脱细胞基质-透明质酸-海藻酸盐混合水凝胶，使多层全层口腔粘膜芯片。

IF 5.5 2区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of dentistry Pub Date : 2025-09-19 DOI:10.1016/j.jdent.2025.106115

Toan V. Phan , Thanaporn Pimpakan , Pipob Suwanchaikasem , Waranyoo Phoolcharoen , Sawang Kesdangsakonwut , Padet Tummaruk , Supansa Yodmuang , Risa Chaisuparat , Oranart Matangkasombut , Yeo-Jun Yoon , Jae-Yol Lim , João N. Ferreira

{"title":"脱细胞基质-透明质酸-海藻酸盐混合水凝胶，使多层全层口腔粘膜芯片。","authors":"Toan V. Phan , Thanaporn Pimpakan , Pipob Suwanchaikasem , Waranyoo Phoolcharoen , Sawang Kesdangsakonwut , Padet Tummaruk , Supansa Yodmuang , Risa Chaisuparat , Oranart Matangkasombut , Yeo-Jun Yoon , Jae-Yol Lim , João N. Ferreira","doi":"10.1016/j.jdent.2025.106115","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><div>Oral mucosa-on-a-chip (OMoC) devices mimic oral epithelium but require continuous nutrient support from underlying lamina propria. However, the common Matrigel or Basement membrane extract (BME) may not effectively support the oral mucosa (OM) native architecture. This study investigated whether a hybrid hydrogel composed of OM decellularized extracellular matrix (OM dECM), hyaluronic acid (HA), and alginate (Alg) surpasses Matrigel and BME for OMoC applications.</div></div><div><h3>Methods</h3><div>Porcine OM tissues were decellularized using 0.1–1 % sodium dodecyl sulfate (SDS), then freeze-dried, and pepsin-digested to produce OM dECM hydrogels (5, 10, and 20 mg/mL). DNA quantification, histological, biochemical and rheological assays, electron microscopy, and mass spectrometry were performed for dECM/hydrogels. Normal oral keratinocytes (NOK), gingival fibroblasts (HGF), and human umbilical vein endothelial cells (HUVEC) cultured in hydrogels were evaluated with proliferation, cytotoxicity, and immunocytochemistry (ICC) assays, and compared to Matrigel/BME. Hybrid hydrogels were used to tri-culture NOK, HGF, and HUVEC in a polydimethylsiloxane (PDMS)-based microfluidic chip for OM modeling.</div></div><div><h3>Results</h3><div>The 1 % SDS decellularization preserved greater ECM components. OM dECM hydrogels enhanced NOK, HGF, and HUVEC viability/proliferation, outperforming Matrigel/BME. Expression of OM-related markers (K14, Ki67, p63, Involucrin, E-cadherin, Von Willebrand factor) increased with hydrogels. Hybrid hydrogel of HA/Alg and 10 mg/mL dECM better supported NOK stratification while maintaining the native phenotype of HGF and HUVEC compared to Matrigel. Within our PDMS-based OMoC, the hybrid hydrogel facilitated the formation of a multi-layered microtissue construct with a stratified epithelial layer, a stromal compartment, and an endothelial network, better replicating the structural features of native OM as compared to other matrices and commercial microfluidic devices.</div></div><div><h3>Conclusions</h3><div>A 1 % SDS decellularization protocol effectively preserved OM ECM, enabling development of an injectable OM dECM hydrogel. The optimized dECM–HA/Alg hybrid hydrogel outperformed Matrigel in supporting oral microtissue stratification, and phenotype maintenance, making it a superior alternative for OMoC applications.</div></div><div><h3>Clinical Significance</h3><div>This study developed a pre-clinical hybrid hydrogel-based OMoC that can more accurately replicate the oral mucosa ECM and tissue architecture when compared to Matrigel/BME. By enhancing the ECM mimicry, the dECM–HA/Alg hybrid hydrogel supported the development of an optimized OMoC for cytotoxicity screening of oral drugs and potential disease modeling of oral mucosal conditions.</div></div>","PeriodicalId":15585,"journal":{"name":"Journal of dentistry","volume":"163 ","pages":"Article 106115"},"PeriodicalIF":5.5000,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Decellularized matrix-hyaluronic acid-alginate hybrid hydrogels to enable a multi-layered full-thickness oral mucosa-on-a-chip\",\"authors\":\"Toan V. Phan , Thanaporn Pimpakan , Pipob Suwanchaikasem , Waranyoo Phoolcharoen , Sawang Kesdangsakonwut , Padet Tummaruk , Supansa Yodmuang , Risa Chaisuparat , Oranart Matangkasombut , Yeo-Jun Yoon , Jae-Yol Lim , João N. Ferreira\",\"doi\":\"10.1016/j.jdent.2025.106115\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objective</h3><div>Oral mucosa-on-a-chip (OMoC) devices mimic oral epithelium but require continuous nutrient support from underlying lamina propria. However, the common Matrigel or Basement membrane extract (BME) may not effectively support the oral mucosa (OM) native architecture. This study investigated whether a hybrid hydrogel composed of OM decellularized extracellular matrix (OM dECM), hyaluronic acid (HA), and alginate (Alg) surpasses Matrigel and BME for OMoC applications.</div></div><div><h3>Methods</h3><div>Porcine OM tissues were decellularized using 0.1–1 % sodium dodecyl sulfate (SDS), then freeze-dried, and pepsin-digested to produce OM dECM hydrogels (5, 10, and 20 mg/mL). DNA quantification, histological, biochemical and rheological assays, electron microscopy, and mass spectrometry were performed for dECM/hydrogels. Normal oral keratinocytes (NOK), gingival fibroblasts (HGF), and human umbilical vein endothelial cells (HUVEC) cultured in hydrogels were evaluated with proliferation, cytotoxicity, and immunocytochemistry (ICC) assays, and compared to Matrigel/BME. Hybrid hydrogels were used to tri-culture NOK, HGF, and HUVEC in a polydimethylsiloxane (PDMS)-based microfluidic chip for OM modeling.</div></div><div><h3>Results</h3><div>The 1 % SDS decellularization preserved greater ECM components. OM dECM hydrogels enhanced NOK, HGF, and HUVEC viability/proliferation, outperforming Matrigel/BME. Expression of OM-related markers (K14, Ki67, p63, Involucrin, E-cadherin, Von Willebrand factor) increased with hydrogels. Hybrid hydrogel of HA/Alg and 10 mg/mL dECM better supported NOK stratification while maintaining the native phenotype of HGF and HUVEC compared to Matrigel. Within our PDMS-based OMoC, the hybrid hydrogel facilitated the formation of a multi-layered microtissue construct with a stratified epithelial layer, a stromal compartment, and an endothelial network, better replicating the structural features of native OM as compared to other matrices and commercial microfluidic devices.</div></div><div><h3>Conclusions</h3><div>A 1 % SDS decellularization protocol effectively preserved OM ECM, enabling development of an injectable OM dECM hydrogel. The optimized dECM–HA/Alg hybrid hydrogel outperformed Matrigel in supporting oral microtissue stratification, and phenotype maintenance, making it a superior alternative for OMoC applications.</div></div><div><h3>Clinical Significance</h3><div>This study developed a pre-clinical hybrid hydrogel-based OMoC that can more accurately replicate the oral mucosa ECM and tissue architecture when compared to Matrigel/BME. By enhancing the ECM mimicry, the dECM–HA/Alg hybrid hydrogel supported the development of an optimized OMoC for cytotoxicity screening of oral drugs and potential disease modeling of oral mucosal conditions.</div></div>\",\"PeriodicalId\":15585,\"journal\":{\"name\":\"Journal of dentistry\",\"volume\":\"163 \",\"pages\":\"Article 106115\"},\"PeriodicalIF\":5.5000,\"publicationDate\":\"2025-09-19\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of dentistry\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0300571225005615\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"DENTISTRY, ORAL SURGERY & MEDICINE\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of dentistry","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0300571225005615","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"DENTISTRY, ORAL SURGERY & MEDICINE","Score":null,"Total":0}

引用次数: 0

摘要

目的：口腔粘膜芯片（OMoC）装置模拟口腔上皮，但需要底层固有层持续的营养支持。然而，普通基质或基底膜提取物（BME）可能不能有效地支持口腔黏膜（OM）的天然结构。本研究研究了由OM脱细胞细胞外基质（OM dECM）、透明质酸（HA）和海藻酸盐（Alg）组成的混合水凝胶是否优于Matrigel和BME用于OMoC。方法：用0.1-1%十二烷基硫酸钠（SDS）对猪OM组织进行脱细胞，冷冻干燥，胃蛋白酶消化，制备OM dECM水凝胶（5、10、20 mg/mL）。对dECM/水凝胶进行DNA定量、组织学、生化和流变学分析、电子显微镜和质谱分析。用水凝胶培养的正常口腔角质形成细胞（NOK）、牙龈成纤维细胞（HGF）和人脐静脉内皮细胞（HUVEC）通过增殖、细胞毒性和免疫细胞化学（ICC）试验进行评估，并与Matrigel/BME进行比较。利用混合水凝胶在聚二甲基硅氧烷（PDMS）微流控芯片中对NOK、HGF和HUVEC进行三种培养，用于OM建模。结果：1% SDS脱细胞保存了较多的ECM成分。OM dECM水凝胶增强了NOK、HGF和HUVEC的活力/增殖，优于Matrigel/BME。om相关标志物（K14, Ki67, p63, Involucrin, E-cadherin, Von Willebrand factor）的表达随着水凝胶的增加而增加。与Matrigel相比，HA/Alg和10 mg/mL dECM混合水凝胶在维持HGF和HUVEC的天然表型的同时，更好地支持NOK分层。在我们基于pdms的OMoC中，混合水凝胶促进了多层微组织结构的形成，包括分层上皮层、间质室和内皮网络，与其他基质和商业微流体设备相比，更好地复制了天然OM的结构特征。结论：1% SDS脱细胞方案有效地保存了OM ECM，从而开发了可注射的OM dECM水凝胶。优化后的dECM-HA/Alg混合水凝胶在支持口腔微组织分层和表型维持方面优于Matrigel，使其成为OMoC应用的优越替代方案。临床意义：本研究开发了一种基于混合水凝胶的临床前OMoC，与Matrigel/BME相比，可以更准确地复制口腔黏膜ECM和组织结构。通过增强ECM模拟，dECM-HA/Alg混合水凝胶支持开发优化的OMoC，用于口服药物的细胞毒性筛选和口腔粘膜状况的潜在疾病建模。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

Decellularized matrix-hyaluronic acid-alginate hybrid hydrogels to enable a multi-layered full-thickness oral mucosa-on-a-chip

Objective

Oral mucosa-on-a-chip (OMoC) devices mimic oral epithelium but require continuous nutrient support from underlying lamina propria. However, the common Matrigel or Basement membrane extract (BME) may not effectively support the oral mucosa (OM) native architecture. This study investigated whether a hybrid hydrogel composed of OM decellularized extracellular matrix (OM dECM), hyaluronic acid (HA), and alginate (Alg) surpasses Matrigel and BME for OMoC applications.

Methods

Porcine OM tissues were decellularized using 0.1–1 % sodium dodecyl sulfate (SDS), then freeze-dried, and pepsin-digested to produce OM dECM hydrogels (5, 10, and 20 mg/mL). DNA quantification, histological, biochemical and rheological assays, electron microscopy, and mass spectrometry were performed for dECM/hydrogels. Normal oral keratinocytes (NOK), gingival fibroblasts (HGF), and human umbilical vein endothelial cells (HUVEC) cultured in hydrogels were evaluated with proliferation, cytotoxicity, and immunocytochemistry (ICC) assays, and compared to Matrigel/BME. Hybrid hydrogels were used to tri-culture NOK, HGF, and HUVEC in a polydimethylsiloxane (PDMS)-based microfluidic chip for OM modeling.

Results

The 1 % SDS decellularization preserved greater ECM components. OM dECM hydrogels enhanced NOK, HGF, and HUVEC viability/proliferation, outperforming Matrigel/BME. Expression of OM-related markers (K14, Ki67, p63, Involucrin, E-cadherin, Von Willebrand factor) increased with hydrogels. Hybrid hydrogel of HA/Alg and 10 mg/mL dECM better supported NOK stratification while maintaining the native phenotype of HGF and HUVEC compared to Matrigel. Within our PDMS-based OMoC, the hybrid hydrogel facilitated the formation of a multi-layered microtissue construct with a stratified epithelial layer, a stromal compartment, and an endothelial network, better replicating the structural features of native OM as compared to other matrices and commercial microfluidic devices.

Conclusions

A 1 % SDS decellularization protocol effectively preserved OM ECM, enabling development of an injectable OM dECM hydrogel. The optimized dECM–HA/Alg hybrid hydrogel outperformed Matrigel in supporting oral microtissue stratification, and phenotype maintenance, making it a superior alternative for OMoC applications.

Clinical Significance

This study developed a pre-clinical hybrid hydrogel-based OMoC that can more accurately replicate the oral mucosa ECM and tissue architecture when compared to Matrigel/BME. By enhancing the ECM mimicry, the dECM–HA/Alg hybrid hydrogel supported the development of an optimized OMoC for cytotoxicity screening of oral drugs and potential disease modeling of oral mucosal conditions.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Journal of dentistry 医学-牙科与口腔外科

CiteScore

7.30

自引率

11.40%

发文量

349

审稿时长

35 days

期刊介绍： The Journal of Dentistry has an open access mirror journal The Journal of Dentistry: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. The Journal of Dentistry is the leading international dental journal within the field of Restorative Dentistry. Placing an emphasis on publishing novel and high-quality research papers, the Journal aims to influence the practice of dentistry at clinician, research, industry and policy-maker level on an international basis. Topics covered include the management of dental disease, periodontology, endodontology, operative dentistry, fixed and removable prosthodontics, dental biomaterials science, long-term clinical trials including epidemiology and oral health, technology transfer of new scientific instrumentation or procedures, as well as clinically relevant oral biology and translational research. The Journal of Dentistry will publish original scientific research papers including short communications. It is also interested in publishing review articles and leaders in themed areas which will be linked to new scientific research. Conference proceedings are also welcome and expressions of interest should be communicated to the Editor.