Mfg/Transport Ex Vivo Prod. Oral Mucosa Equivalents

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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.