Tissue-engineered Oral Mucosa: a Review of the Scientific Literature

Moharamzadeh, Keyvan; Brook, Ian M.; Noort, Richard van; Scutt, A.; Thornhill, Martin H.

doi:10.1177/154405910708600203

Cited by 170 publications

(141 citation statements)

References 80 publications

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“…Culturing keratinocyte sheets on glass or plastic surfaces allowed researchers to analyze the biology of these cells and their responses against different stimuli, e.g., bacteria or mediators of inflammation. However, performing research using non-differentiating cell monolayers does not reflect the real situation in the oral cavity, because oral epithelial cells proliferate at the basal layer of a stratified epithelium, where daughter cells of the basal cells differentiate toward the surface of the epithelium (Moharamzadeh et al 2007). Introduction of keratinocyte cultures with stratification was a milestone in cell biology studies.…”

Section: Introductionmentioning

confidence: 99%

Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model

Gürsoy

Könönen

et al. 2016

Cytotechnology

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In construction of epithelial cells as multilayers, the cells are grown submerged to confluence on fibroblast-embedded collagen gels and, then, lifted to air to promote their stratification. We recently demonstrated that gingival epithelial cells form uniform monolayers on semi-permeable nitrocellulose membranes, supported with a semi-solid growth medium, which allows the cells to grow at an airliquid-solid interface from the beginning of the culturing protocol. In this study, the aim was to further develop our previous model to form a multilayered gingival epithelial culture model. Two different epithelial cell lines (HaCaT from skin and HMK from gingiva) were used in all experiments. Both cell lines were grown first as monolayers for 3 days. After that, keratinocytes were trypsinized, counted and seeded on a sterile semi-permeable nitrocellulose membrane placed on the top of a semi-solid growth medium, forming an air-liquid-solid interface for the cells to grow. At days 1, 4, and 7, epithelial cells were fixed, embedded in paraffin, and sectioned for routine Hematoxylin-Eosin staining and immunohistochemistry for cytokeratin (Ck). At day 1, HMK cells grew as monolayers, while HaCaT cells stratified forming an epithelium with two to three layers. At day 4, a stratified epithelium in the HMK model had four to five layers and its proliferation continued up to day 7. HaCaT cells formed a dense and weakly proliferating epithelium with three to four layers of stratification at day 4 but the proliferation disappeared at day 7. At all days, both models were strongly positive for Ck5, Ck7, and Ck 19, and weakly positive for Ck10. Gingival epithelial cells stratify successfully on semi-permeable nitrocellulose membranes, supported with a semi-solid growth medium. This technique allows researchers to construct uniform gingival epithelial cell multilayers at an air-liquid-solid interface, without using collagen gels, resulting in a more reproducible method.

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Section: Introductionmentioning

confidence: 99%

Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model

Gürsoy

Könönen

et al. 2016

Cytotechnology

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“…Tissue-engineered oral mucosal equivalents have been developed for in vitro biocompatibility studies, as well as for mucosal irritation and oral disease studies with the aim to better understand disease process and discover new treatments [1,2]. In the last decade, research has concentrated on the characterization of human mucosal equivalent by introducing new dermal scaffold and epithelial cell culture methods.…”

Section: Introductionmentioning

confidence: 99%

“…In 1975, Rheinwald and Green introduced a method to grow human keratinocytes in in vitro serial cultures, using a feeder layer composed of irradiated mouse fibroblasts and a specific culture medium [3]. This method is widely used for the culture of keratinocytes and single-layer epithelial sheets, but such sheets are fragile, difficult to handle and tend to contract [1]. Multilayer sheets of cultured epithelium were obtained by culturing oral keratinocytes and fibroblasts, crucial for the production of extracellular matrix, on permeable ethylene terephthalate cell culture membrane (PET) at the air/liquid interface [4] or in polycarbonate cell culture inserts, developed by SkinEthic Laboratories (Nice, France).…”

Section: Introductionmentioning

confidence: 99%

“…Multilayer sheets of cultured epithelium were obtained by culturing oral keratinocytes and fibroblasts, crucial for the production of extracellular matrix, on permeable ethylene terephthalate cell culture membrane (PET) at the air/liquid interface [4] or in polycarbonate cell culture inserts, developed by SkinEthic Laboratories (Nice, France). These models have the characteristic of being very similar to native epithelium and showing signs of differentiation, such as different cytokeratin expression and basement membrane formation, but not to the point of full differentiation because the cells used are derived from oral squamous carcinoma [1]. Moreover, the scaffold that supports the cells is an important element in oral mucosa reconstruction.…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, the scaffold that supports the cells is an important element in oral mucosa reconstruction. The currently available scaffolds can be divided into several categories: naturally derived scaffolds, such as acellular dermis and amniotic membrane [5]; fibroblast-populated skin substitutes [6]; pure collagen scaffolds and collagen-based matrices [7]; gelatin-based scaffolds such as gelatin-hyaluronate and gelatinchitosan-hyaluronic acid; fibrin-based materials; synthetic scaffolds, such as elastin-like recombinant polymer [2] and hybrid scaffolds, which are combinations of natural and synthetic matrices [1]. The benefit of pure or collagen-based matrices is that the collagen gel supports fibroblasts, which provides a suitable substrate for keratinocyte multilayer formation, but it also biodegrades rapidly and has poor mechanical properties.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Medium-term Culture of Normal Human Oral Mucosa: A Novel Three-dimensional Model to Study the Effectiveness of Drugs Administration

Bucchieri

Fucarino²,

Gammazza³

et al. 2012

CPD

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Tissue-engineered oral mucosal equivalents have been developed for in vitro studies for a few years now. However, the usefulness of currently available models is still limited by many factors, mainly the lack of a physiological extracellular matrix (ECM) and the use of cell populations that do not reflect the properly differentiated cytotypes of the mucosa of the oral cavity. For this reason, we have developed a novel three-dimensional culture model reflecting the normal architecture of the human oral mucosa, with the main aim of creating a better in vitro model where to test cellular responses to drugs administration. This novel 3D cell culture model (3D outgrowth) was set up using an artificial extracellular matrix (Matrigel TM ), allowing the interactions required for proper differentiation of the various citotypes which form the mucosal layer. Biopsies of human oral mucosa, in fragments of about 0.5 mm 3 , were placed onto 6.5mm Transwells, covered with Matrigel TM and grown in a specific culture medium. A gradual formation of an architectural structure similar to that of the in vivo oral mucosa was observed. Transmission electron and confocal microscopy were employed to characterize the newly developed model: the cell components (keratinocytes and fibroblasts) differentiated properly within the outgrowth and reconstituted, in vitro, the physiological structure of the human oral mucosa, including a stratified non-keratinized squamous layer composed of four different layers, a proper basal membrane and a lamina propria where fibroblasts produce ECM. Moreover, keratinocytes expressed CK5, CK13, CK19 and E-cadherin, whereas fibroblasts expressed collagen type I and IV, laminin and fibronectin. 3D outgrowths could be considered a valid alternative to animal models, and provide useful information for researchers interested in studying the responses of the human oral mucosa to locally delivered drugs or other exogenous treatments.

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Cutaneous Toxicology

Hermansky¹

2009

General, Applied and Systems Toxicology

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The skin is the largest organ of the body. It serves many functions one of which is protection from the outside world, including chemical and microbiological insults. Skin ailments represent one of the most common complaints of patients in medical settings and account for significant loss of productivity in the workplace. Cutaneous toxicity from chemical exposures is responsible for many of these complaints. Skin reactions to chemicals often appear similar, but have many biological causes, including chemical irritation and sensitization (allergies). A primary goal of the cutaneous toxicologist is to predict the potential irritant and/or sensitization potential of chemicals. This chapter summarizes the varied effects that chemicals may have on the skin, including acute and chronic conditions. The chapter also describes the multiple methods to predict potential cutaneous chemical hazards that may be produced in the workplace or via consumer exposure to topical drugs and cosmetics. Important human, animal and in vitro methods to predict a chemical hazard are described.

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Tissue-engineered Oral Mucosa: a Review of the Scientific Literature

Cited by 170 publications

References 80 publications

Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model

Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model

Medium-term Culture of Normal Human Oral Mucosa: A Novel Three-dimensional Model to Study the Effectiveness of Drugs Administration

Cutaneous Toxicology

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