Development of an Integrated 3D Human Osteo-Mucosal Model

集成 3D 人体骨粘膜模型的开发

• 批准号: 10224467
• 负责人: Lobat Tayebi
• 金额: $ 7.84万
• 依托单位: MARQUETTE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-15 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224467
• 关键词: 3-Dimensional; 3D Print; Address; Adhesions; Adhesives; Air; Alveolar; Amines; Animal Model; Animal Testing; Area; Basement membrane; Biocompatible Materials; Biologic Characteristic; Biological; Biological Assay; Bioreactors; Bone Morphogenetic Proteins; Bone Tissue; Cartilage; Cell-Matrix Junction; Cells; Chemicals; Clinical; Collagen; Communication; Complex; Connective Tissue; Cytokeratin; Data; Defect; Dental Materials; Dental Pulp; Dentin; Desmosomes; Development; Drug Delivery Systems; Drug Evaluation; Employment; Encapsulated; Engineering; Epithelial; Epithelium; Family suidae; Fibrin Tissue Adhesive; Fibroblasts; Formulation; Future; Gelatin; Glues; Growth; Healthcare; Histology; Human; Immunohistochemistry; Implant; In Situ; In Vitro; Incubated; Jaw; Laboratories; Ligaments; Liquid substance; Metabolic; Methacrylates; Methods; Modeling; Modification; Monitor; Monophenol Monooxygenase; Mouth Diseases; Mucous Membrane; Natural regeneration; Nutrient; Oral; Oral Surgical Procedures; Oral health; Oral mucous membrane structure; Osteoblasts; Osteocalcin; Outcome; Penetration; Physiological; Printing; Procedures; Production; Property; Quinones; Reaction; Sampling; Side; Structure; Surface; System; Techniques; Tendon structure; Testing; Thick; Time; Tissue Engineering; Tissue Viability; Tissues; Transmission Electron Microscopy; Transplantation; base; biomaterial compatibility; bone; cell growth; clinically relevant; covalent bond; decorin; design; functional group; improved; in vitro Model; in vitro testing; in vivo; in vivo Model; interfacial; keratinocyte; novel; novel strategies; oral tissue; polycaprolactone; reconstruction; scaffold; screening; soft tissue; three-dimensional modeling; time interval; tricalcium phosphate

SUMMARY: The integration and stability of the hard/soft tissue interface is a major challenge in regeneration and engineering of constructs composed of widely different tissue types. The current approach is employment of biocompatible natural adhesives, such as fibrin glue. However, using such adhesives, the adhesion strength is not strong enough in physiological condition and will loosen in time. Moreover, since they act as an additional layer/material between two tissues, the healthy cellular communication between tissues will be disturbed with adverse influence on interfacial tissue development. In this project, we propose a new method for adhesion of soft/hard tissues, which addresses both of the above drawbacks. We will optimize and apply our technique in oral mucosa/bone adhesion to develop an engineered osteo-mucosal complex as a key example of a construct containing widely dissimilar tissues. Our proposed approach relies on in situ incorporation of adhesive functional groups into a cell-laden soft scaffold based on a biocompatible method, which become readily adherent to a surface treated hard scaffold. More specifically, the photocurable gelatin-based matrix encapsulating cells will be enzymatically treated to have quinone functional groups which can form covalent bonds with the amine groups on the polycaprolactone bone scaffold. The interfacial adhesion will be based on Michael reaction. We claim that this new technique has three major advantages over current methods: 1) It directly integrates two compartments of soft/hard tissues without the need for employing a third material. 2) The adhesion is very strong and will not be diminished over time. 3) The formulation will lead to in depth penetration of cells and enhanced cell growth inside the scaffolds. To examine our hypotheses, the adhesion strength will be fully evaluated in vitro quantitatively and qualitatively over time after development of the osteo-mucosal complex, and the biological characteristics will be inspected in detail. A similar construct treated by fibrin glue as the adhesive agent will be produced in parallel to be used as the control sample. Moreover, the adhesion strength of our osteo-mucosal construct will be compared with the natural oral mucosa/bone interfacial tissue in a freshly slaughtered pig’s jaw. A successful outcome from the proposed adhesion method in the osteo-mucosal complex will suggest that the technique can potentially be applied (after relevant modifications) for engineering of interfacial tissues in periodontal complex, bone/cartilage, bone/ligament, bone/tendon, and dentin/pulp complexes. The applications of the developed osteo-mucosa complex include A) clinical transplantation such as alveolar reconstruction and intraoral grafting, B) production of a clinically relevant in vitro test system and an alternative to animal test models for 1) studying the interaction of biomaterials and oral tissue, and 2) oral disease screening and evaluation of drug delivery systems.

总结：硬/软组织界面的整合和稳定性是再生的主要挑战 以及由广泛不同的组织类型组成的结构的工程化。目前的做法是雇用 生物相容的天然粘合剂，例如纤维蛋白胶。然而，使用这样的粘合剂，粘合强度降低。 在生理条件下不够坚固，并会及时松动。此外，由于它们是一种额外的 当两个组织之间的细胞层/材料被破坏时，组织之间的健康细胞通信将被干扰， 对界面组织发育的不利影响。 在这个项目中，我们提出了一种新的软/硬组织粘附方法，它解决了上述两个问题 缺点.我们将优化并应用我们的口腔粘膜/骨粘附技术，开发一种工程化的 骨-粘膜复合体作为包含广泛不同组织的构建体的关键实例。 我们所提出的方法依赖于在原位掺入粘合剂功能团到细胞负载的软支架 基于生物相容性方法，其变得容易粘附到表面处理的硬支架上。更 具体地，包封细胞的可光固化明胶基基质将被酶促处理， 可与聚己内酯骨架上的胺基形成共价键的醌官能团 脚手架界面粘合将基于迈克尔反应。 我们声称，这种新技术与现有方法相比有三个主要优点：1）它直接集成了两个 在一些实施例中，所述第一材料可以是软/硬组织的隔室，而不需要使用第三材料。2)附着力非常 它是强大的，不会随着时间的推移而减弱。3)该制剂将导致细胞的深度渗透， 增强了支架内的细胞生长。 为了验证我们的假设，将在体外定量和定性地充分评估粘附强度 随着时间的推移，在骨-粘膜复合体的发展后，将检查生物学特性 详细将平行生产使用纤维蛋白胶作为粘合剂处理的类似结构 作为对照样品。此外，我们的骨粘膜结构的粘附强度将与 新鲜屠宰的猪颌骨中的天然口腔粘膜/骨界面组织。 在骨-粘膜复合体中所提出的粘附方法的成功结果表明， 技术可以潜在地应用于（在相关修改之后）界面组织的工程化， 牙周复合体、骨/软骨、骨/韧带、骨/腱和牙本质/牙髓复合体。 所开发的骨-粘膜复合体的应用包括：A）临床移植，如牙槽骨移植， 重建和口内移植，B）生产临床相关的体外测试系统和替代物 用于1）研究生物材料和口腔组织的相互作用，和2）口腔疾病筛查的动物试验模型 和药物输送系统的评价。