Department of Biological Sciences, University of Delaware, Newark, DE 19713-2070, USA.
Otolaryngol Head Neck Surg. 2010 Feb;142(2):191-5. doi: 10.1016/j.otohns.2009.10.039.
Development of an artificial salivary gland will benefit patients with xerostomia after radiation therapy for upper respiratory cancer. The goal is to devise a three-dimensional (3D) culture system in which salivary cells differentiate into polarized acini that express essential biomarkers and directionally secrete alpha-amylase. Differentiated acini-like structures in a 3D biomaterial-based scaffold will mimic salivary gland functions.
Cells were seeded onto HA-based hydrogels containing PlnDIV peptide and allowed to differentiate into acini-like structures. Cell viability and phenotype were examined.
Laboratory-based tissue procurement study.
Salivary gland tissue was obtained from patients undergoing surgery. Marker expression established the phenotype of salivary gland cells. Perlecan/HSPG2, an important component of the basement membrane, was highly expressed in salivary gland tissue. A culture system consisting of hyaluronic acid (HA) hydrogel and a coupled bioactive peptide derived from domain IV of perlecan (PlnDIV) was used. Prior studies demonstrated differentiation of acinar cells into lobular structures that mimicked intact glands when cultured on PlnDIV peptide-coated surfaces.
Lobular acini-like structures formed on hydrogels and expressed tight junction components such as zona occludens 1. Acini-like structures were stained for the presence of alpha-amylase. Live/dead staining revealed the presence of apoptotic cells in the center of the acini-like structures, indicative of lumen formation.
A novel system supporting acini-like assembly in a 3D culture system was established. Presence of biomarkers and secretion of salivary enzymes confirms functionality in vitro. Future experiments will test the 3D system in an animal model.
为了造福接受上呼吸道癌症放射治疗后出现口干的患者，开发人工唾液腺具有重要意义。本研究旨在设计一种三维（3D）培养体系，使唾液细胞分化为具有极性的、表达重要生物标志物并能定向分泌α-淀粉酶的腺泡。基于 3D 生物材料支架的分化的类腺泡结构将模拟唾液腺功能。
将细胞接种到含有 PlnDIV 肽的 HA 水凝胶中，使其分化为类腺泡结构。检测细胞活力和表型。
基于实验室的组织获取研究。
从接受手术的患者中获取唾液腺组织。标志物表达确定了唾液腺细胞的表型。硫酸乙酰肝素蛋白多糖 2（HSPG2）是基底膜的重要组成部分，在唾液腺组织中高表达。我们使用了一种由透明质酸（HA）水凝胶和源自硫酸乙酰肝素蛋白多糖 2（HSPG2）第四结构域的耦合生物活性肽组成的培养体系。先前的研究表明，当在 PlnDIV 肽涂层表面上培养时，腺泡细胞可分化为具有小叶结构的类腺泡结构，从而模拟完整的腺体。
类腺泡结构在水凝胶上形成，并表达紧密连接成分，如封闭蛋白 1。类腺泡结构被α-淀粉酶染色。活/死染色显示，在类腺泡结构的中心存在凋亡细胞，表明腔的形成。
建立了支持 3D 培养体系中类腺泡组装的新系统。生物标志物的存在和唾液酶的分泌证实了其体外功能。未来的实验将在动物模型中测试 3D 系统。
