Testing Wound-healing Activity in T15 Fibroblast Cultures: A Morphometric Analysis

Abstract

The purpose was to evaluate the use of mouse T15 fibroblast cell cultures for the investigation of wound-healing activity. In order to investigate their mechanisms of action, the effects of drugs with wound-healing activities were compared by using morphometric analyses by microscopy after cell staining. A number of parameters were used to evaluate the effects of titrated extracts from Centella asiatica and dexpanthenol (drugs that have been used in medical practice for their wound-healing activities) on cultured mouse T15 fibroblasts. These parameters were: the total number of cells; the number of T15 cells in mitosis; the percentages of fusiform, polygonal, round and vacuole-containing cells; and the number of intracellular collagen granules. The results indicate that these two drugs exhibit wound-healing activities by activating fibroblast cells, and have cytoprotective effects, although their mechanisms of action on mouse T15 fibroblasts were different. On the basis of our findings, mouse T15 fibroblast cell cultures seem to be useful for the pharmacological screening of compounds with wound-healing activity.

Evaluation of the wound healing potentials of two subspecies of Hypericum perforatum on cultured NIH3T3 fibroblasts

“…Fibroblast cell cultures have been proposed as a method for testing wound‐healing activity in vitro (Graham et al ., 1984). However, this approach has been used only in a few studies on the development of new drugs and for the investigation of the mechanism of action (Maquart et al ., 1990; Korkmaz et al ., 2000; Öztürk et al ., 2006; 2007). In this study, it was observed that TECA mainly increases mitotic activity (Table 2) and the number of polygonal cells (Table 3), and slightly stimulates collagen production in the NIH3T3 cells (Table 5).…”

Wound-healing activity of St. John's Wort (Hypericum perforatum L.) on chicken embryonic fibroblasts

“…Fibroblast cell cultures have been proposed as a method for testing wound-healing activity in vitro (Graham et al, 1984). However, this approach has been used in only a few studies on the development of new drugs and for the investigation of mechanism of action (Maquart et al, 1990;Korkmaz et al, 2000;. The data obtained with dexpanthenol and TECA clearly demonstrate that cell culture of chicken embryonic fibroblasts is a suitable model for the investigation of wound-healing activity, since these compounds cause specific morphological changes in the cultured fibroblasts.…”

“…The methods for chicken embryonic fibroblast culture and for administration of DMSO, TECA, dexpanthenol and Hypericum extract into culture media were quite similar to that described earlier for the culture of T15 fibroblast cell lines (Korkmaz et al, 2000). The concentrations for TECA (5 and 10 g/mL) and dexpanthenol (1 and 10 g/mL) were selected on the basis of literature data (Lacroix et al, 1988;Maquart et al, 1990;Korkmaz et al, 2000), whereas the effective concentrations of Hypericum perforatum extract (1, 5, 10 and 100 g/mL) were determined after scanning a wide concentration range in pilot experiments. Dexpanthenol, TECA and Hypericum extract were dissolved in DMSO and the same dose of DMSO at 2 L/mL was used for control and experimental groups were compared to both DMSO and blank controls.…”

“…Staining of cultured chicken embryonic fibroblasts was conducted according to Masson's trichrome technique, as described previously (Jones, 2002;Korkmaz et al, 2000). Cell monolayers grown on cover slips were fixed in 70% ethanol solution for 15 min, and rehydrated in distilled water for 5-10 min.…”

“…Morphometric analysis applied in this study was also quite similar to the procedure described in a previous study (Korkmaz et al, 2000). After staining, controls, extract-and drugtreated fibroblasts were examined under a light microscope (Olympus, BH-2, Tokyo, Japan) with fully automated photographic attachment using 400× and 800× magnification.…”