摘要
目的观察聚乙醇酸(PGA)神经导管降解和生物相容性的相关性能和数据。方法2022年1月至2022年8月,北部战区总医院烧伤整形科将PGA神经导管浸泡在15 ml生理盐水中进行体外降解实验,实验周期为12周,每周测量浸泡液pH值并计算质量损失率。体外生物相容性实验有实验组和对照组,实验组将含有10%胎牛血清的DMEM培养基作为浸提介质,对照组为单纯浸提介质,将PGA神经导管浸泡在浸提介质中,在(37±1)℃下浸提(72±2)h。分别在24 h、48 h、72 h测量实验组与对照组pH值。将培养好的大鼠许旺细胞(RSC 96)分别用实验组和对照组72 h浸提液制成单细胞悬液,接种至96孔板中,进行噻唑蓝(MTT)实验,观察PGA神经导管对RSC 96增殖的影响。将RSC 96分别以实验组和对照组浸提液孵育24 h后进行Transwell实验,观察PGA神经导管对RSC 96迁移的影响。体内降解和生物相容性实验,共计24只成年雄性SD大鼠随机分为实验组和对照组,每组12只,于大鼠右后肢股二头肌和臀大肌之间钝性分离扩大肌间隙,将PGA神经导管(实验组)和Neurotube(对照组)包埋在间隙中,分别在术后2、4、8、12周进行大体观察对比、血常规、肝肾功能检查和组织学检查,评估PGA神经导管降解和生物相容性。采用SPSS 19.0软件对实验数据进行统计学分析,所得采用均数±标准差(Mean±SD)表示,组间比较采用t检验,组内及组间比较采用Turkey法,P<0.05为差异具有统计学意义。结果PGA神经导管体外降解4周时降解率为(1.470±0.026)%,之后降解率逐渐加快,至12周时降解率为(32.180±0.040)%。前2周pH下降较慢,2周时pH值为6.200±0.061,在第3周时pH值明显下降至3.930±0.118,此后pH下降速度较慢,至12周时pH值为2.560±0.003。MTT和Transwell实验结果表明,PGA神经导管浸提液对RSC 96的增殖和迁移无影响,与对照组相比,差异无统计学意义(P>0.05);体内降解包埋实验结果显示,术后2周,两组神经导管均表现出良好的支撑性,术后4~12周,两组神经导管开始逐渐变软、塌陷,但导管完整;血常规及肝、肾功能检查,在实验组和对照组同期两组比较、组内各时间点与术前比较,差异无统计学意义(P>0.05)。术后2、4、8、12周解剖大鼠肝脏和肾脏形态结构无明显异常;组织学检测结果显示,两组大鼠导管周围肌肉组织无明显变性坏死,炎症细胞无明显浸润,肌肉组织形态未见明显异常。结论PGA神经导管生物在本实验条件下,体外可降解性及生物相容性良好,为下一步进行修复神经缺损实验提供依据。
ObjectiveTo explore the characteristics and data related to the degradation and biocompatibility of the domestically procuded polyglycolic acid(PGA)nerve conduit.MethodsThis study was conducted in the Department of Burns and Plastic Surgery,the Northern Theater General Hospital between January and August 2022.PGA nerve conduits were immersed in 15 ml normal saline for in vitro experiments of degradation.The experimental period was 12 weeks.The pH of the immersion solution were tested weekly and the rates of mass loss were calculated.The in vitro experiments for biocompatibility were conducted in both of the experimental group and the control group.In the experimental group,a DMEM solution containing 10%of fetal bovine serum was used as the extraction medium,and domestically produced PGA nerve conduit was immersed in the extraction medium in the control group.An extraction medium was firstly prepared for a controlled extraction time of 72 hours±2 hours at 37℃±1℃.The pH of both experimental and control groups were tested at 24,48 and 72 hours.The 72-hour extracts of both experimental and control groups were used to prepare the single cell suspension.The single cell suspension containing the cultured RSC 96 were separately seeded into 96-well plates for Methylthiazolyldiphenyl-tetrazolium bromide(MTT)assay,to observe the effect of PGA nerve conduits on the proliferation of RSC 96.Then RSC 96 were incubated for 24 hours with the extracts of both experimental and control groups.The effect of PGA nerve conduits on the migration of RSC 96 was observed with Transwell assay.Twenty-four adult male SD rats were used and divided into experimental group and control group,with 12 rats per group,for the in vivo study of degradation and biocompatibility.Rats in the experimental group were implanted with domestically produced PGA nerve conduits,and Neurotubes,a US made nerve conduit,were implanted in the rats of control group.The PGA nerve conduits were implanted in a bluntly prepared gap between the biceps femoris muscle and the gluteus maximus muscle of the right hind limb of rats for the in vivo degradative experient.The degradation and biocompatibility of the PGA nerve conduit were evaluated by means of gross observation,tests of blood routine,liver and kidney functions and histological examinations at 2,4,8 and 12 weeks after implantation.SPSS 19.0 software was used for statistical analysis of the experimental data,and the results were expressed as Mean±SD.T test was used for inter-group comparison and Turkey method was used for intra-group and inter-group comparison.P<0.05 was used to determine whether the difference was statistically significant.ResultsThe rate of in vitro degradation was found at 1.470%±0.026%in week 4,and thereafter,a gradually accelerated degradation rate was observed and at a 32.180%±0.040%of degradation rate in week 12.The pH of the immersion solution decreased slowly in the first 2 weeks,with the pH at 6.200±0.061 in week 2.The pH then suddenly dropped to 3.930±0.118 in week 3,and then decreased slowly,with a pH 2.560±0.003 in week 12.Both MTT and Transwell experiments showed that the extract of PGA nerve conduits had no effect on the proliferation and migration of RSC 96,nor a significant difference existed in comparison with the corresponding control groups(P>0.05).The experiments of in vivo degradation of PGA nerve conduits showed that the nerve conduits in both experimental and control groups had good support at week 2 after implantation.At weeks 4-12 after implantation,the nerve conduits in both groups gradually softened and collapsed,but the conduits were in one piece and not broken.The blood routine,liver and kidney functions showed no statistically significant difference between the 2 groups over the same period,and between each time point within the groups and the groups before implantation(P>0.05).No obvious abnormal appearance of livers and kidneys was found in the rats sacrificed at each observation time point.Also,there was no obvious degeneration and necrosis of the muscle tissue around the conduits in the 2 groups,and no obvious inflammatory cells infiltration was found from the histological examinations.Morphology of the muscle tissue remained normal.ConclusionDomestically produced PGA nerve conduit is fund good in both of biocompatibility and biodegradability.It is safe and reliable,and it provides a basis for the further experients in repair of nerve defects.
作者
孙海玮
白泽明
李春辉
陶凯
SUN Haiwei;BAI Zeming;LI Chunhui;TAO Kai(Department of the First Cadre Ward,the Northern Theater General Hospital,Shenyang 110016,China;Department of Burns and Plastic Surgery,the Northern Theater General Hospital,Shenyang 110016,China)
出处
《中华显微外科杂志》
CSCD
北大核心
2024年第4期443-449,共7页
Chinese Journal of Microsurgery
基金
国家自然科学基金(81971845)。
关键词
聚乙醇酸神经导管
周围神经损伤
降解
生物相容性
大鼠
Polyglycolic acid(PGA)nerve conduit
Peripheral nerve injury
Degradation
Biocompatibility
Rat
