• 1.

    Sawin PD, Traynelis VC, Menezes AH. A comparative analysis of fusion rates and donor-site morbidity for autogeneic rib and iliac crest bone grafts in posterior cervical fusions. J Neurosurg 1998;88:255265.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 2.

    Hu RW, Bohlmann HH. Fracture at the iliac bone harvest site after fusion of the spine. Clin Orthop 1994;309:208213.

  • 3.

    Goulet JA, Senunas LE & DeSilva GL, et al. Autogenous iliac crest bone graft: complications and functional assessment. Clin Orthop 1997;339:7681.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 4.

    Millis DL, Martinez SA. Bone grafts.. In: Textbook of small animal surgery. 3rd ed. Philadelphia: WB Saunders Co, 2003;1876.

  • 5.

    Weibrich G, Kleis WK & Buch R, et al. The Harvest Smart PReP™ system versus the Friadent-Schutze platelet-rich plasma kit. Clin Oral Implants Res 2003;14:233239.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 6.

    Weibrich G, Kleis WK. Curasan PRP kit versus PCCS PRP system. Clin Oral Implants Res 2002;13:437443.

  • 7.

    Kevy SV, Jacobson MS. Comparison of methods for point of care preparation of autologous platelet gel. J Extra Corpor Technol 2004;36:2835.

    • Search Google Scholar
    • Export Citation
  • 8.

    Hauschild G, Merten HA & Bader A, et al. Bioartificial bone grafting: tarsal fusion in a dog using a bioartificial composite bone graft consisting of β-tricalcium phosphate and platelet rich plasma—a case report. Vet Comp Orthop Traumatol 2005;18:5254.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 9.

    Southwood LL, Frisbie DD & Kawcak CE, et al. Delivery of growth factors using gene therapy to enhance bone healing. Vet Surg 2004;33:565578.

  • 10.

    Weibrich G, Kleis WK & Hafner G, et al. Growth factor levels in platelet-rich plasma and correlations with donor age, sex, and platelet count. J Craniomaxillofac Surg 2002;30:97102.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11.

    Okuda K, Kawase T & Momose M, et al. Platelet-rich plasma contains high levels of platelet-derived growth factor and transforming growth factor-beta and modulates the proliferation of periodontally related cells in vitro. J Periodontol 2003;74:849857.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 12.

    Lucarelli E, Beccheroni A & Donati D, et al. Platelet-derived growth factors enhance proliferation of human stromal stem cells. Biomaterials 2003;24:30953100.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 13.

    Fennis JP, Stoelinga PJ, Jansen JA. Mandibular reconstruction: a clinical and radiographic animal study on the use of autogenous scaffolds and platelet-rich plasma. Inter J Oral Maxillofac Surg 2002;31:281286.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 14.

    Fuerst G, Gruber R & Tangl S, et al. Enhanced bone-to-implant contact by platelet-released growth factors in mandibular cortical bone: a histomorphometric study in minipigs. Inter J Oral Maxillofac Implants 2003;18:685690.

    • Search Google Scholar
    • Export Citation
  • 15.

    Marx RE, Carlson ER & Eichstaedt RM, et al. Platelet-rich plasma: growth factor enhancement for bone grafts. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 1998;85:638646.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 16.

    Lowery GL, Kulkarni S, Pennisi AE. Use of autologous growth factors in lumbar spinal fusion. Bone 1999;25:47S50S.

  • 17.

    Kovacs K, Velick N & Huszar T, et al. Histomorphometric and densitometric evaluation of the effects of platelet-rich plasma on the remodeling of β-tricalcium phosphate in beagle dogs. J Craniofac Surg 2005;16:150154.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18.

    Yamada Y, Ueda M & Naiki T, et al. Autogenous injectable bone for regeneration with mesenchymal stem cells and platelet-rich plasma: tissue-engineered bone regeneration. Tissue Eng 2004;10:955964.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 19.

    Babbush CA, Kevy SV, Jacobson MS. An in vitro and in vivo evaluation of autologous platelet concentrate in oral reconstruction. Implant Dent 2003;12:2434.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 20.

    Dugrillon A, Eichler H & Kern S, et al. Autologous concentrated platelet-rich plasma (cPRP) for local application in bone regeneration. Int J Oral Maxillofac Surg 2002;31:615619.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 21.

    Haynesworth SE, Goshima J & Goldberg VM, et al. Characterization of cells with osteogenic potential from human marrow. Bone 1992;13:8188.

  • 22.

    Kadiyala S, Young RG & Thiede MA, et al. Culture expanded canine mesenchymal stem cells possess osteochondrogenic potential in vivo and in vitro. Cell Transplant 1997;6:125134.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 23.

    Bruder SP, Jaiswal N, Haynesworth SE. Growth kinetics, self-renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation. J Cell Biochem 1997;64:278294.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 24.

    Bruder SP, Jaiswal N & Ricalton NS, et al. Mesenchymal stem cells in osteobiology and applied bone regeneration. Clin Orthop 1998;suppl 355:S247S256.

    • Search Google Scholar
    • Export Citation
  • 25.

    Bruder SP, Kraus KH & Goldberg VM, et al. The effect of implants loaded with autologous mesenchymal stem cells on the healing of canine segmental bone defects. J Bone Joint Surg Am 1998;80:985996.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 26.

    Connolly J, Guse R & Lippiello L, et al. Development of an osteogenic bone-marrow preparation. J Bone Joint Surg Am 1989;71:684691.

  • 27.

    Muschler GF, Boehm C, Easley K. Aspiration to obtain osteoblast progenitor cells from human bone marrow: the influence of aspiration volume. J Bone Joint Surg Am 1997;79:16991709.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 28.

    Muschler GF, Nitto H & Matsukura Y, et al. Spine fusion using cell matrix composites enriched in bone marrow-derived cells. Clin Orthop 2003;407:102118.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 29.

    Connolly JF. Clinical use of marrow osteoprogenitor cells to stimulate osteogenesis. Clin Orthop 1998;suppl 355:S257S266.

  • 30.

    Bruder SP, Fink DJ, Caplan AI. Mesenchymal stem cells in bone development, bone repair, and skeletal regeneration therapy. J Cell Biochem 1994;56:283294.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 31.

    Hernigou PH, Poignard A & Beaujean F, et al. Percutaneous autologous bone-marrow grafting for nonunions. J Bone Joint Surg Am 2005;87:14301437.

  • 32.

    Arinzeh TL, Peter SJ & Archambault MP, et al. Allogenic mesenchymal stem cells regenerate bone in a critical-sized canine segmental defect. J Bone Joint Surg Am 2003;85:19271935.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 33.

    Muschler GF, Matsukura Y & Nitto H, et al. Selective retention of bone marrow-derived cells to enhance spinal fusion. Clin Orthop 2005;432:242251.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 34.

    Mueller SM, Glowacki J. Age-related decline in the osteogenic potential of human bone marrow cells cultured in three-dimensional collagen sponges. J Cell Biochem 2001;82:583590.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 35.

    Burstone MS. Histochemical observations on enzymatic processes in bones and teeth. Ann N Y Acad Sci 1960;85:431444.

  • 36.

    Pittenger MF, Alastair MM & Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999;284:143147.

  • 37.

    Martin DR, Cox NR & Hathcock TL, et al. Isolation and characterization of multipotential mesenchymal stem cells from feline bone marrow. Exp Hematol 2002;30:879886.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 38.

    Volk SW, Diefenderfer DL & Christopher SA, et al. Effects of osteogenic inducers on cultures of canine mesenchymal stem cells. Am J Vet Res 2005;66:17291737.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 39.

    Piermattei DL, Flo GL. Brinker, Piermattei, and Flo's handbook of small animal orthopedics and fracture repair. 3rd ed. Philadelphia: WB Saunders Co, 1997;148.

    • Search Google Scholar
    • Export Citation
  • 40.

    Hernigou P, Poignard A & Manicom O, et al. The use of percutaneous autologous bone marrow transplantation in nonunion and avascular necrosis of bone. J Bone Joint Surg Br 2005;87:896902.

    • Search Google Scholar
    • Export Citation
  • 41.

    Ehrhart N. Longitudinal bone transport for treatment of primary bone tumors in dogs: technique description and outcome in 9 dogs. Vet Surg 2005;43:2434.

    • Search Google Scholar
    • Export Citation

Advertisement

Use of a centrifugation-based, point-of-care device for production of canine autologous bone marrow and platelet concentrates

View More View Less
  • 1 Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.
  • | 2 Department of Biomedical Engineering, College of Engineering, Cornell University, Ithaca, NY 14853.
  • | 3 Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853
  • | 4 Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.
  • | 5 Children's Hospital and Center for Blood Research Laboratories, Harvard Medical School, Boston, MA 02115.
  • | 6 Children's Hospital and Center for Blood Research Laboratories, Harvard Medical School, Boston, MA 02115.
  • | 7 Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

Abstract

Objective—To analyze a centrifugation-based, point-of-care device that concentrates canine platelets and bone marrow–derived cells.

Animals—19 adult sexually intact dogs.

Procedures—Anticoagulated peripheral blood (60 mL) and 60 mL of anticoagulated bone marrow aspirate (BMA) were concentrated by centrifugation with the centrifugation-based, point-of-care device to form a platelet and a bone marrow concentrate (BMC) from 11 dogs. Blood samples were analyzed on the basis of hemograms, platelet count, and PCV. The BMA and BMC were analyzed to determine PCV, total nucleated cell count, RBC count, and differential cell counts. The BMC stromal cells were cultured in an osteoinductive medium. Eight additional dogs were used to compare the BMC yield with that in which heparin was infused into the bone marrow before aspiration.

Results—The centrifugation-based, point-of-care device concentrated platelets by 6-fold over baseline (median recovery, 63.1%) with a median of 1,336 × 103 platelets/μL in the 7-mL concentrate. The nucleated cells in BMCs increased 7-fold (median recovery, 42.9%) with a median of 720 × 103 cells/μL in the 4-mL concentrate. The myeloid nucleated cells and mononuclear cells increased significantly in BMCs with a significant decrease in PCV, compared with that of BMAs. Stromal cell cultures expressed an osteoblastic phenotype in culture. Infusion of heparin into the bone marrow eliminated clot formation and created less variation in the yield (median recovery, 61.9%).

Conclusions and Clinical Relevance—Bone marrow–derived cell and platelet-rich concentrates may form bone if delivered in an engineered graft, thus decreasing the need for cancellous bone grafts.

Abstract

Objective—To analyze a centrifugation-based, point-of-care device that concentrates canine platelets and bone marrow–derived cells.

Animals—19 adult sexually intact dogs.

Procedures—Anticoagulated peripheral blood (60 mL) and 60 mL of anticoagulated bone marrow aspirate (BMA) were concentrated by centrifugation with the centrifugation-based, point-of-care device to form a platelet and a bone marrow concentrate (BMC) from 11 dogs. Blood samples were analyzed on the basis of hemograms, platelet count, and PCV. The BMA and BMC were analyzed to determine PCV, total nucleated cell count, RBC count, and differential cell counts. The BMC stromal cells were cultured in an osteoinductive medium. Eight additional dogs were used to compare the BMC yield with that in which heparin was infused into the bone marrow before aspiration.

Results—The centrifugation-based, point-of-care device concentrated platelets by 6-fold over baseline (median recovery, 63.1%) with a median of 1,336 × 103 platelets/μL in the 7-mL concentrate. The nucleated cells in BMCs increased 7-fold (median recovery, 42.9%) with a median of 720 × 103 cells/μL in the 4-mL concentrate. The myeloid nucleated cells and mononuclear cells increased significantly in BMCs with a significant decrease in PCV, compared with that of BMAs. Stromal cell cultures expressed an osteoblastic phenotype in culture. Infusion of heparin into the bone marrow eliminated clot formation and created less variation in the yield (median recovery, 61.9%).

Conclusions and Clinical Relevance—Bone marrow–derived cell and platelet-rich concentrates may form bone if delivered in an engineered graft, thus decreasing the need for cancellous bone grafts.

Contributor Notes

Supported by Harvest Technologies, Plymouth, Mass.

The authors thank Jim Ellsworth and Margaret Vernier-Singer for technical assistance.

Address correspondence to Dr. Thoesen.