Park Jin-Oh, Lee Byung Ho, Kang Young-Mi, Kim Tae-Hwan, Yoon Ji Young, Kim Hyang, Kwon Un-Hye, Lee Kwang-Il, Lee Hwan-Mo, Moon Seong-Hwan
Department of Orthopaedic Surgery, Yonsei University College of Medicine, Brain Korea 21 Project for Medical Science, Seoul, Korea.
J Spinal Disord Tech. 2013 Feb;26(1):E6-12. doi: 10.1097/BSD.0b013e3182698501.
In vitro experiment using degenerated human ligamentum flavum (LF) and various inflammatory cytokines.
To examine the effect of inflammatory cytokines on LF cells and to identify their roles in the pathogenesis of LF hypertrophy and ossification.
Spinal stenosis is caused, in part, by hypertrophy and ossification of the LF, which are induced by the degenerative processes (ie, increased collagen synthesis and chondroid metaplasia) of ligament fibroblasts. Degenerated intervertebral disk spontaneously produces inflammatory cytokines, which might affect the adjacent LF through local milieu of the spinal canal.
The interlaminar portion of the LF was collected during surgical spinal procedures in 15 patients (age range, 49-78 y) with lumbar spinal stenosis. LF fibroblasts were isolated by enzymatic digestion of LF tissue. LF cell cultures were treated with various inflammatory cytokines: interleukin (IL)-1α, IL-6, tumor necrosis factor-α (TNF-α), prostaglandin E2 (PGE2), and nitric oxide (NO). Cytotoxicity was analyzed by MTT assays. DNA synthesis was measured with H-thymidine incorporation, and mRNA expression of types I, III, V, and XI collagen and osteocalcin were performed by reverse transcription-polymerase chain reaction. Histochemical stains such as Von Kossa were also performed to detect bone nodule formation.
There was no cytotoxicity in the LF cells treated with each cytokine. There were significant increases in DNA synthesis and upregulated mRNA expression of types I, V, XI collagen and osteocalcin in LF cultures treated with various cytokines. LF cultures treated with IL-6, TNF-α, PGE2, and NO showed positive Von Kossa staining, indicating bone nodule formation from LF cells.
Inflammatory cytokines (IL-6, TNF-α, PGE2, and NO) seem to play a crucial role in hypertrophy and ossification of LF. Degenerated, herniated intervertebral disks, and facet arthrosis may influence LF through inflammatory cytokines and cause hypertrophy and ossification of LF.
使用退变的人黄韧带（LF）和多种炎性细胞因子进行体外实验。
研究炎性细胞因子对LF细胞的影响，并确定它们在LF肥大和骨化发病机制中的作用。
腰椎管狭窄症部分是由LF肥大和骨化引起的，而LF肥大和骨化是由韧带成纤维细胞的退变过程（即胶原合成增加和软骨样化生）诱导的。退变的椎间盘可自发产生炎性细胞因子，这些细胞因子可能通过椎管局部微环境影响相邻的LF。
在15例（年龄范围49 - 78岁）腰椎管狭窄症患者的脊柱手术过程中收集LF的椎板间部分。通过酶消化LF组织分离出LF成纤维细胞。将LF细胞培养物用多种炎性细胞因子处理：白细胞介素（IL）-1α、IL-6、肿瘤坏死因子-α（TNF-α）、前列腺素E2（PGE2）和一氧化氮（NO）。通过MTT法分析细胞毒性。用H-胸腺嘧啶核苷掺入法测量DNA合成，并通过逆转录-聚合酶链反应检测I、III-V和XI型胶原蛋白以及骨钙素的mRNA表达。还进行了如冯·科萨（Von Kossa）等组织化学染色以检测骨结节形成。
用每种细胞因子处理的LF细胞均无细胞毒性。用多种细胞因子处理的LF培养物中，DNA合成显著增加，I、V、XI型胶原蛋白和骨钙素的mRNA表达上调。用IL-6、TNF-α、PGE2和NO处理的LF培养物显示冯·科萨染色阳性，表明LF细胞形成了骨结节。
炎性细胞因子（IL-6、TNF-α、PGE2和NO）似乎在LF肥大和骨化中起关键作用。退变、突出的椎间盘以及小关节骨关节炎可能通过炎性细胞因子影响LF并导致LF肥大和骨化。
