Oueis A, Yokozeki M, Baba Y, Moriyama K, Kuroda T
Department of Maxillofacial Orthognathics, Maxillofacial Reconstruction, Graduate School, Tokyo Medical and Dental University, Japan.
J Med Dent Sci. 2000 Mar;47(1):95-103.
In order to modulate palatal scar tissue, especially its myofibroblastic component, there is a pressing need for an in vitro model of this tissue. In the present, study we established an organ culture model of the rat palatal scar tissue. After excision of palatal mucoperiosteum, explants from the developing immature scar tissue and from the normal palatal mucosa were used to observe myofibroblasts in vivo and their maintenance in organ culture. Explants were cultured at the gas-liquid interface in serum-free Waymouth's MB 752/1 medium and in a humid atmosphere containing 55% O2/5% CO2 in air at 37 degrees C for 3 days. Viability of the cultured explants was evaluated with morphological and histological criteria and BrdU incorporation. After organ culture, the scar tissue showed good preservation of the in vivo histology. The myofibroblasts and smooth muscle cells of the cultured scar tissues showed continuous expression of alpha-smooth muscle actin (alpha-SMA), mimicking the in vivo situation. In the normal tissues, only smooth muscle cells of the blood vessels expressed alpha-SMA. These results demonstrate that the established model provides a useful in vitro experimental tool for investigating the palatal scar tissue in general and its myofibroblasts in particular.
为了调节腭部瘢痕组织,尤其是其肌成纤维细胞成分,迫切需要该组织的体外模型。在本研究中,我们建立了大鼠腭部瘢痕组织的器官培养模型。切除腭部粘骨膜后,取自发育中的未成熟瘢痕组织和正常腭部黏膜的外植体用于观察体内的肌成纤维细胞及其在器官培养中的维持情况。外植体在无血清的Waymouth's MB 752/1培养基中,于气液界面、37℃、含55% O₂/5% CO₂的潮湿空气环境中培养3天。通过形态学、组织学标准和BrdU掺入法评估培养外植体的活力。器官培养后,瘢痕组织的体内组织学得到良好保存。培养的瘢痕组织中的肌成纤维细胞和平滑肌细胞持续表达α-平滑肌肌动蛋白(α-SMA),模拟了体内情况。在正常组织中,仅血管平滑肌细胞表达α-SMA。这些结果表明,所建立的模型为研究腭部瘢痕组织,尤其是其肌成纤维细胞,提供了一种有用的体外实验工具。