Shi Zhikang,Zhang Shengli,Zhao Minli,et al.Repair of urethral defects in rabbits using adipose stem cells and vascular endothelial cells co-cultured with acellular matrix materials[J].Journal of Clinical Pediatric Surgery,,19():310-316.[doi:10.3969/j.issn.1671-6353.2020.04.006]
Repair of urethral defects in rabbits using adipose stem cells and vascular endothelial cells co-cultured with acellular matrix materials
- Keywords:
- Tissue Engineering; Adipose Derived Stem Cells; Endothelial Cells; Coculture Techniques; acellular matrix; Urethra/IN
- CLC:
- R695;R318
- Abstract:
- Objective To co-culture adipose derived stem cells (ADSCs) and vascular endothelial cells (VEC) in vitro and add acellular matrix materials for reconstructing artificial urinary urethral defect.Methods Isolation,culture and amplification of rabbit ADSCs and VEC were performed.Respectively ADSCs,VEC and both were seeded to acellular matrix.A total of 48 male adult New Zealand rabbits were randomly divided into 4 groups with a length of urethral defect at 2.0 cm.Ccell-free control group was a simple application of acellular matrix; ADSCs:acellular matrix composite surgical repair; VEC:VEC acellular matrix surgical repair; ADSCs & VEC co-culturing group:ADSCs plus VEC co-cultured with acellular matrix composite.Urethral tissues were examined by histology and immunohistochemistry at 1,2,4 and 8 weeks.Urethral fistula and stricture were recorded at 8 weeks.Results ADSCs were fusiform shapes in vitro and cellular morphology of VEC was diverse and slabstone.Both were co-cultured for 7 days,some cells still retained original spindle and other cells were polygonal and nest-like.At 14 days,there were a lot of interconnected synapses and some of them fused into a mass.Urethra of each rabbit was observed at 4 and 8 weeks post-operation.The number of capillary and smooth muscle was higher in ADSCs & VEC co-culturing than that in Cell-free control/ADSCs/VEC group.Vascular endothelial growth factor (VEGF) and smooth muscle actin (SMA) were positive in ADSCs vs. VEC.After 8 weeks,the complications occurred in 9/12 cases (75%) in Cell-free,5/12 (41.7%) in ADSCs,4/12 (33.3%) in VEC and 2/12 (16.7%) in ADSCs & VEC.And the differences were statistically significant (χ2=8.914,P<0.05).Conclusion Cell-free and ADSCs & VEC,the difference was statistically significant (P<0.0083); comparing Cell-free & ADSCs,Cell-free & VEC,ADSCs & VEC,ADSCs & ADSCs & VEC and VEC & ADSCs & VEC,the differences were not statistically significant (P>0.0083).Conclusion After in vitro co-culturing,ADSCs and VEC may be used as seeds for tissue engineering urethra and repairing urethral defect.And VEC transforms ADSCs into epithelial cells and increases the occurrence of blood vessels in urethra.
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收稿日期:2018-05-04。
基金项目:河南省医学科技攻关计划项目（编号：201602055）；河南省医学科技攻关计划项目（编号：201203005）；河南省高等学校重点科研项目计划（编号：18A320046）
通讯作者:范应中,Email:fanyingzhong2@163.com