Mikoshiba Ikuo, Hagikura Kazuhiro, Matsumoto Taro, Kazama Tomohiko, Asai Ryohei, Hao Hiroyuki, Oshima Takeshi
Department of Otolaryngology-Head and Neck Surgery, Nihon University School of Medicine, Tokyo, Japan.
Department of Functional Morphology, Division of Cell Regeneration and Transplantation, Nihon University School of Medicine, Tokyo, Japan.
PLoS One. 2025 Sep 5;20(9):e0324965. doi: 10.1371/journal.pone.0324965. eCollection 2025.
Glottic insufficiency results from impaired vocal fold contact, leading to a gap between the folds and manifesting as hoarseness and respiratory difficulties. Vocal folds injection is a commonly utilized therapeutic approach to rectify this gap by augmenting vocal folds volume; however, the optimal injectable material remains undetermined. Dedifferentiated fat cells (DFATs), derived from mature adipocytes, exhibit robust proliferative capacity and multipotency, establishing them as potential candidates for treating glottic insufficiency. This study investigated the therapeutic efficacy of DFATs in a rat model of recurrent laryngeal nerve paralysis. Twenty-five rats were used for the therapeutic evaluation: the recurrent laryngeal nerve of each rat was resected unilaterally, and 5 weeks later, the atrophied vocal fold muscles were injected with either 10 μl of saline (control), 1.0 × 106 DFATs in 10 μl (DFAT group), or 1.0 × 106 DFATs combined with 500 ng of basic fibroblast growth factor (bFGF) in 10 μl (DFAT + bFGF group). At 4 weeks post-injection, laryngeal endoscopy evaluated the glottic gap, followed by histological analyses of muscle atrophy, collagen deposition. To investigate mechanisms, an independent cohort of 25 rats received identical treatments and was evaluated 2 weeks post-injection for cell proliferation (Ki-67) and apoptosis (TUNEL). To confirm DFAT engraftment, GFP-labeled DFATs were evaluated at 2, 4, and 6 weeks post-injection. Results indicated that both the DFAT and DFAT + bFGF groups exhibited significantly reduced glottic gaps, increased collagen deposition, and decreased TUNEL-positive apoptotic cells compared to controls. Notably, the DFAT + bFGF group displayed superior outcomes, including a greater vocal muscle area and enhanced Ki-67-positive cell proliferation, indicating reduced atrophy. GFP-positive DFATs remained detectable for up to 6 weeks, confirming engraftment. These findings underscore the potential of DFATs, particularly with bFGF, as an innovative and effective therapy for glottic insufficiency secondary to recurrent laryngeal nerve paralysis.
声门功能不全是由声带接触受损引起的，导致声带之间出现间隙，并表现为声音嘶哑和呼吸困难。声带注射是一种常用的治疗方法，通过增加声带体积来纠正这种间隙；然而，最佳的可注射材料仍未确定。去分化脂肪细胞（DFATs）源自成熟脂肪细胞，具有强大的增殖能力和多能性，使其成为治疗声门功能不全的潜在候选者。本研究在大鼠喉返神经麻痹模型中研究了DFATs的治疗效果。25只大鼠用于治疗评估：每只大鼠的喉返神经单侧切除，5周后，将10μl生理盐水（对照组）、10μl中含1.0×10⁶个DFATs（DFAT组）或10μl中含1.0×10⁶个DFATs与500ng碱性成纤维细胞生长因子（bFGF）的混合物（DFAT + bFGF组）注射到萎缩的声带肌肉中。注射后4周，通过喉镜检查评估声门间隙，随后对肌肉萎缩、胶原蛋白沉积进行组织学分析。为了研究机制，另一组25只大鼠接受相同治疗，并在注射后2周评估细胞增殖（Ki-67）和细胞凋亡（TUNEL）情况。为了确认DFAT的植入，在注射后2、4和6周对绿色荧光蛋白标记的DFATs进行评估。结果表明，与对照组相比，DFAT组和DFAT + bFGF组的声门间隙均显著减小，胶原蛋白沉积增加，TUNEL阳性凋亡细胞减少。值得注意的是，DFAT + bFGF组表现出更好的效果，包括更大的声带肌肉面积和增强的Ki-67阳性细胞增殖，表明萎缩减轻。绿色荧光蛋白阳性的DFATs在长达6周的时间内仍可检测到，证实了植入。这些发现强调了DFATs，特别是与bFGF联合使用时，作为治疗喉返神经麻痹继发声门功能不全的一种创新且有效的疗法的潜力。
