Ganey Timothy, Libera Jeanette, Moos Verena, Alasevic Olivera, Fritsch Karl-Gerd, Meisel Hans Joerg, Hutton William C
Atlanta Medical Center, Atlanta, Georgia 30312, USA.
Spine (Phila Pa 1976). 2003 Dec 1;28(23):2609-20. doi: 10.1097/01.BRS.0000097891.63063.78.
Disc degeneration and osteoarthritis are diseases of the matrix. Chondrocytes that have been removed from damaged cartilaginous tissues maintain a capacity to proliferate, produce, and secrete matrix components, and respond to physical stimuli such as dynamic loading. A dog model was used to investigate the hypothesis that autologous disc chondrocytes can be used to repair damaged intervertebral disc.
Given the capacity for the cells in vitro to produce matrix molecules that would be appropriate for disc chondrocytes, the focus of the experiment was to investigate whether the cells would continue to sustain metabolic function after transplantation.
No evidence for long-term integration exists for cell transplantation in species other than rats and rabbits. Furthermore, no controlled studies of 1-year duration have been published.
Disc chondrocytes were harvested and expanded in culture under controlled and defined conditions, returned to the same animals from which they had been sampled (autologous transplantation) via percutaneous delivery. The animals were analyzed at specific times after transplantation by several methods to examine whether disc chondrocytes integrated with the surrounding tissue, produced the appropriate intervertebral disc extracellular matrix, and might provide a formative solution to disc repair.
In the context of degenerative changes in an injury model: (1) autologous disc chondrocytes were expanded in culture and returned to the disc by a minimally invasive procedure after 12 weeks; (2) disc chondrocytes remained viable after transplantation as shown by Bromodeoxyuridine incorporation and maintained a capacity for proliferation after transplantation as depicted by histology; (3) transplanted disc chondrocytes produced an extracellular matrix that displayed composition similar to normal intervertebral disc tissue. Positive evidence of proteoglycan content was supported by accepted histochemical staining techniques such as Safranin O-Fast Green; (4) both type II and type I collagens were demonstrated in the regenerated intervertebral disc matrix by immunohistochemistry after chondrocyte transplantation; and (5) when the disc heights were analyzed for variance according to treatment, a statistically significantcorrelation between transplanting cells and retention of disc height was achieved.
Autologous chondrocyte transplantation is technically feasible and biologically relevant to repairing disc damage and retarding disc degeneration.
椎间盘退变和骨关节炎是基质疾病。从受损软骨组织中分离出的软骨细胞保持增殖、产生和分泌基质成分的能力,并对动态负荷等物理刺激产生反应。本研究采用犬模型来验证自体椎间盘软骨细胞可用于修复受损椎间盘这一假说。
鉴于体外培养的细胞能够产生适合椎间盘软骨细胞的基质分子,本实验的重点是研究这些细胞在移植后是否能继续维持代谢功能。
除大鼠和兔子外,尚无其他物种细胞移植长期整合的证据。此外,尚未发表过为期1年的对照研究。
收集椎间盘软骨细胞并在可控的特定条件下进行体外培养扩增,然后通过经皮给药的方式将其回输到取自的同一只动物体内(自体移植)。在移植后的特定时间,通过多种方法对动物进行分析,以检查椎间盘软骨细胞是否与周围组织整合、是否产生合适的椎间盘细胞外基质,以及是否能为椎间盘修复提供一种有效的解决方案。
在损伤模型的退变背景下:(1)自体椎间盘软骨细胞在体外培养扩增12周后,通过微创手术回输到椎间盘;(2)经溴脱氧尿苷掺入法检测,移植后的椎间盘软骨细胞仍具有活性,组织学检查显示移植后仍具有增殖能力;(3)移植的椎间盘软骨细胞产生了一种细胞外基质,其成分与正常椎间盘组织相似。番红O-固绿等公认的组织化学染色技术证实了蛋白聚糖含量呈阳性;(4)软骨细胞移植后,通过免疫组织化学法在再生的椎间盘基质中检测到Ⅱ型和Ⅰ型胶原蛋白;(5)根据处理方式对椎间盘高度进行方差分析时,移植细胞与椎间盘高度保留之间存在统计学上的显著相关性。
自体软骨细胞移植在技术上是可行的,对于修复椎间盘损伤和延缓椎间盘退变具有生物学意义。