Development of an Ostoechondral Xenograft for Articular Cartilage Repair

用于关节软骨修复的骨软骨异种移植物的开发

• 批准号: 8879748
• 负责人: Steven Howard Elder
• 金额: $ 36.69万
• 依托单位: MISSISSIPPI STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-07 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8879748
• 关键词: Acute; Adult; Affect; Allografting; Appearance; Autologous Transplantation; Behavior; Biological Assay; Blood Vessels; Bone Transplantation; Canis familiaris; Cartilage; Cartilage injury; Cell-Matrix Junction; Cells; Chondrocytes; Cytolysis; DNA; Defect; Degenerative polyarthritis; Detergents; Development; Epigallocatechin Gallate; Evaluation; Excision; Extracellular Matrix; Family suidae; GAG Gene; Galactose; Glycosaminoglycans; Goals; Heterophile Antigens; Histology; Human; Hyaline Cartilage; Implant; In Vitro; Infiltration; Inflammatory Response; Injury; Joint Capsule; Joints; Knee; Knee joint; Lesion; Lung; Measurement; Measures; Mechanics; Medial; Mediating; Methods; Minor; Modeling; Morbidity - disease rate; Oryctolagus cuniculus; Pain; Penetration; Performance; Permeability; Phase; Plants; Ploidies; Porosity; Pre-Clinical Model; Proanthocyanidins; Procedures; Production; Property; Protocols documentation; Research; Resistance; Shear Strength; Site; Source; Stem cells; Stifle joint; Surface; Swelling; Synovial Fluid; Synovitis; System; Techniques; Testing; Thick; Tissues; Transplantation; Weight-Bearing state; Xenograft procedure; alternative treatment; articular cartilage; base; bone; cartilage repair; collagenase; cost; crosslink; fetal bovine serum; genipin; immunogenicity; implantation; improved; injured; interest; migration; novel; novel strategies; nuclease; osteochondral tissue; public health relevance; repaired; treatment duration

 DESCRIPTION (provided by applicant): Articular cartilage of the knee is quite susceptible to injuries which are often painful and which may progress to osteoarthritis. Osteochondral autografting and allografting are currently the only treatment options which immediately restore hyaline cartilage to the injured joint. Drawbacks to these procedures such as donor site morbidity and limited tissue availability have stimulated interest in osteochondral xenografts, which have the potential advantages of low cost and abundant supply. The long-term objective of the proposed research is to develop a decellularized porcine osteochondral xenograft (OCXG) which can be available for implantation as soon as a cartilage lesion is discovered (i.e., "off-the-shelf"). Development includes optimization of decellularization and crosslinking methods to reduce immunogenicity and improve graft durability and integration with host tissue. The specific Aims are as follows: 1. Compare glycosaminoglycan-containing and glycosaminoglycan-free porcine osteochondral dowels in terms of their mechanical properties and ability to support stem cell infiltration and chondroinduction in vitro. 2. Evaluate the effect of four different plant- derived, nontoxic crosslinking agents on the physical and mechanical properties of GAG(+) and GAG(-) OCXGs, as well as their effects on primary human knee chondrocyte behavior in vitro. 3. Evaluate the acute inflammatory response to GAG(+) and GAG(-) decellularized porcine OCXGs, both crosslinked and non- crosslinked using the most promising agent from SA2, in an intraarticular rabbit model. 4. Determine the capacity for crosslinked, decellularized OCXGs to repair full-thickness defects in the trochlear groove of adult dogs. The first aim will provide important information about fundamentally different approaches to decellularization, one which seeks to preserve all extracellular matrix and one which intentionally extracts non- collagenous components (especially glycosaminoglycan) to create more porosity. Evaluations include measurement of the DNA and GAG contents, the graft cartilage's biphasic properties, and the attachment, proliferation, and matrix production of human chondrocytes cultured on the grafts. The first aim also includes development of a novel decellularization protocol. The second aim will reveal the potential for natural, nontoxic crosslinking agents to enhance xenograft mechanical properties and collagenase resistance. It will also determine whether chondrocytes are sensitive to the degree of crosslinking. The third aim involves short-term transplantation of OCXGs into the medial femoral condyle of mature rabbits, mainly to investigate OCXG- induced synovitis. The effects of decellularization method and crosslinking will be examined through synovial fluid analysis and histology. The final aim will evaluate the functionality of porcine osteochondral xenografts used to fill surgically created defects in the canine knee joint. Semiquantitative scoring of macroscopic appearance and histology, in addition to indentation testing, will demonstrate whether osteochondral xengrafts are capable of restoring a healthy joint surface in this preclinical model.

 描述（由适用提供）：膝盖的人工软骨非常容易受到伤害，这通常很痛苦，可能会发展为骨关节炎。骨软骨自体脂肪和同藻目前是唯一立即将透明软骨恢复到受伤关节的治疗选择。这些程序的缺点，例如供体部位的发病率和有限的组织可用性，引起了对骨软骨异种移植的兴趣，这具有低成本和丰富供应的潜在优势。拟议的研究的长期目标是开发脱细胞的猪骨软骨异种移植（OCXG），一旦发现软骨病变（即“现成”），可以立即植入。发展包括优化脱细胞和交联方法，以降低免疫原性并改善移植物的耐用性和与宿主组织的整合。具体目的如下：1。根据其机械性能和支持干细胞浸润和软骨诱导的能力，比较含糖和无糖胺聚糖猪骨软骨销钉。 2。评估四种不同植物衍生的，无毒的交联剂对GAG（+）和GAG（ - ）OCXGS的物理和机械性能的影响，以及它们对体外人膝盖软骨细胞行为的影响。 3。在关节内兔模型中使用来自SA2的最有前途的剂，评估对GAG（+）和GAG（+）和GAG（+）和GAG（ - ）脱细胞型猪OCXGS的急性炎症反应。 4.确定成年犬的交联型OCXG修复全厚度缺陷的能力。第一个目的将提供有关脱细化方法的重要信息，该信息旨在保留所有提取的细胞基质，并有意提取非胶原成分（尤其是糖胺聚糖）以产生更多的孔隙率。评估包括测量DNA和GAG含量，移植软骨的双相特性以及在移植物上培养的人软骨细胞的附着，增殖和基质产生。第一个目的还包括开发一种新型的脱细化协议。第二个目标将揭示自然无毒的交联剂的潜力，以增强异种移植机械性能和胶原酶耐药性。它还将确定软骨细胞是否对交联的程度敏感。第三个目标涉及将OCXG短期移植到成熟兔子的股骨内侧，主要是为了研究OCXG诱导的滑膜炎。脱细胞方法和交联的影响将通过滑液分析和组织学检查。最终目标 评估用于填充犬膝关节中手术产生的缺陷的猪骨软骨异形的功能。除凹痕测试外，宏观外观和组织学的半定量评分还将证明骨软骨异形是否能够在此临床前模型中恢复健康的关节表面。

项目成果

Suitability of EGCG as a Means of Stabilizing a Porcine Osteochondral Xenograft.

• DOI: 10.3390/jfb8040043
• 发表时间: 2017-09-23
• 期刊: Journal of functional biomaterials
• 影响因子: 4.8
• 作者: Elder S;Clune J;Walker J;Gloth P
• 通讯作者: Gloth P

Comparison of natural crosslinking agents for the stabilization of xenogenic articular cartilage.

• DOI: 10.1002/jor.23121
• 发表时间: 2016-06
• 期刊: Journal of orthopaedic research : official publication of the Orthopaedic Research Society
• 作者: Pinheiro A;Cooley A;Liao J;Prabhu R;Elder S
• 通讯作者: Elder S