In vitro engineering of a palatal mucosa equivalent with acellular porcine dermal matrix

Abstract

The objective of this study was to develop a palatal mucosa equivalent composed of multilayered oral keratinocytes grown on the acellular porcine dermal matrix. Acellular porcine dermal matrix was prepared through a series of procedures and assessed by histological, immunohistochemical, and scanning electron microscopy examination. The palatal mucosa equivalent was fabricated by seeding oral keratinocytes, which cultured from human palate mucosa, onto the acellular dermal matrix. After 4 days submerged in medium, this composite was raised to the air-liquid interface for another 7 or 14 days of cultivation. The results demonstrated the processed porcine dermal matrix was totally cell-free. The resultant palatal mucosa equivalent showed a multilayered oral epithelium that had been formed, and the number of cell layers was correlated with the culture period at the air-liquid interface. Oral keratinocytes infiltrated into the empty hair follicles of the acellular porcine dermal matrix and formed an anchor-like structure, which exhibited resemblance to the rete ridges of the native palate mucosa. Immunohistochemical staining for CK10/13, CK19, Ki-67 nuclear antigen, and Heparan sulphate indicated the cultured palatal mucosa equivalent shared the same characteristics with that of the native palate mucosa. In conclusion, our fabricated palatal mucosa equivalent exhibited the characteristics of the native counterpart, and this equivalent might be useful for recovery of the wounds in the palate secondary to palatoplasty.

The influence of elastin-like recombinant polymer on the self-renewing potential of a 3D tissue equivalent derived from human lamina propria fibroblasts and oral epithelial cells

“…In our previous study with our patented collagen-glycosaminoglycan-chitosan scaffold [14], we observed the same pattern: at the end of 3 weeks of culture, only a few proliferative cells (Ki67 positive) remained in the reconstructed oral mucosa [15]. Other studies on tissue-engineered oral mucosa also found that at the end of the culture period (even for cultures of only 11 days) either no proliferative cell [16] or only a few remained in the oral mucosal equivalents [13,17,18], which is much less than found in native oral mucosa. These results led the researchers to investigate into the mechanisms of various regenerative medicine approaches and epithelial stem cell biology [8].…”

Cryopreserved lip mucosa tissue derived keratinocytes can fabricate tissue engineered palatal mucosa equivalent

“…The cultured oral keratinocytes used for the fabrication of mucosa equivalent were in passage 2 and the cell seeding concentration was adjusted to 5 × 10 5 cells/mL. The cell suspension was seeded onto the papillary surface of the acellular porcine dermal matrix, which was made from porcine skin after decellularization with 0.25% trypsin solution and 0.5% Triton X‐100 solution, and cross‐linking with 0.2% glutaraldehyde solution, as previously described 16. Subsequently, these composites were submerged in keratinocyte serum‐free medium for four days cultivation and then, they were lifted to the air‐liquid interface for seven days cultivation.…”

Morphogenesis of Rete Ridges in Human Oral Mucosa: A Pioneering Morphological and Immunohistochemical Study

“…An RR-like structure has been found in microtextured basal lamina analogues [Downing et al, 2005] and in our previous study on the empty hair follicles of an acellular porcine dermal matrix [Xiong et al, 2008]. However, the native morphology, characteristics, and morphogenesis-related factors of RRs at the dermal-epidermal interface are still unclear and warrant further investigation.…”

“…There are several approaches for constructing the microtexture at the epidermal-dermal interface [Pins et al, 2000;Downing et al, 2005;Xiong et al, 2008]. However, the morphological characteristics of native RRs and the expression of ERK1/2, Ki67, and keratin-19 therein are not clear.…”

“…There are several approaches to reconstructing RRs [Jani and Bhargava, 1976;Bale and White, 1982;Moore et al, 1992;Pins et al, 2000;Ahn et al, 2005;Downing et al, 2005;Xiong et al, 2008]. An RR-like structure has been found in microtextured basal lamina analogues [Downing et al, 2005] and in our previous study on the empty hair follicles of an acellular porcine dermal matrix [Xiong et al, 2008].…”