细胞外基质蛋白在胶体微凝胶中的自组装水凝胶在再生牙髓学中的应用

Cell-derived Extracellular Matrix Proteins in Colloidal Microgel as a Self-Assembly Hydrogel for Regenerative Endodontics.

机构信息

Departments of Periodontics and Endodontics, School of Dental Medicine, University at Buffalo, New York.

Department of Biomedical Engineering, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo, New York.

出版信息

J Endod. 2022 Apr;48(4):527-534. doi: 10.1016/j.joen.2022.01.011. Epub 2022 Jan 22.

DOI:10.1016/j.joen.2022.01.011

PMID:35077752

Abstract

INTRODUCTION

This study investigated a colloidal microgel for angiogenic and odontogenic differentiation of cells in the presence of cell-derived extracellular matrix (ECM) proteins using a 3-dimensional culture model.

METHODS

Viscoelastic properties of human dental pulp were determined to understand the native ECM environment. ECM proteins were extracted from dental pulp stem cell (DPSC) cultures, and MaxGel (Millipore Sigma, Burlington, MA) was used as a commercially available ECM protein. DPSCs were incubated in colloidal microgels in the presence of ECM proteins or gelatin methacryloyl (GelMA) as a bulk hydrogel (n = 9/group). The viability and odontogenic differentiation of DPSCs within hydrogels was determined using viability assays, mineralization staining, calcium and alkaline phosphatase assays, and quantitative polymerase chain reaction for odontogenic gene expression. Angiogenic properties of endothelial cells were determined using tubule formation assays and quantitative polymerase chain reaction to detect angiogenic gene expression.

RESULTS

Dental pulp had a higher elastic modulus than the viscous modulus, showing a solidlike response similar to hydrogels. DPSC-derived ECM showed higher collagen and GAG than MaxGel (P < .05). The viability of DPSCs was similar in colloidal microgels, whereas higher cell viability, calcium deposition, and alkaline phosphatase activity were observed in GelMA (P < .05). Colloidal microgels allowed tubule-like structures by endothelial cells, whereas no tubular formation was observed in GelMA. DPSC-derived ECM in colloidal microgel up-regulated odontogenic gene expression, whereas MaxGel up-regulated angiogenic gene expression (P < .05).

CONCLUSIONS

Colloidal microgels allowed cellular organization that can improve penetration and nutritional supply in a full-length root canal system. The bioactivity of cell-derived ECM proteins can be modified depending on the external stimulus.

摘要

简介

本研究使用三维培养模型，研究了在细胞来源的细胞外基质 (ECM) 蛋白存在下，胶体微凝胶对细胞的血管生成和牙源性分化的影响。

方法

为了了解天然 ECM 环境，测定了人牙髓的粘弹性。从牙髓干细胞 (DPSC) 培养物中提取 ECM 蛋白，并将 MaxGel（Millipore Sigma，马萨诸塞州伯灵顿）用作商业上可获得的 ECM 蛋白。将 DPSC 在含有 ECM 蛋白或明胶甲基丙烯酰 (GelMA) 的胶体微凝胶中孵育，作为块状水凝胶（每组 n = 9）。通过细胞活力测定、矿化染色、钙和碱性磷酸酶测定以及牙源性基因表达的定量聚合酶链反应，确定水凝胶内 DPSC 的活力和牙源性分化。通过管形成测定和定量聚合酶链反应检测血管生成基因表达，确定内皮细胞的血管生成特性。

结果

牙髓的弹性模量高于粘性模量，表现出类似于水凝胶的固态响应。DPSC 衍生的 ECM 显示出比 MaxGel 更高的胶原和 GAG（P <.05）。胶体微凝胶中 DPSC 的活力相似，而 GelMA 中观察到更高的细胞活力、钙沉积和碱性磷酸酶活性（P <.05）。胶体微凝胶允许内皮细胞形成管状结构，而 GelMA 中则没有观察到管状形成。胶体微凝胶中的 DPSC 衍生 ECM 上调牙源性基因表达，而 MaxGel 上调血管生成基因表达（P <.05）。