Munguia-Lopez Jose G, Pillai Sangeeth, Zhang Yuli, Gantz Amatzia, Camasao Dimitria B, Nazhat Showan N, Kinsella Joseph M, Tran Simon D
Faculty of Dental Medicine and Oral Health Sciences, McGill University, Montreal, QC, Canada.
Department of Bioengineering, McGill University, Montreal, QC, Canada.
Int J Oral Sci. 2025 May 9;17(1):39. doi: 10.1038/s41368-025-00368-6.
Xerostomia (dry mouth) is frequently experienced by patients treated with radiotherapy for head and neck cancers or with Sjögren's syndrome, with no permanent cure existing for this debilitating condition. To this end, in vitro platforms are needed to test therapies directed at salivary (fluid-secreting) cells. However, since these are highly differentiated secretory cells, the maintenance of their differentiated state while expanding in numbers is challenging. In this study, the efficiency of three reversible thermo-ionically crosslinked gels: (1) alginate-gelatin (AG), (2) collagen-containing AG (AGC), and (3) hyaluronic acid-containing AG (AGHA), to recapitulate a native-like environment for human salivary gland (SG) cell expansion and 3D spheroid formation was compared. Although all gels were of mechanical properties comparable to human SG tissue (~11 kPa) and promoted the formation of 3D spheroids, AGHA gels produced larger (>100 cells/spheroid), viable (>93%), proliferative, and well-organized 3D SG spheroids while spatially and temporally maintaining the high expression of key SG proteins (aquaporin-5, NKCC1, ZO-1, α-amylase) for 14 days in culture. Moreover, the spheroids responded to agonist-induced stimulation by increasing α-amylase secretory granules. Here, we propose alternative low-cost, reproducible, and reversible AG-based 3D hydrogels that allow the facile and rapid retrieval of intact, highly viable 3D-SG spheroids.
头颈部癌症放疗患者或干燥综合征患者经常会出现口干症（口腔干燥），这种使人衰弱的病症目前尚无永久性治愈方法。为此，需要体外平台来测试针对唾液（分泌液体）细胞的治疗方法。然而，由于这些是高度分化的分泌细胞，在数量增加的同时维持其分化状态具有挑战性。在本研究中，比较了三种可逆热离子交联凝胶：（1）藻酸盐 - 明胶（AG）、（2）含胶原蛋白的AG（AGC）和（3）含透明质酸的AG（AGHA），在模拟类似天然环境以促进人唾液腺（SG）细胞扩增和三维球体形成方面的效率。尽管所有凝胶的机械性能都与人SG组织相当（约11kPa）并促进了三维球体的形成，但AGHA凝胶产生了更大（>100个细胞/球体）、有活力（>93%）、增殖且组织良好的三维SG球体，同时在空间和时间上在培养14天内维持关键SG蛋白（水通道蛋白 - 5、NKCC1、ZO - 1、α - 淀粉酶）的高表达。此外，这些球体通过增加α - 淀粉酶分泌颗粒对激动剂诱导的刺激作出反应。在此，我们提出了基于AG的替代低成本、可重复且可逆的三维水凝胶，其能够轻松快速地获取完整、高活力的三维SG球体。
