In-vitro nasal drug delivery studies: comparison of derivatised, fibrillar and polymerised collagen matrix-based human nasal primary culture systems for nasal drug delivery studies
In-vitro nasal drug delivery studies: comparison of derivatised, fibrillar and polymerised collagen matrix-based human nasal primary culture systems for nasal drug delivery studies
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DOI：
10.1211/0022357011777981
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发表时间：
2001-11-01
影响因子：
3.3
通讯作者：
Verbeke, N
中科院分区：
文献类型：
--
作者：
Agu, RU;Jorissen, M;Verbeke, N
本研究的目的是建立一个以胶原基质为基础的鼻腔原代培养系统，用于药物释放的研究。鼻上皮细胞在衍生的（Cellagen膜CD-24）、聚合的（Vitrogen凝胶）和纤维状（Vitrogen膜）胶原基质上培养。通过显微镜评估细胞形态。通过测量细胞暴露于牛磺-24，25二氢夫西地酸钠（STDHF）后的纤毛搏动频率（CBF）、跨上皮电阻（TER）、荧光素钠渗透、线粒体脱氢酶（MDH）活性和乳酸脱氢酶（LDH）释放来进一步表征细胞。在研究的三种胶原基质中，在第4天和第11天之间，在Cellagen膜CD-24上生长的细胞中出现最佳上皮分化表型（单层，具有柱状/立方形态）。与Vitrogen凝胶（70%）和Vitrogen膜（<10%）相比，Cellagen膜CD-24（90%）的细胞培养重现性更好。在Vitrogen凝胶上生长的细胞的TER高于在Cellagen膜CD-24和Vitrogen膜上生长的细胞。在这些条件下，荧光素钠的表观渗透系数（P-app × 10（-7）cm s（-1））为0.45 +/- 0.08（Vitrogen凝胶）和1.91 +/- 0.00（Cellagen膜CD-24）。除LDH外，所有基质的CBF和细胞活力相当。基于MDH活性、LDH释放、释放、CBF、TER和渗透研究，得出结论，基于Cellagen膜CD-24和Vitrogen凝胶的细胞在功能上适用于体外鼻用药物研究。基于Vitrogen薄膜的培养可能仅限于代谢和纤毛毒性研究。
The aim of this study was to establish a Collagen matrix-based nasal primary culture system for drug delivery studies. Nasal epithelial cells were cultured on derivatised (Cellagen membrane CD-24), polymerised (Vitrogen gel) and fibrillar (Vitrogen film) Collagen substrata. Cell morphology was assessed by microscopy. The cells were further characterised by measurement of ciliary beat frequency (CBF), transepithelial resistance (TER), permeation of sodium fluorescein, mitochondrial clehydrogenase (MDH) activity and lactate dehydrogenase (LDH) release upon cell exposure to sodium tauro-24, 25 dihydrofusidate (STDHF). Among the three Collagen substrata investigated, the best epithelial differentiated phenotype (monolayer with columnar/cuboidal morphology) occurred in cells grown on Cellagen membrane CD-24 between day 4 and day 11. Cell culture reproducibility was better with Cellagen membrane CD-24 (90 %) in comparison with Vitrogen gel (70 %) and Vitrogen film (< 10 %). TER was higher in cells grown on Vitrogen gel than on Cellagen membrane CD-24 and Vitrogen film. The apparent permeability coefficient (P-app x 10(-7)cm s(-1)) of sodium fluorescein in these conditions was 0.45 +/- 0.08 (Vitrogen gel) and 1.91 +/- 0.00 (Cellagen membrane CD-24). Except for LDH CBF and cell viability were comparable for all the substrata. Based on MDH activity, LDH release, release, CBF, TER and permeation studies, Cellagen membrane CD-24- and Vitrogen gel-based cells were concluded to be functionally suitable for in-vitro nasal drug studies. Vitrogen film-based cultures may be limited to metabolism and cilio-toxicity studies.