目的 探讨改良脱细胞神经支架复合脂肪干细胞(ADSCs)在裸鼠体内异位构建组织工程化周围神经的可行性,并采用PKH26荧光染料标记及小动物活体荧光成像系统无创性地评估组织工程化周围神经在裸鼠体内的生长.方法 将标记PKH26荧光染料的ADSCs接种于课题组改良制备的脱细胞神经支架中,体外培养1周后植入裸鼠背部皮下,4周后利用小动物活体荧光成像系统对其进行观察;之后处死裸鼠,取材并进行冰冻切片荧光显微镜下观察及组织学检测;分析制备的组织工程化周围神经在裸鼠体内的生长情况.结果 植入物在裸鼠体内培养4周后,小动物活体荧光成像系统观察显示裸鼠背部植入部位可见条状强荧光,表明组织工程化周围神经在裸鼠体内生长情况良好.处死裸鼠后冰冻切片,荧光显微镜下观察可见组织工程化周围神经中细胞均呈红色荧光,表明细胞存活情况良好;组织学苏木素-伊红(HE)染色可见神经样组织形成,且其周围有血管包绕.结论 改良脱细胞神经支架复合脂肪干细胞能够在裸鼠皮下异位构建组织工程化周围神经;PKH26荧光染料标记及小动物活体荧光成像系统能够无创性地评估组织工程化周围神经在体内的生长.
Objective To evaluate the feasibility of ectopic construction of tissue-engineered peripheral nerve in nude mice based on improved acellular nerve scaffold and PKH26-labeled adipose derived-stem cells (ADSCs),and to access the tissue-engineered nerve by small animals living fluorescence imaging system.Methods ADSCs were labeled with PKH-26 and seeded into improved acellular nerve scaffold.The constructs were cultured for 1 week in vitro.Then they were implanted into the dorsal pocket of the nude mice.After cultured for 4 weeks,the constructs were observed by small animals living fluorescence imaging system.After that,the nude mice were executed,the constructs were take out and accessed by fluorescent microscope and hematoxylin and eosin (HE) staining.Results After cultured for 4 week in vivo,the constructs present strip strong fluorescence,which demonstrated that the tissue-engineered nerve can survive well in nude mice.After the nude mice were executed,we access the constructs by fluorescent microscope and HE staining.We can see red fluorescence under fluorescent microscope and nerve-like tissue surrounded by blood vessel under HE staining.Conclusion Tissue-engineered peripheral nerve can be successfully ectopic constructed in nude mice based on improved acellular nerve scaffold and PKH26-labeled ADSCs.By PKH26 and small animals living fluorescence imaging system,we can noninvasive evaluate the tissue-engineered nerve in vivo.