An experimental study of the regeneration of the intervertebral disc with an allograft of cultured annulus fibrosus cells using a tissue-engineering method
An experimental study of the regeneration of the intervertebral disc with an allograft of cultured annulus fibrosus cells using a tissue-engineering method
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DOI：
10.1097/00007632-200303150-00007
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发表时间：
2003-03-15
期刊：
影响因子：
3
通讯作者：
Fujikawa, K
中科院分区：
文献类型：
--
作者：
Sato, M;Asazuma, T;Fujikawa, K
研究设计。用吲哚青绿染料增强激光汽化髓核，将培养的纤维环细胞移植到髓核汽化后的间盘陷窝中。根据同种异体纤维环细胞的活性和组织学状态以及预防椎间隙变窄来评估椎间盘的再生。应用组织工程学方法研究激光腰椎间盘摘除术后的椎间盘再生。椎间盘是人体内最无血管的组织，其再生能力与关节软骨一样低。当进行髓核摘除时，纤维环几乎没有再生，因此，椎间盘退变是不可避免的。从20只日本大白兔分离纤维环细胞，用PKH-26荧光染料标记，种植在带膜密封的环胶原蜂窝状支架中。用ICG染料增强激光汽化兔髓核后，将纤维环细胞种植于人工胶原蜂窝状支架内，并将其移植到髓核汽化的受体兔的椎间陷窝内。分别于术后2周、4周、8周、12周拍摄腰椎软片，测量腰椎间隙，取出腰椎标本。连续冰冻切片检测5-溴-2‘-脱氧尿苷/PKH-26荧光标记的同种异体纤维环细胞的增殖情况，并用藏红O染色进行组织学评价。同种异体纤维环细胞存活，并表现出增殖活性，并形成透明样软骨。如图所示，细胞转移组在术后12周内明显防止了椎间隙的狭窄。将培养在ACHMS支架上的纤维环细胞移植到兔髓核陷窝(激光汽化获得)和纤维环孔(激光纤维植入获得)内。这些细胞在受者的椎间盘组织中存活并显示细胞增殖。
Study Design. Cultured annulus fibrosus cells within an atelocollagen honeycomb-shaped scaffold with a membrane seal were allografted into the lacunas of intervertebral discs of which the nucleus pulposus had been vaporized using an indocyanine green dye-enhanced laser. Regeneration of the intervertebral disc was assessed based on the viability and histologic status of the allografted annulus fibrosus cells, as well as the prevention of narrowing disc space.Objectives. To study the regeneration of intervertebral disc after laser discectomy using tissue-engineering methods.Summary of Background Data. Intervertebral disc is the most avascular tissue in the human body, and its ability to regenerate is as low as that of articular cartilage. When nucleotomy is carried out, little regeneration of the annulus fibrosus is observed; consequently, intervertebral disc degeneration is inevitable.Methods. Annulus fibrosus cells isolated from 20 Japanese white rabbits were labeled with a PKH-26 fluorescent dye and seeded within an atelocollagen honeycomb-shaped scaffold with a membrane seal. Annulus fibrosus cells cultured in atelocollagen honeycomb-shaped scaffold with a membrane seal for 1 week were allografted into the lacunas of intervertebral discs of recipient rabbit of which the nucleus pulposus had been vaporized using an ICG dye-enhanced laser. Soft radiograph photographs of the lumbar spine of these anesthetized rabbits were taken, the disc space measured, and the lumbar spine extracted 2, 4, 8, and 12 weeks after the operation. The proliferation of allografted annulus fibrosus cells with 5-bromo-2'-deoxyuridine/PKH-26 fluorescent labels was assessed using consecutive frozen sections, and safranin-O staining carried out for histologic evaluation.Results. The allografted annulus fibrosus cells were viable and showed proliferation activity with a hyaline-like cartilage being produced. The narrowing of the intervertebral disc space of the cell translation group was significantly prevented, as shown, up to 12 postoperative weeks.Conclusion. The annulus fibrosus cells cultured in an ACHMS-scaffold were allografted into the lacunae of nucleus pulposus ( obtained using laser vaporization), as well as the hole of annulus fibrosus ( obtained by laser fiber insertion) of rabbit intervertebral discs. These cells were viable and showed cell proliferation in the disc tissues of recipients.