접충탈락솔、세포증식솔,역전록-취합매련반응(RT-PCR)법검측Ⅱ형효원(ColⅡ)、취집단백취당(aggrecan )、sox9적mRNA표체,조직학절편관찰각취당지가내세포점부생장정황.결과 세포전평면적、세포증식솔급ColⅡ、aggrecan、sox9적mRNA표체량개위:C조＞B조＞A조(P＜0.05).세포접충탈락솔:A조위C조적3.71배、위B조적2.17배,이B조위C조적1.71배,A조＞B조＞C조(P＜0.05).조직절편현시세포재지가내C조증식왕성、포외기질표체교봉부,A조증식교약、포외기질표체교약,B조개우C여A조간.결론 단항CD44-Biotin-Avidin방정계통능비Biotin-Avidin방정계통경현저적제고연골세포여지가적점부능력,촉진조직공정연골충자세포적증식화연골세포표형적표체.
Objective To oberserve the efficiency of CD44 monoclonal antibody-biotin-avidin binding system for the improvement of cells adhesion to scaffolds in the cartilage tissue engineering.Methods The chondrocytes were cultured in two-dimensional and three-dimensional culture system respectively.And in each system the cells were divided into 3 groups:Group A,the chondrocytes were seeded routinely on the chitosan membrane or scaffolds ; Group B,the chondrocytes were seeded on the chitosan membrane or scaffolds with biotin-avidin binding system; Group C,the chondrocytes were seeded on the chitosan membrane or scaffolds with CD44 monoclonal antibody-biotin-avidin binding system.The spreading area in the two-dimensional culture system,cell exfoliation rate and cell proliferation rate in the three-dimensional culture system were investigated.The histologic analysis was used to evaluate the cell growth in the scaffolds.The mRNA expression level of type Ⅱ collagen,aggrecan and sox9 was detected by using reverse transcription-polymerase chain reaction (RT-PCR).Results Group C showed better results in the cell spreading area,the cell proliferation rate and the mRNA expression level of type Ⅱ collagen,aggrecan and sox9 than group B,and group B more than group A,group C ＞ group B ＞ group A ( P ＜ 0.05 ).The results showed that the cell exfoliation rate in group A was 2.17 times of that in group B which was 1.71 times of that in group C,group A ＞ group B ＞ group C ( P ＜ 0.05 ).The histologic sections showed the proliferation of cells was exuberant and expression of extracellular matrix was stronger in group C,that in group B was between group C and group A in which they were weaker.Conclusion The CD44 monoclonal antibody-biotin-avidin binding system could improve the chondrocytes adhesion,proliferation and phenotypic expression in cartilage tissue engineering.