使用间充质干细胞重建坏死的下颌下唾液腺。
Reconstruction of necrotic submandibular salivary gland using mesenchymal stem cells.
作者信息
Najafi Shamsoulmolouk, Nosrati Haleh, Faraji Zahra, Mohamadnia Abdolreza, Shirian Sadegh, Mortazavi Seyed Mostafa, Bahrami Naghmeh
机构信息
Dental Research Center, Tehran University of Medical Sciences, Tehran, Iran.
Dept. of Oral & Maxillofacial Medicine, School of Dentistry, Tehran University of Medical Sciences, Tehran, Iran.
出版信息
Heliyon. 2020 Oct 8;6(10):e05162. doi: 10.1016/j.heliyon.2020.e05162. eCollection 2020 Oct.
BACKGROUND
The efficacy of mesnchymal stem cells (MSCs) to treat the necrotic tissue of salivary glands (SGs) has yet investigated.
OBJECTIVE
This study was conducted to investigate the potential capacity of MSCs to restore the function and regenerate the necrotic submandiular gland in the rat animal model.
METHODS
Twenty-one Sprague-Dawley rats were provided from a breeding colony and randomly divided into three groups including the positive control or induced SG atrophy without treatment, the treatment group or induced SG atrophy with MSCs isolated transplantation and the negative control group consists of healthy rats. The atrophic and necrotic submandiular gland was induced using intraoral duct ligation of the main duct of submandiular gland for one month. The isolated stem cells were confirmed using flow cytometry for CD90 and CD 105. The isolated MSCs were cultured and injected to submandiular gland and the potential efficacy of MSCs to treat the atrophic submandibular glands was evaluated using histopathology on two weeks post-transplantation. To detect the acinar cell protein secretory granules, Alcian Blue and periodic acid shift (PAS) staining were done. For the demonstration of mitotic index or proliferation rate of the SG epithelia tissue, Ki-67 and Smbg proteins expression were evaluated using immunohistochemistry.
RESULTS
The locally injected MSCs could regenerate the overall histological structure of the necrotic submandibular gland tissue within 2 weeks of post-transplantation. Alcian Blue and PAS staining indicated that the mean amount of serous and mucin secretions in the treatment group was significantly increased compared to the positive control groups. We have also found that the treatment group significantly express higher Ki-67 protein, as a diagnostic marker for cell mitosis and proliferation rate, and lower Smbg protein, as a diagnostic marker, for damage to the submandibular gland than that of control group.
CONCLUSION
This study demonstrates the therapeutic benefits of MSCs isolated from the SG in treating atrophic and necrotic SGs in a rat model. MSCs may be potential candidates for cell-based therapies targeting hypofunction of SG induced by a range of diseases or because of surgery and radiotherapy of head and neck cancers.
背景
间充质干细胞(MSCs)治疗唾液腺(SGs)坏死组织的疗效尚未得到研究。
目的
本研究旨在探讨MSCs在大鼠动物模型中恢复功能并使坏死的下颌下腺再生的潜在能力。
方法
从繁殖群体中提供21只Sprague-Dawley大鼠,随机分为三组,包括不进行治疗的阳性对照组或诱导SG萎缩组、进行MSCs分离移植的诱导SG萎缩治疗组以及由健康大鼠组成的阴性对照组。通过结扎下颌下腺主导管进行口内导管结扎1个月来诱导萎缩和坏死的下颌下腺。使用流式细胞术检测CD90和CD105来确认分离的干细胞。将分离的MSCs进行培养并注射到下颌下腺,在移植后两周使用组织病理学评估MSCs治疗萎缩性下颌下腺的潜在疗效。为检测腺泡细胞蛋白分泌颗粒,进行阿尔辛蓝和高碘酸希夫(PAS)染色。为证明SG上皮组织的有丝分裂指数或增殖率,使用免疫组织化学评估Ki-67和Smbg蛋白表达。
结果
局部注射的MSCs可在移植后2周内使坏死的下颌下腺组织的整体组织结构再生。阿尔辛蓝和PAS染色表明,与阳性对照组相比,治疗组浆液和粘蛋白分泌的平均量显著增加。我们还发现,作为细胞有丝分裂和增殖率诊断标志物的Ki-67蛋白在治疗组中的表达显著高于对照组,而作为下颌下腺损伤诊断标志物的Smbg蛋白在治疗组中的表达低于对照组。
结论
本研究证明了从SG分离的MSCs在大鼠模型中治疗萎缩性和坏死性SGs的治疗益处。MSCs可能是针对一系列疾病或头颈部癌症手术及放疗引起的SG功能减退的基于细胞的治疗的潜在候选者。