Development and Translation of Granulated Human-Derived Biomaterials for Integrative Cartilage Repair

项目摘要/摘要 这项建议的目的是研究一种新的具有颗粒状结构的人类衍生生物材料， NatruLage，刺激成体干细胞分化，用于综合软骨修复。近几年来的研究进展 关节软骨和骨损伤的治疗在解决基质退变方面显示出令人兴奋的潜力。 与骨关节炎(OA)有关，这是一种在美国困扰数百万人的关节疾病。新的 候选疗法利用动物或人类来源的材料来形成独特的微环境 有利于干细胞反应，促进愈合和整合。在健康的骨软骨组织中， 细胞外基质被组织成具有特殊细胞和信号的间质和细胞周域 分子。这些组件共同定义了特定于组织的和结构-功能关系，这些关系是 在疾病发展过程中丢失。不幸的是，综合软骨修复仍然是一种主要的医疗手段。 需要，而且需要新的产品来加强和改进治疗。成功采用 骨软骨组织疗法需要广泛的技术和制造降低风险，以满足严格的 监管审批要求。新的创新必须符合当前的良好制造规范(CGMP) 说明并展示阳性细胞和生物反应的临床前证据以及 在提交研究设备豁免(IDE)包之前和之后执行之前没有毒性 一项临床试验。为了促进软骨的整体修复，我们开发了一种新型的颗粒状细胞外基质。 通过一种名为NatruLage的专有交联技术，这种技术被注入到骨和骨的缺损处 软骨层形成骨软骨组织，与自然的带状组织结构非常相似。我们的设备 是可流动的，以输送紧密堆积的组织颗粒，并在输送后通过 独特的硫醇官能化大分子交联剂。我们已经评估了相同的NatruLage 技术，使用猪而不是人类组织，并发现在12个月内有希望的再生和整合- 月度大动物(山羊)骨软骨修复研究。我们现在专注于发展 NatruLage使用人类来源的同种异体移植组织来输送不同层的骨软骨组织。我们会 确定适当的化学、制造和质量控制以支持cGMP的生产 NatruLage用于未来的临床试验，我们将有助于了解干细胞对 纳特鲁拉奇。我们将追求三个相关的具体目标。在目标1中，我们将调整现有的cGMP设施，以用于 人类天然乳胶的生产。在目标2中，我们将定义NatruLage重复性和供体的规格 可变性。在目标3中，我们将量化NatruLage对成体干细胞增殖和分化的刺激。如果 成功，我们将为原本被丢弃或不可行的骨软骨修复创造一个理想的二级市场 这笔赠款将支持为NatruLage提交一个IDE包。

PROJECT SUMMARY / ABSTRACT The objective of this proposal is to study how a new human-derived biomaterial with granulated structure, NatruLage, stimulates adult stem cell differentiation for integrative cartilage repair. Recent advances in the treatment of articular cartilage and bone injuries have shown exciting potential to address matrix degeneration associated with osteoarthritis (OA), a joint disease afflicting millions of people in the United States. New candidate therapies utilize animal- or human-sourced materials to formulate unique microenvironments favorable to stem cell responses that promote healing and integration. In healthy osteochondral tissue, the extracellular matrix is organized into interstitial and pericellular domains with specialized cells and signaling molecules. Together, these components define tissue-specific and structure-function relationships which are lost during disease progression. Unfortunately, integrative cartilage repair still remains as a major medical need, and new products are required for enhanced and improved therapies. Successful adoption of osteochondral tissue therapies requires extensive technical and manufacturing de-risking to meet stringent regulatory approval requirements. New innovations must meet current good manufacturing practice (cGMP) specifications and demonstrate pre-clinical evidence of positive cellular and biological responses as well as lack of toxicity prior to submission of an investigational device exemption (IDE) package and, later, execution of a clinical trial. To improve integrative cartilage repair, we developed a novel granulated extracellular matrix with a proprietary crosslinking technology, termed NatruLage, that is injected into the defect in bone and cartilage layers to form osteochondral tissue that closely mimics the natural zonal tissue structure. Our device is ‘flowable’ to deliver densely-packed tissue particles, and enable forming and shaping after delivery through unique crosslinking via thiol-functionalized macromolecules. We have evaluated the same NatruLage technology, using porcine instead of human tissue, and found promising regeneration and integration in a 12- month large animal (goat) study of osteochondral repair. We are now focused on the development of NatruLage using human-derived allograft tissue for delivery of distinct layers of osteochondral tissue. We will define the appropriate chemistry, manufacturing, and quality controls to support the cGMP production of NatruLage for future clinical trials, and we will contribute to understanding the stem cell responses to NatruLage. We will pursue three related specific aims. In Aim 1, we will adapt established cGMP facilities for production of human NatruLage. In Aim 2, we will define specifications for NatruLage reproducibility and donor variability. In Aim 3, we will quantify NatruLage stimulation of adult stem cell proliferation and differentiation. If successful, we will create an ideal secondary osteochondral repair market for otherwise discarded or nonviable allograft tissue, and the grant will support the submission of an IDE package for NatruLage.