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Mucosal keratinocyte isolation: a short comparative study on thermolysin and dispase
H A Rakhorst1, W M W Tra, S J Posthumus-van Sluijs
1Department of Plastic and Reconstructive Surgery, Erasmus University Medical Centre Rotterdam, NL-3000CA Rotterdam, The Netherlands. H.Rakhorst@easmusmc.nl
International Journal of Oral and Maxillofacial Surgery
|September 13, 2006
Summary
Dispase is superior to thermolysin for isolating keratinocytes from rabbit buccal mucosa. This method yields more cells, enabling larger grafts for reconstructing oral defects.
Area of Science:
- Regenerative Medicine
- Tissue Engineering
- Oral Surgery
Background:
- Cultured mucosal substitutes are emerging for oral defect reconstruction.
- Efficient keratinocyte isolation is crucial for creating these substitutes.
Purpose of the Study:
- To compare the efficacy of dispase and thermolysin in isolating keratinocytes from rabbit buccal mucosa.
- To determine which enzyme yields a higher quantity and quality of cells for tissue engineering.
Main Methods:
- Rabbit buccal mucosa was treated with either dispase or thermolysin.
- Histology, cytokeratin 13 staining, seeding efficiency, and cell diameter were analyzed.
- Keratinocyte yield per surface area was quantified.
Main Results:
- Thermolysin treatment led to incomplete epidermal-dermal separation.
- Dispase yielded significantly higher keratinocyte numbers per cm², improved cell size, and enhanced seeding efficiency.
- A 3.45-fold larger graft could be produced using dispase-isolated cells.
Conclusions:
- Dispase is a more favorable enzyme than thermolysin for keratinocyte isolation from buccal mucosa.
- This finding supports the use of dispase in developing cultured mucosal substitutes for oral reconstruction.
