应力环境下三维诱导组织工程种植细胞修复关节软骨缺损

Chondrocytes Induced from Autologous Mesenchymal Stem Cell in Three-dimension(3D) Culture System with Dynamic Stress for Repairing Joint Cartilage Defects

目的 :三维立体诱导非软骨来源种植细胞修复关节软骨损伤。方法 :分离培养骨髓间充质干细胞 ,分组诱导 ,进行周期性应力刺激 ,设立对照。 2周后 ,观测大体、组织学形态、生化指标 ,筛选最佳诱导方法修复关节软骨缺损模型 ,分别于 12、 2 4周取材 ,进行组织学观察及评分 ,验证长期修复效果。结果 :与二维培养各组相反 ,立体培养各组出现明显软骨分化 ,并以凝胶微球悬浮细胞进行应力刺激培养组效果最佳。术后动物肢体功能优良 ,12周后软骨缺损被完整修复 ,表面光滑 ,质地坚硬 ,为透明软骨结构 ,与周围软骨结合紧密 ;2 4周后仍然保持透明软骨结构 ,组织学评分无变化。对照各组软骨缺损均未被修复。结论 :立体软骨诱导优于平面诱导 ,应力环境提高诱导质量 ,获得的种植细胞可取得稳定的长期修复效果。

Objective:To derive chondrocytes induced from autologous mesenchymal cell in 3D-culture system for repairing joint cartilage defects.Methods:Mesenchymal stem cells (MSC) were cultured from bone marrow aspirates of rabbits and then induced to cartilage tissue in several in vitro inducing group with cyclic dynamic stress for 2 weeks,and several control groups were also established.Histological examinations were adopted to confirm the repairing effects of induced chondrocytes in joint cartilage defects after 12 and 24 weeks.Results:Two-dimension culture showed a little chondrocyte differentiation,3D inducing experienced obvious chondrocyte differentiation,and the dynamic stress environment improved collagen II and proteoglycan production with the group of MSC in alginate drop as the best.The cartilage defects were smoothly repaired by hard hyaline-cartilage like tissue after 12 weeks,and showed sustained histological appearance until 24 weeks.No repairing was observed in control groups.Conclusions:3D culture inducing is more preferable than two-dimension ones;the dynamic stress environments facilitate the differentiation and could provide excellent non-cartilage derived plant cells for cartilage repairing.