摘要
目的探讨骨基质明胶（bone matrix gelatin，BMG）／聚对苯二甲酸-共-丁二酸丁二醇酯[poly（butylenesucci—nate-co-terephthalate），PBST]双相组织工程纤维环的构建。方法PBST通过静电纺丝的方式制备成薄膜，检测其吸水率、孔隙率。取兔纤维环细胞进行体外培养，并通过番红“O”、Ⅱ型胶原免疫组化染色进行细胞鉴定；鉴定后的细胞种植到PBST薄膜支架上，通过扫描电镜观察细胞在支架上的生长情况。以BMG作为外纤维环支架，以PBST纺丝作为内纤维环支架，构建新型双相组织工程纤维环支架。将细胞种植到双相支架上，体外培养3、7、21d后，分析双相组织工程纤维环的生物学特性及力学性能。结果PBST纺丝薄膜的孔隙率为61．83％±7．33％，吸水率为297．34％±57．13％。纤维环细胞经番红“O”、Ⅱ型胶原免疫组化染色鉴定呈阳性，表现出纤维环细胞的特征。经鉴定后的细胞种植在薄膜支架上培养3、7d后，扫描电镜显示细胞在薄膜支架上黏附并增殖；将纤维环细胞接种到BMG／PBST双相支架上，培养3、7、21d后，HE染色显示细胞随培养时间的增加逐渐渗透到薄膜支架内部；番红“O”、Ⅱ型胶原免疫组化染色呈阳性，表明细胞在支架上分泌出大量糖胺聚糖和Ⅱ型胶原等纤维环细胞特有的细胞外基质。种植细胞后的双相支架体外培养21d后，支架的弹性模量[（17．56±1．47）MPa]明显较无细胞的双相支架的弹性模量[（14．83±1．02）MPa]增加。结论初步构建的BMG／PBST双相组织工程纤维环具有良好的细胞相容性和力学性能，为进一步构建完整组织工程椎间盘奠定了基础。
Objective To construct a novel tissue-engineered bone matrix gelatin （BMG）/poly[butylene succinate-co-terephthalate] （PBST） biphasic scaffold for annulus fibrosus. Methods The PBST spinning fibers were prepared by electrospinning and the porosity and water absorption rate were tested. Rabbit annulus fibrosus cells were isolated, cultured and identified through Safranin ＂O＂ staining, and collagen Ⅱ immunohistochemical staining in vitro. And then annulus fibrosus ceils were implanted on the PBST fiber, whose growth situation was observed by scanning electron microscope （SEM）. Then the BMG/PBST biphasic scaffold was constructed by BMG as the outer annular fibrosus and PBST fiber as the inner annular fibrosus. The annulus fibrosus cells were implanted on the biphasic scafflod and cultured for 3, 7 and 21 days in vitro. The biomechanical and biological property was observed at the predetermined time point. Results The porosity of the fiber was 61.83%-7.33% and its water absorption rate was 297.34%-57.13%. The identified result of annulus fibrosus cells were positive, suggesting that the cells have still kept their annulus fibrosus cells characteristics. The cells growth could be observed through SEM at 3rd and 7th day after implanted on the fibers. After cultured on the BMG/PBST scaffold, HE staining proved that the cells could ingress into the inner of fiber with time. Safranin ＂O＂ staining and collagen Ⅱ immunohistochemical staining proved that the cells can secreted abundant proteoglycan and collagen Ⅱ, the special annulus fibrosus cell extracellular matrix. Compared with the BMG/PBST scaffold without cells, the elastic modulus of biphasic scaffold was increased from 14.83±1.02 MPa to 17.56±1.47 MPa after cultured with cells for 21 days in vitro. Conclusion The novel tissue-engineered biphasic scaffold for annulus fibrosus constructed with BMG and PBST fiber spinning has good cytocompatibility and biomechanical characteristics, which provide a basis for the complete tissue engineered int
出处
《中华骨科杂志》
CAS
CSCD
北大核心
2016年第1期35-42,共8页
Chinese Journal of Orthopaedics
基金
国家自然科学基金（81171472、81201407）
四川省教育厅创新团队资助项目（13TD0030）
四川省教育厅重大培育项目（15CZ0021）
南充市科技支撑计划项目（14A0021）
关键词
组织工程
组织支架
明胶
Tissue engineering
Tissue saffolds
Gelatin
