CAJ | 학술논문

目的采用超声振荡结合化学萃取法制备大鼠脱细胞脊髓支架,观察其三维结构及生物学特性,为脊髓组织工程研究提供理想的支架材料. 方法采用超声振荡结合化学萃取(体积分数2％ Triton X-100+体积分数2％脱氧胆酸钠)方法对大鼠脊髓进行脱细胞处理(脱细胞脊髓组),对照正常大鼠脊髓组织(对照组),观察该脱细胞脊髓支架的大体形态、组织学及超微三维结构特点,并检测该支架材料的孔径大小、孔隙率、含水率、酶解率、在水溶液中稳定性等.结果脱细胞脊髓组原有的细胞成分被有效去除,具有(94.57±3.45)％的孔隙率和(88.62±1.0)％的含水率以及良好的三维空间结构(平均孔径为46 μm),该支架在胰酶溶液中逐步降解,在第20小时达到(69.03±2.19)％,在双蒸水中逐步崩解,在第8天可达(62.55±1.70)％.正常脊髓组织结构紧密,含大量神经细胞及髓鞘,孔隙率和含水量分别为(0.04±0.02)％、(62.4±1.5)％,扫描电镜下未见明显孔隙结构,该支架在胰酶溶液中逐步降解,在第20小时达到(37.62±0.99)％,在双蒸水中逐步崩解,在第8天可达(40.97±0.81)％. 结论超声振荡+化学萃取所制备的脱细胞脊髓支架细胞成分去除彻底,细胞外基质成分保存完整,具有良好三维空间网状结构、良好的孔隙率和含水量,符合组织工程脊髓支架的理论要求,为组织工程脊髓支架提供理想的选择.
목적 채용초성진탕결합화학췌취법제비대서탈세포척수지가,관찰기삼유결구급생물학특성,위척수조직공정연구제공이상적지가재료. 방법 채용초성진탕결합화학췌취(체적분수2％ Triton X-100+체적분수2％탈양담산납)방법대대서척수진행탈세포처리(탈세포척수조),대조정상대서척수조직(대조조),관찰해탈세포척수지가적대체형태、조직학급초미삼유결구특점,병검측해지가재료적공경대소、공극솔、함수솔、매해솔、재수용액중은정성등.결과 탈세포척수조원유적세포성분피유효거제,구유(94.57±3.45)％적공극솔화(88.62±1.0)％적함수솔이급량호적삼유공간결구(평균공경위46 μm),해지가재이매용액중축보강해,재제20소시체도(69.03±2.19)％,재쌍증수중축보붕해,재제8천가체(62.55±1.70)％.정상척수조직결구긴밀,함대량신경세포급수초,공극솔화함수량분별위(0.04±0.02)％、(62.4±1.5)％,소묘전경하미견명현공극결구,해지가재이매용액중축보강해,재제20소시체도(37.62±0.99)％,재쌍증수중축보붕해,재제8천가체(40.97±0.81)％. 결론 초성진탕+화학췌취소제비적탈세포척수지가세포성분거제철저,세포외기질성분보존완정,구유량호삼유공간망상결구、량호적공극솔화함수량,부합조직공정척수지가적이론요구,위조직공정척수지가제공이상적선택.
Objective To observe three-dimensional structure and biological features of rat acellular spinal cord scaffold prepared by sonic oscillation and chemical extraction in order to offer an ideal scaffold for spinal cord tissue engineering research.Methods Rat spinal cord underwent acellular treatment with sonic oscillation and chemical extraction (Triton X-100 at volume fracture of 2％ and sodium deoxycholate at volume fracture of 2％) (acellular spinal cord group).In contrast with spinal cord tissue of normal rats (control group),general morphology,histology and ultramicro three-dimensional structure of acellular spinal cord scaffold were observed and aperture size,factor of porosity,water ratio,enzymolysis ratio and stability in water solution of the scaffold were also detected.Results Acellular spinal cord group showed effective removal of original cell components with factor of porosity for (94.57 ±3.45) ％ and water content for (88.62 ± 1.0) ％,and satisfactory three-dimensional structure with average aperture of 46 μm.Scaffold showed gradual degradation in enzymolysis solution and enzymolysis rate reached (69.03 ± 2.19)％ at 20 hours.Besides,scaffold showed stepwise disintegration in double distilled water and hydrolysis rate was (62.55 ± 1.70) ％ at 8 days.While,normal spinal cord showed close structure,generous neurons and myelin sheath with factor of porosity for (0.04 ± 0.02) ％ and water content for (62.4 ± 1.5) ％,and unobvious pore structure under scanning electron microscope.Normal spinal cord were degraded gradually in enzymolysis solution and enzymolysis rate was (37.62 ± 0.9)％ at 20hours.In the meantime,normal spinal cord were disintegrated gradually in double distilled water and hydrolysis rate was (40.97 ± 0.81) ％ at 8 days.Conclusions Acellular spinal cord scaffold prepared by sonic oscillation plus chemical extraction achieves complete removal of cell components,intact extracellular matrix,and satisfactory results in three-dimensional network structures,factor of porosity and water content.Also,the scaffold meets theoretical demands of tissue-engineered spinal cord scaffold and is an ideal alterative for tissue-engineered spinal cord scaffold.