Effect of Fibroblasts on Tracheal Epithelial Regeneration in Vitro

Kobayashi, K.; Nomoto, Yukio; Suzuki, Teruhisa; Tada, Yasuhiro; Miyake, Masakazu; Hazama, Akihiro; Kanemaru, Shin‐ichi; Nakamura, Tatsuo; Omori, Koichi

doi:10.1089/ten.2006.12.ft-199

Cited by 23 publications

(46 citation statements)

References 49 publications

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“…4 Many researchers are focusing their studies on development of an in vitro model of the respiratory epithelium to study cell interactions, differentiation, and mechanisms of protein production. [12][13][14][15][16] However, in tissue engineering, the study of basic cell molecular level is equally important to maintain cell quality for regenerating a superior quality epithelium, and most importantly, functional for in vivo uses. Three mucins, MUC2, MUC5A, and MUC5B are expressed by the respiratory epithelium.…”

Section: Introductionmentioning

confidence: 99%

Human Nasal Turbinates as a Viable Source of Respiratory Epithelial Cells Using Co‐Culture System Versus Dispase Dissociation Technique

et al. 2007

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Objective: To compare a co‐culture system with a conventional dispase‐dissociation method for obtaining functional human respiratory epithelial cells from the nasal turbinates for tissue engineering application. Methods: Human respiratory epithelial cells were serially passaged using a co‐culture system and a conventional dispase‐dissociation technique. The growth kinetics and gene expression levels of the cultured respiratory epithelial cells were compared. Four genes were investigated, namely cytokeratin‐18, a marker for ciliated and secretory epithelial cells; cytokeratin‐14, a marker for basal epithelial cells; MKI67, a proliferation marker; and MUC5B, a marker for mucin secretion. Immunocytochemical analysis was performed using monoclonal antibodies against the high molecular‐weight cytokeratin 34βE12, cytokeratin 18, and MUC5A to investigate the protein expression from cultured respiratory epithelial cells. Results: Respiratory epithelial cells cultured using both methods maintained polygonal morphology throughout the passages. At passage 1, co‐cultured respiratory epithelial showed a 2.6‐times higher growth rate compared to conventional dispase dissociation technique, and 7.8 times higher at passage 2. Better basal gene expression was observed by co‐cultured respiratory epithelial cells compared to dispase dissociated cells. Immunocytochemical analyses were positive for the respiratory epithelial cells cultured using both techniques. Conclusion: Co‐culture system produced superior quality of cultured human respiratory epithelial cells from the nasal turbinates as compared to dispase dissociation technique.

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Section: Introductionmentioning

confidence: 99%

Human Nasal Turbinates as a Viable Source of Respiratory Epithelial Cells Using Co‐Culture System Versus Dispase Dissociation Technique

et al. 2007

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“…Respiratory epithelial cells and fibroblasts were cultured using the coculture system described in our previous studies [Ziegelaar et al, 2002;Nur Adelina et al, 2007]. Stimulatory effects of fibroblasts facilitated epithelial cell differentiation into mainly basal, ciliated and goblet cells with increased mucin secretion [Goto et al, 1999;Kobayashi et al, 2006Kobayashi et al, , 2007. Respiratory epithelium cells obtained using a coculture system produce significantly more basal cells and stem cell gene expression, suggesting a high potential to regenerate a normal tracheal epithelium in vivo [Hong et al, 2004;Nur Adelina et al, 2007].…”

Section: Discussionmentioning

confidence: 99%

“…Fibroblasts beneath epithelial cells are important for the reconstruction of the basement membrane. The basement membrane was reported to influence differentiation, migration and proliferation of epithelial cells and is im- portant in maintaining structural tissue integrity [Kobayashi et al, 2006[Kobayashi et al, , 2007Nomoto et al, 2008]. Another previous study suggested that epithelial-fibroblast interaction plays a crucial role in the organogenesis, morphogenesis and differentiation of the tracheal epithelial cells [Goto et al, 1999].…”

Section: Discussionmentioning

confidence: 99%

“…Epithelial cell migration, proliferation and differentiation will take place to cover defective regions for the reconstruction of a functional epithelium. However, the slowness of epithelial regeneration of injured tracheal mucosa is a challenging problem [Kobayashi et al, 2006]. The failure of regenerated epithelium to cover the defect caused exaggerated fibrous tissue formation followed by contraction of the scar, which subsequently leads to recurrent tracheal stenosis [Le Visage et al, 2004;Nomoto et al, 2006].…”

Section: Introductionmentioning

confidence: 99%

“…Recently, several studies pertaining to tracheal regeneration using tissue engineering techniques have been reported. Experimental and clinical studies in the development of alloplastic tracheal prostheses have shown that the presence of epithelial lining with excessive ingrowth of granulation tissue on the luminal surfaces may be a significant contributing factor in the failure of these devices [Kanzaki et al, 2006;Nomoto et al, 2006;Yamashita et al, 2007]. The proliferation of granulation tissue which stopped along with the surface epithelialization indicates that the luminal respiratory epithelial lining is needed for quick regeneration of the tracheal epithelium [Tsukada and Osada, 2004;Tan et al, 2006].…”

Section: Introductionmentioning

confidence: 99%

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Autologous Implantation of Bilayered Tissue-Engineered Respiratory Epithelium for Tracheal Mucosal Regenesis in a Sheep Model

Heikal¹,

Aminuddin

Jeevanan

et al. 2010

Cells Tissues Organs

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The objective of this study was to regenerate the tracheal epithelium using autologous nasal respiratory epithelial cells in a sheep model. Respiratory epithelium and fibroblast cells were harvested from nasal turbinates and cultured for 1 week. After confluence, respiratory epithelium and fibroblast cells were suspended in autologous fibrin polymerized separately to form a tissue-engineered respiratory epithelial construct (TEREC). A 3 × 2 cm² tracheal mucosal defect was created, and implanted with TEREC and titanium mesh as a temporary scaffold. The control groups were divided into 2 groups: polymerized autologous fibrin devoid of cells (group 1), and no construct implanted (group 2). All sheep were euthanized at 4 weeks of implantation. Gross observation of the trachea showed minimal luminal stenosis formation in the experimental group compared to the control groups. Macroscopic evaluation revealed significant mucosal fibrosis in control group 1 (71.8%) as compared to the experimental group (7%). Hematoxylin and eosin staining revealed the presence of minimal overgrowth of fibrous connective tissue covered by respiratory epithelium. A positive red fluorescence staining of PKH26 on engineered tissue 4 weeks after implantation confirmed the presence of cultured nasal respiratory epithelial cells intercalated with native tracheal epithelial cells. Scanning electron microscopy showed the presence of short microvilli representing immature cilia on the surface of the epithelium. Our study showed that TEREC was a good replacement for a tracheal mucosal defect and was able to promote natural regenesis of the tracheal epithelium with minimal fibrosis. This study highlighted a new technique in the treatment of tracheal stenosis.

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Development of resorbable nanocomposite tracheal and bronchial scaffolds for paediatric applications

Teoh

Crowley²,

Birchall

et al. 2015

British Journal of Surgery

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Effect of Fibroblasts on Tracheal Epithelial Regeneration in Vitro

Cited by 23 publications

References 49 publications

Human Nasal Turbinates as a Viable Source of Respiratory Epithelial Cells Using Co‐Culture System Versus Dispase Dissociation Technique

Human Nasal Turbinates as a Viable Source of Respiratory Epithelial Cells Using Co‐Culture System Versus Dispase Dissociation Technique

Autologous Implantation of Bilayered Tissue-Engineered Respiratory Epithelium for Tracheal Mucosal Regenesis in a Sheep Model

Development of resorbable nanocomposite tracheal and bronchial scaffolds for paediatric applications

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