CAJ | 학술논문

目的探讨软骨组织工程方法中修复软骨缺损理想的种子细胞和支架材料. 方法免疫磁珠分离纯化新生大鼠前软骨干细胞(PSCs),分别采用KLD-12自组装肽纳米凝胶三维培养(实验组)和普通培养瓶平面培养(对照组),用转化生长因子β3(TGF-β_3)诱导两组PSCs向软骨细胞特性方向分化.利用RT-PCR、免疫组化等方法测定其向软骨细胞特性方向分化过程中特异性细胞外基质Ⅱ型胶原(collagen Ⅱ)和聚集蛋白聚糖(aggrecan)表达的情况. 结果 RT-PCR和免疫组化检测结果表明KLD-12自组装肽纳米凝胶组细胞在诱导7、14 d后均有collagen Ⅱ和aggrecan表达,且RT-PCR检测结果表明KLD-12自组装肽凝胶组细胞的collagen Ⅱ和aggrecan mRNA表达水平较对照组高,其差异有统计学意义(均P＜0.05). 结论 TGF-β_3诱导后的PSCs可向成软骨方向分化,诱导后的PSCs与KLD-12自组装肽凝胶复合有望构建组织工程化软骨,KLD-12自组装肽纳米凝胶是较好的软骨组织工程细胞支架.
목적 탐토연골조직공정방법 중수복연골결손이상적충자세포화지가재료. 방법 면역자주분리순화신생대서전연골간세포(PSCs),분별채용KLD-12자조장태납미응효삼유배양(실험조)화보통배양병평면배양(대조조),용전화생장인자β3(TGF-β_3)유도량조PSCs향연골세포특성방향분화.이용RT-PCR、면역조화등방법 측정기향연골세포특성방향분화과정중특이성세포외기질Ⅱ형효원(collagen Ⅱ)화취집단백취당(aggrecan)표체적정황. 결과 RT-PCR화면역조화검측결과표명KLD-12자조장태납미응효조세포재유도7、14 d후균유collagen Ⅱ화aggrecan표체,차RT-PCR검측결과표명KLD-12자조장태응효조세포적collagen Ⅱ화aggrecan mRNA표체수평교대조조고,기차이유통계학의의(균P＜0.05). 결론 TGF-β_3유도후적PSCs가향성연골방향분화,유도후적PSCs여KLD-12자조장태응효복합유망구건조직공정화연골,KLD-12자조장태납미응효시교호적연골조직공정세포지가.
Objective To evaluate the effects of implanted cell-scaffold composites in the repair of the cartilage defects.Methods Precartilaginous stem cells(PSCs)were isolated and purified by immunomagnetic beads.PSCs were seeded into self-assembling peptide KLD-12 hydrogel and culture flask,then induced into a chondrogenic pathway by TGF-β_3.The expression of cartilage-specific extracellular matrix Collagen Ⅱ and Aggrecan during chondrogenesis was detected by RT-PCR,and immunohistochemistry.Results Collagen Ⅱ and Aggrecan staining was positive at the 7th and 14th day in KLD-12 hydrogel culture.RT-PCR revealed that the expression of collagen Ⅱ and Aggrecan mRNA in KLD-12-PSCs was significantly higher than in the PSCs that seeded in culture flask(P＜0.05).Conclusion PSCs can be induced into chondrogenic differentiation by TGF-β_3.The self-assembling peptide KLD-12 hydrogel loaded with PSCs may be well used for cartilage tissue engineering,and self-assembling peptide KLD-12 hydrogel could be better cell-seeded scaffolds for repair of articular cartilage defects.