Epithelia biochemical barrier

Passive transcellular transport across the intestinal epithelium involves three discrete steps (1) uptake across the apical membrane, (2) diffusion through the cytoplasm, and (3) efflux across the basolateral membrane. Occasionally, drug molecules without favorable physicochemical properties traverse the intestinal epithelium using endogenous membrane transporters.6-8 In addition, the intestinal mucosa, with its numerous drug-metabolizing enzymes and efflux transporters, such as P-glycoprotein (Pgp), functions as a biochemical barrier.9... 

Gan, L.-S. L., Thakker, D. R., Applications of the Caco-2 model in the design and development of orally active drugs elucidation of biochemical and physical barriers posed by the intestinal epithelium, Adv. Drug Deliv. Rev. 1997, 23, 77-98. 

Because of the possible effects of active and carrier-mediated processes and metabolic biotransformation, the issue of tissue viability is important for in vitro buccal mucosal experiments. The barrier nature of the buccal mucosa resides in the upper layers of the epithelium, where unlike in the stratum corneum, the cells contain a variety of functional organelles [119, 122, 125, 150], and so tissue viability may be an important component of the barrier function of the tissue. Various methods have been employed to assess the viability of excised buccal mucosa, including measurement of biochemical markers, microscopic methods, and linearity of transport data [42], While biochemical methods, including measurement of adenosine 5 -triphosphate (ATP) levels and utilization of glucose, provide information on the metabolic activity of the tissue, this does not necessarily relate to the barrier function of the tissue. In excised rabbit buccal mucosa, levels of ATP were measured and found to decline by 40% in 6 h, and this correlated well with transmission electron microscopic evaluation of the tissue (intact superficial cells) [32], In addition, the permeability of a model peptide was unaltered up to 6 h postmortem, but at 8 h, a significant change in permeability was observed [32], These investigators therefore claimed that excised rabbit buccal mucosa could be used for diffusion studies for 6 h. 

Cell lines of both airway and alveolar epithelial origin are available (immortal cells, transformed or carcinoma-derived), the most commonly used being the CALU-3 cell line derived from airway epithelium [37], and the A549 cell line derived from a bronchioalveolar carcinoma [38]. However, these cultured cell lines share only limited similarity in morphology, biochemical characteristics, and barrier properties (low epithelial resistance due to the lack of tight junctions) with the epithelia in vivo. Consequently, the results of absorption and metabolism studies based on cultured cell lines should be interpreted with caution. 

The primary cell culture model is a more valid model for the study of absorption and transport processes of a drug via the pulmonary route. It provides a tight epithelial barrier with morphological and functional properties resembling those of the in-vivo condition. Primary alveolar epithelial cells from rats [39], rabbits [40] and humans [41] which display morphological and biochemical characteristics similar to the native epithelium have been isolated and can be used for drug transport studies. 

The epithelium is supported underneath by lamina propria and a layer of smooth muscle called muscularis mucosa (3-10 cells thick). These three layers, i.e., the epithelium, lamina propria, and muscularis mucosa, together constitute the intestinal mucosa.On the apical surface, the epithelium along with lamina propria projects to form villi. The cell membranes of epithelial cells that comprise the villi contain uniform microvilli, which give the cells a fuzzy border, collectively called a brush border. These structures, although greatly increase the absorptive surface area of the small intestine, provide an additional enzymatic barrier since the intestinal digestive enzymes are contained in the brush border. In addition, on the top of the epithelial layer lies another layer, the UWL, as previously described. The metabolic and biochemical components of the epithelial barrier will be discussed. 

Gan LSL and Thakker DR. Applications of the Caco-2 Model in the Design and Development of Orally Active Drugs Elucidation of Biochemical and Physical Barriers Posed by the Intestinal Epithelium. Adv Drug Del Rev 1997 23 77-98. 

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