骨科组织工程动物模型汇编课件

,Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,骨科组织工程动物模型,骨科组织工程动物模型,Tissue Engineering,Cato T. Laurencin et al (1999):,“,The application of biological, chemical, and engineering principles towards the,r,epair,r,estoration and,r,egeneration of living tissue using biomaterials, cells, and factors alone or in combination,”,The “3Rs”,Tissue EngineeringCato T. Lau,Main Subtitles:,I. How to Choose Animal ModelsII.Commonly Used Models III.Project Designing IV.Methods of Evaluation,Main Subtitles:I. How to Ch,Researchers should consider the following issues when selecting a research animal:,Ethics,Availability,Housing Requirements,Ease of Handling,Costs,Diseases,Background Data of the Animal,I. How To Choose Animals,Researchers should consider th,Ethics / Animal Research,Balance -,Public concerns rabbits, pigs, and goats dogs, cats,(Rats, pigs, rabbits, or goats should be considered first. Dogs and cats are used if it is essential.),- Directly applicable to patients Basic science- Less pain and suffering more pain- Less number large number,Ethics / Animal Research Bal,Ethics “3Rs”,In 1959, Russell and Burch, two English scientists, introduced,The Principles of Humane Experimental Technique,. - “The 3Rs”:,Replacement,of animals, by,in vitro,methods or more primitive animals,Reduction,of the number of animals required,Refinement,of experimental methods to reduce pain and stress of the animal,Ethics “3Rs”In 1959, Russel,Availability,Securing animals of a particular species, strain, type, or age is very important for a specific project.,In developed countries (,United States, Canada, Japan, some European countries,), it is not a problem.,In developing countries it can be a major problem, especially for healthy research animals.,AvailabilitySecuring animals,Housing Requirements,Your facility may not have the capacity for certain animals (such as primates or goats). It is better to choose another species at your institution if the project allows because:,it avoids traveling to another place;,you can have close and easy observation, animal treatment, and data management; and,it is generally cheaper.,Housing RequirementsYour fa,Ease of handling,All based on the weight:,Mice and Rats (20-500 g) Rabbits (2-5 kg) Dogs (20-30 kg) Pigs (20-50 kg) Goats or sheep (50-80 kg),Ease of handlingAll based on,Costs,The costs for animal purchasing, transportation, housing, unexpected loss, surgical supply and services, radiography, mechanical testing, and histology are basic.,Financial limitations: the size of the grant, the money available.,Use a cheaper animal species if it is valid for verifying the hypothesis,Costs are not decisive factors for animal selection.,Costs The costs for ani,Diseases,Spontaneous diseases in animals can seriously compromise the experimental plan, confuse research data, and raise the cost. Researchers should avoid using species known to have high incidence of particular diseases.,Examples:,Conventl. rats - chronic respiratory disease,Conventl. rabbits - Pasteurella multocida infection,Dogs canine hepatitis,Goats Q-fever,Pigs a good choice (healthy, good size, ease of handling),Transmissible to human,DiseasesSpontaneous diseases,Background Data of the Animal,Including the,anatomy,physiological features,biomechanical data, histological data, or,drug responses,and certainly the,model (surgical procedures), which can:,help researchers to repeat or extend the work.,help caretakers to better maintain animals.,have control values for evaluation.,Goats, pigs, rats, mice, becoming valuable subjects,(more easily accepted by the public too).,Background Data of the Animal,Trend:,Small: Nude Mice, Rats,Medium: Rabbits,Large: Pigs or Goats,II. Commonly Used Animals,Trend:II. Commonly Used Animal,Mouse,(regular and nude),Small rodents, popular in research because of the well-researched and easily manipulated genome (regular mice), limited immune response (for nude mice).,Widely used in the studies of,Osteogenesis and chondrogenesis,Inflammatory arthritis,OsteoporosisBone tumorsNerve repair,Mouse (regular and nude) Sma,Nude Mouse SC Model of Osteogenesis,A calcified graft,Through injection or implantation. Time period: 4 to 8 weeks,Porous PLGA + Bovine Osteoblasts,6 weeks,Up to 6 small implants,Nude Mouse SC Model of Osteoge,Nude Mouse Model,of SC Osteogenesis,Alginate gel + bovine osteoblasts,6 weeks,Nude Mouse Model Alginate gel,Nude Mouse SC Model,of Chondrogenesis,S&F stain,6 weeks,12 weeks,Alginate gel + bovine chondrocytes,Nude Mouse SC Model of Chondr,Rat,(a universal research animal),Rodent,Lifespan: 21-24 months,Lower level vertebrate,Low cost (still costs $20-30 / SD rat),Easy handling,Dying hard,Due to its advantages, it is not unreasonable to consider the rat first for a new project.,Remember:,Use SPF rats !,Rat skeletons made by Dr. An and a medical student (GM),Rat (a universal research an,Rats,Commonly used for the studies of,Fracture, bone defect repair,Biocompatibility of biomaterials,Osteoporosis,Bone ingrowth to implant,Bone or joint infections,Prosthetic debris,Nerve repair,RatsCommonly used for the st,Rat Tibial Fracture Model,(An YH, et al 1994),The tibia,Model is,easier than,femur,model,due to,1. less muscle,2. percutaneous pining,Rat Tibial Fracture ModelThe t,Rats,Nerve repair,RatsNerve repair,Rat Models for Testing Biocompatibility,Direct Contact,Gap,Titanium in SC tissue,Rat Models for Testing Biocomp,Rat Tibial IM Nail Model,Percutaneous insertion,a,level - more cancellous bone,b,level - more cortical bone,a,a,b,b,Rat Tibial IM Nail ModelPercut,Rat Inflammatory Arthritis Model,(Papain injection),Papain (a proteinase) injection,arthritis develops,in 1-2 weeks,Rat Inflammatory Arthritis Mod,Rabbits,Relatively high level vertebrate,Good size which enables easy surgical and evaluation procedures,Less expensive dogs, goats,Rabbit is the most commonly used animal in orthopaedic research,(,J. Orthop. Res.,survey 45/171 = 26%).,Jan and Rabbit on Dec 15, 2003,Rabbits Relatively high le,Rabbits,(“Universal” Research Animal),Rabbits are commonly used for:,Articular cartilage repair,Bone defect, fracture, or osteotomy,Bone ingrowth to implant,Osteoarthritis or inflammatory arthritis,ACL, MCL reconstruct, meniscal replacement,Osteonecrosis,(Femoral head),Bone and joint infections, and prosthetic infection,Rabbits (“Universal” Research,Rabbits,Shortcomings,(Compared to dogs and rats):,More wound infections,More complications,More unexpected deaths,Become a lesser issue due to the use of SPF rabbits,Rabbits Shortcomings (Compar,Osteochondral Defect Model,Rabbit Femoral Intercondylar Groove,Using trephines; 3 - 4 mm diam., 3 - 4 mm deep; if too large, can cause arthritis,Osteochondral Defect Model Usi,Osteochondral Defect Model,Rabbit Femoral Intercondylar Groove,Using trephines (better than drill bits); 3 mm diam., 3 mm deep; if too large, can cause arthritis,Osteochondral Defect Model Usi,Rabbit,Meniscus,Defect,Models,for,Tissue,Engineering,Goats, Sheep,And Dogs Can,Also Be used.,RabbitGoats, Sheep,Rabbit Radial Defect Models,(Herold HZ, et al, 1976),Critical size: 15-20 mm. 23 mm above the wrist. 8-month old rabbit. It is wise to use one side.,Rabbit Radial Defect Models (H,Rabbit Radial Defect Models,(Herold HZ, et al, 1976),Critical size: 15-20 mm. 23 mm above the wrist. 8-month old rabbit. It is wise to use one side.,Rabbit Radial Defect Models (H,Rabbit Femoral Condyle Defect Model,5 mm in diameter, 10 mm in depth,12 weeks,DBX,Collagraft,Rabbit Femoral Condyle Defect,Critical size,15 mm,Multiple,7 mm,Rabbit Calvarial Defect Model,Critical sizeMultiple Rabbit C,First by Scott Boden, 1995L5 and L6 (or L4 and L5)Vol: 2.5 cc per sidedTime period: 6 weeks,Rabbit Lumbar Transverse Process Model,June 2003, Charleston, SC,Rats,Dogs,Seep,Goats,First by Scott Boden, 1995L5,Rabbit Femoral Condyle,Plug Model,(Keller, 1987),Implant,Porous Plugs,Ceramic Plugs,Spool Plugs,Rabbit Femoral Condyle Implant,Rabbit,Calvarial On-lay,Graft Model,Titanium disk,Titanium foam coated with HA,Rabbit Titanium diskTitanium f,Rabbit Distraction Osteogenesis Model,(Meffert RH, et al 2000),External Fixator: M-03, Orthofix0.5 mm per time, 2 times per day, total 10 days = 10 mm,It is not a easy model,Rabbit Distraction Osteogenesi,Dogs,Higher level vertebrate.,The closest,in vivo,condition to human except for large primates.,Relatively easy for handling.,Not easily accepted by the public.,The goat is a good alternative.,Dogs Higher level vertebrate,Dogs,Commonly used to study:,Fracture, bone defect repair,Bone ingrowth,Prosthetic prototype testing,Osteoarthritis,ACL reconstruction,Osteonecrosis, osteoporosis,Implant infections, osteomyelitis,Meniscal repair, cartilage repair,Spinal procedures,Bioabsorbable screw,DogsCommonly used to study:,Canine Femoral Plug Model for Testing Bone Ingrowth,Other Species for the same purpose:,Rats, rabbits, pigs, goats, primates,Canine Femoral Plug Model for,Goats,Higher level vertebrate,Becoming a popular and valid animal subject in recent years, especially for its role in the research of ACL, cartilage repair, and bone graft materials,Also for biocompatibility, joint replacement, fracture, limb lengthening, or meniscal repair,Goats Higher level vertebra,Goats,A newcomer compared to dogs and lacks basic data,Due to higher level vertebrate and the non-pet status, goats will play a significant role in orthop-aedic research in the future,Shortcomings,Difficult handling Large housing space,Goats A newcomer compared t,Goat ACL Anchor Models,Young, FA,An, YH,1995,Goat ACL Anchor ModelsYoung, F,Sheep,Large animal, similar to goats, which has both growing and adult skeletal phases,There is less information about sheep as a subject for orthopaedic research compared to dogs and goats,Sheep have been used for:,-,Bone defect repair,-,Osteoporosis,-,Meniscal repair,-,Cartilage repair,-,Osteoarthritis,-,ACL reconstruction,-,Vascular grafting,-,Nerve repair,-,Limb lengthening,Sheep Large animal, similar,Pigs,Have both growing and adult skeletal phases.Have been used for studying:,Cartilage and bone repair Osteoporosis Femoral head osteonecrosis Bone ingrowth to implant,PigsHave both growing and adu,Miniature Pig,Osteochondral,Defect Model,Project at MUSC:,In vivo testing of,cryopreserved cartilage graft,C,Miniature Pig Project at MUSC:,Primates,Primates have both growing and adult skeletal phases and the peak bone mass: at age 10-11 years,Ideal for research because they are the closest to human among animals,Has been used in the studies of,OsteoporosisBone ingrowthBone defect repairCartilage repair,Osteoarthritis,Because of the lack of availability and high cost, the use of primates has been limited to the research projects for which they are definitely essential, such as the evaluation of new drugs for osteoporosis.,Primates Primates have both,Horses,The largest animal used as an experimental subject,Mainly used for the studies of,cartilage or joint conditions,since the horse has rich cartilage tissue,Reported as an experimental animal for -,articular defect repair,-,experimental synovitis,-,septic arthritis,Horses The largest animal u,Other Animals,Guinea pigs - popular for studying osteoarthritis,Cats - osteoarticular transplantation,Hamsters - implant infection,Chickens - scoliosis or tendon repair,Turkeys - bone remodeling.,Jan and Linda on Dec 15, 2003,Other Animals Guinea pigs -,Bone or Cartilage Tissue Engineering,The first step in generating an animal research project is the generation of a new idea or question to be investigated.,Small animal - Nude mice, rats,Medium size animal: Rabbits,Large animals: Pigs, goats/sheep, dogs,III. Project Design,Rat skeletons made by Y. H. An and his student G. Macky, 1992,Bone or Cartilage Tissue Engin,IV. Methods of Evaluation,Although many sophisticated and expensive methods are available, the most useful techniques are those which are,simple, reliable, efficient and economical,.,Most importantly, the test method,must be valid,it must be capable of effectively measuring the parameter intended to measure.,IV. Methods of EvaluationAlt,Basic Methods of Evaluation,Clinical observation,Radiography,Macro-observation at necropsy,Histological evaluation,Mechanical testing,Basic Methods of Evaluation ,Clinical Evaluation,Animal behavior,Temperature,Food/Feces,Function,Incision,Swelling,Redness,Range of motion,.,Clinical EvaluationAnimal beha,Macro Observation,Shoot pictures!,Macro Observation Shoot pictur,Macro Observation,Shoot pictures!,Macro Observation Shoot pictur,Measurement of Length and Area,1. Direct Measurement2. Measurement on X-ray Images3.,In Vivo,Measurement,4. Dissecting Microscopy,Measurement of Length and Area,Radiography,1. Plain Radiography2. High Resolution Radiography3. Micro-radiography,What are these?,Radiography1. Plain Radiogr,Dissecting Microscope,Dissecting Microscope,Histological Evaluation,Descriptive histology,SD,Histological EvaluationDescri,Histological Evaluation,Descriptive histology,Histological EvaluationDescr,Histomorphometry,Histomorphometric analysis has been performed using,histological sections, microradiographs,and,backscattered electron microscopic (BSEM) images,(on plastic embedded surfaces).,Common Parameters:,Length, distance, area,and,numbers,HistomorphometryHistomorphom,Histomorphometry,Histomorphometry,Mechanical Testing,Mechanical properties are basic parameters of living tissues.,Indentation test,Push-out test,Mechanical Testing Mechanica,Special Techniques,Autoradiography, Bone Scan, and Scintigraphy,Computed Tomography,Magnetic Resonance Imaging,Single-Photon Absorptiometry, Single X-Ray Absorptiometry, Dual-Energy Absorptiometry,Ultrasound (US),Arthroscopy,Measuring Tissue Blood Flow,Special Techniques Autorad,Electron Microscopy,SEM,(commonly used),TEM,(less frequently),Electron MicroscopySEM (comm,Confocal Laser Scanning Microscopy,Laser penetrates in tissue: 300-500 m2D images to 3D or cross-sectional viewGood for tissue specimen or cells in wet condition.,Confocal Laser Scanning Mic,Biochemical Methods,Detecting Biochemicals in Tissues,Biochemical stains:,Safranin O and Alcian blue, ALP or ACP in bone and cartilage,Quantitative methods:,DNA content, Hydroxyproline content (a measure of collagen), Calcium, ALP, osteocalcin, collagen content, PGs or GAG synthesis and content,Biochemical Methods Detectin,Acknowledgment,Robert Draughn, DSc,Richard Friedman, MD,Christopher Hill, VMD,Qian Kang, MD,Jackie LaBreck, HT,Kylie Martin, HT,Frank Young, DSc,Post-doc Fellows,Residents,Students,AO Foundation,Arthritis Foundation,Biomet,ETEX Corp,Howmedica Osteonics,NIH,Marine Polymer Technologies,Oncore Orthopaedics,OREF,Sulzer,AcknowledgmentRobert Draughn,此课件下载可自行编辑修改，仅供参考！感谢您的支持，我们努力做得更好！谢谢,此课件下载可自行编辑修改，仅供参考！感谢您的支持，我们努力,