脂肪源性干细胞联合血管束植入对体内血管化组织工程支架的影响

[Impact of adipose-derived stem cells combined with vascular bundle implantation on vascularized tissue engineering scaffolds in vivo].

作者信息

Ji Wenchen, Yang Pei, Zhang Yuelin, Wang Chunsheng, Ni Jianlong, Zhang Yongtao, Wang Kunzheng

机构信息

No. 1 Department of Orthopedics, the Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an Shaanxi, 710004, PR China.

出版信息

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2012 Feb;26(2):129-34.

PMID:22403872

Abstract

OBJECTIVE

To discuss the impact of adipose-derived stem cells (ADSCs) combined with vascular bundle implantation on vascularized tissue engineering scaffolds in vivo so as to provide a theoretical basis for the repair of avascular necrosis of the femoral head.

METHODS

ADSCs were isolated from 4-month-old Sprague Dawley (SD) rats and cultured, then were induced to osteogenesis and identified. ADSCs at the 3rd passage were seeded on the nano-hydroxyapatite/polyamide-66 (nHA/PA66) to prepare the composite scaffolds. The compound condition of cells and scaffold materials were observed under scanning electronic microscope (SEM). Twenty-four 4-month-old SD rats (weighing 350-400 g) were randomly divided into 3 groups (n=8). In group A and group B, the inferior epigastric artery and vein of rats were implanted into composite scaffold cultured for 10 days or simple nHA/PA66 scaffold, respectively. In group C, two composite scaffolds cultured for 10 days were embedded into quadriceps femoris muscle of both thighs, respectively. After 2 and 4 weeks of operation, angiogenesis was observed by HE staining and CD34 immunohistochemical staining.

RESULTS

Cells isolated from adipose were identified as ADSCs. SEM showed that the number of cells increased after being cultured for 10 days, cell morphology stretched fully with a shape of long spindle. HE staining and immunohistochemical staining showed that a large number of angiogenesis was observed around the implanted artery and vein in group A, which was superior to groups B and C in the number of blood vessels and the maturity of blood vessel wall. After 2 and 4 weeks of operation, the blood vessel density and blood vessel diameter were significantly higher in group A than in group B and group C, and in group B than in group C (P < 0.05).

CONCLUSION

Combined application of ADSCs and vascular bundle implantation can promote the degree of vascularization, which could make the scaffold vascularization reliable.

摘要

目的

探讨脂肪来源干细胞（ADSCs）联合血管束植入对体内血管化组织工程支架的影响，为股骨头缺血性坏死的修复提供理论依据。

方法

从4月龄Sprague Dawley（SD）大鼠分离脂肪干细胞并培养，然后进行成骨诱导及鉴定。将第3代ADSCs接种于纳米羟基磷灰石/聚酰胺-66（nHA/PA66）上制备复合支架。在扫描电子显微镜（SEM）下观察细胞与支架材料的复合情况。将24只4月龄SD大鼠（体重350-400 g）随机分为3组（n=8）。A组和B组分别将大鼠腹壁下动静脉植入培养10天的复合支架或单纯nHA/PA66支架中。C组将两个培养10天的复合支架分别植入双侧大腿股四头肌内。术后2周和4周，通过HE染色和CD34免疫组织化学染色观察血管生成情况。

结果

从脂肪中分离的细胞鉴定为ADSCs。SEM显示，培养10天后细胞数量增加，细胞形态充分伸展呈长梭形。HE染色和免疫组织化学染色显示，A组植入的动静脉周围观察到大量血管生成，血管数量和血管壁成熟度均优于B组和C组。术后2周和4周，A组血管密度和血管直径明显高于B组和C组，B组高于C组（P<0.05）。