CAJ | 학술논문

目的探讨构建组织工程椎间盘的可行性及其生物学性能. 方法利用组织工程技术构建复合椎间盘.将纤维环和髓核细胞分别接种到复合椎间盘的相应区域;然后将复合椎间盘植入裸鼠背部皮下,分别于植入后4,12周进行大体、组织形态学观察,检测复合椎间盘纤维环和髓核区域羟脯氨酸(Hyp)、糖胺聚糖(GAG)、DNA含量及生物力学性能. 结果构建的复合椎间盘大体形态与正常椎间盘组织相似.HE、番红0组织染色显示,复合椎间盘细胞具有分泌细胞外基质的功能,有进展性组织形成,同时纤维环和髓核区域得到良好的整合.扫描电镜显示,复合椎间盘纤维环和髓核区域均有丰富的新生细胞外基质形成.纤维环区域Hyp含量植入前、植入后4,12周分别为(7.1±2.3)μg/mg、(10.1±1.1) μg/mg、(12.6 ±2.1)μg/mg (P＜0.05),髓核区域Hyp含量分别为(5.9±2.5) μg/mg、(6.6±2.0)μg/mg、(12.5±3.3) μg/mg (P ＜0.05);纤维环区域GAG含量分别为(3.3±1.3)μg/mg、(10.9±2.7)μg/mg、(12.4±3.7) μg/mg(P ＜0.05),髓核区域分别为(13.4±1.7) μg/mg、(18.4±2.5) μg/mg、(22.7±3.6) μg/mg(P＜ 0.05);纤维环区域DNA含量分别为(52.2±5.8) ng/mg、(99.7±17.0) ng/mg、(149.5 ±12.0)ng/mg(P ＜0.05),髓核区域DNA含量分别为(66.5±9.2) ng/mg、(148.6±9.8) ng/mg、(187.6±16.6) ng/mg (P＜0.05).植入后12周,最大载荷、抗压强度、屈服强度及弹性模量与植入前比较差异有统计学意义(P＜0.05).各时相点生物力学指标均显著低于正常椎间盘(P＜0.05). 结论组织工程复合椎间盘具有与正常椎间盘相似的组织形态和生物学特性,为组织工程椎间盘的临床应用提供了一种可能.
목적 탐토구건조직공정추간반적가행성급기생물학성능. 방법 이용조직공정기술구건복합추간반.장섬유배화수핵세포분별접충도복합추간반적상응구역;연후장복합추간반식입라서배부피하,분별우식입후4,12주진행대체、조직형태학관찰,검측복합추간반섬유배화수핵구역간포안산(Hyp)、당알취당(GAG)、DNA함량급생물역학성능. 결과 구건적복합추간반대체형태여정상추간반조직상사.HE、번홍0조직염색현시,복합추간반세포구유분비세포외기질적공능,유진전성조직형성,동시섬유배화수핵구역득도량호적정합.소묘전경현시,복합추간반섬유배화수핵구역균유봉부적신생세포외기질형성.섬유배구역Hyp함량식입전、식입후4,12주분별위(7.1±2.3)μg/mg、(10.1±1.1) μg/mg、(12.6 ±2.1)μg/mg (P＜0.05),수핵구역Hyp함량분별위(5.9±2.5) μg/mg、(6.6±2.0)μg/mg、(12.5±3.3) μg/mg (P ＜0.05);섬유배구역GAG함량분별위(3.3±1.3)μg/mg、(10.9±2.7)μg/mg、(12.4±3.7) μg/mg(P ＜0.05),수핵구역분별위(13.4±1.7) μg/mg、(18.4±2.5) μg/mg、(22.7±3.6) μg/mg(P＜ 0.05);섬유배구역DNA함량분별위(52.2±5.8) ng/mg、(99.7±17.0) ng/mg、(149.5 ±12.0)ng/mg(P ＜0.05),수핵구역DNA함량분별위(66.5±9.2) ng/mg、(148.6±9.8) ng/mg、(187.6±16.6) ng/mg (P＜0.05).식입후12주,최대재하、항압강도、굴복강도급탄성모량여식입전비교차이유통계학의의(P＜0.05).각시상점생물역학지표균현저저우정상추간반(P＜0.05). 결론 조직공정복합추간반구유여정상추간반상사적조직형태화생물학특성,위조직공정추간반적림상응용제공료일충가능.
Objective To construct a novel tissue-engineered intervertebral disc complex and investigate the biological properties of the complex.Methods By the technique of tissue engineering,the anulus fibrosus scaffolds and nucleus pulposus cells were seeded in corresponding regions in the composite intervertebral discs.Then,the cell-scaffold hybrids were implanted in the subcutaneous space of the dorsum of athymic mice.At 4,8,and 12 weeks after implantation,gross observation and histological morphology were performed and hydroxyproline,glycosaminoglycan,DNA content and biochemical properties were evaluated.Results Gross morphology and histology of the engineered discs resembled those of native intervertebral discs.Morphological studies revealed progressive tissue formation and junction integration between anulus fibrosus and nucleus pulposus cells.Scanning microscopy showed new extracellular matrix was rich in anulus fibrosus and nucleus pulposus cells.Content of hydroxyproline in anulus fibrosus and nucleus pulposus cells increased with time,i.e.before the implantation [(7.1 ± 2.3) μg/mg,(5.9 ± 2.5) μg/mg],at 4 weeks [(10.1 ± 1.1) μg/mg,(6.6 ± 2.0) μg/mg] and at 12 weeks [(12.6 ± 2.1) μg/mg,(12.5 ± 3.3) μg/mg] (P ＜ 0.05).Content of proteoglycan and DNA showed similar variation trends,i.e.in anulus fibrosus [(3.3 ± 1.3) μg/mg,(10.9 ± 2.7) μg/mg,(12.4 ±3.7) μg/mg;(52.2 ±5.8) ng/mg,(99.7 ± 17.0) ng/mg,(149.5 ± 12.0) ng/mg respectively],in nucleus pulposus cells [(13.4 ± 1.7) μg/mg,(18.4 ± 2.5) μg/mg,(22.7 ± 3.6) μg/mg;(66.5 ± 9.2) ng/mg,(148.6 ± 9.8) ng/mg,(187.6 ± 16.6) ng/mg respectively] (P ＜ 0.05).At 12 weeks,the maximum loads,compressive strength,yield strength and elastic modulus differed significantly from those before implantation (P ＜ 0.05).Biomechanical markers of the composite were substantially lowered with time compared to those of native tissues (P ＜ 0.05).Conclusion Tissue-engineered discs with similar morphological and biochemical properties to native discs provides a portability in tissue engineering.