Plastic Surgery: Research Prospectus 2002-03

The Section of Plastic Surgery has completed a highly successful year in both clinical and basic science research. Projects focused on the basic aspects of wound healing, muscle and nerve biology, transplant immunology, outcomes of breast reconstruction, and craniofacial development have been successfully supported by the extramural funding of the National Institutes of Health and other agencies. The diversity of research interests and projects arise from the faculty's enthusiasm to develop and understand basic biological methods and the technology of the future.

The laboratory of Cynthia Marcelo, PhD investigates the control of keratinocyte growth and differentiation by cell membrane phospholipid fatty acids. The fatty acid lipids affect membrane fluidity/viscosity second messenger systems and the expression of lipid metabolism enzymes and the nuclear receptor family of PPARS. She collaborates with several members of Biophysical Research Division and William Richard Dunham, PhD, Emeritus Distinguished Research Scientist in Plastic Surgery, in these NIH-funded studies. Dr. Marcelo is Co-PI with members of the Oral Surgery Department, the Dental School, and Internal Medicine on an NIH-funded award to study the development oral mucosal skin equivalents for oral reconstruction (Drs. Stephen Feinberg and Blake Roessler).

With Dr. Ambati Reddy, a visiting research scientist, the Marcelo laboratory is continuing studies into the role of peroxisome-proliferator activated receptors (PPARs) in fatty acid and eicosanoid induced keratinocyte function.

Dr. Marcelo's laboratory has a joint effort with Stephen Feinberg, DDS, PhD from the University of Michigan to grow permanent oral tissue. Junhi Song, MD, Takayosti Tobital, DDS and Kenji Izumi, DDS have just joined the Marcelo/Feinberg research effort. These investigators, from China and Japan, are working to form oral mucosal keratinocyte composites for use in oral surgery. These composites will carry genetic information to accelerate wound healing. The Phase I clinical trials are about to begin.

In a continuing funded effort with Keracure, LLC, Dr. Marcelo is developing methods to stabilize the living keratinocyte product for maximal shelf-life. Apoptosis, cell aging, and cell stem cell biology are the basis of these studies.

In 1997, Dr. Marcelo as PI and the faculty of the Section of Plastic Surgery were awarded a Training Grant in Burn, Trauma and Wound Healing Research. This NIH training grant supports the training of academic surgeons/scientists in basic research related to trauma and burn injuries. Over the past five years, this grant has supported the training of Drs. Marlene Calderon, Sam Rhee, Sameer Jejurikar, and Christi Cavaliere. Dr. Calderon is obtaining her PhD degree in the Department of Physiology, mentoring under Dr. Faulkner (and Kuzon). Dr. Rhee completed his training with Dr. Buchman, and Dr. Jejurikar completed his training with Drs. Kuzon and Marcelo. Dr. Christie Cavaliere is working with Dr. Steve Goldstein (Department of Surgery and Associate Dean of Research and Graduate Studies) and Dr. Steve Buchman. Dr. Gregory Borschel, who is working with Dr. David Mooney (Dept. of Biological and Material Sciences) and Dr. William Kuzon is a trainee also associated with the training grant.

Kevin C. Chung, MD, MS, continues his outcomes research in hand surgery and plastic surgery. He is the principal investigator for an RO1 grant, "An Outcome Study of Rheumatoid Hand Arthroplasty," which is under review by the NIH. Dr Chung was recently selected by the NIH to review rheumatoid arthritis center grants as a part of the NIAMS study section. He is the principal mentor for Amy Alderman, MD, a plastic surgery resident currently in the Robert Wood Johnson Clinical Scholars Program. Dr. Alderman is studying outcomes of hand surgery for rheumatoid patients and national variations in practice for rheumatoid hand procedures. She is also working with Dr. Edwin Wilkins on issues involving access to reconstruction for women with breast cancer. She won the 2002 Plastic Surgery Educational Foundation Scientific Essay contest, taking first place in the Clinical Science-Junior Category. In addition to Dr. Alderman, Dr Chung is mentoring Catherine Curtin, MD, a plastic surgery resident, who is also a first-year Robert Wood Johnson Clinical Scholar. Dr. Curtin will study outcomes of upper extremity surgery for spinal cord injury patients. Her project examines factors that lead to the underutilization of surgery in this population. Dr. Curtin's collaborators also include members of the Spinal Cord Center at the University of Michigan.

The Plastic Surgery Faculty is an integral part of The Muscle Mechanics Laboratory (MML) in the Institute of Gerontology. The laboratory provides a multidisciplinary environment for over 30 investigators, post-doctoral fellows, pre-doctoral students, undergraduate students, and research technicians. David Brown, MD, Paul Cederna, MD, William Kuzon, Jr., MD, PhD, Melanie Urbanchek, PhD, Jack Van der Meulen, PhD, and Dennis Claflin, PhD continue to work as principal investigators in the laboratory. John Faulkner, PhD, the director of the laboratory, holds an appointment as Professor of Plastic Surgery. Ongoing work in the laboratory is aimed at determining the mechanisms responsible for the contractile dysfunction in skeletal muscle observed after denervation and reinnervation, injury, microneurovascular transfer, and aging. Whole muscle, single motor unit, and single muscle fiber contractile property evaluations are all performed to measure outcomes from various experimental manipulations.

The tissue engineering initiative has played a prominent role in the scientific advances of the MML over the past year. Work has focused on tissue engineering of skeletal muscle and peripheral nerve as relevant to reconstructive surgery. Acellularized constructs of both muscle and nerve have been developed and have been used to produce functional nerve and muscle. Current work is aimed at determining the basic mechanisms that generate cell-matrix interactions and at optimizing function in engineered tissue. A new focus on brain mechanism interfaces has gotten underway. Drs. Dennis, Larkin, Kipke, Goldstein, and Mooney are collaborators integral to this work.

Substantial scientific advances have been made in the area of peripheral nerve allotransplantation. The basic immunology of peripheral nerve transplantation has been evaluated through collaborative efforts with Keith Bishop, PhD in transplant immunology. Short-term monoclonal antibody administration at the time of nerve allograft transportation has successfully induced tolerance with the need for long-term immunosuppression reconstruction of long peripheral nerve gaps with nerve allografts in animal models. Through the use of genetically altered animal models, the immunology has been extensively evaluated. This work has been submitted in multiple scientific research awards at regional and national meetings.

M. Asim Aydin, MD, a visiting plastic surgeon from Suleyman Damiral University, Ispecta, Turkey, contributed to the Muscle Mechanics Lab studies. His contributions included studies of FK506, muscle denervation and nerve repair on muscle junction.

Gregory Borschel, MD continues in the laboratory as a post-doctoral research fellow. He has reported the successful in vitro generation of 3-dimensional contractile skeletal muscle scaffold. His work has been presented at the Plastic Surgery Research Council and at the American College of Surgeons.

David Brown, MD has recently joined the research consortium of the Muscle Mechanics Lab as a Principle Investigator. He is working on peripheral nerve allotransplantation and how tolerance can be induced through co-stimulative pathway blockade. He is studying the functional outcomes of peripheral nerve allograft hindlimb reconstruction through neurophysiologic evaluation and muscle isometric force measurements. He has also continued his fellowship at the O'Brien Institute of Microsurgery in Melbourne, Australia. This effort is focused around angiogenesis and the development of tissue engineered blood vessels.

Marlene Calderon, MD continues to be a full-time post-doctoral trainee in the laboratory. She has achieved candidacy for a PhD in the Department of Physiology with Drs. Faulkner and Kuzon as her mentors. Dr. Calderon served as a Plastic Surgery Education Foundation Research Fellow, one of only four fellowships awarded nationwide. Dr. Calderon was given an award for outstanding research at this year's Plastic Surgery Research Council Meeting.

Paul Cederna, MD recently was recognized as the first John E. Hoopes Academic Scholar of the American Association of Plastic Surgeons. He has also joined the faculty of the Institute of Gerontology as an Assistant Research Scientist to pursue the mechanisms responsible for skeletal muscle force deficits with aging. Dr. Cederna has been named to the editorial board of the Journal of Reconstructive Microsurgery and is a journal referee for 9 different journals. His research funding includes being Principal Investigator on grants from the American Association of Plastic Surgeons and the Plastic Surgery Education Foundation. A new award from the Michigan Life Science Initiative will focus on the adaptive response in skeletal muscle to training and aging. Dr. Cederna is Co-investigator on NIH and VA Merit Grants. He is the supervisor for Dr. Anil Mungara.

Dennis Claflin, PhD was the recipient of two pilot grants, one from the Center for Biomedical Engineering Research and one from the Nathan Shock Center for the Biology of Aging, to support his work concerning force deficits in skeletal muscle due to compromised excitation-contraction coupling.

Douglas Dow, PhD has completed his doctoral thesis investigating the effects of functional electrical stimulation on muscle mass and force through mentoring from Drs. Carlson, Cederna, Dennis and Faulkner. Dr. Dow has performed novel research to delineate the mechanisms responsible for maintaining physiologically healthy muscle prior to reinnervation. His work has been presented regionally and nationally and has been recognized with awards. At the 2002 Meeting of the American Society of Peripheral Nerve, he was awarded with the Outstanding Nerve Paper Award. He will continue to work in the Muscle Mechanics Laboratory as a postdoctoral research fellow.

William Kuzon, MD, PhD, continues to function as the Associate Chair for Research in the Department of Surgery, as an Assistant Research Scientist in the Institute of Gerontology, and as Section Head of Plastic Surgery. Dr. Kuzon is on the editorial board of 3 prominent journals and is a journal referee for 16 different journals. Dr. Kuzon has been honored as the American College of Surgeons Traveling Fellow to Australia and New Zealand for 2001. His research funding includes being the Principal Investigator on NIH and VA Merit Awards, as well as Co-investigator on 2 additional NIH grants. Dr. Kuzon participates as a NIH Training Training Grant Faculty member on 2 separate training grants. Dr. Kuzon is currently the President-Elect of the American Society For Peripheral Nerve.

Jennifer Lynch, MD joined the laboratory in July 2002 and will perform studies to determine the effect of mandibular distraction osteogenesis on the structure and function of adjacent skeletal muscles.

Anil Mungara, MD has joined the Muscle Mechanics Laboratory as a Visiting Research Fellow. He is a general surgeon from Toronto, Canada with fellowship training in Laparosegsie General Surgery. Dr. Mungara has been studying the immunology of peripheral nerve allotransplantation through use of various monoclonal antibodies and knockout mice. Working with Drs. Bishop and Cederna, he has completed work resulting in numerous abstracts and publications. He was awarded a Basic Science Award at the recent Michigan Academy of Plastic Surgeons Meeting. He was also selected as one of the outstanding nerve papers at the upcoming meeting of the American Society For Peripheral Nerve.

Melanie Urbanchek, PhD continues to participate actively in the MML consortium, studying muscle neurotization, and effects of aging, FK-506, and reinnervation on muscle. Dr. Urbanchek has been very active mentoring students in the laboratory. Under the guidance of Dr. Urbanchek, Diana Ganz was honored to present at the National Conference for Undergraduate Research. Ms. Ganz, John Piersma and Steven Rayappa have all received research training in the MML over the past year, securing funding from the University of Michigan Summer Biomedical Research Training Program and the Undergraduate Research Opportunity Program.

Jack Van der Meulen, Ph.D., continues to be an active participant in the MML consortium, investigating the mechanisms responsible for skeletal muscle force deficits following denervation and reinnervation. His expertise in single motor unit evaluations and single muscle fiber contractile property measurements have allowed great strides to be made in these investigations. Dr. Van der Meulen's work is funded through NIH and VA Merit Awards.

During the past year, members of the MML consortium have presented their work at following scientific meetings: The Plastic Surgery Research Council, The American Society for Peripheral Nerve, The American Society of Plastic Surgeons, The American College of Sports Medicine, The Surgical Forum of the American College of Surgeons, The Michigan Academy of Plastic Surgeons, The Michigan Medical Society and the American Society of Reconstructive Microsurgery. Members of the MML have published over 30 manuscripts in peer-reviewed journals in the past year.

Research in the laboratory is funded by grants from numerous sources, including the National Institutes of Health, the VA Merit Review Board, the Department of Surgery Research Advisory Committee, the Plastic Surgery Educational Foundation, the American Association of Plastic Surgeons, the University of Michigan Biomedical Technology Development Fund, the Center For Biomedical Engineering Research, The Michigan Life Sciences Initiative, and the Nathan Shock Center For the Biology of Aging. Outside the Muscle Mechanics Consortium, significant collaborators in this work include Drs. Steven Buchman (Plastic Surgery), Bruce Carlson (Anatomy and Cell Biology), Daniel Goldman (Mental Health Research Institute), Steven Goldstein (Department of Surgery), Susan Mackinnon (Washington University at St. Louis), Michael Longaker (Stanford University), Keith Bishop (General Surgery/Immunology), Robert Gilmont (Plastic Surgery), Cynthia Marcelo (Plastic Surgery), Michael Ignelzi (Dentistry), Robert Dennis (Mechanical Engineering), Lisa Larkin (Internal Medicine) William Walsh (University of New South Wales, Australia), Mark Gianostsous (South Wales, Australia).

Steven R. Buchman, MD continues his work in the area of craniofacial research. The main research initiatives in his laboratory are the analysis of the relationship between biomechanical environment and bone graft behavior, the etiology of craniosynostosis and cranial suture morphogenesis, and the underlying molecular mechanisms of distraction osteogenesis. Dr. Buchman is a faculty participant of the University of Michigan Multipurpose Arthritis and Musculoskeletal Diseases Center (UM-MAC), a National Institutes of Health funded program providing support and funding for interdisciplinary projects. Sam Rhee, MD a NIH trainee, presented his studies on apoptosis and mandibular distraction osteogenesis at the 2000 Moses Gunn Conference. A multitude of undergraduate and medical students have also been active in several research projects. Programmatic support for these initiatives has been won through the Undergraduate Research Opportunities Program, the Student Biomedical Research Training Program, and the Howard Hughes Medical Research Scholarship Program. Work from the laboratory has been presented at The International Society of Craniofacial Surgery, The Michigan Academy of Plastic Surgery, The American Society of Plastic and Reconstructive Surgery, The American College of Surgeons, The Plastic Surgery Research Council, and the American Cleft Palate-Craniofacial Association in the past year.

Dr. Buchman continues his close liaison with Dr. Steve Goldstein in the Orthopedic Research laboratory. Dr. Buchman is a Co-Principle Investigator on a Collaborative RO1 NIH Grant entitled "Mechanically Guided Tissue Regeneration" with PI S. Goldstein and Co-PI M. Ignelzi. This study looks at the relationship between biomechanical forces and new bone formation. Another award, entitled "Distraction Osteogenesis: Endogenous Tissue Engineering" with Investigators: M. Longaker (Stanford), S. Buchman, W. Kuzon, S. Goldstein, and M. Ignelzi is also funded by the NIH. This study develops a method of endogenous tissue engineering for the reconstruction and replacement of pediatric mandibular deficiency that would help to alleviate the significant psychosocial and biomedical burden of this common problem. In addition, research in the laboratory is funded by grants from numerous other sources including The American Society of Maxillofacial Surgeons, The Maxillofacial Surgeons Foundation, and the Plastic Surgery Research Council.

Christi Cavaliere, MD continues in the laboratory as a post-doctoral research fellow. She is utilizing a rat model of mandibular distraction osteogenesis to study the molecular and mechanical signals involved in bone formation during distraction. In addition, she continues to investigate the role of force acting on cranial sutures using an in vitro cranial loading device.

Robert R. Gilmont PhD studies a primary mechanism underlying normal dermal wound healing, that is, contraction. Wound contraction is mediated by fibroblast contraction of the wound edge and the granulation tissue. Determining the molecular mechanism of fibroblast mediated wound contraction is the primary goal of his research effort. Dr. Gilmont uses fibroblast populated collagen lattice (FPCL) contraction as a tissue culture model of wound contraction and studies mitogen activated protein kinase (MAP kinase) activation dependent contraction. Using a series of inhibitors to MAP kinase, he and his trainees have identified hsp27 as one of the target molecules for MAP kinase dependent phosphorylation. Cell lines transfected to over express or under express hsp27 show a quantitative relationship between hsp27 expression and FPCL contraction. These studies have resulted in the submission to the NIH of a proposal designed to identify other MAP kinase substrates involved in contraction and determine the molecular mechanisms for hsp27 mediated contraction. Drs. Welsh and Benndorf from the Department of Cell and Developmental Biology and Dr. Marcelo from Plastic Surgery are collaborators.

Dr. Sahoko Tanigawa Hirano, a visiting plastic surgeon from Japan, has just finished her studies with Dr. Gilmont and Dr. Marcelo and is continuing at the University in the Department of Cell and Molecular Biology. She worked with Dr. Gilmont on hsp27 function in skin and with Dr. Marcelo on PPRAP and lipids in skin keratinocytes.

Riley S. Rees, MD has started a new initiative for the treatment of melanoma. He has received funding from the Surgery Research Advisory Committee (RAC) and the Cancer Center to study a new treatment for head and neck melanoma. The studies in mice are designed to provide data for an FDA trial if they are successful. Other ongoing activities include Dr. Rees' treatment and management of chronic wounds. He directs the Wound Care Program. Dr. Riley Rees is a founder of a University of Michigan start-up wound care company resulting from the collaboration of a group of basic scientists and clinical employees of the University of Michigan and Wayne State University. The efforts of the group have resulted in four issued patents to date.

Edwin G. Wilkins, M.D., M.S. continues his work in the area of health services research. He is a clinical investigator at the Center for Outcomes Research and Practice Management in the Ann Arbor V.A. Medical Center. He is collaborating with other health services researchers in evaluating patient outcomes for post mastectomy breast reconstruction. He is also involved in ongoing efforts to develop and implement an internet based telemedicine system for evaluation and treatment of pressure ulcers and other chronic wounds. Development of a multicenter trial of this technology is currently underway. Dr. Wilkins is also working with colleagues at the School of Public Health and in the University of Michigan Breast Care Center on a study evaluating the impact of shared decision making in the treatment of early breast cancer. Finally, Dr. Wilkins is also continuing his work to develop, implement, and study web based shared decision making technologies for an array of Plastic Surgery procedures. The goal of this latter effort is to improve patient education in decision-making in Plastic Surgery.

The research areas of the Section of Plastic Surgery include:

  • Clinical investigation of growth factors in wound healing
  • Inflammation and tissue injury in skin flaps and chronic wounds
  • Autogenous tissue reconstruction following mastectomy
  • Alterations and potential manipulations of molecular genetic activity in normal and diseased or injured nerves
  • Cytokines in the pathogenesis of progressive skin injury
  • Skeletal muscle reinnervation
  • Use of photophores to sensitize melanoma cells
  • Outcomes following breast reconstruction
  • Bone remodeling with craniofacial development
  • Bone graft behavior
  • Force induced craniosynostosis
  • Molecular mechanisms of Distraction Osteogenesis
  • Biomechanical performance of normal and abnormal human skin
  • Control of epidermal cell function using adult human cells in culture
  • Defining epidermal stem cells
  • Mechanism of fatty acid metabolism
  • Validation of the Michigan Hand Outcomes Instrument
  • Outcomes of Rheumatoid Hand Arthroplasty
  • Mechanisms responsible for diminished mechanical function in skeletal muscle after reinnervation, transplantation, injury, and aging.
  • The alterations in skeletal muscle structure and function as a result of distraction osteogenesis.
  • Alternative methods for nerve reconstruction.
  • Tissue engineering of skeletal muscle and peripheral nerve.

Plastic & Reconstructive Surgery Researc

Michigan Breast Reconstruction Outcome Study (MBROS)

Collaborators: E.G. Wilkins, Julie Lowery PhD, School of Public Health, W. Kuzon, Richard Lichtenstein PhD, School of Public Health, Randy Roth PhD, School of Public Health, J. William Thomas PhD, School of Public Health, James A. Leonard MD, Physical Medicine and Rehabilitation
Funding: U.S. Department of the Army

The objective of MBROS is to compare the long-term outcomes of two common techniques of post-mastectomy breast reconstruction: tissue expansion/breast implants and transverse rectus abdominis musculocutaneous (TRAM) flaps. A four year prospective study, the project will adapt existing instruments and formulate new methodologies to assess outcomes in five categories: complication rates, aesthetic results, functional results, psychosocial status and costs. Study results will provide much needed information to patients, providers, and payers for determining the procedure of choice. In addition, the research will establish standardized methods for evaluation of breast reconstruction results in future studies. Finally, initial data assembled by this research can also be used for long-term analysis of breast reconstruction outcomes.

Subjects are being recruited from the practices of 25 plastic surgeons across four states and Canada and will include women undergoing expander/implant or TRAM procedures. Patients will be followed for a minimum of two years from the time of reconstruction. Measurements of outcome variables will be obtained prior to reconstruction and at annual intervals starting one year after the procedure. Instruments used will include questionnaires, physical testing, evaluation of digitized photographs, and analysis of billing and hospital record data. This approach will identify the cumulative effects of reconstruction on some outcome measures (e.g., complications and costs) and will help determine the earliest time point at which other outcomes stabilize (e.g., functional, aesthetic, and psychosocial status).

Michigan Breast Reconstruction Outcome Study (MBROS)

Collaborators: E.G. Wilkins, Julie Lowery PhD, School of Public Health, W. Kuzon, Richard Lichtenstein PhD, School of Public Health, Randy Roth PhD, School of Public Health, J. William Thomas PhD, School of Public Health, James A. Leonard MD, Physical Medicine and Rehabilitation

Funding: U.S. Department of the Army

The objective of MBROS is to compare the long-term outcomes of two common techniques of post-mastectomy breast reconstruction: tissue expansion/breast implants and transverse rectus abdominis musculocutaneous (TRAM) flaps. A four year prospective study, the project will adapt existing instruments and formulate new methodologies to assess outcomes in five categories: complication rates, aesthetic results, functional results, psychosocial status and costs. Study results will provide much needed information to patients, providers, and payers for determining the procedure of choice. In addition, the research will establish standardized methods for evaluation of breast reconstruction results in future studies. Finally, initial data assembled by this research can also be used for long-term analysis of breast reconstruction outcomes.

Subjects are being recruited from the practices of 25 plastic surgeons across four states and Canada and will include women undergoing expander/implant or TRAM procedures. Patients will be followed for a minimum of two years from the time of reconstruction. Measurements of outcome variables will be obtained prior to reconstruction and at annual intervals starting one year after the procedure. Instruments used will include questionnaires, physical testing, evaluation of digitized photographs, and analysis of billing and hospital record data. This approach will identify the cumulative effects of reconstruction on some outcome measures (e.g., complications and costs) and will help determine the earliest time point at which other outcomes stabilize (e.g., functional, aesthetic, and psychosocial status).

Effect of Fatty Acid Supplementation on Keratinocyte Function A technique to grow epidermal keratinocytes in culture causes the cells to become essential fatty acid depleted. The restoration of normal fatty acid composition results in significant changes in cell replication and differentiation rates. Studies to determine the changes in cellular function associated with these fatty acid alterations are ongoing. This project is the basis of the work currently being done by Drs. Hirano, Corpron, and Gilmont, in Dr. Marcelo's laboratory.

Modulation of Keratinocyte Growth and Specialization

Investigators: C. Marcelo, W.R. Dunham, R. Sands
Funding: National Institutes of Health

The ongoing research is an interdisciplinary study to define epidermal cell function. The hypothesis driving this proposal is that the fatty acid content of membrane phospholipids controls the viscosity of the cell membrane and that this parameter, in turn, affects membrane mechanisms that control cell function. In vitro EFAD keratinocytes can be grown in essential fatty acid supplemented medium to yield cells with "normalized" membranes. Thus, this characterized keratinocyte culture is being used in electron paramagnetic resonance (EPR) and enzyme kinetic experiments to: a) Test the hypothesis that changes in membrane fluidity caused by altered phospholipid fatty acid content alters the kinetic rates of membrane associated enzymes. b) Understand the effect of altered fatty acid phospholipid content (GC) on membrane viscosity resulting from supplementing the growth medium with various fatty acids. c) Determine the effect of dermatological effective drugs on EFA deficient and normalized keratinocytes. d) Analyze second messenger and signaling systems in EFA deficient and fatty acid normalized cells. The role of PPARs in this interaction among lipids and cell function will be studied.

Training Grant in Burn, Trauma and Wound Healing Research

PI and Program Director: C. Marcelo Faculty: D. Smith Jr., W. Kuzon, R. Rees, R. Gilmont, S. Buchman, J. Faulkner, D. Remick, S. Goldstein, D. Mooney, P.Cederna, and W.R. Dunham
Funding: National Institutes of Health

This training grant will support the training of potential academic medical surgeon/scientist in basic research related to trauma and burn injuries. Over the last 6-10 years, a group of researchers with similar interest in trauma and burn research have been interacting and collaborating, and training students in this type of research. We have formalized the training by being awarded a NIH T-32 training grant for a 5-year period. There are twelve members of the program faculty, from the Section of Plastic Surgery, Department of Surgery, the Department of Pathology and the Biophysics Research Division. These are Dr. Cynthia L. Marcelo, a cell physiologist, her collaborator, Dr. Richard Dunham, and Dr. David J. Smith, Dr. Daniel Remick, member of the Department of Pathology, who are well-funded scientists in the area of tissue damage in trauma. Dr. William Kuzon who works in collaboration with Dr. John Faulkner in the Muscle Mechanics Laboratory Research Consortium, and finally, Dr. Riley Rees and his collaborator Dr. Robert Gilmont, who are currently investigating ischemic tissue injury. Dr. Steve Buchman will be added to the faculty in year 2, and his research is in the area of cranial facial injury. The Grant supports two postdoctoral fellows per year, plus supplies and travel allowances. This grant will support the extensive training of medical doctors to enter research/academic careers.

Behavioral, Neuropsychological, and Social Outcomes of Children Treated with Distraction Osteogenesis

Investigators: S. Buchman, Seth Warschausky, PhD, Mary Berger, MS
Funding: National Institutes of Health

Distraction osteogenesis (DO) is a relatively new technique used to restore structural deficiencies of the oral-facial complex and is presumed to have positive effects on children's quality of life, in fact, early surgical intervention of much of craniofacial surgery is predicated on the assumption of an improvement of psychosocial development. Outcome analysis substantiating these assumptions have been woefully inadequate and yet significant numbers of complex craniofacial reconstructive procedures and overwhelming health care expenditures are considered to be the standard of care for children afflicted with these maladies. A rationale basis for reconstructive intervention in these patients is imperative, however, the lack of specificity of outcome measures has been a fundamental problem in determining the quantity, quality, and timing of care. In this multicenter study, we will examine the changes in psychosocial and communicative status in children with and without a DO procedure. We hypothesize that DO will result in specific and positive change in psychosocial and communicative development. Results are expected to have significant implications for biorestorative treatment planning by identifying predictive relations among psychosocial and communicative factors and by establishing verifiable outcome measures.

Molecular and Cellular Mechanisms of Mandibular Regeneration Using Distraction Osteogenesis

Investigators: M. Ignelzi, S. Buchman
Funding: National Institutes of Health

Distraction Osteogenesis, the stimulation of new bone formation by gradual separation of two osteogenic fronts, has been used for decades in long bones. Although distraction osteogenesis to correct mandibular deficiencies has gained popularity recently, the molecular mechanisms that regulate these events are unknown. The global hypothesis to be tested in this project is that mechanical strains, produced by distraction forces, induce changes in the cytoskeletal architecture, which function to regulate patterns of gene expression and ultimately, patterns of cell differentiation. This study will identify genes that regulate mandibular regeneration in vivo using distraction osteogenesis. The long-term goals of this study are to understand how distraction force regulates gene expression and to identify genes that can be used in therapeutic interventions to promote regeneration for complete biorestoration of oral health.

Mechanical Forces Regulate Molecular Mediators in Skeletal Articulations

Investigators: M. Ignelzi, S. Buchman
Funding: National Institutes of Health

This research project investigates the regulatory role of force on the cellular and molecular genetic mechanisms of craniosynostosis in vivo and in vitro. We postulate that compressive forces, such as intrauterine compression, or tensional forces, such as abnormal forces exerted by the cranial base, lead to a change in gene expression at the osteogenic front of the cranial suture and that these changes lead to abnormal suture fusion and craniosynostosis. The central hypothesis of this project is that mechanical forces can change the pattern of gene expression in a course of normal suture fusion and craniosynostosis. This hypothesis will be tested by applying compressive and tensile forces to mouse calveria and performing subsequent analyses using micro-computed tomography, immunohistochemistry, in situ hybridization, as well as differential display techniques. The specific aims of this study will provide valuable insight into the basic mechanisms that regulate the development and maintenance of cranial sutures, and to utilize these mechanisms to reduce the morbidity associated with craniosynostosis.

Mechanically Guided Tissue Regeneration

Investigators: S. Goldstein, M. Ignelzi, S. Buchman
Funding: National Institutes of Health

The purpose of this project is to test the global hypothesis that the incorporation and function of a tissue engineered construct for bone regeneration is significantly influenced by the transmission of physical forces through its extracellular matrix. Utilizing a series of in vivo rat models, an analysis is performed of the synergistic effects of device mediated mechanical forces and delivery of biologic factors stimulating the repair and incorporation of tissue engineered bone substitutes. The models in both the craniofacial and long bone sites utilize specially designed external fixators, which enable the implementation of experimentally controlled mechanical loading conditions across surgically created osteotomy gaps. The effects of the mechanical and biologic factors on tissue repair and regeneration will be assayed by documenting the spatial patterns of gene expression, matrix synthesis, and bone morphology as a function of time and anatomic location.

Distraction Osteogenesis: Endogenous Tissue Engineering

Investigators: M. Longaker (NYU), S. Buchman, W. Kuzon, S. Goldstein, M. Ignelzi
Funding: National Institutes of Health

A method of endogenous tissue engineering for the reconstruction and replacement of pediatric mandibular deficiency would help to alleviate the significant psychosocial and biomedical burden of this common problem. Distraction osteogenesis (DO), the creation of new bone by the gradual separation of two bony fronts, is the ultimate method of tissue engineering because it provides an anatomical and functional replacement of deficient tissue with endogenous bone generated from local substrate. The central hypothesis to be tested in this proposal is that the creation of newly engineered bone generated by mandibular DO is the result of an adaptive response by bone and the surrounding soft tissue envelope. We further hypothesize that there is a fundamental difference in the mechanism of this adaptive response, characterized by the creation of new bone, and a maladaptive response, characterized by injury and fibrous non-union. It is further hypothesized that under adaptive conditions the response of adjacent masticatory muscles will be adaptive, and that under maladaptive conditions, the response will be injury to the muscles of mastication. To test this hypothesis, we will utilize a novel rodent model to determine the cellular, molecular, and biomechanical mechanisms that distinguish the adaptive from the maladaptive response in mandibular DO. This interdisciplinary proposal is timely and relevant because mandibular DO is rapidly becoming the treatment of choice for reconstructing pediatric mandibular deficiencies.

The Michigan Hand Outcomes Questionnaire

Investigator: K. Chung

Emphasis on the evaluation of patient outcomes as a part of quality improvement initiatives in health care has sparked debate among physicians, researchers, and third-party payers regarding the appropriate measurement of health status for specific populations. While researchers have developed instruments such as the Short Form-36 (SF-36) to measure and quantify overall health status, these general questionnaires may not capture the outcomes of interest to hand surgeons. Hand surgeons, like other specialized practitioners, need reliable and valid methods to quantify functional performance and overall satisfaction among their patients.

Dr. Chung has developed and validated an outcomes instrument - the Michigan Hand Outcomes Questionnaire (MHQ) - to measure health state domains important to hand-injured patients. He has shown it to be a valid and reliable instrument in a survey of 200 hand-injured patients, measuring six domains: 1) overall hand function; 2) activities of daily living (ADLs); 3) pain; 4) work performance; 5) aesthetics; and 6) patient satisfaction with hand function. Dr. Chung has utilized the MHQ in several of his studies, including an outcomes study of thumb reconstruction using microvascular toe transfer and a prospective study on the outcomes of minimal incision carpal tunnel release.

An Outcome Study of Rheumatoid Hand Arthroplasty

Investigator: K. Chung
Funding: Pending NIH funding

Rheumatoid arthritis (RA) is a common condition in the United States. Approximately 25% of patients with RA experience pain and disability of the hand. A common hand deformity results from the destruction of the metacarpophalangeal (MCP) joints with dislocation of the MCP joints and ulnar deviation of the fingers. Silastic implants are a surgical option to replace destroyed MCP joints in the rheumatoid hand. Dr. Chung is conducting a study to determine the outcomes and effectiveness of Swanson Metacarpophalangeal Joint Arthroplasty (SMPA), a type of silastic joint implant. Although Swanson implants have been used for over 30 years world wide, controversy still exists among hand surgeons and rheumatologists and in the literature about the effectiveness of this procedure in improving hand function for RA patients with MCP joint disease. This study will utilize a prospective design along with a multi-center and international collaboration. Surgical cases as well as medically treated (non-surgical) control patients will be enrolled for comparison. This study will use functional and health-related quality of life measures, such as the MHQ, to evaluate outcomes. All of these aspects reflect the unique design of this study which will aim to resolve the controversy regarding this surgical procedure.

Distraction Osteogenesis: Endogenous Tissue Engineering

Investigators: M. Longaker (NYU), S. Buchman, W. Kuzon, S. Goldstein, M. Ignelzi
Funding: National Institutes of Health

A method of endogenous tissue engineering for the reconstruction and replacement of pediatric mandibular deficiency would help to alleviate the significant psychosocial and biomedical burden of this common problem. Distraction osteogenesis (DO), the creation of new bone by the gradual separation of two bony fronts, is the ultimate method of tissue engineering because it provides an anatomical and functional replacement of deficient tissue with endogenous bone generated from local substrate. The central hypothesis to be tested in this proposal is that the creation of newly engineered bone generated by mandibular DO is the result of an adaptive response by bone and the surrounding soft tissue envelope. We further hypothesize that there is a fundamental difference in the mechanism of this adaptive response, characterized by the creation of new bone, and a maladaptive response, characterized by injury and fibrous non-union. It is further hypothesized that under adaptive conditions the response of adjacent masticatory muscles will be adaptive, and that under maladaptive conditions, the response will be injury to the muscles of mastication. To test this hypothesis, we will utilize a novel rodent model to determine the cellular, molecular, and biomechanical mechanisms that distinguish the adaptive from the maladaptive response in mandibular DO. This interdisciplinary proposal is timely and relevant because mandibular DO is rapidly becoming the treatment of choice for reconstructing pediatric mandibular deficiencies.

Force Deficit in Neurovascular Muscle Transfers

Investigators: W. Kuzon, P. Cederna, J. Faulkner, D. Goldman, B. Carlson, M. Urbanchek Funding: National Institutes of Health

The goal of this work is to determine the mechanisms responsible for the unexplained specific force deficit in neuromuscular transfers. The working hypothesis is that, after vascularized transfer, the deficit in specific force occurs when regenerating motor axons fail to reinnervate all the fibers in a muscle. The specific aims are to test the hypotheses that, after neurovascular transfer: 1) there is in a population of individual muscle cells that remain denervated; 2) that this population of denervated fibers explains all or part of the force deficit; and 3) that incomplete reinnervation occurs, in part, because motor unit innervation ratio can increase only up to a maximum value as a response to a reduced number of innervating motor axons. To test these hypotheses, an orthotopic neurovascular transfer of the extensor digitorum longus muscle in adult rats has been designed so that the number of axons supplied to the muscle for reinnervation can be reduced dramatically. Whole muscle force and single motor unit contractile properties are measured in each muscle 120 days after the transfer procedure. The number of motor units in each muscle is estimated from peripheral nerve histomorphometry, by using the manual incremental electromyographic method, and indirectly from motor unit force and muscle fiber histomorphometric data. Large scale computer simulations are used to evaluate the effect of sample size on estimates of innervation ratio and to examine the assumptions used in the indirect estimation of innervation ratio. Molecular markers of muscle cell denervation are identified via immunohistochemical demonstration of neural cell adhesion molecule and in situ hybridization to localize mRNA coding for the embryonic subunit of the nicotinic acetylcholine receptor. These markers are used to demonstrate directly individual muscle fibers that remain denervated after vascularized transfer.

Mechanisms Responsible For Skeletal Muscle Weakness In Aging

Investigators: P. Cederna, W. Kuzon, J. Faulkner
Funding: Plastic Surgery Education Foundation

Skeletal muscle atrophy and weakness lead to frailty and impaired mobility in the elderly. This progressive decline in skeletal muscle mass and strength directly impacts the quality of life, and limits the effectiveness of functional reconstructions for the treatment of central or peripheral paralysis in the elderly. The mechanisms responsible for this progressive decline in muscle strength have not been determined. The purpose of this investigation is to determine the effect of aging on whole muscle and single motor unit contractile properties, and to examine the contribution of myofiber denervation on muscle force production. We hypothesize that in muscles from extremely old animals: 1) whole muscle force deficits exist which are partly accounted for by a reduction in motor unit numbers; 2) motor unit innervation ratios are reduced which contribute to the force deficits seen; and 3) increased populations of denervated myofibers are present in old versus adult animals.

In the aged animal, 35% reductions in maximum force production have been demonstrated. In humans, declining maximum performance and loss of strength has been shown even in highly motivated older athletes, demonstrating that physical conditioning cannot prevent the loss of muscle mass or strength. The declining force production with aging, in contrast to physical inactivity, appears to be related to some alternative mechanism including denervation of muscle fibers and loss of fast motor units. The evaluation of single motor unit contractile properties will provide significant insights into the mechanisms responsible for skeletal muscle weakness and atrophy in old animals through the determination of motor unit forces, numbers, and composition in extremely old versus adult animals; significant changes at the motor unit level may not be reflected by changes at the whole muscle level. Our specific hypotheses will be tested by comparing whole muscle force production, single motor unit mechanical properties, motor unit numbers, innervation ratios, and the proportion of denervated fibers in muscles from extremely old rats with the same variables in muscles of adult rats.

Force Deficit in Reinnervated Skeletal Muscle

Investigators: W. Kuzon, P. Cederna, M. Urbanchek, J. Van der Meulen
Funding: VA Merit Review

The objective of this work is to identify and quantify the mechanisms that result in the unexplained deficit in specific force in reinnervated skeletal muscles. This study examines the hypothesis that reduced force production by individual, innervated muscle fibers accounts for a portion this specific force deficit. We also hypothesize that contraction-induced injury occurs more frequently in reinnervated muscles, and that muscles reinnervated with a reduced number of motor axons will have an even greater susceptibility to contraction-induced injury than muscles reinnervated by an adequate number of axons. To test these hypotheses, rat extensor digitorum longus (EDL) muscles are denervated by transection of the motor nerve and reinnervated via repair of the nerve with either an adequate or a reduced number of motor axons included in the repair. At various times following reinnervation, muscles are subjected to standardized protocols of stretch-induced injury. Maximal tetanic force and specific force are measured in both whole EDL muscles and in single, permeabilized muscle fibers. Muscles are studied at two specific time points after the contraction-induced injury: 3 days (maximum injury) and 2 months (maximum recovery). Our hypotheses will be supported if: 1) reduced specific force values are observed for single, permeabilized fibers from reinnervated compared with control muscles, 2) the whole muscle specific force deficits induced by lengthening contractions are greater in reinnervated compared with control EDL muscles, 3) acute contraction-induced injury results in greater specific force deficits in single, permeabilized fibers from reinnervated muscles than in single fibers from control muscles, 4) the recovery from contraction-induced injury is delayed and less complete in reinnervated compared with control muscles, and 5) reinnervation of the EDL muscle by a reduced number of motor axons increases the susceptibility to, and impairs the recovery from, contraction-induced injury when compared with reinnervation by a greater number of axons.

A Commercial Biodevelopmental Emulator for Skeletal Muscle

Investigators: R. Dennis, W. Kuzon, J. Faulkner
Funding: Biomedical Technology Development Fund, University of Michigan

We have developed a method to generate a three-dimensional, skeletal muscle construct, termed a myooid, from primary mammalian satellite cell and fibroblast co-culture. A prerequisite to the commercial application of myooids is technology to allow production of myooids on a large-scale basis, to allow both physical and chemical stimuli to be applied during in vitro myogenesis, and to allow the automated recovery of data on muscle excitability and contractility. Our current, first-generation bioemulator allows small-scale myooid production and the application of electrical and mechanical stimuli during myooid formation. We propose to increase the commercial value of these core technologies by development of a second-generation bioemulator with the following features: 1) flow through perfusion for the automated control of the chemical and humoral environment during myooid formation, 2) automated control of mechanical and electrical stimuli during myooid formation, 3) automated monitoring of myooid development, 4) automated, continuous or pulsed application of pharmaceuticals to developing and mature myooids, and 5) automated, online collection of muscle excitability and contractility data. This system will have architecture scalable to large volume production. Our objective is to demonstrate that this bioemulator can be used, on a commercial scale, to evaluate the teratogenic effects of pharmaceuticals on muscle development and to generate dose-response curves for the effects of drugs on muscle excitability and contractility.

Motor Unit Reorganization in Aging Muscle

Investigators: P. Cederna, W. Kuzon, M. Urbanchek, J. Van der Meulen
Funding: Research Advisory Committee, University of Michigan

Muscle atrophy and weakness lead to frailty and impaired mobility in the elderly. Declines in skeletal muscle mass and strength impact the quality of life and directly effect the potential for independent living. In addition, the progressive loss of muscle mass and force output as a result of aging limits both the options for and the efficacy of reconstructions for facial paralysis or other motor deficits in the elderly. The age-related deficits in muscle strength are well documented in both humans and laboratory animals. Reduced muscle mass does not completely account for the force deficit observed in muscles of old animals, because when muscle force is normalized to the physiologic cross sectional area (CSA), a specific force deficit is observed in humans, rats, and mice. The mechanisms responsible for the age-related deficit in muscle strength have not been fully determined. The aim of this work is to determine the effect of aging on whole muscle and single motor unit contractile properties, and examine the effect of myofiber denervation on force deficits in muscles from old animals. We will test the hypothesis that the age-associated specific force deficits can be fully or partially attributed to a reduction in motor unit numbers; this progressive loss of motor units will result in a subpopulation of denervated muscle fibers in old compared with middle-aged and young animals. This hypothesis is tenable because previous studies have clearly demonstrated an age-related motor neuron loss that engenders ongoing denervation of muscle fibers with advancing age. Therefore, a principle mechanism responsible for the age-related impairment in muscle function may be the development of a population of denervated or poorly innervated muscle fibers. Using the extensor digitorum longus (EDL) muscle of Brown/Norway rats as our experimental model, we will test these hypotheses by first measuring the whole muscle force (Fo) production and calculating the specific force (sFo) output in adult, middle-age, and old animals. The effect of muscle atrophy on force production in middle-aged and extremely old animals can then be determined by examination of the sFo deficit. Single motor unit contractile properties will then be evaluated to determine the number of motor units per muscle and permit calculations of innervation ratios. These data will allow us to directly calculate the proportion of the age-related whole muscle force deficit that can be accounted for by reductions in motor unit numbers and innervation ratios.

Detection of Detubulation in Skeletal Muscle Fibers

Investigator: D. Claflin
Funding: Center for Biomedical Engineering Research

The purpose of this study is to develop techniques to detect "detubulation", the loss of patent transverse tubules (t-tubules) in skeletal muscle fibers. The t-tubules are a membrane system of tubular conduits that arise from the outer membrane (sarcolemma) of skeletal muscle fibers and conduct electrical depolarizations from the sarcolemma to the fiber's interior. Depolarization of the t-tubules causes activating calcium (Ca2+) to be released from the sarcoplasmic reticulum into the myoplasm, initiating contraction. The cytoskeletal protein dystrophin is located between the myofibrillar matrix and the sarcolemma in control fibers, but is absent in patients with Duchenne muscular dystrophy and in the mdx mouse. Dystrophin-deficient skeletal muscle fibers are susceptible to injury during contraction, particularly when activated fibers are stretched. Indirect evidence suggests that dystrophin is an essential component in a mechanical link between the force generating structures of the fiber and the extracellular matrix. The working hypothesis is that dystrophin preserves Ca2+-activation by protecting transverse tubules from detubulation by shear forces associated with movement of the sarcolemma with respect to the underlying myofibrillar matrix. Predictable consequences of detubulation include a reduction in membrane capacitance, a change in the action potential, and an increase in the stimulation parameters required to elicit maximum tension. Development of techniques for detecting detubulation are necessary to test mechanistic hypotheses based on the working hypothesis.

Role of Activation in Skeletal Muscle Injury

Investigator: D. Claflin
Funding: Nathan Shock Center for the Biology of Aging

The purpose of this study is to determine whether damage to the contractile activation mechanism contributes to the increased susceptibility of skeletal muscle from old animals to contraction-induced injury. Muscle weakness is a significant problem among the elderly, yet the mechanisms responsible for the reductions in force and power output with aging are not fully understood. Increased susceptibility to injury during pliometric (lengthening) contractions is one contributing factor. A hallmark of contraction-induced injury is mechanical disruption of the ultrastructure of sarcomeres. The transverse tubules (t-tubules) and sarcoplasmic reticulum (SR) are intracellular membrane systems that play a central role in the activation of contractile activity. Both t-tubules and SR are intimately associated with the sarcomeres. The working hypothesis is that, during pliometric contractions, the forces that cause disorder of sarcomere ultrastructure also disturb the critical spatial relationship between t-tubule and SR membranes, disrupting the molecular interactions required for activating calcium (Ca2+) to be released from the SR into the myoplasm. Consequently, regions in which ultrastructural damage to sarcomeres is apparent would also exhibit evidence of impairment of the Ca2+-activation system. Specific hypotheses to be tested are that pliometric contractions result in impaired intracellular Ca2+ transients, that the regions in which impaired Ca2+ transients appear undergo lengthening during fixed-end contractions, and that the cumulative size of these impaired regions is larger in fibers from old mice than those from young or adult mice. Improving our understanding of the mechanisms underlying the increase in muscle weakness with age will increase the probability that interventions can be developed to avoid or reverse the problem.

Acellularized Peripheral Nerve Allograft: A Non-Immunogenic Construct To Support Axonal Regeneration

Investigators: P. Cederna, J. Faulkner, J. Rovak
Funding: Howard Hughes Medical Institute

The purpose of this experiment is to determine the efficacy of acellularized peripheral nerve allografts for reconstruction of peripheral nerve gaps. In the clinical setting, peripheral nerve gaps may occur after traumatic injuries or oncologic resections; autologous nerve grafting is currently the standard technique for the reconstruction of these defects. However, the prime limitation to the reconstruction of long or multiple nerve gaps is the limited quantity of nerve autograft that can be harvested from any patient. To overcome the limited supply of nerve autograft, nerve allografting, synthetic nerve conduits, and various alternative biologic nerve conduits have been used. While these alternatives hold promise for some clinical situations, each presents limitations or disadvantages compared with autologous nerve grafts. Our overall objective is to develop a tissue-engineered peripheral nerve construct for the reconstruction of clinical nerve gaps that: 1) is readily available; 2) supports axonal regeneration over long gaps in a manner equivalent to nerve autografts; and 3) can be used without immunosuppressive therapy. Our strategy is to eliminate the immunogenicity of nerve allografts while: 1) retaining the structural integrity of the endoneurial conduits; and 2) restoring the appropriate cellular and molecular signals for optimal axonal regeneration. We propose to eliminate the immunogenicity of peripheral nerve allografts using a process of chemical acellularization that we have developed in our laboratory. Repopulating the acellular endoneurial conduits with autologous Schwann cells will be used to optimize the cellular and molecular signals for axonal regeneration.

Peripheral Nerve Reconstruction Utilizing Chemically Acellularized Nerve Allografts Repopulated With Autogenous Schwann Cells.

Investigators: P. Cederna, W. Kuzon, K. Bishop
Funding: American Association of Plastic Surgeons

The purpose of this experiment is to determine the efficacy of peripheral nerve reconstruction utilizing acellularized peripheral nerve allografts repopulated with autogenous Schwann cells. This technique would potentially provide a non-immunogenic construct that could support axonal regeneration across long nerve gaps. In the clinical setting, peripheral nerve gaps may occur after traumatic injuries or oncologic resections; autologous nerve grafting is currently the standard technique for the reconstruction of these defects. However, the prime limitation to the reconstruction of long or multiple nerve gaps is the limited quantity of nerve autograft that can be harvested from any patient. To overcome the limited supply of nerve autograft, nerve allografting, synthetic nerve conduits, or various alternative biologic nerve conduits have been used. While these alternatives hold promise for some clinical situations, each presents limitations or disadvantages compared with autologous nerve grafts. Our overall objective is to develop a tissue-engineered peripheral nerve construct for the reconstruction of clinical nerve gaps that 1) is readily available, 2) supports axonal regeneration over long gaps in a manner equivalent to nerve autografts, and 3) can be used without immunosuppressive therapy. Our strategy is to eliminate the immunogenicity of nerve allografts while 1) retaining the structural integrity of the endoneurial conduits, and 2) restoring the appropriate cellular and molecular signals for optimal axonal regeneration. We propose to eliminate the immunogenicity of peripheral nerve allografts using a process of chemical acellularization that we have developed in our laboratory. The cellular and molecular signals for appropriate axonal regeneration will be optimized by repopulating the acellular endoneurial conduits with autologous Schwann cells. We specifically hypothesize that: 1) A peripheral nerve allograft can be rendered completely non-antigenic by chemical acellularization; 2) For short nerve gaps (1-2 cm in the rat hindlimb model), acellularized peripheral nerve allografts can support axonal regeneration and end-organ recovery in vivo in a manner indistinguishable from fresh peripheral nerve autografts; 3) After repopulation of acellularized nerve allografts with autologous Schwann cells, the composite, tissue engineered nerve graft can support axonal regeneration and end organ recovery in vivo in a manner indistinguishable from fresh nerve autografts, regardless of the length of the nerve gap.

Improving Muscle Power and Mobility of Elderly Men and Women; Project 3: Muscle Fiber Analysis

Investigators: J. Faulkner, B. Carlson, P. Cederna, D. Claflin
Funding: Michigan Life Sciences Corridor

The Michigan Life Sciences Corridor has awarded $3.7 million to a group of investigators including Drs. Bruce Carlson, John Faulkner, Paul Cederna, and Dennis Claflin, all experienced in the study of the biology of aging muscle and mobility, to develop a multilevel program with a long-term goal of improving muscle power and mobility in the elderly. Broad aims of the project are: 1) to conduct research and development on a specific project designed to apply a novel technology directed toward improving muscle power, 2) to coalesce a multidisciplinary aggregate of researchers on problems of muscle power and mobility in the elderly into a recognizable unit that is nationally recognized as a center for such research, 3) To create an environment of effective collaboration between academic research and commercial development. The proposed research project involves both young and elderly human subjects and involves a multilevel comparison of the effects of a progressive resistance training program on muscle power and mobility. In addition, a group of younger subjects will test the effects of a "Thermal Hand" device that is designed to improve the efficiency of muscle training programs by rapidly lowering body core temperature. If the device proves to be safe and effective, future studies will be conducted on elderly subjects. Analysis of the data will be carried out at levels from whole body motion analysis to single muscle fiber contractile power analysis, biochemistry and morphology.

Techniques

  • Gel electrophoresis
  • Northern Blot
  • Western Blot
  • Primary tissue culture
  • Conjugated-dienes
  • Neutrophil histochemistry
  • Xanthine oxidase assay
  • Light and electron microscopy
  • High performance liquid chromatography
  • Gas chromatography
  • Thin layer chromatography
  • In situ hybridization
  • Lipid biochemistry and analysis
  • Confocal microscopy
  • Electron spin resonance spectroscopy
  • Fatty acid enzyme kinetics
  • Immunohistochemistry
  • Muscle Contractile Functioning
  • Micro-Computed Tomography
  • Muscle contractile properties (force, power, sustained power): in vivo (muscle groups), in situ (isolated whole muscles), in vitro (isolated whole muscles, muscle fiber bundles), single, permeabilized muscle fibers
  • Single motor unit contractile properties
  • Muscle and nerve electrophysiology (EMG, NCV, etc.)
  • Muscle and nerve histomorphometry and immunohistomorphometry
  • Myosin isozyme determinations
  • Muscle biochemical analysis
  • In situ hybridization
  • Computer modeling
  • Retrograde Axonal Tracing
  • Molecular Biology
  • Myoblast and Schwann cell culture

Equipment

  • Bioquant Image Analysis System
  • Zoom-stereo Microscope for Bright and Dark Field
  • Stage Microtome
  • Sorvall Centrifuge
  • Ultra-low freezer; and refrigerator
  • Combi-cold cabinet
  • Fraction collectors
  • HPLC
  • CO2 incubators
  • Hybridization Oven
  • Meditec Laser
  • Jena microscopes, regular and inverted
  • WaterÕs HPLC
  • -70 freezers
  • Kelvinator cold cabinet
  • Beta RAM instrument and computer
  • -20 C freezers
  • Cryostorage cell systems
  • Media filtering systems
  • LKB laser densitometer
  • Cryostat
  • EMG Biologic Traveler
  • Cell Imaging
  • Water's 2690 HPLC
  • INUS Systems Beta-Ram Detector
  • Shimadzu Gas Chromatograph
  • Composite apparati for quantifying mechanical function in single muscle fibers, single motor units, and whole muscles (in vivo, in situ, and in vitro).
  • Zeiss operating microscopes
  • Sorvall Discovery M150 Ultracentrifuge
  • Biorad Smart Spec 3000
  • Labsystems Multiscan plus reader
  • Freezers; and refrigerators
  • Jena microscopes, regular and inverted
  • Laminar flow hoods
  • Mettler balances
  • pH meters
  • Water baths
  • -20 C refrigerator and chest freezer
  • Water purification system
  • Cryostat
  • EMG Biologic Traveler
  • Electrophoresis equipment
  • Spectrophotometer, fluorometer, etc. for muscle metabolic studies

Investigators

  • David L. Brown, MD, 936-8668
  • Steven R. Buchman, MD, 763-8063
  • Paul S. Cederna, MD, 615-3435
  • Kevin C. Chung, MD, MS, 936-8925
  • Dennis R. Claflin, PhD, 936-2817
  • John A. Faulkner, PhD, 936-2817
  • Robert Gilmont, PhD, 936-8679
  • William M. Kuzon, Jr., MD, PhD, 936-5890
  • Julie C. Lowery, PhD, 936-5890
  • Cynthia L. Marcelo, PhD, 763-6721
  • M. Haskell Newman, MD, 763-8063
  • Riley S. Rees, MD, 615-3435
  • David J. Smith, Jr., MD, 936-8925
  • Melanie Urbanchek, PhD, 936-2817
  • Jack Van der Meulen, PhD, 936-2817
  • Edwin G. Wilkins, MD, 936-5890

Location/Contact

Muscle Mechanics Laboratory
Institute of Gerontology, Room 1060
300 North Ingalls Bldg.
Box 2007VA Medical Center
Medical Research Service (11R)
2215 Fuller Road