Krishnan Illayaraja, Vijakumaran Ubashini, Hwei Ng Min, Xian Law Jia, Mohd Yusof Mohd Rafizul, Thangarajah Thavachelvi, Chin Tan Geok, Wong Yin Ping, Kalyanasundaram Anusha, Mahmood Zalina, Rajamanickam Shathiya, Subramani Baskar, Lokanathan Yogeswaran
Department of Tissue Engineering and Regenerative Medicine (DTERM), Faculty of Medicine, Universiti Kebangsaan Malaysia (UKM), Cheras, Kuala Lumpur 56000, Malaysia.
National Pharmaceutical Regulatory Agency (NPRA), Lot 36, Jalan Prof Diraja Ungku Aziz, Petaling Jaya 46200, Malaysia.
Int J Mol Sci. 2025 Jul 16;26(14):6806. doi: 10.3390/ijms26146806.
Umbilical cord mesenchymal stem cells (UCMSCs)-derived small extracellular vehicles (sEVs) are reported to offer therapeutic effects in regenerative medicine, but they lack safety and biodistribution profiles to support smooth translation at the clinical stage and regulatory requirements. Our study aimed to determine the safety and biodistribution profile in a healthy animal model before application in the metabolic syndrome model. Method: Healthy male Sprague Dawley (SD) rats were given an intravenous (IV) injection of normal saline (control group) or pooled fetal UCMSCs-derived sEVs (treated group) every three weeks for 90 days. Morbidity and mortality observation (daily), physical measurements (weekly), selected serum biochemistry (every three weeks), and hematology (every three weeks) were performed for 90 days. Acute toxicity (on day 14) and sub-chronic toxicity (on day 90) were assessed for gross necropsy, relative organ weight, and histopathological assessment of lungs, liver, spleen, kidney, and lymph nodes. Separately, a biodistribution study was conducted with the sEVs preparations labeled with PKH26 fluorescent dye, given intravenously to the rats. The organs were harvested 24 h post-injection. There were no drastic changes in either group's morbidity or mortality, physical, hematological, and biochemistry evaluation. The histopathological assessment concluded moderate (focal) inflammation in the treated group's kidneys and signs of recovery from the inflammation and vascular congestion in the liver. A biodistribution study revealed a higher accumulation of sEVs in the spleen. Multiple IV injections of the pooled fetal UCMSCs-derived sEVs in healthy male SD rats were deemed safe. The sEVs were abundantly distributed in the spleen 24 h post-injection.
据报道，脐带间充质干细胞（UCMSCs）衍生的小细胞外囊泡（sEVs）在再生医学中具有治疗作用，但它们缺乏安全性和生物分布特征，无法支持在临床阶段顺利转化并满足监管要求。我们的研究旨在确定在应用于代谢综合征模型之前，其在健康动物模型中的安全性和生物分布特征。方法：每三周给健康雄性Sprague Dawley（SD）大鼠静脉注射生理盐水（对照组）或汇集的胎儿UCMSCs衍生的sEVs（治疗组），持续90天。在90天内进行发病率和死亡率观察（每日）、身体测量（每周）、选定的血清生化检测（每三周）和血液学检测（每三周）。在第14天评估急性毒性，在第90天评估亚慢性毒性，进行大体尸检、相对器官重量测定以及对肺、肝、脾、肾和淋巴结进行组织病理学评估。另外，用PKH26荧光染料标记的sEVs制剂对大鼠进行静脉注射，开展生物分布研究。注射后24小时采集器官。两组的发病率、死亡率、身体、血液学和生化评估均无剧烈变化。组织病理学评估得出，治疗组肾脏有中度（局灶性）炎症，肝脏有炎症和血管充血的恢复迹象。生物分布研究显示，sEVs在脾脏中的积累量更高。在健康雄性SD大鼠中多次静脉注射汇集的胎儿UCMSCs衍生的sEVs被认为是安全的。注射后24小时，sEVs在脾脏中大量分布。
