Structural Characterization and Safety Validation of Cultured Skin Cells Grown on Routinely Used Dressing in Pediatric Incubators: A Preliminary Report

Kleintjes WG and Prinsloo TK

Department of Surgical Sciences, Stellenbosch University Medical School, Francie van Zijl Avenue, South Africa WC Provincial Adult Tertiary Burns Centre, Tygerberg Hospital, South Africa Department of Biomedical Sciences, Cape Peninsula University of Technology (CPUT), South Africa Department of Emergency Medical Sciences, Cape Peninsula University of Technology (CPUT), South Africa

^*Correspondance to: Wayne George Kleintjes 

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Abstract:

Cultured Epithelial Autografts (CEA), have improved wound treatment outcomes and combining it with temporary coverage reportedly increases graft take. In this study, a modified composite technique was established that mitigated several drawbacks experienced in resource-limited settings. The aim was to report on the CEA’s preliminary histological findings and demonstrate its safety for routine use. Biopsy-retrieved keratinocytes in autologous plasma were seeded onto routinely-used bacteria- and fungi-binding dressing pads. Incubation in pediatric incubators at 37°C then followed and fresh autologous plasma was applied daily while amorphous hydrogel every third or fourth day. Confluence was reached (2 weeks), and culture samples were histologically evaluated using Light microscopy (4X and 10X magnification) with two stains (Hematoxylin and eosin, and Masson’s Trichrome) and transmission electron microscopy (800-4000X magnification) with Reynold’s lead nitrate stain. Bacteriological safety was externally assessed by analyzing wound swabs (n=10; 2 each =20 samples) of CEA grown in both pediatric and standard culture incubators before transplant. Light microscopy demonstrated densely packed keratinocytes, collagen fibers, and classical epidermal and dermal layer differentiation. Electron microscopy also displayed the differentiated cellular nature including basal epithelial and dermal cells, keratinocytes and melanocytes. No pathogenic organisms were observed in any of the cell culture specimens tested. The differentiated nature of this CEA emulated the epidermis to a large degree which was not only indicative of functional integrity, but also mechanical integrity outcomes of the technique. No differences were observed between the incubators in terms of the pathogenic bacteria grown or cell culture growth.

Keywords:

Burns; Cultured epithelial autografts; Light microscopy; Transmission electron microscopy; Biosafety

Cite the Article:

Kleintjes WG, Prinsloo TK. Structural Characterization and Safety Validation of Cultured Skin Cells Grown on Routinely Used Dressing in Pediatric Incubators: A Preliminary Report. World J Surg Surgical Res. 2024; 7: 1569..