Development and characterization of an in vitro gingival epithelial model.
Development and characterization of an in vitro gingival epithelial model.
复制标题
体外牙龈上皮模型的开发和表征。
DOI：
--
复制
发表时间：
1995
影响因子：
3.5
通讯作者：
D. Stevens
中科院分区：
文献类型：
--
作者：
L. L. Odioso;M. Doyle;K. W. Quinn;R. Bartel;M. Zimber;D. Stevens
一个三维牙龈上皮模型已经开发和特点。从人牙龈组织中分离口腔上皮细胞和结缔组织成纤维细胞，并用于建立体外口腔粘膜共培养模型。将成纤维细胞接种在尼龙网支架上，使其增殖并分泌胶原蛋白和细胞外基质蛋白，以形成能够支持上皮细胞生长的基质。上皮细胞接种在融合的基质层上，增殖并分化形成复层鳞状上皮。通过操作生长培养基和培养条件刺激驻留上皮细胞，以形成多层口腔粘膜样组织。组织学分析显示细胞结构表现出支持上皮层生长和分化的基质-上皮相互作用。免疫组织化学分析证实了I型和III型胶原蛋白的产生。基质层的免疫荧光鉴定IV型胶原和纤连蛋白。纤维连接蛋白也在表面上皮细胞上被检测到。观察基底细胞、棘细胞和颗粒细胞的分化，并使用广谱细胞角蛋白抗体AE 1和AE 3确认分化标记物酸性（K10，14-16，19）和碱性（K1-8）细胞角蛋白的存在。电镜观察到不连续的基膜区，半桥粒的发展。共培养物具有代谢活性，通过噻唑蓝（MTT）试验测定线粒体功能和[3 H]胸苷掺入DNA。人牙龈上皮共培养模型在上皮接种后长达35天是可行的。该模型可能为牙周组织反应性的有限研究提供机会。
A 3-dimensional gingival epithelial model has been developed and characterized. Oral epithelial cells and connective tissue fibroblasts were isolated from human gingival tissue and used to create an in vitro oral mucosa co-culture model. Fibroblasts were seeded on a scaffold of nylon mesh, allowed to proliferate and secrete collagen and extracellular matrix proteins to form a stroma capable of supporting the growth of epithelial cells. Epithelial cells were seeded on top of a confluent stromal layer, proliferated and differentiated to form a stratified squamous epithelium. Resident epithelial cells were stimulated, by manipulation of growth medium and culture conditions, to form a multi-layered oral mucosa-like tissue. Histologic analyses revealed cellular architecture exhibiting stromal-epithelial interaction which supports the growth and differentiation of an epithelial layer. Immunohistochemical analyses confirmed production of types I and III collagen. Immunofluorescence of the stromal layer identified type IV collagen and fibronectin. Fibronectin was also detected on surface epithelium. Differentiation of basal, spinous and granular cells was observed, and the presence of differentiation markers, acidic (K10, 14-16, 19) and basic (K1-8) cytokeratins were confirmed using broad spectrum cytokeratin antibodies, AE1 and AE3. Development of a discontinuous basal lamina zone, with hemidesmosomes, was observed by electron microscopy. The co-culture was metabolically active, as measured by the thiazoyl blue (MTT) assay for mitchondrial function and [3H] thymidine incorporation into DNA. The human gingival epithelial co-culture model was viable up to 35 days post-epithelial seed. This model may offer opportunities for limited study of periodontal tissue responsiveness.
纤连蛋白：其结构、功能和生理学的简要概述。
DOI：
10.1093/clinids/9.supplement_4.s317
发表时间：
1987
期刊：
Reviews of infectious diseases
影响因子：
--
作者：
Proctor,RA
通讯作者：
Proctor,RA