Gelatin sponge-supported histoculture of human nasal mucosa

Abstract

Considerable progress has recently been made in the understanding of airway inflammation by cell culture assays and in vivo provocation studies. Inasmuch as ethical considerations limit experimental work in humans, physiologically relevant in vitro models are required to better understand cellular and molecular tissue interactions in human nasal mucosa. Here we describe a human nasal mucosa culture model utilizing a simple gelatin sponge-supported histoculture system at the air-liquid interface. Viable mucosa was preserved for at least 48 h, as shown by morphology and immunohistochemical staining with Ki-67 as marker for proliferation. Pro-inflammatory mediators (kinins, histamine, thromboxane B2, prostaglandin F2 alpha, and substance P) are detectable in serum-containing as well as serum-free culture medium. Incubation with 10(-8) M substance P increases the number of degranulated mast cells after 48 h by 26% (P < 0.01). In this model, biochemical responses can be correlated with histologic alterations of the target tissue. Inflammatory parameters can be examined and compared in various patient groups and different stimulators/inhibitors. This culture method provides a valuable research tool for analyzing all compartments present in nasal mucosa under physiologically relevant conditions, and for studying complex interactions and responses of mucosal cell populations in their natural tissue environment.

Ozone-Induced Augmentation of Eicosanoid Metabolism in Human Nasal Mucosa in vitro

“…Recently we found significantly in creased concentrations of histamine, IL-1|3, 1L-6, IL-8 and TNFa following ozone exposure [6], To investigate the hy pothesis that ozone can augment cicosanoid metabolism in the airway tissues, human nasal mucosa was cultured using a previously described histoculture technique [7] exposed to 0.1 ppm ozone for 24 h. The concentrations o f cyclooxy genase and lipoxygenase metabolites were measured in the supernatant of full-thickness nasal mucosa in organ culture.…”

Substance-P-lnduced Histamine Release from Human Nasal Mucosa in vitro

“…Under the conditions used for short-term culture, human nasal mucosa remained viable for at least 4 h and at least 48 h when kept in culture on collagen-containing gelatin sponges, as demonstrated by positive Ki-67 staining [11].…”

“…Previously, we described a culture system o f human na sal mucosa [11], based on the work of Leighton [12], to histoculture human nasal mucosa on gelatin sponges at the air/ liquid interface [13][14][15]. We could show that cells remain proliferative and viable for up to 48 h as evaluated by immu nohistochemical staining for proliferation (Ki-67).…”

Antioxidative enzymes in human nasal mucosa after exposure to ozone. Possible role of GSTM1 deficiency

“…Mucosa surface control was assessed by a stereomicroscope (Zeiss, Oberkochen, Germany). As culture medium we used HEPES buffer containing 4 mM KCl, 1,1 mM MgCl 2 (Sigma), 1 mM CaCl 2 (Merck), and 50 µg gentamicine [21].…”