Aginko - Your Partner of Choise for Projects in Bone and Joint Diseases

AGINKO Research AG

Route de l’Ancienne Papeterie

1723 Marly - SWITZERLAND - Email: [email protected] -Tel: +41 31 331 76 79 - Fax: +41 41 560 80 53

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TheOsteoarticularCROYourPartnerofChoiceforProjectsinBoneandJointDiseases

AGINKO Research is an osteo-articular and inflammation focusedpreclinical and clinical research organization with an unrivaledreputation for conducting global preclinical and clinical developmentprogramsofthehighestintegrity.Ourexpertise:

• Preclinical–Clinical–Regulatories• Preclinical&ClinicalHistologyServices• BiocompatibilityTesting• ToxicityTesting• CartilageBioassays • CartilageRepairModels• FractureRepair• OsteoporosisModels• PainInflammationAnimalModel• InVivoArthritisModels• ExpertiseinArthritisDiseaseModels• SpinalFusionAnimalModels• DiscDegenerationAnimalModels

Our scientificnetworks integrate researchers and clinicians, resultingin an efficient process from lab bench to clinic and reducing time tomarketfordeveloppersofpharmaceuticalbiotechnology.

Preclinical - Clinical - Regulatories

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Medical device clinical trials require a host of resources that many companies do not have in house. As a full-service contract research organization (CRO) focused on the osteoarticular medical device industry, AGINKO Research is uniquely able to support you through all phases of clinical trial development and testing. Working with your team, we will help you to design and implement a clinical trial that will best meet your business needs, whether you need clinical data to support a regulatory pre-market submission, drive product adoption, support product reimbursement, or monitor post-market product use. Strategic Advice AGINKO Research can be your clinical partner, providing comprehensive assistance from IND to post-submission activities, or supporting you in specific areas when an extra hand is needed. AGINKO Research assists its clients by providing strategic advice on many aspects of their drug development and medical devices programs. This includes: • Clinical study design • Advice on regulatory strategy • Selection of the best countries in which to conduct a clinical development program • Obtaining advice from both EMEA and FDA • Organizing clinical expert groups Strategic advice is provided by in-house personnel supplemented by our network of external consultants as appropriate.

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Data Management AGINKO Research offers complete data management services to meet your specific project needs. From database design to data analysis, AGINKO Research assists clients in achieving reliable, verifiable statistical results: • Flexible design • EDC or paper solutions • Fast database lock • Cost effective data solutions • Data Entry • CRF Design and Annotation • Database Design and Validation • Electronic Edit Checks • Tracking, Query Processing

Medical Writing AGINKO can be your medical writing partner, whatever documentation you require. We have prepared protocols, as well as New Drug and Product Licensing applications, including clinical, statistical, safety and efficacy summaries, CPMP dossiers, expert reports, and clinical procedure write-ups. We have also written investigator brochures, SOPs and annual reports, designed case record forms, and drafted manuscripts and monographs. After completion of the clinical and data work, the success of your project rests upon the way in which that work is explained and described. To gain regulatory approval, it is essential to make the most compelling case for your project, and we have the experience to help you achieve that. Our team of medical and technical writers is integrated within the regulatory affairs department offering particularly strong expertise in complex and demanding medical therapeutic areas such as bone and inflammatory diseases.

Preclinical & Clinical Histology Services

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AGINKO is a CRO providing dedicated histological processing Services. Our strengths are in the areas of regulatory pre-clinical toxicology studies and clinical trials. We have a lot of experience utilizing the most widely used specific techniques leading to production of the highest quality sections and stains for pathological examination.

Focused on quality and service At AGINKO, we have established a reputation for quality, reliability and flexibility. We work hard to maintain this reputation by focusing on: QUALITY - our success is founded on the consistent high quality of our work. Our technicians understand the importance of quality and consistency and ensure our high standards are maintained.

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SERVICE - we strive to provide a flexible and responsive service. We build close partnerships with our clients and have a culture that is driven by placing the needs of our clients first. FLEXIBILITY - we have the resources and flexibility to complete studies to your schedule and to deliver results according to your requirements.

Standard Histology Service

• Slide production from wet tissue or paraffin block • Standard or customized client trimming planes • Specific tissue orientation Standard or customised blocking codes • Standard H&E staining and a wide range of special stains, including:

- Alcian Blue - PAS - Masson Trichrome - Safranin O - …

• Shipment of residual wet tissues to sponsors • International transportation and documentation

Specialized Histology In addition to routine histology, AGINKO specializes in histology for the medical device industry. Our facilities are fully equipped and include x-ray and digital imaging systems for high quality microradiographs of both large and small specimens, microtomes for cutting paraffin and plastic sections, precision trimming, as well as automated tissue processor and staining and equipment to ensure high quality and reproducible results. Even with biomaterials that are “difficult” or “impossible” to process, our extensive knowledge of materials chemistry enables preservation of intact tissue/material interfaces and even recovery of data from poorly embedded specimens obtained from external histology laboratories. Stains for special histology include Toluidine blue, H&E, Goldner’s trichrome, safranin O and others modified in-house specifically designed stains for plastic-embedded specimens.

Immunohistochemistry AGINKO immunohistochemistry laboratory can provide the support needed for the smallest research projects through large follow-up investigative works on toxicity studies. Our selection of commercial antibodies have been reliably used on both frozen and fixed tissues and in tissues embedded in paraffin, methylmethacrylate polymer or Spurr resin.

Pathology Capabilities

• GLP toxicology studies with all species including contract necropsy • Pharmacology and efficacy studies

Biocompatibility Testing

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European and North American standards for biocompatibility testing are similar but not identical. European requirements are based on ISO 10993. The US FDA has substantially adopted ISO with some differences in specific testing requirements. For tissue and bone implants, including engineered tissues and scaffold, a series of evaluation tests are required to assess biological effects. All testing in support of regulatory submissions must meet GLP standards.

Test ISO FDA Description

Cytotoxity x x In vitro assay

Sensitization x x Murine local lymph node assay (LLNA), Megnuson-Klingman test

Irritation x x Intracutaneous injection of test material

SystemicToxicity

x x

Acute

(<24h)

Intravenous

Rabbit endotoxin andpyrogen test

Subacute

Subchronic

(more than 24h and less than10% of animallifespan)

Intravenous

Intraperitoneal

Single ormultiple exposure

Chronic

(>10% animal lifespan)

Supplemental test

SubchronicToxicity

x x IP dosing, long follow-up

Genotoxicity x x Ames test, ChromosomalAberration Assay, Mouse MicronucleusAssay

Implantation x x Test for local effectsafter implantation

Carcinogenicity x x Supplemental test

Biodegradation x x

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Toxicity Testing AGINKO propopses a serie of tests for general toxicology assessment. These can be either acute or chronic toxicology studies. We also conduct studies with very specific adapted designs (adapted to the tested product and specific national regulatory requirements if needed). The routes of administration can be diverse: - Oral: gavage, capsules, dietary admixtures, drinking water - Parenteral:

• Intravenous: bolus, slow injection, continuous infusion, cycles (vascular acces port) • Others: subcutaneous, intradermal, intramuscular, intraperitoneal �

- Dermal: open, semi-occluded and occluded dressing, patches and other devices, with or without rinsing, with or without collar. Each study is adapted to the specificity of the product

- Ocular: including intra-vitreous - Intra-nasal - Inhalation: in non-rodents - Intra-vaginal - other routes: we have experience in a number of very specific routes of

administrations in different species. Please contact us for any specific request.

Code Test Type of Study Method Turnover (Week)

AGR1 Acute Toxicity (p.o, i.v., i.m., s.c.,i.p., dermal), rodents

Full study

OECD 420, 423, 425, 402, EU B.1.tris, , EU B.1bis, EU B.3, OPTTS 870-1200

8 - 10

Limit Test 8

AGR2 Acute injection toxicity/pathogenicity, rodents

Full study OPTTS 885.3200 8 - 10

AGR4 Acute oral toxicity/ pathogenicity, rodents Full study OPTTS

885.3300 8 - 12

AGR5 Maximum tolerated dose, rodents

Full study (3 - 5 dose levels), clinical and clinical-laboratory observation, gross pathology

CHMP/SWP/302413/08 8 - 12

Toxic ity Testing

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AGR6 Extended single dose toxicity study, rodents

Full study, clinical and clinical-laboratory observation, gross and histopathology

CPMP/ICH/286/95, M3 (R2)

8 - 12

AGR7

Maximum tolerated dose, non-rodents (rabbits, ferrets, dogs, non-human primates)

Full study (3 - 5 escalated dose levels), clinical and clinical-laboratory observation, gross pathology

CHMP/SWP/302413/08 8 - 12

AGR8 Dose range finding study (p.o, i.v., i.m., s.c.,i.p., dermal), rodents

2 weeks of administration, 7d/wk exposure, clinical and clinical-laboratory examination, gross pathology, optional histopathology of selected organs

CPMP/SWP/1041/99, OPTTS 870-3050,

8 - 12

AGR9

Dose range finding study (p.o, i.v., i.m., s.c.,i.p.), non-rodents, (rabbits, ferrets, dogs, non-human primates)

2 weeks of administration, 7d/wk exposure, clinical and clinical-laboratory examination, gross pathology, optional histopathology of selected organs

CPMP/SWP/1041/99, OPTTS 870-3050,

8 - 12

AGR10

14-21 days repeated dose toxicity study (p.o, i.v., i.m., s.c.,i.p., dermal), rodents

2 - 3 weeks of administration, 7d/wk exposure, clinical and clinical-laboratory examination, gross pathology, full set of histopathology

CPMP/SWP/1041/99, OECD 407, EU B.7, OPTTS 870-3050,

10 - 16

AGR11

14-21 days repeated dose toxicity study (p.o, i.v., i.m., s.c.,i.p.), non-rodents, (rabbits, ferrets, dogs, non-human primates)

2 - 3 weeks of administration, 7d/wk exposure, clinical and clinical-laboratory examination, gross pathology, full set of histopathology

CPMP/SWP/1041/99, OECD 407, EU B.7, OPTTS 870-3050,

10 - 16

AGR12

28-day repeated dose toxicity study (p.o, i.v., i.m., s.c.,i.p., dermal), rodents, primates)

7d/wk exposure, clinical and clinical-laboratory examination, functional observation battery, gross pathology, full set of histopathology

OECD 407, 410, EU B.7. OPTTS 870-3050

18 - 22

AGR14

28-day repeated dose toxicity study (p.o, i.v., i.m., s.c.,i.p., dermal), non-rodents

7d/wk exposure, clinical and clinical-laboratory examination, functional observation battery, gross pathology, full set of histopathology

OECD 407, EU B.7. OPTTS 870-3050

18 - 22

AGR16 6-month repeated dose toxicity study, rodents

7d/wk exposure, clinical and clinical-laboratory examination, gross pathology, full set of histopathology

OECD 452, CPMP/SWP/1041/99

40 - 42

AGR17

6-month repeated dose toxicity study, non-rodents (rabbits, ferrets, dogs, non-human primates)

7d/wk exposure, clinical and clinical-laboratory examination, gross pathology, full set of histopathology

OECD 452, CPMP/SWP/1041/99

40 - 42

Cartilage Bioassays

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Osteoarthritis (OA) is a joint disease characterized by progressive degeneration of articular cartilage and changes in other joint tissues. No disease-modifying therapies are currently available. OA severely impacts the quality of life of the elderly and is one of the most important disease that leads to disabilities.

Test Systems

• Healthy and ostearthritis cadaveric primary chondrocytes of human, porcine and bovine origin (Cell isolation according to AGINKO SOP’s or customized)

• Cultures of cartilage explants (Human/Porcine/Bovine) • Human synoviocyte cell cultures (Healthy donor/OA/RA) • Chondrocyte alginate cultures from various origin • Cultures of stem cells isolated from bone marrow, fat, or muscle for

differentiation potential screening • Chondrocyte, synovial cell, and bone marrow stem cell co-cultures • Supply of controlled and characterized tissues and cells (cell

sourcing/disease stage)

C artilage Bioassays

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Contract Research

• In Vitro Cultures and Bioassays • Molecular and cellular analysis, gene

expression, RT-PCR, northern and western blotting, silencing and heterologous expression of proteins, MMP and protease quantification (ELISA), cytokine characterization

• Proliferation assay, co-culture, cell characterization (FACS)

• Histology, IHC, IF, morphological characterization of cells with specific staining

• Biochemistry, matrix analysis, synthesis and degradation of collagen and Proteoglycans

Cell Culture Screening System using stem cells and chondrocytes. Courtesy of : Population Doublings and Percentage of S100-Positive Cells as Predictors of In Vitro Chondrogenicity of Expanded Human Articular Chondrocytes SAMOA GIOVANNINI, JOSE DIAZ-ROMERO, THOMAS AIGNER, PIERRE MAINIL-VARLET, AND DOBRILA NESIC from J. Cell. Physiol. 222: 411–420, 2010.

(*) Cellular, Tissue and Gene Therapies Advisory meeting, May 14-15, 2009 (*) Cellular, Tissue, and Gene Therapies Advisory Committee meeting, March 3, 2005 (http://www.fda.gov/ohrms/dockets/ac/05/transcripts/2005-4093T1.htm); March 4, 2005 (http://www.fda.gov/ohrms/dockets/ac/05/transcripts/2005-4093T2_01.htm).

Cartilage Repair Models

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Injury of articular cartilage due to trauma or pathological conditions is the major cause of disability worldwide, especially in North America. The increasing number of patients suffering from joint-related conditions leads to a concomitant increase in the economic burden. Several strategies to repair and replace knee joint cartilage are needed since knee-associated disabilities are more prevalent than any other joint. Because of inadequacies associated with widely used approaches, the orthopedic community has an increasing tendency to develop biological strategies, which include transplantation of autologous (i.e., mosaicplasty) or allogeneic osteochondral grafts, autologous chondrocytes (chondrocyte transplantation), tissue-engineered cartilage substitutes or the local application of a compound. Aginko’s vast experience in the study of human and animal bone and cartilage biology allows Aginko to provide a wide range of services tailored to individual needs that support product development intended for clinical human applications.

Cartilage Repair Preclinical Strategies The FDA recognizes that choosing and determining the suitability of an animal model(s) for evaluation of any specific product is difficult because there is no perfect animal model of articular cartilage injury. As discussed at the March 2005 CTGTAC meeting (*):

• the scientific literature contains descriptions of numerous methods for evaluating the nonclinical behavior of native cartilage and, consequently, articular cartilage repair or replacement products;

• not all of these methods may apply to a specific articular cartilage repair or replacement product; and goats, sheep and horses are the most frequently used large animal models for cartilage repair.

Chondral and Osteochondral Defect Models Species available include: rabbit, dog, goat, sheep, and horse. Rabbit models are well described in the literature and are recommended by the FDA for preliminary, short-term, and proof – of - concept studies. These rabbit model permit an easy screening of compounds.

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Osteochondral defect model in the rabbit repaired with a tissue engineered implant. Large animal models, recommended for long-term studies, can incorporate multiple defects and are amenable to arthroscopy and MR imaging.

Osteochondral defect superficial (Full thickness defect)

covered with the application of a membrane.

Outcome Measures •Histology: morphology, proteoglycan and cell loss, cartilage damage, osteophyte

formation. Aginko has developed special protocols to samples cartilage defects. All slides are graded by expert histopathologists.

•Radiology: X-ray, micro-CT •Serum, joint fluid, and urine biomarkers •Cartilage biochemistry •Chondrocyte metabolism •Arthroscopy •MRI (morphology, quantitative analysis)

Courtesy of Osteoarthritis and Cartilage Volume 17, Issue 10, October 2009, Pages 1341-1349 A. Watanabe , C. Boesch, S.E. Anderson , W. Brehm and P. Mainil Varlet

Fracture Repair

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Bone Healing Fracture healing is a complex physiological process where the bone and its neighboring tissues play important roles. Thus, fracture healing can only be assessed in animal models. However, the choice of the most appropriate animal model for fracture repair remains an unanswered question as no animal model accurately reproduces the human bone physiology, biology, structure and biomechanics. Despite these limitations, small and large animal models have been developed to study the effects of bone substitutes, scaffold, biologics or cell-based products on bone fracture repair. Fracture bone healing is usually an optimal biological process. However, delayed healing or non-union can occur in patients for multiple reasons. In addition, bone fractures are more frequent and more problematic in osteoporotic patients as the bone structure is compromised. Fracture, segmental and critical-size defect models have been developed in several species.

Animal Models Critical-size defects:

• Calvaria critical-size defect in rats, rabbits • Mandibular critical-size defect in mini-pigs, in dogs • Femur critical-size defects in rats • Ulna critical-size defects in rabbits • Tibial critical-size defects in sheeps and mini-pigs

New-Zealand White (NZW) rabbit ulna critical-size defect.

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Osteotomy (fracture and segmental bone defects):

• Tibial osteotomy in mini-pigs, sheeps and goats • Femoral osteotomy in mini-pigs, sheeps and goats • Mandibular osteotomy in sheeps

Outcome Measurements A wide range of measurements can be performed such as:

• In vivo and ex vivo BMD and BMC measurements of cortical and trabecular bones by DEXA, pQCT and µCT

• Biomechanical testing • Physiological bone turnover markers (blood, urine) • Ash chemical analysis • Histomorphometry, histology

Radiography of New-Zealand White (NZW) rabbit ulna critical-size defect.

Histological staining of (NZW) rabbit ulna critical-size defect.

Osteoporosis Models

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Senile, post-menopausal or secondary osteoporosis weakens long bones by decreasing their mineral content and altering their structure. Animal models of osteoporosis allow assessing the efficacy and safety of novel anti-osteoporotic drugs.

Animal Models Primary osteoporosis models:

• Ovariectomy in female rats • Orchidectomy in male rats

Outcome Measurements A wide range of measurements can be performed such as:

• In vivo and ex vivo BMD and BMC measurements of cortical and trabecular bones by DEXA, pQCT and µCT

• Biomechanical testing • Physiological bone turnover markers

(blood, urine) • Ash chemical analysis • Histomorphometry, histology

Three-dimensional reconstruction of rat femoral structure. Courtesy of B-Cube.

MicroCT imaging of lumbar vertebral cancellous bone loss in the ovariectomized (OVX) rat. Left: pre-OVX. Right: 16 weeks post-OVX.

Secondary osteoporosis models: • Glucocorticoid treatment in rats • Bone immobilization in rats

Pain Inflammation Animal Model

AGINKO Research AG



Pain is a critical problem and is the most common reason for medical consultation. It often results in disability and psychological distress in patients. Improvement in the management of pain is a high priority. The use of preclinical pain models allows the identification and development of novel drugs and offer new opportunities of pain relieving therapies for patients. According to the nature and the origin of the pain, appropriate pre-clinical models are useful to study the mechanisms and efficacy of novel pain relieving drugs. A variety of preclinical models are available to study persistent and chronic pains. More specific models for chronic osteoarticular and neuropathic pains are of great value to assess the efficacy of novel drugs targeting bone cancer, rheumatoid arthritis, osteoarthritis and neuropathic pains such as post-surgical pain syndrome, traumatic nerve injury, spinal cord injury and diabetic neuropathy. Osteoarthritis-related pain can be modeled by chemical or surgical alteration of the joint. A robust preclinical model consists in intra-articular injection of monosodium acetate, which induces similar pathological changes and pain as observed in human osteoarthritis. Preclinical models of inflammatory and rheumatoid arthritis pain involve the injection of immunogenic or non-immunogenic adjuvants in the joints or in the systemic system. Neuropathic pain models involve partial nerve injury such as ligature of the spinal or sciatic nerves. Inflammatory pains of high to mild amplitude and with short to long duration can be induced by formalin or by immune-stimulating substances such as Freund’s Complete Adjuvant or Carrageenan. Endpoints assessment of quality of life in a preclinical model of arthritis:

- Paw withdrawal test (latency and duration) - Static weight-bearing - Validated pain scores for several animal species - Validated protocols for gait analysis in goat and sheep - Video-monitoring of animals (behavioural studies) - Algometry - Electrophysiology

Pain

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Models Species Endpoints

Name (Abbreviation)

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Bone and Joint Pain Bone Cancer Pain

(BCP) x x x Monosodium Iodoacetate-

ankle (MIA ankle) x x

Monosodium Iodoacetate-

knee (MIA knee)

x x x x

Freund’s Complete

Adjuvant-ankle (FCA Joint)

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x x

Neuropathic Pain Spinal Nerve Ligation (SNL) x x x x Streptozotocin

Induced Diabetic Neuropathy (STZ)

x x x Chronic

Constriction Injury (CCI)

x x

Inflammatory Pain Freund’s

Complete Adjuvant (FCA)

x x x x x

Formalin x x x x

Carrageenan x x x x

In Vivo Arthritis Models

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Transection of the anterior cruciate ligament to induce OA in a rabbit model.

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Aginko offers a wide range of OA models. Our expertise allows us to perform state-of-the-art studies to test new therapies, and to advise on study design

and data interpretation. Surgically-induced Osteoarthritis Models

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Surgical models use a variety of methods (e.g. ACL transection, meniscal tear, meniscectomy) in small and large species to induce joint instability. While etiologically similar to human secondary OA, the resulting lesions also resemble primary OA. Depending on the model, lesions may be more reproducible, more rapidly induced, or more severe than in the spontaneous models. The chosen model should be carefully selected to match the expected therapeutic result.

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Spontaneous Osteoarthritis Models Several species and laboratory rodent strains spontaneously develop OA with age, including the STR/ort mouse, Dunkin Hartley guinea pig, cynomolgus monkey, and Syrian hamster. The Dunkin Hartley guinea pig model is one of the few preclinical OA models recommended by the FDA Guidance for Industry (1999). While these models display some morphological and biochemical similarities to human OA, they are also characterized by slow progression, mild degeneration, and high variability. These models are best suited to study disease modifying agents, but require large group sizes to achieve statistical significance. Inflammatory Arthritis Models Inflammatory arthritis models range from mild disease to highly destructive and rapidly progressing lesions, and are performed almost exclusively in rodents. The inflammatory component may also range from mild to severe and varies in character and pathogenesis between models. More information can be found in the Aginko Pain and Inflammation Models fact sheet.

Outcome Measures In most arthritis models, the primary outcome measures are morphologic characterization and quantification of lesions. Pain and other behaviours, cartilage biomechanics, and biochemical markers may also be of interest.

For preclinical arthritis models in the CRO industry and the expertise to select and interpret them.

• Morphology: plain film radiography, arthroscopy, µCT, MRI, histology, histological grading (e.g. modified Mankin’s score), histomorphometry

• Biochemistry: serum, synovial fluid and urinary biomarkers, cartilage biochemistry

• Behaviour: quantification of hyperalgesia, allodynia, and motor behaviours

• Biomechanics: mechanical characterization of cartilage, friction and lubrication testing.

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Expertise in Arthritis Disease Models

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BIOMARKERSMODELSCellular & Molecular Testing

Cell proliferation, phenotype

mRNA, DNA

Proteins, proteoglycans

Enzymes, cytokines

Matrix biochemistry, function

Imaging

Radiology, DXA, pQCT

VivaCT, microCT, MRI

Digital macroscopic imaging

Histology, IHC, ISH, IF, CLSM

Histomorphometry

Pain assessment / Behavioral testing

Motor function / gait analysis

Static weight-bearing

Allodynia, hyperalgesia

Electrophysiology

Biomechanical TestingExplant / in-situ cartilage mechanical tests

Friction and lubrication tests

Stem cell cultures

Chrondrocyte cultures

Cartilage explants

Alginate cultures

In Vivo

Inflammatory arthritis / RACollagen-induced

Iodoacetate

Collagenase

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Bradykinin / PGE2

Kaolin / carageenan

Osteoarthritis

Spontaneous

Meniscal tear

Partial / total meniscectomy

ACL transection

ACLT + meniscectomy

All models +/ - exercise

In Vitro

Spinal Fusion Animal Models

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Spine fusion is a frequent surgical procedure consisting in fusionning 2 vertebrae to stabilize the spine through intertransverse or vertebral body fusion. It is indicated in several pathological cases such as: - Vertebral fractures - Degenerative disc diseases and herniation - Scoliosis or kyphosis - Spinal tumors The success of spinal fusion depends on the capacity of the bone graft to form new bone. The use of auto graft, the “gold standard”, has limitation due to limited availability and problem at the donor site. Thus, the development of alternatives to auto graft can improve the quality and the rate of success of bone fusion. Additionally, the use of fixation devices such as screws, plates or cages improve the fusion by stabilizing the vertebrae. Novel bone graft substitutes, including tissue-engineering constructs, growth factors and gene therapies, aim to improve vertebrae fusion. Establishment of feasibility, efficacy and proof of concept of these novel technologies require validation in an animal model. Validated small and large animal models are available to study spinal fusion at the cervical, thoracic and lumbar levels using anterior or posterior approaches. Although animal studies may be necessary to address specific questions prior to human use, the FDA recognize that choosing and validating an animal model is difficult because there is no perfect animal model. In choosing an animal model, the sponsor should consider the morphology, histology, biomechanics, and kinetics as compared to the human situation. Species Available:

• Rabbit • Rat • Dog • Goat and Sheep • Pig

The following are some examples of questions that animal studies might be used to address: - For an intervertebral body fusion device used in conjunction with a biological component (e.g., bone morphogenic protein), animal studies may provide information as to whether bony fusion occurred or is progressing and the quantity and quality of bone formed inside the device (a stress protected environment) as compared to an appropriate control group. Animal studies are used to evaluate dilution, dose, and concentration factors for certain systems.

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Animal studies may be used to evaluate a novel system design in earlier development stages (e.g., different design concepts of vertebral disc replacement devices). - Animal studies may be used to support the biocompatibility of a totally new material, a new material for use in orthopedics, or a new material for spinal exposure. - Additionally, these studies may be used to address the probability of generating abrasion or wear particles of the new materials. Types of Model: Procedures are usually performed at one level but multilevel fusions are also possible: • Anterior cervical fusion • Anterior cervical discectomy (Sheep, Goat) • Posterolateral lumbar fusion (Rabbit, Primates) • Posterior lumbar fusion (Rat) • Posterior thoracic fusion (Rat) • Posterolateral muscle splitting intertransverse fusion (Dog) Outcome Measures: The success of fusion can be assessed following established procedures: • Radiography: intervertebral angle • CT MicroCT, • DXA: bone mineral density • Histology and histomorphometry: bone quality • Mechanical testing

Representative A) histology, B) CT images, and C) CT color-mapped images. For all images, the upper region is the ventral side and the lower region is the dorsal side. The left side is the cranial region and the right side is the caudal region. From: Evaluation of early tissue reactions after lumbar intertransverse process fusion using CT in a rabbit. Shinbo J, Mainil-Varlet P, Watanabe A, Pippig S, Koener J, Anderson SE. Skeletal Radiol. 2010 Apr;39(4):369-73. Epub 2009 Jun 25.

Disc Degeneration Animal Models

AGINKO Research AG


1723 Marly 1 - SWITZERLAND - Email: [email protected] -Tel: +41 31 331 76 79 - Fax: +41 41 560 80 53

Degeneration of intervertebral discs (IVD) is one main cause of back pain and can also lead to other complications such as disc herniation, spinal stenosis and degenerative spondylolisthesis. Although IVD degeneration presents similar features as natural aging, IVD degeneration is present in all classes of age in the adult population. The etiology and development of IVD degeneration is poorly understood and may involve complex interactions of genetic, biological and mechanical factors. In complement to in vitro and ex-vivo models, small and large animal models offer a physiological model to study IVD repair, where the mechanical and biochemical environment of the IVD is preserved. However, these animal models do not accurately picture human IVD biology. Animal models of IVD degeneration present specific features, such as better nutrient and metabolite transport, the presence of notochordal cells within the mature IVD core and quadruped spine load, account for a facilitated IVD repair process in these animal models. In spite of these limitations, small and large animal models of IVD degeneration are of great value for studying the progression of the disease and for validating the safety and the efficacy of novel therapeutic treatments. Reproducibility, progression and severity of the induced degeneration vary according to the nature of the injury and to the animal species. IVD degeneration can be created by using chemical (papain), physical (nucleus puncture, annulus fibrosus stab) or mechanical injury to the IVD. Repair of the IVD can be assessed using in vivo and ex-vivo imaging, immunohistology, biochemical and molecular analyses and biomechanical testing.

Species available:

• Rabbit • Mouse • Rat • Dog • Goat and Sheep • Pig

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Aginko - Your Partner of Choise for Projects in Bone and Joint Diseases