评估使用脱细胞人羊膜与细胞疗法相结合治疗局部放射损伤的效果

A. Rastorgueva, T. Astrelina, V. Brunchukov, I. Kobzeva, Y. Suchkova, V. Nikitina, S. Lishchuk, E. Dubova, K.A. Pavlov, T. Malivanova, D. Usupzhanova, O.G. Mikhadarkina, A.D. Kobzev, V.I. Bulygina, A. Samoilov
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Material and methods: The study included 42 laboratory animals (white male rats of the Wistar line), randomly divided into 6 groups: K – control group without therapy, on the 21st day after irradiation, groups using: Am+LD – application of lyophilisate of the conditioned medium of MSCs of the human gingival mucosa (LD) under the decellularized amniotic membrane, Am+LP – application of lyophilisate of the conditioned medium of MSCs of human placental tissue (LP) under the decellularized amniotic membrane, Am – application of the decellularized amniotic membrane, AmCl – application of the decellularized amniotic membrane with medical glue BF-6, Kl – application medical glue BF-6. 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引用次数: 0

摘要

细胞疗法是一种前景广阔的医学方向,它能显著改善包括局部辐射损伤(LRI)在内的各种疾病的治疗效果。本研究的目的是比较使用脱细胞人羊膜分离出的间充质干细胞与来自人牙龈粘膜的间充质干细胞和来自人胎盘组织的间充质干细胞的条件培养基冻干物的结果及其对局部辐射损伤组织再生的影响。材料和方法:研究包括 42 只实验动物(Wistar 系白色雄性大鼠),随机分为 6 组:K - 对照组,照射后第 21 天,不进行治疗;使用Am+LD组--在脱细胞羊膜下应用人牙龈粘膜间充质干细胞条件培养基冻干物(LD),Am+LP组--在脱细胞羊膜下应用人胎盘组织间充质干细胞条件培养基冻干物(LP),Am组--应用脱细胞羊膜,AmCl组--应用脱细胞羊膜与医用胶BF-6,Kl组--应用医用胶BF-6。在 LNK-268 X 射线装置上以 110 Gy 的剂量对动物的 LRI 进行建模,并观察至照射后第 112 天,监测皮肤溃疡面和 LRI 的炎症过程,用 ImageTool 软件计算整个皮肤变化的面积。进行了平面和组织学分析。结果显示结果表明,与对照组相比,在照射后第77天和第112天使用脱细胞羊膜治疗LRI,伤口愈合率分别提高了4.6倍和18倍;使用胎盘间充质干细胞条件培养基冻干物的脱细胞羊膜,伤口愈合率分别提高了13.5倍和27倍;使用人牙龈粘膜间充质干细胞冻干物的脱细胞羊膜,伤口愈合率达到100%。在 Am 组中,83.5% 的动物 LRI 溃疡表面愈合,这表明即使在没有人类间充质干细胞冻干物的情况下,使用分离的脱细胞羊膜也是有效的。与此同时,AmCl 组的 LRI 溃疡表面愈合率为 20%,其中脱细胞羊膜是用 BF-6 医用胶固定的,与其他组相比效果较差。结论建议将脱细胞羊膜与胎盘间充质基质细胞(MSCs)条件培养基冻干物和人体牙龈粘膜组织联合使用,以及单独使用脱细胞羊膜,可使溃疡面完全上皮化、实验动物的 IIIb-IV 级 LRI 病程明显改善,愈合时间缩短,无局部或全身并发症,可视为皮肤 LRI 的一种有前途的保守治疗方法,也是治疗不愈合和慢性伤口的一种新疗法。
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Evaluation of the Effectiveness of The Use of Decellularized Human Amniotic Membrane in Combination with Cell Therapy for Local Radiation Injuries
Cell therapy is a promising direction in medicine, which can significantly improve the results of treatment of various diseases, including local radiation injuries (LRI). The purpose of the study was to compare the results of using decellularized human amniotic membrane isolated and together with lyophilisates of conditioned media of MSCs from the human gingival mucosa and MSCs from human placental tissue and their effect on tissue regeneration in LRI. Material and methods: The study included 42 laboratory animals (white male rats of the Wistar line), randomly divided into 6 groups: K – control group without therapy, on the 21st day after irradiation, groups using: Am+LD – application of lyophilisate of the conditioned medium of MSCs of the human gingival mucosa (LD) under the decellularized amniotic membrane, Am+LP – application of lyophilisate of the conditioned medium of MSCs of human placental tissue (LP) under the decellularized amniotic membrane, Am – application of the decellularized amniotic membrane, AmCl – application of the decellularized amniotic membrane with medical glue BF-6, Kl – application medical glue BF-6. Modeling of LRI in animals was carried out on an LNK-268 X-ray unit at a dose of 110 Gy and observed until the 112th day after irradiation, monitoring of the ulcerative surface of the skin and the course of the inflammatory process of LRI was carried out with calculation of the area of ​​the total changed skin in the ImageTool software. Planimetric and histological analyzes were performed. Results: It was demonstrated that, when compared with the control, the use of decellularized amniotic membrane in LRI on days 77 and 112 after irradiation increased the healing rate by 4.6 and 18 times, respectively; decellularized amniotic membrane with lyophilisate of placental MSC conditioned medium increased the rate of wound healing by 13.5 and 27 times, respectively; and a dellularized amniotic membrane with lyophilisate of MSCs from the human gingival mucosa to 100 % healing. Healing of the ulcer surface area of the LRI in 83.5 % of animals in the Am group demonstrated the effectiveness of using isolated decellularized amniotic membrane even in the absence of human MSC lyophilisates. At the same time, healing of the ulcer surface area of the LRI in the AmCl group was 20 % of cases, where the decellularized amniotic membrane was fixed with BF-6 medical glue, which was less effective compared to other groups. Conclusion: The proposed combined use of decellularized amniotic membrane with lyophilisates of the conditioned medium of mesenchymal stromal cells (MSCs) of the placenta and mucous tissue of the human gums and the isolated use of decellularized amniotic membrane led to complete epithelization of the ulcer surface, a pronounced favorable course of grade IIIb–IV LRI in laboratory animals and a reduction in healing time , the absence of local or systemic complications can be considered as a promising method of conservative treatment for LRI of the skin and a new therapeutic approach in the treatment of non-healing and chronic wounds.
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Medical Radiology and Radiation Safety
Medical Radiology and Radiation Safety Medicine-Radiology, Nuclear Medicine and Imaging
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