Tsuyoshi Kojima

Regeneration of Radiation Damaged Salivary Glands with Adipose-Derived Stromal Cells

Radiotherapy is one of the most effective treatments for head and neck cancer. However, the development of dry mouth syndrome is an unavoidable side effect because, in addition to the tumor, the normal salivary glands are included in the irradiation field. Previously, we investigated the protective efficacy of basic fibroblast growth factor (bFGF) in radiation‐damaged salivary glands. In this study, we investigated the efficacy of adipose‐derived stromal cell (ADSC) transplantation for the regeneration of radiation damaged salivary glands.

Increased Expression of Phosphatidylcholine (16:0/18:1) and (16:0/18:2) in Thyroid Papillary Cancer

A good prognosis can be expected for most, but not all, cases of thyroid papillary cancer. Numerous molecular studies have demonstrated beneficial treatment and prognostic factors in various molecular markers. Whereas most previous reports have focused on genomics and proteomics, few have focused on lipidomics. With the advent of mass spectrometry (MS), it has become possible to identify many types of molecules, and this analytical tool has become critical in the field of omics. Recently, imaging mass spectrometry (IMS) was developed. After a simple pretreatment process, IMS can be used to examine tissue sections on glass slides with location information. Here, we conducted an IMS analysis of seven cases of thyroid papillary cancer by comparison of cancerous with normal tissues, focusing on the distribution of phospholipids. We identified that phosphatidylcholine (16:0/18:1) and (16:0/18:2) and sphingomyelin (d18:0/16:1) are significantly higher in thyroid papillary cancer than in normal thyroid tissue as determined by tandem mass (MS/MS) analysis. These distributional differences may be associated with the biological behavior of thyroid papillary cancer.

Implantation of an atelocollagen sponge with autologous bone marrow-derived mesenchymal stromal cells for treatment of vocal fold scarring in a canine model.

Objectives: Vocal fold scarring remains a therapeutic challenge. A new regenerative approach is needed to restore disorganized extracellular matrix. Tissue regeneration requires appropriate cells and a scaffold. Bone marrow–derived mesenchymal stromal cells (BMSCs) are multipotent and secrete many kinds of growth factors to regenerate tissues. Atelocollagen sponges have many large pores that permit cell entry. The present study was performed to evaluate whether implantation of an atelocollagen sponge plus BMSCs is effective for the treatment of vocal fold scarring. Methods: Twelve beagles underwent implantation of an atelocollagen sponge or of an atelocollagen sponge with autologous BMSCs (1.0 × 106 cells) in the subepithelial pockets of scarred vocal folds. Six months after the operation, vibratory examinations and histologic examinations were performed. Results: Mucosal vibrations improved significantly for the atelocollagen sponge–implanted vocal folds. Histologic analyses revealed favorable restoration of the extracellular matrix in the lamina propria. Increased distribution of hyaluronic acid and decreased dense collagen deposition were also noted. These improvements were enhanced by implantation of BMSCs. Conclusions: Implantation of atelocollagen sponges with autologous BMSCs into scarred vocal folds significantly increased hyaluronic acid distribution and decreased dense collagen deposition in the lamina propria, leading to better mucosal vibration.

Implantation of an Atelocollagen Sheet for the Treatment of Vocal Fold Scarring and Sulcus Vocalis

Objectives: The management of vocal fold scarring and sulcus vocalis is challenging. These disorders are thought to be fibroplastic anomalies in the cover portion of the vocal fold that cause deterioration of the vibratory properties of the vocal fold mucosa. Histologic studies have revealed disorganization of extracellular matrix that needs to be addressed in the treatment of scarred vocal folds. Replacement of scar tissues with an appropriate implant may lead to regeneration of the vocal fold mucosa and its tissue properties. This retrospective case study examined the feasibility of using an atelocollagen sheet as a regenerative implant. Methods: Six patients with a post-cordectomy scar or sulcus vocalis underwent implantation of an atelocollagen sheet into the lamina propria of the vocal folds. The procedure consisted of elevation of a microflap, dissection and removal of scar tissue, implantation of the material, and wound closure. Vocal function was evaluated before and after surgery by stroboscopic examination and by aerodynamic and acoustic analyses. Results: The postoperative changes of aerodynamic and acoustic parameters varied among patients; however, gradual improvement was seen in most cases over a year. Stroboscopic findings also revealed gradual improvement of vibratory properties in most cases. Conclusions: Implantation of an atelocollagen sheet may have restorative effects on vocal fold scarring and sulcus vocalis in terms of tissue properties and function of the mucosa.

Recruitment patterns of side population cells during wound healing in rat vocal folds

Despite big advances in understanding the mechanisms of wound healing in vocal fold injury, it still remains unclear which are the decisive factors that lead to a complete restoration or to scarring. Among several other factors, stem cells are believed to play an important role in vocal fold restoration. Side population (SP) cells are considered to contain high numbers of stem cells and have gained great interest in the tissue engineering community. The aim of the following study was to investigate the recruitment pattern of SP cells in a rat vocal fold injury model.

Atelocollagen Sponge as a Stem Cell Implantation Scaffold for the Treatment of Scarred Vocal Folds

Objectives: Treatment of vocal fold scarring remains a therapeutic challenge. Our group previously reported the efficacy of treating injured vocal folds by implantation of bone marrow—derived stromal cells containing mesenchymal stem cells. Appropriate scaffolding is necessary for the stem cell implant to achieve optimal results. Terudermis is an atelocollagen sponge derived from calf dermis. It has large pores that permit cellular entry and is degraded in vivo. These characteristics suggest that this material may be a good candidate for use as scaffolding for implantation of cells. The present in vitro study investigated the feasibility of using Terudermis as such a scaffold. Methods: Bone marrow—derived stromal cells were obtained from GFP (green fluorescent protein) mouse femurs. The cells were seeded into Terudermis and incubated for 5 days. Their survival, proliferation, and expression of extracellular matrix were examined. Results: Bone marrow—derived stromal cells adhered to Terudermis and underwent significant proliferation. Immunohistochemical examination demonstrated that adherent cells were positive for expression of vimentin, desmin, fibronectin, and fsp1 and negative for beta III tubulin. These findings indicate that these cells were mesodermal cells and attached to the atelocollagen fibers biologically. Conclusions: The data suggest that Terudermis may have potential as stem cell implantation scaffolding for the treatment of scarred vocal folds.

Quantification of acute vocal fold epithelial surface damage with increasing time and magnitude doses of vibration exposure.

Because the vocal folds undergo repeated trauma during continuous cycles of vibration, the epithelium is routinely susceptible to damage during phonation. Excessive and prolonged vibration exposure is considered a significant predisposing factor in the development of vocal fold pathology. The purpose of the present study was to quantify the extent of epithelial surface damage following increased time and magnitude doses of vibration exposure using an in vivo rabbit phonation model. Forty-five New Zealand white breeder rabbits were randomized to nine groups and received varying phonation time-doses (30, 60, or 120 minutes) and magnitude-doses (control, modal intensity phonation, or raised intensity phonation) of vibration exposure. Scanning electron microscopy and transmission electron microscopy was used to quantify the degree of epithelial surface damage. Results revealed a significant reduction in microprojection density, microprojection height, and depth of the epithelial surface with increasing time and phonation magnitudes doses, signifying increased epithelial surface damage risk with excessive and prolonged vibration exposure. Destruction to the epithelial cell surface may provide significant insight into the disruption of cell function following prolonged vibration exposure. One important goal achieved in the present study was the quantification of epithelial surface damage using objective imaging criteria. These data provide an important foundation for future studies of long-term tissue recovery from excessive and prolonged vibration exposure.

Transforming growth factor β3 for the prevention of vocal fold scarring

Vocal fold scarring poses a therapeutic challenge. It causes hoarseness and decreases the quality of life. Transforming growth factor β3 (TGFβ3) is highly expressed in fetal wounds that heal without scarring, and administration of TGFβ3 has been reported to prevent scarring of the skin and the buccal mucosa. Thus TGFβ3 is considered to be a key molecule in scar‐free healing. This study aimed to examine the ability of TGFβ3 to prevent vocal fold scarring, with particular attention paid to the distribution of extracellular matrices and functional outcomes.

The protective efficacy of basic fibroblast growth factor in radiation‐induced salivary gland dysfunction in mice

Radiotherapy is one of the most effective treatments for head and neck cancer. However, in addition to the target tumor, normal salivary glands are also included in the irradiation field. This unavoidably results in dry mouth syndrome as a side effect. In this study, the protective efficacy of basic fibroblast growth factor (bFGF) was investigated in radiation‐damaged salivary glands.

Effects of phonation time and magnitude dose on vocal fold epithelial genes, barrier integrity, and function.

To investigate the effects of increasing time and magnitude doses of vibration exposure on transcription of the vocal folds junctional proteins, structural alterations, and functional tissue outcomes.