NOVEL THERAPEUTICS TO TREAT NECROTIC TEETH: THE COMBINATORIAL EFFECT OF A 3D DRUG DELIVERY SYSTEM AND SPATIALLY DESIGNED STEM CELL NICHES

治疗坏死牙齿的新型疗法：3D 药物输送系统和空间设计的干细胞生态位的组合效应

• 批准号: 10231139
• 负责人: Marco C Bottino
• 金额: $ 33.56万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10231139
• 关键词: 3-Dimensional; Adhesions; Adolescent; Affinity; Age; Antibiotics; Apoptosis; BMP2 gene; Bacteria; Biological Assay; Biophysics; Blood Vessels; Canis familiaris; Cell Survival; Cells; Chemicals; Child; Clinical; Clinical Trials; Collagen; Collagen Fibril; Confocal Microscopy; Couples Therapy; Data; Dental; Dental Pulp; Dental Pulp Necrosis; Dental caries; Dentin; Dentin Formation; Development; Diagnostic radiologic examination; Disease model; Disinfection; Drug Delivery Systems; Electrospinning; Encapsulated; Endodontics; Endothelium; Engineering; Environment; Evaluation; Extracellular Matrix; Future; Goals; Growth Factor; Growth and Development function; Health; Hemorrhage; Histologic; Histology; Human; Immunocompetence; Implant; In Vitro; Infection; Infection Control; Inflammation; Injectable; Injections; Injury; Lead; Left; Location; Measurement; Mediating; Methods; Microbial Biofilms; Mission; Modeling; Morphogenesis; Nanostructures; National Institute of Dental and Craniofacial Research; Natural regeneration; Necrosis; Odontoblasts; Oral; Oral health; Paste substance; Patients; Periapical Diseases; Plant Roots; Polymers; Population; Procedures; Property; Public Health; Pulp Canals; Regenerative capacity; Regulation; Reporting; Research; Role; Schools; Severe Combined Immunodeficiency; Slice; Spatial Design; Standardization; Stem cell transplant; System; Therapeutic; Time; Tissues; Tooth structure; Toxic effect; Translations; Trauma; Tubular formation; Vascular Endothelial Growth Factors; Work; antimicrobial; base; cell behavior; chemical property; combinatorial; craniofacial; deciduous tooth; experience; implantation; improved; in vivo; in vivo Model; innovation; mRNA Expression; mouse model; nanofiber; neovascularization; novel; novel strategies; novel therapeutics; patient variability; permanent tooth; physical property; professor; protein expression; regenerative; regenerative approach; regenerative therapy; restoration; stem cell differentiation; stem cell growth; stem cells; success; traditional therapy

Abstract/Summary: Caries and dental trauma are major oral health burdens. Globally, 21% of children (age 6 to 11 years) have caries in their permanent teeth. In the US, 18% of school children experience dental trauma. Dental pulp injury due to caries or trauma, leads to inflammation, which if left untreated, results in necrosis. Traditional therapeutics of necrotic immature permanent teeth allows for infection control, but support neither root development nor restoration of the immunocompetence of the pulp. To date, no clinical therapy exists that promotes root canal disinfection and can consistently guide the growth and development of pulp and dentin in necrotic teeth. Thus, there is a pressing need to develop a strategy for predictable pulp-dentin regeneration in a bacteria-free environment which may ultimately lead to the establishment of novel therapeutics to treat immature teeth with pulpal necrosis. The objective of this application is to develop a novel strategy to stimulate pulp and dentin regeneration by engineering an injectable collagen-fibril matrix system with heterogeneous stiffness and selected growth factors (GFs), which will first require the attainment of a bacteria-free niche. Our first hypothesis is that electrospinning can be used to develop non-toxic and antimicrobially effective 3D tubular drug delivery constructs for root canal disinfection that release initially high amount of antibiotics and sustain its effects for several days. The proposed construct will be evaluated for its release properties and cell compatibility in vitro. Antimicrobial properties will be determined using an in vitro infected tooth slice model and an in vivo model of immature dog teeth with periapical disease (Aim 1). Our second hypothesis is that dental pulp stem cell transplantation within a stiffer collagen matrix added with bone morphogenetic protein-2 (BMP-2) or within a more compliant matrix added with vascular endothelial growth factor (VEGF), when concentrically injected into a disinfected root canal, will lead to dentin and pulp regeneration, respectively. We propose to optimize the novel self-assembling collagen-fibril matrix system by evaluating the cell viability, proliferation, apoptosis and differentiation to endothelial and odontoblast cells using in vitro cell-based assays and a well- established in vivo tooth slice SCID mice model (Aim 2). Finally, the regenerative capacity of the optimal and standardized injectable collagen-fibril matrix system will be evaluated using an in vivo model of immature dog teeth with periapical disease after disinfection with the drug delivery construct (Aim 3). This application is highly innovative as we propose, for the first time, the clinical role of a cell-friendly electrospun 3D tubular drug delivery construct for root canal disinfection. Further, we propose to couple this therapy with a unique regenerative strategy using injectable and highly tunable collagen-fibril matrices to amplify dental pulp stem cell differentiation to form pulp and dentin in the appropriate locations. The proposed research is significant because it will expedite the establishment of a reliable regenerative therapeutics to treat necrotic immature permanent teeth.

摘要/总结： 龋齿和牙外伤是主要的口腔健康负担。全球 21% 的儿童（6 至 11 岁）患有 恒牙有龋齿。在美国，18% 的学童遭受过牙齿外伤。牙髓损伤 由于龋齿或外伤，会导致炎症，如果不及时治疗，会导致坏死。传统的 坏死的未成熟恒牙的治疗可以控制感染，但不支持牙根 牙髓免疫能力的发展或恢复。迄今为止，尚无临床疗法可以 促进根管消毒，持续引导牙髓和牙本质的生长发育 坏死的牙齿。因此，迫切需要开发一种可预测的牙髓-牙本质再生策略 无菌环境，最终可能导致建立新的治疗方法 未成熟的牙齿，伴有牙髓坏死。该应用程序的目的是开发一种新颖的策略来刺激 通过设计具有异质性的可注射胶原纤维基质系统来再生牙髓和牙本质 硬度和选定的生长因子（GF），这首先需要获得无菌环境。我们的 第一个假设是静电纺丝可用于开发无毒且抗菌有效的 3D 管状材料 用于根管消毒的药物输送结构，最初释放大量抗生素并维持其作用 效果持续数日。将评估所提出的构建体的释放特性和细胞 体外相容性。将使用体外感染的牙齿切片模型来确定抗菌特性，并 患有根尖周疾病的未成熟狗牙齿的体内模型（目标 1）。我们的第二个假设是牙科 添加骨形态发生蛋白-2 (BMP-2) 的较硬胶原基质内的牙髓干细胞移植 或在添加有血管内皮生长因子（VEGF）的更顺应的基质内，当同心 注射到消毒的根管中，将分别导致牙本质和牙髓再生。我们建议 通过评估细胞活力、增殖、 使用体外基于细胞的测定和良好的方法进行内皮细胞和成牙本质细胞的凋亡和分化 建立体内牙片SCID小鼠模型（目标2）。最后，最佳和的再生能力 将使用未成熟狗的体内模型来评估标准化的可注射胶原纤维基质系统 使用药物输送结构消毒后患有根尖周疾病的牙齿（目标 3）。这个应用程序是 高度创新，我们首次提出细胞友好型电纺 3D 管状药物的临床作用 用于根管消毒的输送结构。此外，我们建议将这种疗法与独特的 使用可注射和高度可调的胶原纤维基质来放大牙髓干的再生策略 细胞分化，在适当的位置形成牙髓和牙本质。拟议的研究意义重大 因为它将加速建立可靠的再生疗法来治疗坏死性未成熟细胞 恒牙。

项目成果

Chlorhexidine-modified nanotubes and their effects on the polymerization and bonding performance of a dental adhesive.

氯己定改性纳米管及其对牙科粘合剂聚合和粘合性能的影响。

• DOI: 10.1016/j.dental.2020.03.007
• 发表时间: 2020-03-30
• 期刊: Dental materials : official publication of the Academy of Dental Materials
• 作者: Sara Kalagi;Sara Kalagi;S. Feitosa;E. Münchow;V. M. Martins;Ashley Karczewski;N. B. Cook;K. Diefenderfer;G. Eckert;S. Geraldeli;M. Bottino
• 通讯作者: M. Bottino

Bioactive amorphous magnesium phosphate-polyetheretherketone composite filaments for 3D printing.

用于3D打印的生物活性非晶态磷酸镁-聚醚醚酮复合长丝。

• DOI: 10.1016/j.dental.2020.04.008
• 发表时间: 2020-05-22
• 期刊: Dental materials : official publication of the Academy of Dental Materials
• 作者: P. Sikder;J. Ferreira;Ehsan Akbari Fakhrabadi;K. Z. Kantorski;M. Liberatore;M. Bottino;S. Bhaduri
• 通讯作者: S. Bhaduri

Extracellular Matrix/Amorphous Magnesium Phosphate Bioink for 3D Bioprinting of Craniomaxillofacial Bone Tissue.

用于颅颌面骨组织 3D 生物打印的细胞外基质/无定形磷酸镁生物墨水。

• DOI: 10.1021/acsami.0c05311
• 发表时间: 2020-04-30
• 期刊: ACS applied materials & interfaces
• 影响因子: 9.5
• 作者: Nileshkumar Dubey;J. Ferreira;J. Malda;S. Bhaduri;M. Bottino
• 通讯作者: M. Bottino

Self-assembling peptide-laden electrospun scaffolds for guided mineralized tissue regeneration.

自组装负载肽的电纺支架，用于引导矿化组织再生。

• DOI: 10.1016/j.dental.2022.09.011
• 发表时间: 2022-11
• 期刊: Dental materials : official publication of the Academy of Dental Materials
• 作者: Araujo, Isaac J. de Souza;Ferreira, Jessica A.;Daghrery, Arwa;Ribeiro, Juliana S.;Castilho, Miguel;Puppin-Rontani, Regina M.;Bottino, Marco C.
• 通讯作者: Bottino, Marco C.

Three-Dimensional Printing and Biomaterials for Periodontal Regeneration

用于牙周再生的三维打印和生物材料

• DOI: 10.21203/rs.3.rs-368145/v1
• 发表时间: 2024-09-14
• 期刊: Al-Azhar Assiut Dental Journal
• 作者: S. S. Kumar