Clinical and Histopathological Analysis of Healing Process of Intraoral Reconstruction with ex vivo Produced Oral Mucosa Equivalent
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概要
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We fabricated ex vivo produced oral mucosa equivalent (EVPOME) from patients'oral mucosal keratinocytes without using animal-derived serum or feeder layer cells.To confirm the clinical benefits of 1) early initiation of epithelialization, 2) a shortperiod until complete healing and 3) negligible scar contracture, the mechanism ofwound healing after EVPOME transplantation for oral mucosal defects was analyzedhistopathologically. Transplantation was performed on 15 patients (eight men andseven women; aged between 51 and 76 years, mean, 66.6 years). Two patients hadsquamous cell carcinoma of the tongue, nine had leukoplakia (four in the tongue only,two in the gingiva only, one in the buccal mucosa, and two in two or more areas), andfour had hypoplasia in the alveolar ridge. The mean interval between punch-biopsy forthe fabrication of EVPOME and its transplantation for the reconstruction of oralmucosal defects was 28.5 days, by which time EVPOME with a mean size of 6.5 cm^2and a cell count of 8.6 × 10^5 could be obtained. The underlying disease, past history,and smoking history of the patients did not constitute negative factors for EVPOMEfabrication. About 10 days after transplantation, EVPOME began uniting with thesurrounding epithelium. The mean duration required for the wound to be completelycovered (28.2 days) was much shorter than after transplantation of only an acellularallogenic dermal matrix (AlloDerm○!R), and showed only slight scar formation, similarto that observed after artificial dermis (Terdermis○!R) transplantation. Presence oflaminin-1, 5 and type IV collagen in the basement membrane of EVPOME wasconfirmed, and the arrangement and positioning of keratinocytes were preservedduring the degradation of perlecan and anchoring fibrils (type VII collagen) forremodeling, i.e., the period of the most active remodeling of EVPOME transplantation.Only a few fibroblasts were observed in the lamina propria during this period,suggesting that keratinocyte-derived cytokines, rather than fibroblast-derivedcytokines, play an important role in the early stages of mucosal wound healing afterEVPOME transplantation. The efficacy of EVPOME is associated with closely relatedto the presence of the keratinocyte-derived system and the usefulness of AlloDerm○!R thatsustains keratinocytes.
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