Cell lineage in vascularized bone transplantation

Authors

  • Wouter F. Willems M.D.,

    1. Microvascular Research Laboratory, Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN
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  • Mikko Larsen M.D., Ph.D.,

    1. Microvascular Research Laboratory, Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN
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  • Patricia F. Friedrich A.A.S.,

    1. Microvascular Research Laboratory, Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN
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  • Allen T. Bishop M.D.

    Corresponding author
    1. Microvascular Research Laboratory, Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN
    • Correspondence to: Allen T. Bishop, M.D., Department of Orthopedic Surgery, Microvascular Research Laboratory, Mayo Clinic, 200 First Street SW, Rochester, Minnesota 55905. E-mail: bishop.allen@mayo.edu

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Abstract

Background

The biology behind vascularized bone allotransplantation remains largely unknown. We aim to study cell traffic between donor and recipient following bone auto-, and allografting.

Methods

Vascularized femoral transplantation was performed with arteriovenous bundle implantation and short-term immunosuppression. Twenty male Piebald Virol Glaxo (PVG; RT1c) rats received isotransplants from female PVG (RT1c) rats and 22 male PVG rats received allografts from female Dark Agouti rats (DA, RT1a), representing a major histocompatibility mismatch. Both groups were randomly analyzed at 4 or 18 weeks. Bone remodeling areas (inner and outer cortical samples) were labeled and laser capture microdissected. Analysis of sex-mismatch genes by real-time reverse transcription-polymerase chain reaction provided the relative Expression Ratio (rER) of donor (female) to recipient (male) cells.

Results

The rER was 0.456 ± 0.266 at 4 weeks and 0.749 ± 0.387 at 18 weeks (p = 0.09) in allotransplants. In isotransplants, the rER was 0.412 ± 0.239 and 0.467 ± 0.252 at 4 and 18 weeks, respectively (p = 0.21). At 4 weeks, the rER at the outer cortical area of isotransplants was significantly lower in isotransplants as compared with allotransplants (0.247 ± 0.181 vs. 0.549 ± 0.184, p = 0.007). Cells in the inner and outer cortical bone remodeling areas in isotransplants were mainly donor derived (rER < 0.5) at 18 weeks, whereas allotransplants contained mainly recipient-derived cells (rER > 0.5) at 18 weeks.

Conclusions

Applying novel methodology, we describe detailed cell traffic in vascularized bone transplants, elaborating our comprehension on bone transplantation. © 2013 Wiley Periodicals, Inc. Microsc. Res. Tech. 34:37–43, 2013.

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