Department of Orthopedic Surgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, People's Republic of China.
Department of Minimally Invasive Spine Surgery, Qingdao Municipal Hospital, East Branch, Qingdao, Shandong, People's Republic of China.
Kaohsiung J Med Sci. 2019 Aug;35(8):486-492. doi: 10.1002/kjm2.12084. Epub 2019 May 15.
The present study determines whether the in vivo injection of TGFβ1 and CTGF mediated by AAV2 to transfect nucleus pulposus cells in degenerative lumbar discs can reverse the biological effects of rhesus lumbar disc degeneration. A total of 42 lumbar discs obtained from six rhesus monkeys were classified into three groups: experimental group, control group, and blank group. Degenerative lumbar discs were respectively injected with double gene-transfected human nucleus pulposus cells using minimally invasive techniques. Immumohistochemical staining, RT-PCR, and western blot were performed to observe the biological effects of double gene-transfected human nucleus pulposus cells in degenerative lumbar discs on rhesus lumbar disc degeneration. At 4, 8, and 12 weeks after the transplantation of nucleus pulposus cells, the expression levels of TGF-ß1, CTGF, proteoglycan mRNA, and type-II collagen were detected by RT-PCR. The values of immumohistochemical staining and RT-PCR in the experimental group increased at 8 weeks, decreased with time at 12 weeks, and remained greater than the values in the control group, and the differences were statistically significant (P < .05). The western blot revealed that the values in the experimental group decreased with time, but remained greater than those in the PBS control group and blank control group, and the differences were statistically significant (P < .05). The double gene-transfection of human nucleus pulposus cells in degenerative lumbar discs mediated by rAAV2 can be continuously expressed in vivo after transplantation in lumbar discs of rhesus monkeys, and promotes the synthesis of proteoglycan and type II collagen, achieving the treatment purpose.
本研究旨在探讨通过 AAV2 介导 TGFβ1 和 CTGF 向退变腰椎间盘核细胞体内转染,能否逆转恒河猴腰椎间盘退变的生物学效应。共 42 个来自 6 只恒河猴的腰椎间盘分为实验组、对照组和空白组。采用微创技术向退变的腰椎间盘分别注射双基因转染的人髓核细胞。免疫组化染色、RT-PCR 和 Western blot 检测双基因转染的人髓核细胞对恒河猴退变腰椎间盘的生物学效应。在细胞移植后 4、8 和 12 周,通过 RT-PCR 检测 TGF-β1、CTGF、蛋白聚糖 mRNA 和 II 型胶原的表达水平。实验组的免疫组化染色和 RT-PCR 值在 8 周时增加,在 12 周时随时间下降,但仍大于对照组,差异具有统计学意义(P<.05)。Western blot 显示实验组的值随时间下降,但仍大于 PBS 对照组和空白对照组,差异具有统计学意义(P<.05)。rAAV2 介导的人退变髓核细胞双基因转染可在恒河猴腰椎间盘移植后在体内持续表达,并促进蛋白聚糖和 II 型胶原的合成,达到治疗目的。