Index 571

INDEX

ABC immunostaining method, 307–308 Ankle cartilage Accelerating voltage autoradiographs, 427f TEM, 523–524 Apoptosis, 433, 434 Acid decalcifiers, 168 Aragonite, 514 Acid fuchsin Araldite/Embed-812, 386 mineralized bone, 317f Arthritis, 546, 546f osteoblasts, 316f degenerative titanium implant, 440 radiological examination, 535–541, woven bone, 316f 536f, 538f, 539f, 540f–541f Acid phosphatase (ACP) Arthroplasty histochemical staining, 316–319 total hip ACP radiological examination, 535–542 histochemical staining, 316–319 Articular cartilage, 83f, 415–416 Activation frequency, 395–396 animal models, 427–431 remodeling transients, 397 decalcification, 428–429 Adamantinoma, 483–484 lesion severity, 429–431 Adapted window, 70 sectioning and staining, 429 Adhesive, 10 tissue collection and fixation, 428 Adobe Photoshop, 381, 383 composition, 73–78 Agarose, 85 evaluation, 416 Aggrecan, 75, 75f freeze-fracture SEM, 50f Alcian blue, 16 functional aspects, 78–83 Alcian blue/PAS stain, 300 growth factors, 86 Alcohols, 160, 162–163 H&E stain, 46f, 49f Aldehydes, 160 human Alizarin complexone, 103 analysis, 423–427 Alizarin red S, 103 injuries, 83–88 Alizarins, 103–104 mechanical forces, 86–87 Alkaline phosphatase (ALP), 284, 443 mechanical properties, 79 histochemical staining, 316–319 variations, 79–83 osteoblasts, 318f micrograph, 418f Alkaline phosphatase-anti-alkaline phosphatase repair, 412–420 technique, 308f canine model, 417–418 ALP, 284, 443 failed, 418–420 histochemical staining, 316–319 histology, histochemistry, immunohis- osteoblasts, 318f tochemistry, 412 Aluminum intoxication, 400f histomorphometric evaluation, 412–413 Anatomical planes methods, 412–417 schematic, 145f resurfacing, 84 Anatomical terminology, 144 safranin O stain, 291f Aneurysmal bone cyst, 487–488 scaffold, 85 Ankle structure and function, 73–88 Collins grade, 426f tissue engineering, 85–86 t: table f: figure 571 572 Index

tissue types histogenesis, 65–70 light micrographs, 414f macroscopic structure, 59–62 transplantation, 84–85 microscopic structure, 62–64 trauma, 431–434 organization and constituents, 39–42 calcified tissue damage, 431 porosity, 41 cell viability, 431–434 SEM, 501 early matrix damage, 431 ultrastructure, 64–65 zonal arrangement, 76–77, 77f Bone balance, 67 Artifacts, 561–570 irreversible bone loss/gain, 69 autolysis, 563f Bone biopsy, 119–122 frozen sections, 568 arrangements, 121–124 hydroxyapatite coating, 569f–570f closed-needle transilial, 123 sectioning, 567f evaluation prior to, 135–136 routine decalcified paraffin-embedded inappropriate histology, 566–567 consequences, 129–130 Aspirin, 123 limitations, 120–121 Assay biopsy, 131–132, 140 outline, 138–141 Autolysis artifacts, 563f patient instructions, 122t Automatic microtome, 557f patient position, 125f Autoradiographs site, 125f ankle cartilage, 427f specimen handling, 126–127 Avascular necrosis technical aspects, 136–138 radiological examination, 541–546, 542f, site, 136–138 544f–545f timing, 120 transilial Backscattered electron microscopic (BSEM) specimen, 127f images, 19–21 types, 132–135 Band saw, 5f uses, 130–132 Barium sulfate Bone cement microangiography, 549–551 cement penetration study, 367–370 Barium sulfate and Chinese ink cement thickness study, 365–367 microangiography, 551 histomorphometric analysis, 361–372 Bending interface contact study, 362–363 decalcification, 169 interface pores, 365f Benign fibro-histiocytic tumors, 474 interface porosity study, 363–365 Benign lipomas, 485 radio-opaque, 367 Benign vascular tumors, 482–483 Bone dust, 565f BIAS image analyzer, 339 as artifact, 564 Biomet hand-driven core needle, 133 Bone-forming tumors, 463–473 Biopsy incision Bone healing. See Fracture healing proper vs improper, 138 Bone histomorphometry, 331–349 BioQuant, 238 histological methods, 332–336 Biphasic theory, 79 microanatomical methods, 336–349 Bisphosphonates physical methods, 331–332 postmenopausal osteoporosis, 396 software, 10 Blastic metastases, 485–486 Bone-implant interface Bone biocompatibility evaluation, 353–354 cell structure and biology, 35–42 histological analysis, 353–359 cellular components, 35–39 histomorphometric analysis, 356–359 Index 573

SEM, 505–510 Calcified cartilage examples, 507–508 evaluation, 416 sample preparation, 506–507 Calcified tissue TEM, 513–526 radiological examination, 531–546 analytical facilities, 525 case reports, 535–537 bone bonding to biomaterials, 514–515 methods, 531–535 evaluation, 523–526 Calcium implant materials, 513–515 stains, 150t interpreting micrographs, 525–526 Calcium phosphate (CaP) specimen grids and support films, bone interface 521–522 TEM, 520f ultrathin intact interfaces, 515–523 coated titanium fiber mesh, 261–262, 262f Bone-labeling techniques, 99–114 implants, 255–256, 256f Bone-lining cells, 36 synthetic, 513–514 Bone loss Cal-Ex, 211 primary hyperparathyroidism, 401 Calipers Bone marrow biopsy, 178 electronic, 10 Bone mass reversible changes Camera postmenopausal osteoporosis, 395–396 Sony DFW-V300 Digital Interface, 234 Bone mineral, 44–45 Cancellous bone, 337f Bone remodeling unit (BRU), 68f compact desktop micro-CT scan, 23f Bone sectioning vs cortical bone, 41 Exakt system, 243–251 decalcified histology, 562f modified inner diamond saw, 253–263 DEXA scanner Precise 1 automated cutting system, 265–269 CT, 41 staining techniques, 281–285 disintegration Bone tumors postmenopausal osteoporosis, 391–393 classification, 449 fluorochrome common associations, 451t animal studies, 110–111 grading and staging, 448–449 formation periods, 107f laboratory handling, 447–448 H&E stain, 37f primary, 449t loss mechanisms radiology, 449–450 postmenopausal osteoporosis, 393–396 Bone turnover modeling and remodeling, 67 reversible bone loss/gain, 68–69, 69f organization, 60f Bouin's solution, 163–164 photomicrographs, 348f Bromodeoxyuridine injections, 386 radiograph, 40f BRU, 68f remodeling, 393f BSEM images, 19–21 three-dimensional method, 392–393 Buehler Isomet 2000 Precision saw, 237f two-dimensional method, 392 Buffered formalin (BF) CaP. See Calcium phosphate (CaP) neutral, 11, 144, 159 Carbohydrates stains, 150t Calcein blue, 103 Carpal bone undecalcified iliac bone, 335f celloidin-embedded decalcification, 550f Calcein double-labeling, 100f Chinese ink, 550f Calcification front Cartilage, 45–51 histomorphometry and fluorochrome aggregate modulus, 82f staining, 334 alkaline phosphatase, 318f 574 Index

ankle sectioning, 554 autoradiographs, 427f specimen holder, 554 articular. See Articular cartilage staining, 554–555 calcified Cellular antigens, 309–310 evaluation, 416 Cemented femoral implant, 371f cellular components, 46–47 Cemented metal implant constituents and organization, 47–48 cutting technique, 272–273 decalcified. See Decalcified bone and Cemented prosthesis, 366f cartilage Cemented stem, 364f histological analysis, 423–434 Cemented total hip prosthesis, 238f hyaline Cement mantle, 369f articular cartilage repair, 412–413 thickness, 370f van Gieson's stain, 302f Cement-metal interface mechanical properties histomorphometric analysis, 361–372 diabetes mellitus, 84f Cement penetration study, 367–370 permeability, 50f Cement thickness study, 365–367 SEM, 501 Chelators, 169, 174, 177 semiquantitative grading scales, 291–292 Chemical fixation, 146 shear modulus, 82f Chemical storage, 25 Cartilage-forming tumors, 450–463 Chemical testing Cartilage matrix, 48f decalcification, 170 Cartilage sections Chinese ink basic staining techniques, 287–292 carpal bone, 550f histochemical and immunohistochemical epiphysis, 551f staining, 295–312 microangiography, 551 paraffin embedding, 296–297 Chondral defect tissue-embedding procedure, 296–297 deepening, 84 Cartilage stains, 290–291 Chondroblastoma, 453–454 Cartilage tissue Chondrocytes, 46–47, 74 histochemical findings, 301–304 hydrostatic pressure, 86–87 histochemical staining, 297–301 superficial zone, 78 immunohistochemical observations, 311 TEM, 499f immunohistochemical staining, 305–309 van Gieson's stain, 303f section pretreatment, 308–309 Chondroma, 452–453, 481–482 targets, 309–311 periosteal, 452f types, 295–296 Chondromyxoid fibroma, 454–455, 455f Celloidin Chondrosarcoma, 455–463 characteristics, 553 clear-cell, 459, 460f embedding, 191–192 dedifferentiated, 459–461, 461f infiltration, 191–192 differential diagnosis, 457–458 Celloidin-embedded decalcification grading, 456–457 carpal bone, 550f low grade, 457f hip, 550f management, 458 radius, 550f mesenchymal, 461–462, 462f ulna, 550f vs Ewing's sarcoma, 478 Celloidin-embedded specimens, 553–555 Circular diamond saws, 4–6, 6f embedding, 553–554 materials and methods, 254–255 infiltration, 553–554 schematic view, 254f mounting microvascular slices, 554 Clear-cell chondrosarcoma, 459, 460f Index 575

Clearing, 11–15, 188–189 Cutting and grinding, 233–239 Closed-needle transilial bone biopsy, 123 fixation, 234–235 CLSM, 22–23 interpretation, 236–239 Cole's hematoxylin 17, 17 Cutting and polishing Collagen, 47, 74, 85, 514 bone-implant interface, 507 superficial zone, 78 Cutting machine, 221 Collagenous connective tissue Cysts, 486–488 staining, 298 Collins grade, 425f Decalcification, 167–183, 287–290, 552–553 Collins score, 425 components/action, 168–169 Compact bone, 59 end point determination, 169–170 Compact desktop micro-CT scanners, 22, 23f Gill's II H&E stain, 181f, 182f Composite bone/soft tissue, 180 human tibia, 171f Composite modeling, 45 improper Compression, 87 routine decalcified paraffin-embedded Computer-based image analysis histology, 564–565 threaded implant, 359 Masson's trichrome stain, 181f Computerized tomography (CT), 22 processing protocol, 190, 191t cancellous bone radiographs, 212f compact desktop scanners, 22, 23f in situ hybridization, 323 DEXA scanner, 41 specimen preparation/management, 170–172 quantitative, 19, 22, 44 TEM, 497–499 Condenser lens aperture Decalcified bone and cartilage TEM, 524 completion testing, 211–213 Confirmatory biopsy, 132, 140 decalcification, 210–211 Confocal laser scanning microscopy (CLSM), further processing, 213–214 22–23 histological techniques, 209–218 Connective tissue rough cut and fixation, 209–210 stains, 150t section cutting and precautions, 214–216 ConnEuler method, 344 Decalcified cartilage. See Decalcified bone and Contrast media cartilage microangiography, 549–551 Decalcified preparations, 282 Copper Decalcified tissue block, 215f grids, 521–522 thicker sections, 216f stains, 151t Decalcifiers Core needle biopsy, 133 chemical end point determination, 175–176 Cortical bone fluid volume, 174–175 vs cancellous bone, 41 selection, 172–174 DEXA scan, 40f suspension, heat, agitation, vacuum, 175 fluorochrome Decorin, 76 animal studies, 110–111 Dedifferentiated chondrosarcoma, 459–461, modeling and remodeling, 67 461f photomicrograph, 283f Defatting, 553 radiograph, 40f bone-implant interface, 506–507 Coumadin, 124 Degenerative arthritis Craig Core Needle, 133, 139 radiological examination, 535–541, 536f, Craig-type closed needle, 447 538f, 539f, 540f–541f CT. See Computerized tomography (CT) Dehydration, 11–15, 187–188, 188t, 200, 553 Custom Prostheses Magnification Marker, 380 bone-implant interface, 506–507 576 Index

inadequate ECM, 42–45, 64 routine decalcified paraffin-embedded antigens, 310 histology, 565 Ecomet III, 386 PMMA embedding, 235t EDTA, 169, 174, 210 in situ hybridization, 323 Elastica-van Gieson's stain, 299 Spurr's embedding, 236t Elastic fibers Demeclocycline, 101 stains, 150t, 298–299 DEPC Electronic calipers, 10 in situ hybridization, 322 Electronic microtomes, 10 Desiccators, 10 Electron Spectroscopy for Chemical Analysis Desmoplastic fibroma, 473, 474f (ESCA), 444 DEXA, 22 Elongation Diabetes mellitus fluorescence photomicrograph, 112f–113f articular cartilage, 81 fluorochrome cartilage animal studies, 111 mechanical properties, 84f Embed-812, 386 Diamond-coated wire saw unit, 14 Embedding, 185–196, 287–290 Diamond knife, 518 bone-implant interface, 507 SEM, 518f cartilage sections, 296–297 Diamond-plated lapping disks, 376 celloidin, 191–192 Diamond saws, 4–5 celloidin-embedded specimens, 553–554 bone sectioning, 253–263 inadequate materials and methods, 254–255 routine decalcified paraffin-embedded results, 255–262 histology, 565 schematic view, 254f paraffin, 188–190 Diamond wire saws, 6, 7f, 14f plastics, 192–196 Diaphysis containers, 193f microphotograph, 556f resin Diazepam cartilage sections, 297 for bone biopsy, 124 in situ hybridization, 323–324 Diethyl pyrocarbonate (DEPC) Spurr's in situ hybridization, 322 dehydration, 236t Digital camera TEM, 497–499 Sony DFW-V300 Digital Interface camera, 234 ultrathin intact interfaces, 516–517 DIG RNA labeling and detection kit undecalcified bone tissue sectioning, 224–225 in situ hybridization, 322 Emergency deluge shower, 25 Directional terms, 145t Enchondroma protuberans, 452 Dissection Endochondral ossification, 65–66 in situ hybridization, 323 Endocortical surface Dissection and processing form, 153–154 trabecularization, 69–70 Dissection techniques, 143–144 Enzyme histochemistry, 284 Disuse window, 70 Enzyme staining procedures, 16–17 Doxycycline, 123 Epiphysis D-profile knife, 332 celloidin-embedded decalcification, 551f Dremel rotary tool, 4–6 Chinese ink, 551f Dual-energy X-ray absorptiometry (DEXA), 22 Epon, 204–205, 386 EPTFE membranes, 442, 443f Erosion Easy-Cast BN-72, 366 measurement, 395 Ecchordosis physaliphora, 481 ESCA, 444 Index 577

Estrogen cemented implant, 363f articular cartilage, 81 gross slab, 179f, 180f postmenopausal osteoporosis, 396–397 immersion fixed vs perfusion fixed, 149f Ethanol, 234 Masson’s-trichrome stain Ethylenediaminetetraacetic acid (EDTA), 169, photomicrograph, 282f 174, 210 microphotograph, 555f Ewing's sarcoma, 476–478, 477f microradiography, 22f differential diagnosis, 478 Fibrin Exakt 401, 247 stains, 151t Exakt band-cutting system, 244–248 Fibrocartilage, 302 Exakt band system, 250 articular cartilage repair, 412 Exakt 400 CL grinding system, 246f meniscus surface preparation, 249 Elastica-van Gieson's stain, 303f Exakt 400 CL microgrinding system, 245–247 Fibrogenic tumors, 473–475 Exakt 300 CP band system, 245f Fibro-histiocytic tumors, 473–475 Exakt 400 CS grinding system, 246f, 250–251 benign, 474 surface preparation, 249 Fibroma Exakt 400 CS microgrinding system, 245–247 desmoplastic, 473, 474f Exakt 402 precision adhesive press, 247–248, Fibronectin, 37f 247f Fibrosarcoma, 473–474 Exakt System, 6–7, 243–251 Fibrous dysplasia, 489 disposable materials, 248 Fibrous tissue pre-bone sectioning, 243–244 articular cartilage repair, 412 procedure, 248–251 Film, 533 steps, 244 Fine-needle aspiration, 447 Exakt Vacuum Adhesive Unit, 249 Fixation, 144–151, 162f, 287–290, 552 Excisional biopsy, 140 articular cartilage, 428 Expanded polytetrafluoroethylene (ePTFE) bone-implant interface, 506–507 membranes, 442, 443f chemical, 146 Explanted biomaterials cutting and grinding, 234–235 fixation, 151 decalcified bone and cartilage, 209–210 Extracellular ground substance explanted biomaterials, 151 staining, 298–299 factors affecting, 160–161 Extracellular matrix (ECM), 42–45, 64 heat, 146 antigens, 310 methods Eyewash station, 25 TEM, 497–499 perfusion, 149, 149f Fading, 106 lower extremities, 154–155 Failed articular cartilage repair, 418–420 routine decalcified paraffin-embedded Fast green stain, 217–218, 218f, 300 histology, 563–564 Fats in situ hybridization, 323 stains, 150t time, 148 Faxitron, 212 ultrathin intact interfaces, 515–523 Faxitron 43855A Cabinet X-Ray System, 4, 4f, undecalcified bone tissue sectioning, 222 376 Fixatives, 145–151, 150t, 159–164 Faxitron Cabinet X-Ray System, 3 advantages and disadvantages, 147t Faxitron Specimen Radiography System Model components, 147t MX-20, 4 concentrations, 146–147 Femur, 178–180 formulations, 155–157 biopsy tract, 140f groups, 160 penetration, 160f Fresh frozen specimens primary, 160 histomorphometry, 332–333 classification, 161t Frozen sections safety, 151 artifacts, 568 Fixed specimen cartilage sections, 296 cutting, slabbing, radiography, 552 Fume hood, 25 Fluoresceins, 103 Fungi Fluorescent microscopy stains, 151t staining and new bone formation, 377–379 Fluorochrome GAG, 74–78, 514 animal studies, 110–111 Geometrically Deformable Model (GDM), 367 human studies, 110 Giant-cell tumor, 478–481, 479f subcutaneous administration, 355t differential diagnosis, 480–481 Fluorochrome-labeled specimens vs osteosarcoma, 469 histological processing, 105 Giemsa stain, 16, 238t Fluorochrome labels, 354–355 Gill's II H&E stain epifluorescent microscopy, 105–106 decalcification, 181f, 182f Fluorochromes, 99–106, 102f Glass bottles administration route, 104–105 for tissue dehydration, 188f Formalin, 161–162 Glass scintillation vial, 224f Formalin-fixed tissue Glove powder TEM, 497 as artifact, 568 Formalin pigment, 566f Glucocorticoid-induced osteoporosis, 398 as artifact, 564 Glutaraldehyde, 163, 234 Formation period (FP), 108 Glycol methacrylate, 194–195 MI ratio, 108, 109f Glycosaminoglycans (GAG), 74–78, 514 Formic acid, 174, 176–177 Gnathic osteosarcomas, 466–467 Formvar, 522 Gold Fractal analysis, 344 stains, 151t Fracture callus Goldenhar's trichome stain Goldner's stain iliac crest bone, 400f photomicrograph, 289f Goldner's Masson trichrome stain, 17 Fracture gap, 378f Goldner's method, 340 Fracture healing Goldner's trichrome stain, 16 callus area, 380 bone remodeling, 16f cell kinetics, 385–387 iliac crest bone, 400f DNA denaturation, 386 Good Laboratory Practice, 23–24 fracture union, 381–382 Grinding, 233–239 histologic analysis, 375–377 Ground sections, 284 specimen preparation, 375 Growth factors immunohistochemical staining, 386–387 articular cartilage, 86 microangiographic analysis, 384–385, 385f Growth hormone, 404 microradiography, 375–377, 376f section preparation, 386 tissue type, 380–381 HA. See Hydroxyapatite (HA) trabecular bone orientation within fracture Hard-tissue sectioning gap, 383–384 Precise 1 system, 268 Fracture union Haversian canals, 62–63 bone healing, 381–382 Healing fracture Frank Read mechanism, 519 tetracycline, 379f Heat fixation, 146 bone interface, 524f Helical scanner Hyperparathyroidism HiSpeed Advantage, 367 primary, 398–401, 400f Hemangioendothelioma, 483 bone loss, 401 Hemangiomas, 482–483 bone mass and structure, 399 Hematin bone remodeling, 399–401 stains, 151t Hematoxylin & eosin (H&E) staining, 16, 17 Iliac bones, 61 High-grade surface osteosarcoma, 472 Iliac crest bone High-pressure water cutter, 271–276 Goldenhar's trichome stain, 400f schematic, 273f MMA-embedded specimen Hip Goldner's stain, 282f celloidin-embedded decalcification, 550f photomicrograph, 283f prosthesis, 239f Image for Windows, 381 HiSpeed Advantage helical scanner, 367 ImageQUEST, 234 Histiocytomas Immunochemistry, 284–285 malignant fibrous, 475 Immunohistochemical staining, 17 Histological evaluation, 19 Impaired wound healing, 257–258 Histologic grading scales, 356–357, 357t Improper decalcification Histomorphometric analysis, 19 routine decalcified paraffin-embedded Histomorphometric grading histology, 564–565 inter-observer error during, 416–417 Inadequate dehydration Histomorphometry, 19–20 routine decalcified paraffin-embedded Histostain-SP mouse universal kit, 386 histology, 565 Holey Formvar films, 522 Inadequate embedding Homer-Wright pseudo-rosettes, 478 routine decalcified paraffin-embedded Human articular cartilage histology, 565 analysis, 423–427 Inappropriate bone biopsy evaluation rationale, 423–424 consequences, 129–130 gene expression assessment, 426–427 Incisional biopsy, 133–135 histological assessment, 424–426 India ink, 16 joint grading scale, 424 Infiltration, 188–189, 200–201 Human tibia celloidin, 191–192 decalcification, 171f celloidin-embedded specimens, 553–554 Humerus plastics, 192–196 biopsy tract, 137f techniques, 199–206 Hyaline cartilage In situ hybridization, 321–326 articular cartilage repair, 412–413 decalcification, 323 osteoarthritic equipment and reagents, 322 safranin O stain, 305f human articular cartilage, 426–427 van Gieson's stain, 302f hybridization and wash, 324–325 Hydrochloric acid, 173, 176 hybridized probe detection, 325–326 Hydrostatic pressure method selection, 322t chondrocytes, 86–87 probe preparation, 324 Hydroxyapatite (HA), 64, 514 specimen preparation, 323–324 coatings, 258–259 Intermittent parathyroid hormone treatment, PS implant, 260f 402–403 SEM, 520f Intervertebral disc powder, 517f degeneration, 304–305, 311 TEM, 519f, 523f, 526f TEM, 498f 580 Index

Intramembranous ossification, 65 Lymphoma, 475–476 Intraosseous lesion vs Ewing's sarcoma, 478 biopsy, 139f Lytic metastases, 485–486 Intraosseous schwannomas, 485 Intra osseous well-differentiated osteosarcoma, 471 Macrophage colony-stimulating factor (M- Iron CSF), 38 stains, 151t Maffucci's syndrome, 452 Isomet low-speed saw, 4–6, 5f Magnification calibration TEM, 525 Jawbones Malignant fibrous histiocytomas, 475 osteosarcoma, 466–468 Manipulation Joint grading scale decalcification, 169 human articular cartilage, 424 Mankin score, 424, 425 MAR, 108, 377 Keratinocytes Marker interval (MI) SEM, 444f FP ratio, 108, 109f Kilovoltage, 533 Marrow space star volume, 344 Knee Masson's trichrome stain, 16, 298 Collins grade, 426f decalcification, 181f femur Label escape error, 107–108, 109f photomicrograph, 282f Labeling regimens, 106–114 Goldner's, 17 Laboratory safety, 24–25 physis Lamellar bone, 42, 62 photomicrograph, 289f renewal, 68f Matrix flow Lamina splendens, 77 articular cartilage, 415 Lead M-CSF, 38 stains, 151t Media Lead citrate, 522 Spurr's Leica SM2500 heavy-duty sliding microtome, 9f embedding protocol, 195t Leica SM2500 microtome, 8 Megacassettes, 10 Leica SP1600 microtome, 8, 10f Meniscus fibrocartilage Lipid deposits Elastica-van Gieson's stain, 303f staining, 300–301 Meperidine Lipids for bone biopsy, 122–123, 124 stains, 150t Mercurials, 160 Lipogenic tumors, 485 Mesenchymal chondrosarcoma, 461–462, 462f Lipomas vs Ewing's sarcoma, 478 benign, 485 Mesenchymal stem cells (MSC), 85 LKB PMV 2258 heavy-duty cryomicrotome, Metabolic bone conditions 332 histomorphometry, 391–405 London Resin (LR) White Resin, 205 radiograph, 535 Long bone Metaphyseal fibrous defect, 488 diaphysis, 60–61 Metastases, 485–486 Longitudinal growth, 66 blastic, 485–486 Lower extremities to bone, 485 perfusion fixation, 154–155 systemic effects, 486 Low grade chondrosarcoma, 457f Metastatic neuroblastoma LR White Resin, 205 vs Ewing's sarcoma, 478 Lymphangioma, 482–483 Methacrylates, 193 Index 581

Methylene blue/basic fuchsin stain, 16, 217 Mitutoyo Surftest 211 Profilometer, 444 Methyl methacrylate (MMA), 555–556 ML-521D, 376 embedding MMA, 555–556 modified Goldner's trichrome method, embedding 334t modified Goldner's trichrome method, undecalcified bone, 333t 334t Methylprednisolone undecalcified bone, 333t articular cartilage, 81 Modeling, 51, 66–70 MI Model Micro 50, 234 FP ratio, 108, 109f Modified Goldner's trichrome method Microangiography, 549–552 methylmethacrylate embedding, 334t amount of material injected, 552 Modified inner circular sawing technique, 14 artery selection, 552 Modified inner diamond saw barium sulfate, 549–551 bone sectioning, 253–263 barium sulfate and Chinese ink, 551 materials and methods, 254–255 Chinese ink, 551 results, 255–262 contrast media, 549–551 Moisture chamber fracture healing, 385f in situ hybridization, 322 Microgrinding Molds final results, 251f paraffin embedding, 190f Microradiography, 355–356, 533 small glass bottles, 194f bone healing, 375–377, 376f Multifocal osteosarcoma, 470 femur, 22f Multiple labeling, 113–114 fracture healing, 375–377, 376f Multiply connected trabecular system, 347f tibia, 381f Myeloma, 475 Microscopic images digitization, 381 National Institutes of Health (NIH) Image, Microslice 2, 346 369, 377, 380, 383 Microtome, 8–9 Necrosis, 433 automatic, 557f Needle biopsy, 139 blade, 179f Neuroblastoma cutting metastatic schematic illustration, 230f vs Ewing's sarcoma, 478 electronic, 10 Neurogenic tumors, 485 knives, 10, 227–229 Neutral buffered formalin, 11, 144, 159 rotary, 8–9, 9f, 226, 227f New bone formation rotating disc, 8 tetracycline, 380f sliding, 8 xylenol orange, 380f undecalcified bone tissue sectioning, 226–231 NF-KB, 38 Microvascular quantification, 556–557 NIH Image, 369, 377, 380, 383 Microvascular slices Nitric acid, 173, 176 celloidin-embedded specimens, 554 Non-coated Ti implants, 256–257 Mild overload window, 70 Nuclear factor kappa B (NF-KB), 38 Millonig's fixative, 126 Nucleus pulposus, 302 Mineral apposition rate (MAR), 108, 377 Mineralization defect Objective lens aperture osteomalacia, 402 TEM, 524–525 Mineralized bone Ollier's disease, 452 acid fuchsin, 317f Open biopsy, 133, 139 silver nitrate, 317f Open incisional biopsy, 447 582 Index

OPGL, 38 vs Ewing's sarcoma, 478 Optomax V AMS, 339 telangiectatic, 470–471 Orcein stain, 298 Osteotomy Osteoarthritic hyaline cartilage light microscopy, 383f safranin O stain, 305f Overfixation, 148–149 Osteoarthritis, 305f, 311 Overlapping, 14–15 radiological examination, 535–541, 536f, Oxidizing agents, 160 538f, 539f, 540f–541f Osteoarthrosis, 304 Paraffin Osteoblast-like cell embedding, 188–190, 555 SEM, 444f cartilage sections, 296–297 Osteoblastoma, 464–465 molds, 190f vs osteosarcoma, 469 melting points, 189 Osteoblasts, 36, 38 Paraformaldehyde, 234 acid fuchsin, 316f Paraostial osteosarcoma, 472–473 alkaline phosphatase, 318f Paraplast, 189 silver nitrate, 318f Parathyroid hormone treatment toluidine blue stain, 316f intermittent, 402–403 Osteocalcin, 443 Pathologic overload window, 70 Osteochondroma, 450–452 Patient instructions Osteoclast differentiation factor, 38 bone biopsy, 122t Osteoclastoma. See Giant-cell tumor Patient position Osteoclasts, 38 bone biopsy, 125f Osteocytes, 36, 38 PBS, 18 H&E stain, 36f Peacock Micrometer, 376 Osteoid osteoma, 463–464, 464f Pedicles, 367–368 Osteomalacia, 400f, 401–402 Peel-A-Way plastic mold, 10, 11f, 193, 193f mineralization defect, 402 Percutaneous implant, 259f OsteoMeasure system, 335–336, 337f, 338t, Percutaneous transilial closed needle bone 339, 378, 379, 381 biopsy, 124 Osteons, 62 Perfusion fixation, 149, 149f cancellous, 63f lower extremities, 154–155 cross section, 63f Periodic acid-Schiff, 16 Osteopontin Periosteal chondroma, 452f in situ hybridization, 326f Periosteal osteosarcoma, 471–472 Osteoporosis, 400f Periprosthetic bone glucocorticoid-induced, 398 backscatter SEM, 43f Osteoprotegerin (OPGL), 38 Perkin-Elmer PHI 5500 ESCA System Osteosarcoma, 465–473, 467f–468f instrument, 444 differential diagnosis, 469 Permacol type 370, 255 following chemotherapy, 470f Peroxidase techniques, 307–308 gnathic, 466–467 Personnel high-grade surface, 472 laboratory safety, 24–25 intra-osseous well-differentiated, 471 Phosphate-buffered saline (PBS), 18 jawbones, 466–468 Photomicrograph management, 469 iliac crest bone, 283f multifocal, 470 Physis paraostial, 472–473 Masson's trichrome stain periosteal, 471–472 photomicrograph, 289f small-cell, 471 PAS stain, 290f Index 583

Picrates, 160 hard-tissue sectioning, 268 Plane parallel block problems and precautions, 268 mounting, 249 Primary bone tumors, 449t producing, 248 conditions simulating, 485–489 Plane parallel plexiglas slide Primary fixatives, 160 affixing on block surface, 249–250 classification, 161t Plasma sprayed (PS) implants Primary hyperparathyroidism, 398–401, 400f HA coated, 260f bone loss, 401 Plastic-embedded specimens bone mass and structure, 399 histomorphometry, 332–333 bone remodeling, 399–401 300 mm thick slices, 348t Probing staining techniques, 315–318 decalcification, 169 Plastic resins, 201–206 Profile D knife, 229f Plastics Prostaglandin E2, 443 embedding, 192–196 Proteinase K, 434 containers, 193f Proteoglycans, 47–48, 75–76, 77 infiltration, 192–196 anions, 76f Plastic support films, 522 Prussian blue stain, 301 PMMA Pseudocapsule tissue, 39f embedded dehydration, 235t PS implants embedding HA coated, 260f dehydration, 235t gas bubbles in, 225f QCT, 19, 22, 41 Polishing Quantitative analysis, 383 bone-implant interface, 507 Quantitative CT (QCT), 19, 22, 41 Polycut E 5000, 386 Quantitative elemental analysis, 501–502 Poly(DL-lactic acid) (PLA), 85 Quantitative interfacial analysis, 358–359 Poly(glycolic acid) (PGA), 85 Quenching, 106 Polylactic acid, 442f Polymethylmethacrylate (PMMA), 148–149 Radiographic union score, 382t embedded dehydration, 235t Radiographs embedding avascular necrosis, 541–546, 542f, 544f–545f dehydration, 235t decalcification, 169, 212f gas bubbles in, 225f degenerative arthritis, 537–541, 538f, 539f, Porosity, 377 540f–541f Post-decalcification, 180–183 interpretation, 534 Postmenopausal osteoporosis, 391–398 metabolic bone conditions, 535 anti-resorptive treatment, 396–397 osteoarthritis, 537–541, 538f, 539f, 540f– bone mass reversible changes, 395–396 541f cancellous bone disintegration, 391–393 total hip arthroplasty, 535–542, 536f, 538f, cancellous bone loss mechanisms, 393–396 539f, 540f–541f, 542f missing data and potential bias, 397 Radio-opaque bone cement, 367 sample size and design, 397 Radius Potter's Plaster, 369 celloidin-embedded decalcification, 550f Precise 1 automated cutting system, 266f Random undulating parameter (RUP), 370, bone sectioning, 265–269 372f Precise 1 grinding system, 266–267, 266f RANKL, 38 Precise system RANK ligand (RANKL), 38 advantages, 267–268 Rapid undecalcified cryosections, 333t Precise 1 system Refrigerant tanks 584 Index

TEM, 525 band, 5f Remodeling, 66–70, 432 circular diamond, 4–6 fluorochrome materials and methods, 254–255 animal studies, 110–111 schematic view, 254f Goldner's trichrome stain, 16f Isomet low-speed, 4–6, 5f histomorphometry, 335–336 Scaffold primary hyperparathyroidism, 399–401 articular cartilage, 85 Remodeling balance Scanning electron microscopy (SEM) cancellous bone, 393–394 bone, 501 postmenopausal osteoporosis, 394–395 bone-implant interface, 505–510 Resin, 201f cartilage, 501 Resin embedding images, 19–21 cartilage sections, 297 methods, 499–501 Resorption Schwannomas fluorochrome intraosseous, 485 animal studies, 111–113 Scion Image, 381, 383 SEM, 500f Secondary hyperparathyroidism, 400f Reticular fibrils Section staining, 298 thickness of, 14–15, 15f Reticulin stain, 299 Sectioning Rotary microtomes, 8–9, 9f, 226, 227f celloidin-embedded specimens, 554 Rotating disc microtome, 8, 9f Sectioning artifacts, 567f Rough titanium-implant surface routine decalcified paraffin-embedded fluorescence micrograph, 355f histology, 566–567 Routine decalcified paraffin-embedded Sectioning bones histology, 561–568 microtome types, 226–227 fixation, 563–564 in situ hybridization, 323–324 improper decalcification, 564–565 Sectioning planes, 186f inadequate dehydration or embedding, 565 Selective estrogen receptor modulators obtaining sample, 562–563 postmenopausal osteoporosis, 396–397 sectioning artifacts, 566–567 SEM. See Scanning electron microscopy staining, 567–568 (SEM) RTV silicone rubber, 366 Serial selection technique, 344–345 Ruffled membrane, 38 videorecording, 346f Ruthenium hexamine trichloride, 498 Shear, 87 Shoulder prosthesis Safety water cutter, 276f fixatives, 151 Silver nitrate Safranin O/fast green stain, 217–218, 218f, 300 mineralized bone, 317f Safranin O stain, 16 osteoblasts, 318f articular cartilage, 291f Simple bone cyst, 486–487 osteoarthritic hyaline cartilage, 305f Skewed sampling error, 107–108 Säge-Schliff technique, 243 Sliding microtomes, 8 Saltykov's method of directed secants, 383 SMA Sarcoma immunohistochemical micrograph, 419f soft-tissue Small-cell osteosarcoma, 471 growth pattern, 134f vs Ewing's sarcoma, 478 Sawing methods, 14 Small needle biopsy, 132–133 Sawing system, 8 Smooth muscle actin (SMA) Saws, 14f, 237f immunohistochemical micrograph, 419f Index 585

Soft-tissue biomaterial interface Strong acid decalcifiers, 173, 176 cell cultures and biocompatibility tests, Subchondral bone 443–445 evaluation, 416 histological analysis, 439–445 Subchondral (synovial) cyst, 487 Soft-tissue sarcoma Subjective light microscopy, 354 growth pattern, 134f Sulcular epithelium, 439 Sony CCD-IRIS Video, 238 Superficial zone, 49 Sony DFW-V300 Digital Interface camera, 234 chondrocytes, 78 Sony DXC-390, 381 collagen, 78 Specimen Super glue, 10 accessioning, 233–234 Support films, 521–522 bone biopsy, 126–127 Surface decalcification, 215f celloidin-embedded, 553–555 Surgical dissecting instruments, 3–10 detachment, 202f Synovial cyst, 487 dissection, 234 Synthetic calcium phosphates, 513–514 fixed cutting, slabbing, radiography, 552 Talus fluorochrome-labeled gross photograph, 425f histological processing, 105 Tartrate-resistant acid phosphatase localization, grids, 521–522 284 holder TAS, 339–344, 340f celloidin-embedded specimens, 554 artifacts, 341f orientation, trimming, fixation, decalcifica- data printout, 342f tion, 186–187 Technovit 4000, 249 plastic-embedded Technovit 3040 glue, 10 histomorphometry, 332–333 Technovit 7200 Kulzer, 201–203 300 mm thick slices, 348t Technovit 8100 Kulzer, 203–204 staining techniques, 315–318 Technovit 9100 Kulzer, 204 post-fixation treatment, 164 Technovit 7200 VLC, 248 preparation, 552–553 Telangiectatic osteosarcoma, 470–471 stages, 200–201 TEM. See Transmission electron microscopy storage, 164 (TEM) transilial bone biopsy, 127f Tetracycline, 101–103 Speed-lapping machine, 376 allergic to, 123 Spongy bone, 59 healing fracture, 379f Spurr's embedding labeling, 16 dehydration, 236t new bone formation, 377, 380f Spurr's media undecalcified iliac bone, 335f embedding protocol, 195t Therapeutic biopsy, 132, 140–141 preparation, 195t Thermal artifact, 563f Spurr's plastic, 236 Thick-slice technique, 345–349 Spurr's resin, 195–196, 205–206, 237f Threaded implant Staining computer-based image analysis, 359 celloidin-embedded specimens, 554–555 3-triethoxylosilylpropylamine procedures, 16–17 in situ hybridization, 322 bone sections, 281–285 Thyroid hormones, 404–405 routine decalcified paraffin-embedded Ti. See Titanium (Ti) histology, 567–568 Tibia Stains. See individual stains decalcification, 171f Stains-All, 16 longitudinal section, 61f 586 Index

microradiography, 381f Trabecular system Tidemark, 49, 432 multiply connected, 347f Timed immersion Transforming growth factor beta, 443 decalcification, 169–170 Transilial bone biopsy Tissue cassettes, 189f specimen, 127f Tissue section Transilial closed needle bone biopsy, 124 transferring onto microslide, 216f Transitional zone, 49 Titanium (Ti), 514 Transmission electron microscopy (TEM) alloy, 514 accelerating voltage, 523–524 bone interface bone-implant interface, 513–526 TEM, 521f condenser lens aperture, 524 fiber mesh decalcification, 497–499 porosity, 259f embedding, 497–499 implant, 257f filling refrigerant tanks, 525 acid fuchsin, 440 fixation methods, 497–499 light micrograph, 357f formalin-fixed tissue, 497 toluidine blue, 440 hydroxyapatite-bone interface, 524f mesh implant, 261f, 442f magnification calibration, 525 osteogenic activity, 261–262 method, 497–499 oxygen grit-blasted implants, 255 objective lens aperture, 524–525 Toluidine blue stain, 16, 315 optimal operation, 523–526 osteoblasts, 316f specimen retrieval from formalin and procedure, 333t paraffin, 502 titanium implant, 440 Transverse biopsy incision, 137f woven bone, 316f Tree rings, 123f Torrex Cabinet X-Ray System, 3 Triethoxylosilylpropylamine Total hip arthroplasty in situ hybridization, 322 radiological examination, 535–542, 536f, Trimming, 553 538f, 539f, 540f–541f, 542f Tru-cut closed needle, 447 Total new bone determination, 378–379 Trylon coloring paste, 345 Trabecular Analysis System (TAS), 339–344, TUNEL method, 434 340f 2D histomorphometric analysis, 19 artifacts, 341f Type I collagen, 64–65 data printout, 342f Trabecular architecture Ulna three-dimensional image, 344–349 celloidin-embedded decalcification, 550f two-dimensional image, 336–344 Ultramicrotomy Trabecular bone ultrathin intact interfaces, 517–518 orientation within fracture gap, 383–384 Undecalcified bone Trabecular bone volume methylmethacrylate embedding, 333t SEM, 18f Undecalcified bone tissue sectioning Trabecular elements block trimming, 225–226 irreversible loss, 69–70 bone preparation, 222–237 Trabecular indices dehydration, 223 mathematical equations, 343t embedding, 224–225 Trabecular perforations fixation, 222 cancellous bone microtomes, 226–231 postmenopausal osteoporosis, 393–394 microtome sectioning, 226–231 Trabecular star volume resin infiltration, 223 measurement, 392, 392f section preparation, 229–230 Index 587

Undecalcified histology, 569 schematic, 273f Undecalcified iliac bone Water jet cutter, 271–272, 272f calcein blue, 335f cemented metal implant tetracycline, 335f cutting technique, 272–273 Undecalcified preparations, 283–284 cutting technique, 271–272 Underfixation, 148–149 Weak acid decalcifiers, 174, 176–177 Unicameral bone cyst (UBC), 486–487 Well 4240, 7f Uranyl acetate, 522 Well diamond wire saw, 6 Well 3241 diamond wire saw, 14f Vacuum-adhesive system, 247 Well 4240 diamond wire saw, 14f Vacuum pumps, 10, 202f Well 3242 Precision diamond wire saw, 7f Van Gieson's stain, 298 Well 3241 Precision Wire Saw, 14 Vascular tumors Well 4240 Precision Wire Saw, 14 benign, 482–483 Wheel grinder/polisher, 6, 8f Vasoformative tumors, 482–483 Wound healing Vertebral bones, 61 impaired, 257–258 Video Woven bone, 42, 62 Sony CCD-IRIS Video, 238 acid fuchsin, 316f VIP image analyzer, 340 backscatter SEM, 43f Vital staining, 17–18 H&E stain, 42f Vitronectin, 37f toluidine blue stain, 316f Von Kossa/Eosin method, 333t Von Kossa stain, 16, 301 X-ray machine, 3–4 X-ray Photoelectron Spectroscopy (XPS), Wall thickness 443–444 measurement, 395 Xylenol orange, 104 Water cutting new bone formation, 380f cemented stem, 274f characteristics, 273–276 Zeiss MOP 3 digitizer, 339 high-pressure, 271–276 Zimmer counter