摘要
目的将脂肪干细胞(adipose-derived stem cells,ADSCs)作为种子细胞,以透明质酸(hyaluronic acid,HA)作为载体制备复合物,观察分析其对放射性复合损伤创面愈合的影响。方法取10只近交系SD大鼠双侧腹股沟部脂肪组织,采用酶消化法分离培养ADSCs,取第6代细胞与HA(10 mg/mL)混合均匀,制备密度为5×106个/mL的ADSCs-HA复合物。取30只近交系SD大鼠,雌雄各半,随机分为3组(n=10)。首先在3组大鼠背部制备2 cm×2 cm的皮肤全层缺损创面并行20 Gy 60Co辐照,制备放射性复合损伤模型。模型制备后1周,创面彻底清创后,A组覆盖凡士林纱布,B组创面滴加HA 0.4 mL并覆盖凡士林纱布,C组创面滴加ADSCs-HA复合物0.4 mL并覆盖凡士林纱布。于治疗后观察各组创面情况,并于第1、2、3、4周取创面标本行HE染色观察微血管密度,免疫组织化学染色观察CD90阳性细胞的分布情况。结果 A组创面愈合缓慢,第4周时仍以肉芽组织为主;B组创面愈合较A组快,第4周时创面中央呈凹陷样,未完全愈合;C组创面第4周时完全愈合,无明显凹陷,边缘见增厚的表皮。组织学观察示,C组第1、2、3周微血管密度均高于A、B组(P<0.05),第3周时B组明显高于A组(P<0.05)。第4周时,C组与B组比较差异无统计学意义(P>0.05),但这两组仍明显高于A组(P<0.05)。免疫组织化学染色示各时间点A、B组创面均未见CD90阳性细胞,C组CD90阳性细胞随时间延长逐渐减少。结论 ADSCs-HA复合物通过促进并调控大鼠放射性复合损伤创面微血管的新生,加速创面修复。
Objective To observe and explore the effects of adipose-derived stem cells（ADSCs）-hyaluronic acid（HA） composite on healing of wound combined with radiation injury.Methods The ADSCs were harvested from the fat tissue in groin of 10 inbred Sprague Dawley（SD） rats and were isolated and cultured by enzyme digestion.The ADSCs-HA composite was prepared with ADSCs（5 × 106 cells/mL） at passage 6 and HA（10 mg/mL）.Thirty inbred SD rats,15 males and 15 females,were randomly divided into groups A（n=10）,B（n=10）,and C（n=10）.A 2 cm × 2 cm full-thickness skin defect was made on the rat back before 20 Gy 60Co radiation exposure.One week after debridement,wounds were treated by petrolatum gauze in group A as the control group,by HA（0.4 mL） and petrolatum gauze in group B,and by ADSCs-HA composite（0.4 mL） and petrolatum gauze in group C.The microvessel density（MVD） and the distribution of CD90 positive cells were observed at 1st,2nd,3rd,and 4th weeks.Results The wound healing was slower,and wound did not heal at 4th week and still filled with granulation tissue in group A;the wound healing of group B was faster than that of group A,and the wound did not heal completely with depression in the center at 4th week;the wound healed completely with epidermilizated surface and no obvious depression at 4th week in group C.The histological observation showed that MVD was significantly higher in group C than in groups A and B at the 1st,2nd,and 3rd weeks（P 0.05）,and in group B than in group A at the 3rd week（P 0.05）;MVD was significantly higher in groups B and C than in group A（P 0.05）,but no significant difference was found between groups B and C（P 0.05） at 4th week.No CD90 positive cell was found in groups A and B;CD 90 positive cells were observed in group C and gradually decreased with time.Conclusion ADSCs-HA composite can accelerate healing of wound combined with radiation injury by promoting and controlling wound angiogenesis.
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2011年第12期1499-1503,共5页
Chinese Journal of Reparative and Reconstructive Surgery
关键词
脂肪干细胞
透明质酸
放射性复合损伤
创面愈合
大鼠
Adipose-derived stem cells Hyaluronic acid Radiation combined injury Wound healing Rat
