CBI Thermal Dermal Burn Modeling in Rats and Minipigs

Slides:



Advertisements
Similar presentations
2) Closed wound: Skin is intact (not opened) include crushing injury and contusions. Wounds A) Skin involvement: 1) Open wound: when the whole thickness.
Advertisements

Burns Heat, electricity, radiation, certain chemicals  Burn (tissue damage, denatured protein, cell death) Immediate threat: –Dehydration and electrolyte.
Pressure Ulcer Management By Susan Yap, PT. Anatomy of the Skin Epidermis Dermis Subcutaneous Tissue Fascia Muscle Tendon and Bone.
Comparative Biosciences, Inc. 786 Lucerne Drive Sunnyvale, CA Telephone: A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH.
Wounds 2 categories: - surgical - traumatic Wound examples Closed surgical Open surgical Closed traumatic Open traumatic.
Activity Burn Unit Treatment Options
The anatomy of the skin, depth of burns and Jackson burn wound model
Calciphylaxis Induced Ulcerations. John M. Lavelle, 1 DO; Paul Liguori MD 2 1. Boston University Medical Center, Rehabilitation Department 2. Whittier.
Burn Injuries Adaobi Okobi, M.D.. Learning Objectives Epidemiology Pathophysiology Classification of burns Red flags Treatment.
Burns By Matthew & Ivan. Anatomy of the Skin The anatomy of the skin is complex, and there are many structures within the layers of the skin. There are.
Skin Integrity and Wound Care
 Head and neck cancer is the 6 th leading cancer world wide.  Oral Mucositis is one of the most frequent complications seen in the treatment of head.
Introduction The repair of an epithelial wound is merely a normal physiological process. Wound healing depends on elimination of any source of infection.
Pediatric Burns.
Burns Degree of Burns 1 st superficial partial-thickness burn 2 nd deep partial- thickness burn 3 rd full-thickness burn.
Pressure Ulcers & Nutritional Deficits in Elderly Long-Term Care Patients: Effects of a Comprehensive Nutritional Protocol on Pressure Ulcer Healing, Length.
The Integumentary System
Healing of Wounds and Burns & the Aging of Skin Chapter 6 Sections 5 & 6Chapter 6 Sections 5 & 6.
1 Yellowish skin coloration which is many times caused by liver disease “Bili” lights are used to treat this condition in newborns. Jaundice.
Soft Tissue Injuries Chapter 10. Soft Tissue The skin is composed of two primary layers:  Outer (epidermis)  Deep (dermis) The dermis layer contains.
Copyright © 2010 Wolters Kluwer Health | Lippincott Williams & Wilkins Timby/Smith: Introductory Medical- Surgical Nursing, 10/e 01/25 PG 1054 Chapter.
The Integumentary System Chapter Organs are two or more tissues which together perform a specialized function. Epithelial membranes are thin structures.
The Integumentary System
Body Membranes & Skin. Epithelial Membranes  3 Types  Cutaneous  Mucous  Serous.
Lecture # 32 TISSUE REPAIR: REGNERATION, HEALING & FIBROSIS - 4 Dr
Preclinical model: Oxygen-induced retinopathy in mice
Co 5.
Burns and scalds Burns = dry burn scald = wet or moist burn
And the Epithelium Skin
EFFECTIVENESS OF INDONESIAN HONEY TOWARD ACCELERATION
A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH
Local Insulin-Zinc Injection Accelerates Skin Donor Site Wound Healing
Dori R. Germolec, Judson Spalding, Hsin-Su Yu, G. S. Chen, Petia P
Skin and the Integumentary System
Fundamentals of Anatomy & Physiology
Diseases of the Pulp.
Evaluation and Management of Burns
Pressure Ulcers/Injuries
Principles of Wound Management
Maintaining Homeostasis
BURNS Dr.Ishara Maduka M.B.B.S. (Colombo)
A Novel Mouse Model for Frostbite Injury
Topical Application of A Novel Immunomodulatory Peptide, RDP58, Reduces Skin Inflammation in the Phorbol Ester-Induced Dermatitis Model  Christopher G.
Prevention and Treatment of Athletic Injuries
CUTS, SCRAPES, BURNS and BRUISES
The Integumentary System
CBI UV-light induced dermal inflammation in hairless mice
NURSING CARE MANAGEMENT OF BURNS IN ER
Severely Impaired Wound Healing in the Collagenase-Resistant Mouse
The OARSI histopathology initiative – recommendations for histological assessments of osteoarthritis in the horse  C.W. McIlwraith, D.D. Frisbie, C.E.
The short-term therapeutic effect of recombinant human bone morphogenetic protein-2 on collagenase-induced lumbar facet joint osteoarthritis in rats 
P. Coutinho, C. Qiu, S. Frank, C. M. Wang, T. Brown, C. R. Green, D. L
Figure S4 A C Vehicle control AC0010 (300mg/kg) Gefitinib (50mg/kg) D
Volume 125, Issue 6, Pages (December 2003)
Use of a Canine Model of Atopic Dermatitis to Investigate the Efficacy of a CCR4 Antagonist in Allergen-Induced Skin Inflammation in a Randomized Study 
The Loss of MCP-1 Attenuates Cutaneous Ischemia–Reperfusion Injury in a Mouse Model of Pressure Ulcer  Yuki Saito, Minoru Hasegawa, Manabu Fujimoto, Takashi.
Blockade of poly(ADP-ribose) synthetase inhibits neutrophil recruitment, oxidant generation, and mucosal injury in murine colitis  Basilia Zingarelli,
Warm Up Objective: Scientists will describe wounds and burns by analyzing and creating a model. What is the topic? What will you be doing? Why is this.
Molecular Imaging-Assisted Optimization of Hsp70 Expression during Laser-Induced Thermal Preconditioning for Wound Repair Enhancement  Gerald J. Wilmink,
Eric A.G. Blomme, Charles C. Capen, Thomas J. Rosol 
Weight bearing as a measure of disease progression and efficacy of anti-inflammatory compounds in a model of monosodium iodoacetate-induced osteoarthritis 
Jun Asai, Hideya Takenaka, Norito Katoh, Saburo Kishimoto 
By: M. Rustom Plastic Surgeon
First Aid.
Martina I. Okwueze, Nancy L. Cardwell, Alonda C. Pollins, Lillian B
Pressure Injuries: Just the facts!
Activity Burn Unit Treatment Options
Role of TGFβ-Mediated Inflammation in Cutaneous Wound Healing
Nurses Can & Should Stage Wounds!
Presentation transcript:

CBI Thermal Dermal Burn Modeling in Rats and Minipigs A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH CBI Thermal Dermal Burn Modeling in Rats and Minipigs Comparative Biosciences, Inc. 786 Lucerne Drive Sunnyvale, CA 94085 Telephone: 408.738.9261 www.compbio.com

Comparative Biosciences, Inc. A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Comparative Biosciences, Inc. Premier Preclinical Contract Research Organization Specializing in Dermal and Ocular Pharmacology, Toxicology and Histopathology 17 Years in business Located in the heart of Silicon Valley State of the art, purpose-built facility ~30 employees with 7 PhDs, 2 pathologists Experienced study directors and scientists GLP, OECD, FDA, USDA, OLAW AAALAC Accreditation

Model induction in MiniPigs A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Model induction in MiniPigs CBI offers either full or partial thickness dermal burn modeling in minipigs that demonstrates dermal healing and is also humane and well tolerated by the rat and pig Uniform full or partial thickness burns are surgically-induced under strict aseptic conditions with intense post surgical care, systemic antibiotics and pain control. Device used is a stainless steel or brass block of defined weight (generally 300 gm) approximately 3x3 cm that is heated to a defined temperature (either 100 or 200C for partial or full thickness respectively) and held onto the skin for a defined time (generally 1 min for partial and 30 sec for full thickness). Alternatively, for partial thickness burns, hot water method (scald) may be used. Test article and bandaging is administered appropriately Grafting, escharectomy and other procedures available Lesions are measured and photographed Biopsy or histopathology for histology, IHC or bioanalysis may be performed

Model induction in Rats A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Model induction in Rats CBI offers either full or partial thickness dermal burn modeling in rats that demonstrates dermal healing and is also humane and well tolerated by the rat Uniform full or partial thickness burns are surgically-induced under strict aseptic conditions with intense post surgical care, systemic antibiotics and pain control. Device used is a small aluminum rod that is heated to a defined temperature (either 100 or 200C for partial or full thickness respectively) and held onto the skin for a defined time (generally 30 sec min for partial and 45-60 sec for full thickness). Alternatively, hot water method may be used. Test article and bandaging is administered appropriately Lesions are measured and photographed Biopsy or histopathology for histology, IHC or bioanalysis may be performed

Typical Parameters for both pigs and rats A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Typical Parameters for both pigs and rats Daily to weekly body weights Daily clinical observations Food consumption Clinical pathology Toxicokinetics or pharmacokinetics Measurements and photography Necropsy or site biopsy Histopathology with special staining Sponsor specific bioassays

Typical findings for both pigs and rats for standard parameters A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Typical findings for both pigs and rats for standard parameters For typical burn studies in which focal burns are placed on the dorsum: Procedure is well tolerated with adequate pain relief Daily to weekly body weights: There is mild body weight loss over the first week with return to weight gain after 1 week Daily clinical observations: Animals may be slightly lethargic Food consumption: No change or slight qualitative decrease Clinical pathology: Within normal limits For extensive burns (>50%) changes are seen in clinical pathology including glucose, total protein, albumin, LDH, insulin, BUN

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Histopathology Surgical biopsies during course of study as well as necropsy Tissue stained with HE, trichrome, other stains upon request IHC for various markers upon request Digital image analysis for wound size, cellular infiltrates, inflammatory markers or other markers Photomicroscopy

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Full thickness dermal wound area Days 7-10 Dermal burn area following placement of 2 dorsal full thickness burns and treated with various topical test articles. (N=8)

Full thickness dermal wound area Days 11-14 A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Full thickness dermal wound area Days 11-14 Dermal burn area following placement of 2 dorsal full thickness burns and treated with various topical test articles. (N=8)

Sample data from a minipig burn study A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Sample data from a minipig burn study Data from several studies Burn study using 2-4 pigs per group, treated with no treatment, vehicle and 5 topical compounds for 4 weeks Six dorsal full thickness burns Body weights, clinical observations Wound area Histopathology

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Surgical procedures Under strict aseptic conditions, burns are created. Pig receives appropriate anesthesia, post operative pain control and antibiotics. Sterile wound dressing with test article is applied as per protocol.

Post-procedure bandaging A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Post-procedure bandaging Stockinet covering Bandage completed

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Burn research Partial and full thickness burns are typically created Mini pigs-skin is most similar to human and are a robust laboratory animal for this model Healing and re-epithelialization are of interest Biomarkers and cytokines are currently of interest and not well understood Histology is well characterized at CBI

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Burn research Burn progression is also of research interest Following the initial burn, the lesion often worsens with time and is a significant adverse event Progression is not well understood, but appears to be related to vascular injury, tissue oxygenation and mediate release.

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Appearance of three types of burns: Full thickness burns extend to the underlying muscle layer

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Deep partial thickness burns or second degree burns in patient and in pig model. Burn extends into the dermis, but not to the muscle layer.

Partial thickness burn Day 1: A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Partial thickness burn Day 1: Day 1-8 Focal area of blister and rim of hyperemia and edema. Day 11-21 Progressive re-epithelialization and healing

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Deep partial thickness burns or second degree burns in patient and in pig model. The epithelium is still intact, but the superficial underlying collagen is bluish and amorphous. Vascular and cellular events have not yet occurred. Deeper collagen is not as effected.

Partial thickness burns-Day 1 A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Partial thickness burns-Day 1 Day 1: Acute thermal damage to the epidermis post surgically. Note hyperemic zone at burn-dermis interface.. Day 1: Acute thermal damage to the epidermis. Note hyperemic zone at burn-dermis interface.

Partial thickness burn-Day 8 and 21 A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Partial thickness burn-Day 8 and 21 Day 8-Dense scab formation. There is little re-epithelialization but dense new granulation tissue with inflammation is present in the wound bed Day 8-Dense scab formation. There is little re-epithelialization but dense new granulation tissue with inflammation is present in the wound bed

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Full thickness or 3rd degree burns. Damage extends to the underlying muscle.

Full thickness burns-increased heat and time exposure A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Full thickness burns-increased heat and time exposure Day 8: The surface is necrotic with a deep escharotic crust overlying the lesion Day 1: Acute thermal damage to the epidermis post surgically. Note hyperemic zone at dermis-burn interface

Histology, full thickness, with loss of epithelium-2 days A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Histology, full thickness, with loss of epithelium-2 days Full thickness burn, 2 days. There is a blister with separation of the epithelium from the underlying dermis. There are large, fluid fill blisters containing large numbers of neutrophils. The underlying dermis is exposed with inflammation, edema, hemorrhage and congestion. 40x HE Full thickness burn, 2 days. There is a blister with separation of the epithelium from the underlying dermis. There are large, fluid fill blisters containing large numbers of neutrophils. The underlying dermis is exposed with inflammation, edema, hemorrhage and congestion. 40x HE

Histology, full thickness, with loss of epithelium-3 days A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Histology, full thickness, with loss of epithelium-3 days Full thickness burn, 3 Days. There is complete loss of epithelium 40x HE Full thickness burn, 3 days, with complete loss of epithelium. The underlying dermis is exposed with inflammation, edema, hemorrhage and congestion. 40x HE

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Full thickness burns-increased heat and time exposure-Day 3 biopsy with histology Histology Day 3: Full thickness acute thermal damage to the epidermis post -surgically. Note that thermal damage extends through the dermis into the deep adipose tissue. HE 10x Day 3: Acute thermal damage to the epidermis post surgically. Note hyperemic zone at dermis-burn interface. The center of the burn has been biopsied under brief general anesthesia

Lesion development-Full thickness; Day 14 A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Lesion development-Full thickness; Day 14 Day 14 Epithelium partially complete or nearly complete covering Epithelium is thickened in comparison to normal epithelium (2-3x as thick) at edges, but may be very thin in the center Epithelial hyperplasia is common at edges Underlying collagen is consistent with granulation tissue Thick eschar and crust present

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Full thickness Day 14 There is crust formation with a thickened epithelium, There is edema and inflammation primarily of the superficial dermis and formation of more mature granulation tissue filling the wound bed.

Lesion development-Full thickness; Day 28 A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Lesion development-Full thickness; Day 28 Day 28+ Epithelium complete or nearly complete covering Epithelium is thickened in comparison to normal epithelium (2-3x as thick) Epithelial hyperplasia is common Underlying collagen is more mature with normal orientation Thick eschar and crust present

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Full thickness Day 28 Day 28 Histologically, the epithelium is covering all or most of the wound bed and is hyperplastic. There is a dense fibrous wound bed with collagen, neovascularization and mild inflammation. Macroscopically, there is re-epithelializaton and healing of the wound with some small scabbing. Day 28 The epithelium is covering all or most of the wound bed and is hyperplastic. There is a dense fibrous wound bed with collagen, neovascularization and mild inflammation

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Histologic scoring Semi-quantitative Industry standard 0-4 severity for Inflammation Re-epithelialization Collagen formation Epithelial hyperplasia Quantitative Measure entire wound length and measure amount of new epithelial covering and determine percentage

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Histologic scoring TA 1 Mean Histological Scores following partial thickness burns in 4 minipigs treated with vehicle, silver sulfadiazine and two test articles. The test articles, vehicle and positive control all positively effected the burn resolution.

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Histologic scoring CD31 and KI67 IHC demonstrating marked increases in the burn site at 20 days consistent with tissue healing

A TRANSLATIONAL APPROACH TO PRECLINICAL RESEARCH Summary At CBI, we have developed and validated a consistent and reproducible model of partial and full thickness burns in minipigs The lesion is uniform and consistent between sites on the same animal and between animals both macroscopically and microscopically.