Department of Orthopaedic Surgery and Hyperbaric Oxygen, Therapy Center, Chang Gung Memorial Hospital, College of Medicine, Chang Gung University, Kwei-Shan Tao-Yuan, Taiwan.
Spine (Phila Pa 1976). 2011 Nov 1;36(23):1925-31. doi: 10.1097/BRS.0b013e3181feebde.
An in vitro study with degenerated human lumbar intervertebral disc specimens cultured under hyperbaric oxygenation (HBO).
To observe the changes in interleukin (IL)-1β, prostaglandin (PG)-E2, nitric oxide (NO), cell growth, and apoptosis of the human nucleus pulposus cell (NPC) after HBO.
Intervertebral disc degeneration has been demonstrated as related to IL-1β, PG-E2, NO, and O2 concentration but the actual mechanism is not clear. HBO also has also been reported in the literature to influence changes in IL-1β, prostaglandin E2, NO, and O2 concentration. However, the direct effect of HBO on the disc cells has not been previously reported.
We collected 12 human lumbar degenerated disc specimens and evaluated the effects of HBO on the cultured NPCs. The amounts of IL-1β, PG-E2, and NO in the conditioned medium were quantified by enzyme-linked immunosorbent assay and high performance liquid chromatography. Cell growth was measured by increase in cell number. Cell viability and proteoglycan content were evaluated by histologic study using safranin O staining. In situ analysis of apoptosis was performed using Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining.
Our data indicated that HBO treatment inhibited IL-1β, PG-E2, and NO production but increased cell number and matrix synthesis of cultured NPCs. TUNEL staining showed that HBO treatment suppressed the apoptosis of cultured NPCs.
HBO provides a potential treatment modality for disc degeneration.
体外研究使用高压氧(HBO)培养退变的人腰椎间盘标本。
观察人椎间盘髓核细胞(NPC)在 HBO 后白细胞介素(IL)-1β、前列腺素(PG)-E2、一氧化氮(NO)、细胞生长和凋亡的变化。
椎间盘退变已被证明与 IL-1β、PG-E2、NO 和 O2 浓度有关,但具体机制尚不清楚。文献中也有报道 HBO 影响 IL-1β、前列腺素 E2、NO 和 O2 浓度的变化。然而,HBO 对椎间盘细胞的直接影响尚未有报道。
我们收集了 12 个人腰椎退变椎间盘标本,并评估了 HBO 对培养的 NPC 的影响。酶联免疫吸附试验和高效液相色谱法定量测定条件培养基中 IL-1β、PG-E2 和 NO 的含量。通过细胞计数增加来测量细胞生长。通过番红 O 染色的组织学研究评估细胞活力和蛋白聚糖含量。通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记(TUNEL)染色进行原位分析凋亡。
我们的数据表明,HBO 治疗抑制了 IL-1β、PG-E2 和 NO 的产生,但增加了培养的 NPC 的细胞数量和基质合成。TUNEL 染色显示 HBO 治疗抑制了培养的 NPC 的凋亡。
HBO 为椎间盘退变提供了一种潜在的治疗方法。