Mfg/Transport Ex Vivo Prod. Oral Mucosa Equivalents

描述（由申请人提供）：我们的临床试验数据拨款的主要目的是获得初步数据，以帮助我们评估细胞装置制造过程的均匀性和体外生产口腔黏膜等效物（EVPOME）在制造过程中细胞成分活力的一致性。目前，我们已成功获得CBER/FDA批准，启动5例患者的概念验证临床试验，BB-IND #10118，“体外生产口腔粘膜复合材料的口腔内移植”。我们预计在2005年夏天招募第一位患者。这项研究将在美国国立卫生研究院资助的密歇根大学普通临床研究中心（GCRC）进行。EVPOME将在人类应用实验室（HAL）生产，该实验室已获得良好生产规范（GMP）的批准，允许体外制备人体组织，用于移植回人体。一旦我们成功地完成了涉及5名患者的原理证明研究，我们计划在密歇根大学进行一项i - i期临床试验，以评估我们的非转基因或“初始”EVPOME移植物的口内移植的安全性。具体目标：开发非侵入性技术，以确定EVPOME在制造过程中的活力和代谢活性以及细胞成分的形态。假设：“体外产生的口腔黏膜等效物（EVPOME），或“原始”移植物，在确定的组织培养基中培养，不含血清，转化辐照饲养细胞，垂体提取物可以监测，无创，不丧失细胞成分的活力或功能”。这将通过使用以下非侵入性技术来完成：葡萄糖利用试验评估细胞活力和代谢活性；基于表面增强激光解吸/电离（SELDI）概念的Ciphergen Biosystems ProteinChip(r)系统，用于评估口腔角质形成细胞的组成产物（如与细胞活力和功能相关的人b-防御素1）的释放。3 .利用反射共聚焦显微镜获得原生状态下EVPOME的实时高分辨率光学切片，以评估其在制造过程中的形态发展；荧光寿命光谱和成像开发，优化和验证一种技术和方法，快速，无创，定量地感知组织中的内源性荧光团。这项调查的数据将允许继续开发“仿生尺度”来评估EVPOME，将其放回人体自体宿主的适用性，以及在其制造过程中形成设备/产品质量保证的基础。

DESCRIPTION (provided by applicant): The primary objective of our Clinical Pilot Data Grant is to obtain preliminary data to assist us in assessing uniformity of the manufacturing process of the cellular device and consistency in viability of the cellular component of the Ex Vivo Produced Oral Mucosa Equivalent (EVPOME) during its fabrication. At present, we have successfully received CBER/FDA approval to initiate a clinical trial for a proof of concept of 5 patients, BB-IND #10118, "Intraoral Grafting of Ex Vivo Produced Oral Mucosa Composites". We anticipate recruiting our first patient the summer of 2005. This study will be done in the University of Michigan's NIH-funded General Clinical Research Center (GCRC). The EVPOME will be produced in the Human Applications Laboratory (HAL), approved for Good Manufacturing Practice (GMP) that allows preparation of human tissue, ex vivo, for grafting back into humans. Plans have been initiated for a phase l-ll clinical trial at the University of Michigan to assess the safety of intraoral grafting of our non-genetically-modified or "naive" EVPOME grafts once we successfully completed the proof of principle study involving the 5 patients. Specific Aim: Development of non-invasive techniques, to determine viability and metabolic activity, and morphology of the cellular component of the EVPOME during its manufacturing process. Hypothesis: "Ex vivo produced oral mucosa equivalent (EVPOME), or "naive" grafts", developed in a defined tissue culture medium, free of serum, transformed irradiated feeder cells, and pituitary extract can be monitored, non-invasively, without loss of viability or function to the cellular component". This will be accomplished by using the following non-invasive techniques: 1. Glucose utilization assay to assess cellular viability and metabolic activity, 2. Ciphergen Biosystems ProteinChip(r)System based on the concept of surface-enhanced laser desorption/ionization (SELDI) to assess release of constitutive products of oral keratinocytes such as human b-defensin 1 that correlate with cell viability and function, 3. Reflectance confocal microscopy to yield real-time high resolution optical sections of EVPOME in its native state to assess its morphologic development through the manufacturing process, and 4. Fluorescence lifetime spectroscopy and imaging to develop, optimize, and validate a technology and method to rapidly, non-invasively, and quantitatively sense endogenous fluorophores in tissues. Data from this investigation will allow continued development of a "biomimetic scale" to assess EVPOME, suitability to be placed back into its human autogenous host as well as form a basis for quality assurance of the device/product during its fabrication process.