Intraoperative bioprinting of composite tissues with zonal stratification for craniomaxillofacial reconstruction

用于颅颌面重建的带状分层复合组织的术中生物打印

• 批准号: 10322402
• 负责人: Ibrahim Ozbolat
• 金额: $ 57.04万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322402
• 关键词: 3-Dimensional; Adipocytes; Adipose tissue; Allografting; Anatomy; Area; Autologous Transplantation; Biological Products; Biology; Blood - brain barrier anatomy; Blood Vessels; Bone Regeneration; Bone Tissue; Brain; CD34 gene; Calvaria; Cell Differentiation process; Ceramics; Choristoma; Clinical; Collaborations; Complex; Cosmetics; Debridement; Defect; Dermal; Dermis; Development; Diffuse; Epithelial; Esthetics; Excision; Extracellular Matrix; Fracture; Future; Gene Delivery; Growth; Heterogeneity; Histologic; Human; Immune response; In Situ; In Vitro; Infection; Injury; Intervention; Knowledge; Lead; Manuals; Measures; Mechanics; Methods; MicroRNAs; Modeling; Morbidity - disease rate; Natural regeneration; Nude Rats; Operating Rooms; Operative Surgical Procedures; Pathogenicity; Pathway interactions; Patients; Plastic Surgical Procedures; Polymers; Preparation; Procedures; Process; Production; Property; Public Health; Rattus; Regenerative Medicine; Rodent Model; Role; Scanning; Series; Shapes; Site; Skin; Skin Tissue; Skin repair; Sterility; Stratification; Technology; Testing; Tissue Engineering; Tissue constructs; Tissues; Transfection; Translations; Vascularization; Work; analog; base; bench to bedside; bioink; bioprinting; bone; bone quality; craniomaxillofacial; cranium; cytokine; dosage; functional restoration; gene therapy; immunogenicity; implantation; improved; in vivo; injured; injury and repair; lipid biosynthesis; morphogens; novel; operation; osteogenic; progenitor; reconstruction; repaired; scaffold; skin regeneration; soft tissue; stem; stem cell differentiation; stem cells; subcutaneous; tissue regeneration

PROJECT SUMMARY/ABSTRACT Current approaches in repairing craniomaxillofacial (CMF) defects possess several limitations and the reconstruction of CMF defects seamlessly is highly challenging, as precise layer-by-layer stacking of multiple tissue compartments is not trivial. Such compartmentalization necessitates the precision and effective use of stem cells and differentiation factors, and differentiating stem cells into multiple lineages is crucial in order to recapitulate the native tissue anatomy. With the advances in three-dimensional (3D) bioprinting, reconstruction of composite tissues in situ for CMF repair has recently become feasible as 3D bioprinting enables complex tissue heterogeneity in an anatomically accurate and cosmetically appealing manner. Intraoperative bioprinting, which can be defined as bioprinting directly into the defect area for repairing injuries in a surgery setting, is a highly effective process for CMF reconstruction, where the defect information can be rapidly acquired with minimum manual interventions, enabling accurate personalized reconstructions immediately after characterization of the defect. In this project, we hypothesize that intraoperatively bioprinted multi-layer composite tissues loaded with differentiation factors including microRNA(miRNA)-transfected human progenitor cells and adipose-derived extracellular matrix components (adECM) induce compartmentalization of soft and hard tissues that recapitulates CMF tissue anatomy on an athymic rat model. In order to test our hypothesis, Specific Aim I will use intraoperative bone tissue bioprinting to reveal the impact of miR-148b- transfected human adipose-derived stem cells (ADSCs) at respective dosages on bone tissue regeneration. In Specific Aim II, we will intraoperatively bioprint multi-layer skin tissues, including adipose and dermis layers, in order to explore the impact of localized delivery of human ADSCs and adECM components at different dosages and concentrations on skin tissue regeneration, respectively. Particularly, we will observe if ADSCs differentiate into adipocytes and also understand the impact of the presence of adipose layer on dermis regeneration. In Specific Aim III, we will intraoperatively bioprint three-layer composite tissues, including cranium, adipose and dermis layers, in order to understand the role of a vascularization on soft and hard tissue regeneration. In addition, we will explore the role of miR-210 in vascularization. In this regard, we have formed a complementary collaboration that merges essential domain knowledge in bioprinting, regenerative medicine, CMF surgery, plastic surgery, gene therapy, gene delivery, bone mechanics, and bone and skin biology with the depth necessary to propel the proposed work towards meaningful advances that would otherwise not be possible. Successful completion of the proposed work is anticipated to give rise to an advanced bioprinting technology revealing the complex interactions between stratified layers of engineered tissues in an immunodeficient rodent model and thereby provide a novel understanding of how localized delivery of differentiation factors impacts craniomaxillofacial reconstruction.

项目概要/摘要 目前修复颅颌面（CMF）缺损的方法存在一些局限性，并且 无缝地重建 CMF 缺陷非常具有挑战性，因为需要精确地逐层堆叠多个 组织隔室并非微不足道。这种划分需要精确和有效地使用 干细胞和分化因子，将干细胞分化成多个谱系对于 概括原生组织解剖结构。随着三维（3D）生物打印、重建技术的进步 由于 3D 生物打印能够实现复杂的复合组织原位修复以进行 CMF 修复，最近已变得可行 以解剖学上准确且美观的方式呈现组织异质性。术中 生物打印，可以定义为直接在缺损区域进行生物打印，用于修复手术中的损伤 设置，是CMF重建的高效过程，可以快速获取缺陷信息 通过最少的手动干预获得，从而能够在之后立即进行准确的个性化重建 缺陷的表征。在这个项目中，我们假设术中生物打印多层 装载有分化因子（包括 microRNA（miRNA）转染的人）的复合组织 祖细胞和脂肪来源的细胞外基质成分（adECM）诱导 软组织和硬组织，再现无胸腺大鼠模型上的 CMF 组织解剖结构。为了测试我们的 假设，具体目标 I 将使用术中骨组织生物打印来揭示 miR-148b- 的影响 不同剂量的转染人脂肪干细胞（ADSC）对骨组织再生的影响。在 具体目标 II，我们将在术中生物打印多层皮肤组织，包括脂肪和真皮层。 为了探索人类 ADSC 和 adECM 成分在不同条件下局部递送的影响 分别对皮肤组织再生的剂量和浓度。特别是，我们将观察 ADSC 是否 分化为脂肪细胞，并了解脂肪层的存在对真皮的影响 再生。在特定目标 III 中，我们将在术中生物打印三层复合组织，包括 颅骨、脂肪和真皮层，以了解血管化对软组织和硬组织的作用 再生。此外，我们将探讨miR-210在血管化中的作用。对此，我们成立了 一种互补的合作，融合了生物打印、再生医学、 CMF 手术、整形手术、基因治疗、基因传递、骨力学以及骨和皮肤生物学 推动拟议工作取得有意义的进展所需的深度，否则这些进展将无法实现 可能的。成功完成拟议工作预计将产生先进的生物打印 揭示工程组织分层之间复杂相互作用的技术 免疫缺陷啮齿动物模型，从而为如何局部递送提供了新的理解 分化因素影响颅颌面重建。