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M M Ghosh

Publications and source records attributed to M M Ghosh.

12 recordsLinked to original sources
PubMed1997

A comparison of methodologies for the preparation of human epidermal-dermal composites.

The purpose of this study was to compare methodologies for the preparation of human skin composites based on deepidermized acellular dermal matrix, epidermal keratinocytes, and dermal fibroblasts with the aim of preparing a clinically useful skin substitute. Dermal matrices were prepared from normal human skin and we compared methods of sterilization (glycerol treatment, ethylene oxide treatment, and gamma irradiation), methods of removing the epidermis (sodium chloride, phosphate buffered saline, and dispase), and methods of seeding the composites with fibroblasts and keratinocytes. We report protocols for reproducibly preparing composites that share many of the features of normal skin after 7 days culture at an air-liquid interface. Such composites can be based on allodermis pretreated with either glycerol or ethylene oxide (although the latter gave less consistent results than the glycerol treatment). Fibroblast penetration into the dermis could be achieved by culture of cells on the reticular or papillary surface of the dermis. However, we report for the first time that fibroblast entry from the papillary surface only occurred when keratinocytes were also present.

Cell Differentiation↗
PubMed1995

Effect of glycerol on intracellular virus survival: implications for the clinical use of glycerol-preserved cadaver skin.

Glycerol has long been used for the preservation of skin allografts. The antimicrobial activity of glycerol has not been fully documented. This paper reports the results of an investigation of a model studying the effect of glycerol on the inactivation of intracellular viruses. Two viruses--herpes simplex type I (HSV-1) and poliovirus--were cultured within human dermal fibroblasts. These intracellular viruses were incubated with 50 per cent, 85 per cent and 98 per cent glycerol at 4 degrees C and 20 degrees C for 4 weeks. Each week, the cultures in glycerol and controls in fibroblast maintenance medium were assayed for virus infectivity by examining the ability of harvested viruses to infect further fibroblasts. At 4 degrees C, 85 per cent glycerol could not fully inactivate intracellular HSV-I or poliovirus even after 4 weeks; 98 per cent glycerol inactivated intracellular HSV-I (after 3 weeks) but could not fully inactivate intracellular poliovirus after 4 weeks. At 20 degrees C, 85 per cent glycerol inactivated intracellular HSV-I (within 1 week) but could not fully inactivate intracellular poliovirus after 4 weeks; 98 per cent glycerol inactivated intracellular HSV-I (within 1 week) and inactivated intracellular poliovirus (after 2 weeks). It is suggested that, on the basis of this study, glycerol can reduce intracellular virus infectivity but that its effects are very dependent on concentration, time and temperature such that we would recommend that allograft skin be exposed to 98 per cent glycerol for a minimum of at least 4 weeks at a minimum temperature of 20 degrees C before clinical use.

Cadaver↗
PubMed1981

Echocardiography.

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Echocardiography↗
PubMedDate not supplied

A simple human dermal model for assessment of in vitro attachment efficiency of stored cultured epithelial autografts.

This study investigated the effect of short-term storage on the viability and in vitro attachment efficiency of cultured epithelial autograft sheets. Four storage protocols were investigated: overnight at 37 degrees C in keratinocyte culture medium, overnight at 4 degrees C in phosphate-buffered saline solution, overnight at -80 degrees C in cryopreservation medium (containing 10% dimethyl sulphoxide), and 1 week at -80 degrees C in cryopreservation medium. Viability was assessed before and after storage by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. All the storage conditions significantly reduced viability compared with fresh sheets, and no significant decrease was seen when the sheets stored under the different protocols were compared with each other. The best viability obtained was 60% of that of the fresh sheets. The in vitro viability of these stored sheets was then compared with that of the fresh sheets by culturing them on deepidermized acellular allodermis and assessing the composites formed by light microscopy and the MTT assay. The fresh sheets attached and formed a histologically demonstrable composite with the dermal substrate, whereas none of the stored sheets formed demonstrable composites. The MTT assay demonstrated that composites formed with the stored sheets had less than 10% viability compared with composites formed with fresh sheets. It is concluded that under the conditions of storage examined, the viability of cultured epithelial autograft sheets was significantly reduced, but up to 60% of viability could be retained in some cases. However, the subsequent in vitro attachment and proliferation of such preserved sheets on allogeneic deepidermized dermis was poor compared with that of fresh cultured epithelial autograft sheets.

Cells, Cultured↗