Our Research

Traumatic Fractures and Sports Injuries

Blast/Burn Injury

Traumatic Spinal Cord Injury

Some of our models

Increased Vascularity around HO

Our Problem

Patients are affected by trauma induced heterotopic ossification from a variety of injuries and surgical procedures. HO is found in >65% of blast/burn injury, 20-80% of joint replacement, 20% of traumatic fractures and sports, and 20% of traumatic spinal cord injury patients.

This formation of bone in places where it is not normally found can lead to problems with:

  • pain
  • wound healing
  • decreased joint range of motion
  • drastic reduction in quality of life
  • There is NO effective treatment or prevention of this disease in contemporary medicine. Furthermore, surgeons have their hands tied, as cutting the growth out with surgery can cause even more bone to form.

Strategies & Results

Our variety of models have and continue to facilitate our elucidation of key steps along a convoluted HO pathway. We have demonstrated that the formation of HO is due to dysregulation of inflammation and normal stem cells needed for proper repair going awry. Given many amazing scientists before us have carefully studied normal bone development, we are applying those lessons into clarifying the behind-the-scenes of HO formation to treat and prevent it from forming.

We’ve developed a mouse model that closely resembles the disease in humans, both post-traumatic and genetic varieties of HO (fibrodysplasia ossificans progressiva), to tackle the key questions behind the disease from many contexts. These investigations include the role of vascularity, mechanotransduction, and nerves just to name a few. To answer these questions, we rely on a myriad of state-of-the-art imaging techniques including micro CT, immunohistochemistry, and immunofluorescence.

Burn + Tenotomy Model

Our fundamental trauma model consists of a concomitant dorsal burn and achilles tendon tenotomy. This model reliable produces HO at the local tenotomy site and nearby environments.

Inducible Geneotype

Many of our experiments rely heavily on inducible gene technology. Using Cre recombinase and lox-P site technology, we can create robust models whose genes of interest can be turned on and off at will.

Genetic Recombination

Stemming from our inducible gene schema, combined permutations expand our scope and types of interrogations we can apply to our HO pathway.

Collaborative Spaces

Michigan Experts

Second Harmonic Generation

Cutting-edge microscopy modalities allow us to probe what is going on at the cellular level. For instance, we can see what the extracellular matrix in the region of injury looks like, giving us a better idea about what cells encounter when they get there.

3D in vivo

Super magnification of immunolabeled tissues can show us what cells are expressing and making in the actual animal’s tissues, allowing us to better understand how to bring therapeutics from the bench to bedside.

Micro CT Reconstructions

Micro CT reconstruction is a foundational component of much of our research allowing us visualization for exploration of our models and quantifying HO growth.

Heterotopic Ossification

Early diagnosis and Prevention strategies
As many as 65% of our severely combat-injured service members will go on to develop heterotopic ossification (HO), a musculoskeletal disorder characterized by the formation of mature bone in soft tissues including muscle, tendon, ligaments and fascia. As a complication of trauma, HO presents the most important barrier to functional recovery and independence.Furthermore, over 60% of civilian major burn patients and over 50% of joint replacement surgery patients develop HO, with risks that only increase after subsequent operations. The pervasive nature of HO extending across several tissue types suggests that treatment directed to a specific tissue may be ineffective, whereas therapy directed towards centrally acting pathways may be more effective. Once bone forms around a joint, patients develop debilitating joint contractures and loss of mobility. Surgical excision of HO can be attempted to restore function, but patients with periarticular HO rarely regain complete range of motion, with contractures due to persistent or recurring HO. Current medical strategies to prevent de novo or recurrent HO including glucocorticoids, bisphosphonates, and non-steroidal anti-inflammatory medications either have significant side effects and/or uncertain impact, possibly because they fail to target key pathways involved in HO formation. In addition to effective therapies, methods of identifying potential HO patients prior to radiographic diagnosis are needed to target early intervention to a population that is likely to benefit. Given the burden of HO disease in these populations, its high morbidity and suboptimal treatments, there is substantial need for early diagnosis and therapy to inhibit HO by targeting its causative processes.

Fibrodysplasia Ossificans Progressiva

One of the Earliest Clues
Fibrodysplasia ossificans progressive (FOP) is a rare autosomal dominant disorder of skeletal malformations and progressive extraskeletal ossification, a congenital disorder that mirrors our topic of focus. Subsequent studies of this parallel disease have elucidated biochemical pathway steps and agents such as the ALK2 receptor. ALK2 mutations have now been well described to play a role in patients with (FOP), and recent studies have implicated ALK2 as playing a more central role in cartilage condensation that can lead to subsequent bone formation in unwanted areas. Establishing more links of this nature is a powerful strategy in localizing relevant agents.

Levi Lab 

Equipment List
fluorescent and confocal microscopy
MicroCT scanners
IVIS imaging devices
10X RNA sequencing facilities
a mouse genomics core.
Leica confocal microscope and Bruker
SKYSCAN 1272 CMOS EDITION – High-resolution 3D X-ray Microscopy
Cytation 5- digital microscope and microplate reader
4D-Nuclefector System
PARAMETER Cabinet X-ray System

 

Injury Models
Burn
Tenotomy
Muscle Crush
Volumentric Muscle Loss
Fracture
Calvarial Defect

Advanced Imaging Research Center for Mouse Imaging:
An IVIS Imaging System 200 Series (Caliper Life Sciences) for bioluminescence imaging
The Mouse Core Facility, run and maintain by the Advanced Imaging Research Center has Micro CT and small animal SPECT In vivo fluorescence and BLI (Xenogen IVIS Spectrum and Lumina units) and Ultrasound machine (Vevo 770, Visual Sonics). The CRI and Center for Mineral Metabolism both have MicroCT imagers.

Molecular Pathology Core
The Molecular Pathology Core maintains infrastructure and expertise for (i) embedding tissue samples; (ii) preparing plastic and paraffin sections, including hard-tissue processing and resin thinning (Buehler IsoCut Slow Speed Saw, PetroThin Grinder, and AutoMet 300/Ecomet 250 Polisher, Leica RM2255 Rotary Microtome outfitted for Tungsten-Carbide resin microtomy).

Bioinformatics
The hardware includes a PowerWulf Compute Engine, which contains 208 total processor cores, 800 GB system RAM, 84 TB Raw RAID6 storage with 10 GigE access to switch, and an Overland Tape Backup System, a Windows server for software, a database server, and a web server.

Confocal Microscopes: The Levi Laboratory has a live-cell compatible Leica Stellaris DM8 Confocal microscope with Leica LASX+ (NL8.122A) capable of super resolution and fluorescence lifetime analytics. The lab also maintains access to multiple Zeiss LSM 880 confocal/multiphotons (NL7.134A, K1.224 with Airyscan), an Andor spinning disk confocal with FRAP and TIRF (NL5.120R), and a Perkin Elmer Ultraview spinning disk confocal – BSL2 (Y9.326). Zeiss LSM 780 confocal/multiphoton (NL5.120N) Zeiss LSM 780 confocal/multiphoton (NL11.125A) Zeiss LSM 880 confocal/multiphoton (NL7.134A). Zeiss LSM 880 Airyscan confocal microscope (K1.224).Andor spinning disk confocal with FRAP and TIRF (NL5.120R) Perkin Elmer Ultraview spinning disk confocal – BSL2 (Y9.326)

Confocal Microscopes

The Levi Laboratory has a live-cell compatible Leica Stellaris DM8 Confocal microscope with Leica LASX+ (NL8.122A) capable of super resolution and fluorescence lifetime analytics. The lab also maintains access to multiple Zeiss LSM 880 confocal/multiphotons (NL7.134A, K1.224 with Airyscan), an Andor spinning disk confocal with FRAP and TIRF (NL5.120R), and a Perkin Elmer Ultraview spinning disk confocal – BSL2 (Y9.326). Zeiss LSM 780 confocal/multiphoton (NL5.120N) Zeiss LSM 780 confocal/multiphoton (NL11.125A) Zeiss LSM 880 confocal/multiphoton (NL7.134A). Zeiss LSM 880 Airyscan confocal microscope (K1.224).Andor spinning disk confocal with FRAP and TIRF (NL5.120R) Perkin Elmer Ultraview spinning disk confocal – BSL2 (Y9.326)

 

Mouse Genotypes
Hox TSP2
LysMCre TSP1
TSP1/TSP2
AviliCre
DDR2 flox
DDR2-CreER2
DDR2LacZ
Fgf18 flox
Fgfr1,2,3 flox
Hoxa11 GFP
Hoxa11CreER
PV cre
C57BL/6J
PDGFRaCreER
Rosa Luseap/Luseap (LUC)
Arf6-floxed
Arkadia-floxed
FKbp10-floxed
GPR81 KO
GPR81-Floxed
Hif1a-floxed
Hif1a-floxed;TdTom
Hoxa11CreER; Arf6-floxed
Hoxa11CreER; Arkadia-floxed; tdTom
Hoxa11CreER; GLS-floxed
Hoxa11CreER; LdhA-floxed
Hoxa11CreER; SLC1A5-floxed
Hoxa11CreER; Srf-floxed
Hoxa11CreER; Tak1-floxed
Hoxa11CreER; Vhl-floxed;tdTom
Hoxa11CreER;Hif1a-floxed; tdTom
PDGFRaCreER; LdhA/B-floxed
LdhA/B-floxed
LdhA-floxed
LysMCre; GPR81 Flox
MCT4 KO
PDGFRaCreER; FKbp10-floxed
PDGFRaCreER; GPR81-Floxed
PDGFRaCreER; LdhA/B-Floxed
PDGFRaCreER;Hif1a-floxed;TdTom
Plod2-floxed
Snai1 flox Snai2 flox
Srf-floxed
Tak1-floxed
VHL-floxed
Hoxa11CreER; TGFbR2-floxed; tdTom
LysMCre
Mrp8Cre
Mrp8CreTLR9 flox
PDGFRaCreER Alk5 flox
PDGFRaCreER DDR2 flox
PDGFRaCreER FAK flox
PDGFRaCreER TGFBR2 flox
PDGFRaCreER Yap/Taz flox
TGFB1 flox
TGFBR 2 flox
TLR9 flox
Yap flox / Taz flox
PDGFRaCreER TdTom
Mrp8Cre tdTom
CD169iCre tdTom
DDR2-CreER2 x Ai14
FLT4Cre iDTR
FLT4Cre Pik3Ca flox
FLT4Cre Pten flox
K14CreER
PDGFRaCreER Rainbow
Prox1eGFP
Rainbow
TSP2 Reporter
Acod1 KO
Cthrc1 
LysMCre Irg1 flox
Mrp8Cre Irg1 flox
PDGFRaCreER TSP1 flox
PDGFRaCreER TSP2 flox
TSP2 flox
Hoxa11CreER tdTom
TSP1 flox
P53 flox / Rb flox
BMP2 fn
Arg1 flox
CD169iCre
CD169iCre tdTom Piezo1 flox
CD4Cre Piezo1 flox
Hoxa11CreER Jmjd3 flox
LysMCre Arg1 flox
LysMCre Jmjd3 flox
LysMCre MLL flox
LysMCre Piezo1 flox
LysMCre SetDB2 flox
LysMCre TLR9 flox
LysMCre TMEM16F Flox
LysMCre Yap/Taz flox
Osx Cre
PDGFRaCreER Jmjd3 flox
PDGFRaCreER Piezo1 flox
Piezo 1 flox
Piezo 2 flox
TMEM 16 KO
TMEM16F Flox
Trem2
LysMCre Notch flox
FAK flox
Vegfa flox

Nomellini Lab

Equipment List
Fisher scientific Isotemp circulators/Baths
Bio-Rad power pac-200
Olympus CH microscope
Fisher scientific Isotemp
Scientific Industries Vortex Genie
Sheldon Manufacturing H2O Bath
Denver Instrument APX 200 Weighing Balance
Corning Stirrer/Hotplate
Dade Vortex Mixer
Denville Vortexer 59
Aldinger Weighing Balance
AWEL Mf48-R Refrigerated Centrifuge
Denville Scientific 260d Brushless Microcentrifuge
VWR Scientific Model 1235 Water Bath
Eppendorf Centrifuge 5424r
Buxco Fine Pointe Whole Body Plethysmography

Mouse Genotype

CD1

Tower Lab

Equipment List
PH Reader
PIPETMAN G 4-PIPETTE KIT  
VIP ECO ULT FREEZER UPRIGHT 26 CU.FT. 115V
Scientific Industries Vortex-Genie 2 Mixer
Mettler Toledo™ Standard ME Analytical Lab Balance
Fisherbrand™ Isotemp™ Shaking Water Baths
Fisherbrand™ Multi-Platform Shaker
Shaker platform (accessory)
ThermoMixer F1.5, with thermoblock for 24 reaction vessels 1.5 mL
Mastercycler® X50a, 110–230V, aluminium block  
OPEN AIR ROCKER
DWK Life Sciences Wheaton™PIPET-PAL Pipet Controller
OHAUS™ Navigator™ Portable Balance
Corning™ Pyroceram™ Hot Plate Stirrer, 550°C, Glass Ceramic
Flat Platform with Nonslip Rubber Mat (for Fisher shaker)
Thermo Scientific™ Sorvall™ Legend™ Micro 21R Microcentrifuge
Corning™ Mini Microcentrifuge, 100-240V
Mouse Genotype
acod1
tsp1
pdgfraCreER Irg1 
cthrc1
LysMCre Irg1
PdgfraCreER tsp2

Dellinger Lab

Equipment
Leitz 1512 microtome for tissue sectioning
Nikon Eclipse E600 with Nikon DS-Qi2 camera for fluorescent microscopy
Nikon Eclipse E600 with Amscope MU1000 camera for bright field microscopy
Nikon Eclipse TS100 for imaging cells from in vitro experiments
Amscope dissecting microscope
Mouse Strain Information
Vegfr3wt/Chy In activating point mutation in Vegfr3
Vegfc-flox  
Vegfcwt/null  
LSL-KrasG12D Oncogenic form of Kras
LSL-Pik3caH1047R Oncogenic form of Pik3ca
LSL-rtTA  
TetO-GFP  
TetO-KrasG12D Oncogenic form of Kras
TetO-Vegfd  
TetO-Vegfc  
Pik3ca-RBD Point mutation in RAS binding domain
Prox1-tdTomato  
mTmG  
Prox1CreERT2  
VegfcCreERT2  
Lyve1-Cre  

 

Wang Lab

Equipment List
Nikon Eclipse Si (Nikon digital sight 1000 camera)
Eppendorf Centrifuge 5910Ri
Bio-Rad  Trans-Blot Turbo
Bio-RAD T100 Thermal Cycler
Miltenyi GentleMacs Dissociator

Brekken Lab

Equipment List
BioRad C1000 Touch Thermal Cycler
AKTA Pure FPLC

 

Mouse Genotype
Gas6-/-
Fibulin5 -/-
Axl-/-
VEGFR2; CSF1R-Cre
DDR1-/-
KIC (Kras G12D;Cdkn2a f/f; PTf1a cre/+)
KPC (Kras G12D;TRP53+/-; pdx1Cre)
KPCS (Kras G12D;TRP53+/-; pdx1Cre; SMAD4-/-)
KFC (Kras G12D; TRP -/- ; pdx1Cre)
KFCS (Kras G12D;TRP53 -/-; pdx1Cre; SMAD4-/-)
KPCS+/-  (Kras G12D;TRP53 +/-; pdx1Cre; SMAD4+/-)
PTN -/-; PYMT +. (pleiotrophin, MMTV-PyMT)
PYMT+ (MMTV-PyMT)
ALK-/- ( anaplastic lymphoma kinase)
Pleiotrophin-/- B6
Pleiotrophin-/-  FVB
Smad4-/- (Smad4tm2.1Cxd/J)
KpyC (p53 point mut) 
Pdgfra-Cre/ERT;  SMAD4-/-
TLR2-/-
Wt1CreGFP; DDR11-/-
Axl lacz/lacz
Beta2GP1 -/-
B2 het
KP  (Kras G12D; pdx1Cre)

The Levi Lab Team

The BWR Laboratory is grounded on a foundation of teamwork, bringing together doctors, professional scientists, medical students, graduate students, and undergraduates from a variety of backgrounds.

Support Our Research

Burn/trauma and regenerative medicine researchers in plastic surgery seek to prevent the formation of bones in abnormal locations. Gifts help alleviate this condition affecting burn, auto accident, orthopaedic surgery and blast injury patients by providing additional resources to employ state of the art technologies, recruit talented researchers, and disseminate our scientific findings widely.

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