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J Reconstr Microsurg 2019; 35(01): 046-056
DOI: 10.1055/s-0038-1667048
Original Article
Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Irradiation Delays Tissue Growth but Enhances Osteogenic Differentiation in Vascularized Constructs

Ahmad Eweida
1   Department of Hand, Plastic and Reconstructive Surgery, Burn Center, BG Trauma Center Ludwigshafen, University of Heidelberg, Ludwigshafen, Germany
2   Department of Head, Neck and Endocrine Surgery, Faculty of Medicine, University of Alexandria, Egypt
,
Olga Rosenbauer
1   Department of Hand, Plastic and Reconstructive Surgery, Burn Center, BG Trauma Center Ludwigshafen, University of Heidelberg, Ludwigshafen, Germany
,
Oliver Frisch
1   Department of Hand, Plastic and Reconstructive Surgery, Burn Center, BG Trauma Center Ludwigshafen, University of Heidelberg, Ludwigshafen, Germany
,
Frank A. Giordano
3   Department of Radiation Oncology, University Medical Center, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
,
Jens Fleckenstein
3   Department of Radiation Oncology, University Medical Center, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
,
Frederik Wenz
3   Department of Radiation Oncology, University Medical Center, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
,
Stephanie Kirschner
4   Department of Neuroradiology, University Medical Center, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
,
Marc A. Brockmann
4   Department of Neuroradiology, University Medical Center, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
5   Department of Neuroradiology, University Medical Center Mainz, Mainz, Germany
,
Matthias Schulte
1   Department of Hand, Plastic and Reconstructive Surgery, Burn Center, BG Trauma Center Ludwigshafen, University of Heidelberg, Ludwigshafen, Germany
,
Ulrich Kneser
1   Department of Hand, Plastic and Reconstructive Surgery, Burn Center, BG Trauma Center Ludwigshafen, University of Heidelberg, Ludwigshafen, Germany
,
Leila Harhaus
1   Department of Hand, Plastic and Reconstructive Surgery, Burn Center, BG Trauma Center Ludwigshafen, University of Heidelberg, Ludwigshafen, Germany
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Publikationsverlauf

05. Februar 2018

21. Mai 2018

Publikationsdatum:
11. Juli 2018 (online)

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Abstract

Background Regenerative medicine is still deficient in the reconstruction after cancer due to impaired vascularization after radiotherapy and due to the need to substitute larger defects after tumor excision. Aiming at introducing regenerative medicine for reconstruction after cancer, we tested an axially vascularized bone construct in an experimental setting that mimics the clinical situation after tumor resection and adjuvant radiotherapy.

Methods Twenty bone constructs were axially vascularized using microsurgically created arteriovenous loops and were implanted subcutaneously in Lewis rats. After 2 weeks, the animals were randomly allocated either to receive a clinically relevant single dose of external beam irradiation or not (n = 10 for each group). The animals were sacrificed either after 1 week or 10 weeks after irradiation (n = 5 for each time point). The constructs were tested for vascularization, tissue growth, cellular proliferation, cellular apoptosis, and osteogenic differentiation via histomorphometric, immunohistochemical, and polymerase chain reaction (PCR) analysis. One construct per group was subjected at 10 weeks to qualitative micro-computed tomography (CT) imaging.

Results Tissue generation and cellular proliferation were significantly reduced at 1 week after irradiation, but no longer significantly different after 10 weeks.

No significant differences in vascularization were detected at any time point. Apoptosis did not show any statistically significant differences between both groups at both time points. At the late time point, mature bone was considerably more in the irradiated group, but the results were not statistically significant. PCR analysis showed a significantly enhanced expression of osteocalcin in the irradiated group at 1 week. Micro-CT imaging showed that both constructs were adequately vascularized with no evident morphologic differences regarding vascular density or vascular distribution.

Conclusions Axially vascularized bone constructs can withstand clinically relevant doses of irradiation and retain their angiogenic and osteogenic potential in the long term. Irradiation led to a delayed tissue generation with a comparatively enhanced osteogenic differentiation within the constructs.

Keywords

tissue engineering - bone regeneration - axially vascularized construct - arteriovenous loop - adjuvant radiotherapy
 

  • References

  • 1 Reuther T, Schuster T, Mende U, Kübler A. Osteoradionecrosis of the jaws as a side effect of radiotherapy of head and neck tumour patients--a report of a thirty year retrospective review. Int J Oral Maxillofac Surg 2003; 32 (03) 289-295
    CrossrefPubMedGoogle Scholar
  • 2 Jegoux F, Malard O, Goyenvalle E, Aguado E, Daculsi G. Radiation effects on bone healing and reconstruction: interpretation of the literature. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109 (02) 173-184
    CrossrefPubMedGoogle Scholar
  • 3 Wei FC, Celik N, Yang WG, Chen IH, Chang YM, Chen HC. Complications after reconstruction by plate and soft-tissue free flap in composite mandibular defects and secondary salvage reconstruction with osteocutaneous flap. Plast Reconstr Surg 2003; 112 (01) 37-42
    CrossrefPubMedGoogle Scholar
  • 4 Hanasono MM. Reconstructive surgery for head and neck cancer patients. Adv Med 2014; 2014: 795483
    PubMedGoogle Scholar
  • 5 Eweida AM, Horch RE, Marei MK. , et al. Axially vascularised mandibular constructs: is it time for a clinical trial?. J Craniomaxillofac Surg 2015; 43 (07) 1028-1032
    CrossrefPubMedGoogle Scholar
  • 6 Eweida AM, Nabawi AS, Elhammady HA. , et al. Axially vascularized bone substitutes: a systematic review of literature and presentation of a novel model. Arch Orthop Trauma Surg 2012; 132 (09) 1353-1362
    CrossrefPubMedGoogle Scholar
  • 7 Kneser U, Schaefer DJ, Polykandriotis E, Horch RE. Tissue engineering of bone: the reconstructive surgeon's point of view. J Cell Mol Med 2006; 10 (01) 7-19
    CrossrefPubMedGoogle Scholar
  • 8 Schmidt VJ, Hilgert JG, Covi JM. , et al. Flow increase is decisive to initiate angiogenesis in veins exposed to altered hemodynamics. PLoS One 2015; 10 (01) e0117407
    CrossrefPubMedGoogle Scholar
  • 9 Lokmic Z, Stillaert F, Morrison WA, Thompson EW, Mitchell GM. An arteriovenous loop in a protected space generates a permanent, highly vascular, tissue-engineered construct. FASEB J 2007; 21 (02) 511-522
    CrossrefPubMedGoogle Scholar
  • 10 Eweida AM, Nabawi AS, Abouarab M. , et al. Enhancing mandibular bone regeneration and perfusion via axial vascularization of scaffolds. Clin Oral Investig 2014; 18 (06) 1671-1678
    CrossrefPubMedGoogle Scholar
  • 11 Horch RE, Beier JP, Kneser U, Arkudas A. Successful human long-term application of in situ bone tissue engineering. J Cell Mol Med 2014; 18 (07) 1478-1485
    CrossrefPubMedGoogle Scholar
  • 12 Eweida A, Frisch O, Giordano FA. , et al. Axially vascularized tissue-engineered bone constructs retain their in vivo angiogenic and osteogenic capacity after high-dose irradiation. J Tissue Eng Regen Med 2018; 12 (02) e657-e668
    CrossrefPubMedGoogle Scholar
  • 13 Eweida A, Schulte M, Frisch O, Kneser U, Harhaus L. The impact of various scaffold components on vascularized bone constructs. J Craniomaxillofac Surg 2017; 45 (06) 881-890
    CrossrefPubMedGoogle Scholar
  • 14 Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl 1968; 97: 77-89
    PubMedGoogle Scholar
  • 15 Tchanque-Fossuo CN, Monson LA, Farberg AS. , et al. Dose-response effect of human equivalent radiation in the murine mandible: Part II. A biomechanical assessment. Plast Reconstr Surg 2011; 128 (05) 480e-487e
    CrossrefPubMedGoogle Scholar
  • 16 Pfister DG, Ang KK, Brizel DM. , et al; National Comprehensive Cancer Network. Head and neck cancers, version 2.2013. Featured updates to the NCCN guidelines. J Natl Compr Canc Netw 2013; 11 (08) 917-923
    CrossrefPubMedGoogle Scholar
  • 17 Kneser U, Polykandriotis E, Ohnolz J. , et al. Engineering of vascularized transplantable bone tissues: induction of axial vascularization in an osteoconductive matrix using an arteriovenous loop. Tissue Eng 2006; 12 (07) 1721-1731
    CrossrefPubMedGoogle Scholar
  • 18 Schambach SJ, Bag S, Groden C, Schilling L, Brockmann MA. Vascular imaging in small rodents using micro-CT. Methods 2010; 50 (01) 26-35
    CrossrefPubMedGoogle Scholar
  • 19 Kirschner S, Felix MC, Hartmann L. , et al. In vivo micro-CT imaging of untreated and irradiated orthotopic glioblastoma xenografts in mice: capabilities, limitations and a comparison with bioluminescence imaging. J Neurooncol 2015; 122 (02) 245-254
    CrossrefPubMedGoogle Scholar
  • 20 Sønstevold T, Johannessen AC, Stuhr L. A rat model of radiation injury in the mandibular area. Radiat Oncol 2015; 10: 129
    CrossrefPubMedGoogle Scholar
  • 21 Maeda A, Leung MK, Conroy L. , et al. In vivo optical imaging of tumor and microvascular response to ionizing radiation. PLoS One 2012; 7 (08) e42133
    CrossrefPubMedGoogle Scholar
  • 22 Mouthon MA, Vereycken-Holler V, Van der Meeren A, Gaugler MH. Irradiation increases the interactions of platelets with the endothelium in vivo: analysis by intravital microscopy. Radiat Res 2003; 160 (05) 593-599
    CrossrefPubMedGoogle Scholar
  • 23 Polykandriotis E, Tjiawi J, Euler S. , et al. The venous graft as an effector of early angiogenesis in a fibrin matrix. Microvasc Res 2008; 75 (01) 25-33
    CrossrefPubMedGoogle Scholar
  • 24 Liu Q, Douglas T, Zamponi C. , et al. Comparison of in vitro biocompatibility of NanoBone(®) and BioOss(®) for human osteoblasts. Clin Oral Implants Res 2011; 22 (11) 1259-1264
    CrossrefPubMedGoogle Scholar
  • 25 Buehrer G, Balzer A, Arnold I. , et al. Combination of BMP2 and MSCs significantly increases bone formation in the rat arterio-venous loop model. Tissue Eng Part A 2015; 21 (1–2): 96-105
    CrossrefPubMedGoogle Scholar
  • 26 Amini AR, Laurencin CT, Nukavarapu SP. Bone tissue engineering: recent advances and challenges. Crit Rev Biomed Eng 2012; 40 (05) 363-408
    CrossrefPubMedGoogle Scholar
  • 27 Pham PV, Vu NB, Pham VM. , et al. Good manufacturing practice-compliant isolation and culture of human umbilical cord blood-derived mesenchymal stem cells. J Transl Med 2014; 12: 56
    CrossrefPubMedGoogle Scholar
  • 28 Rennert RC, Sorkin M, Garg RK, Gurtner GC. Stem cell recruitment after injury: lessons for regenerative medicine. Regen Med 2012; 7 (06) 833-850
    CrossrefPubMedGoogle Scholar
  • 29 Tsai TL, Li WJ. Identification of bone marrow-derived soluble factors regulating human mesenchymal stem cells for bone regeneration. Stem Cell Reports 2017; 8 (02) 387-400
    CrossrefPubMedGoogle Scholar
  • 30 Mieloch AA, Suchorska WM. The concept of radiation-enhanced stem cell differentiation. Radiol Oncol 2015; 49 (03) 209-216
    CrossrefPubMedGoogle Scholar
  • 31 Chen FH, Chiang CS, Wang CC. , et al. Vasculatures in tumors growing from preirradiated tissues: formed by vasculogenesis and resistant to radiation and antiangiogenic therapy. Int J Radiat Oncol Biol Phys 2011; 80 (05) 1512-1521
    CrossrefPubMedGoogle Scholar
  • 32 Chen FH, Chiang CS, Wang CC. , et al. Radiotherapy decreases vascular density and causes hypoxia with macrophage aggregation in TRAMP-C1 prostate tumors. Clin Cancer Res 2009; 15 (05) 1721-1729
    CrossrefPubMedGoogle Scholar
  • 33 Ceradini DJ, Kulkarni AR, Callaghan MJ. , et al. Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1. Nat Med 2004; 10 (08) 858-864
    CrossrefPubMedGoogle Scholar
  • 34 François S, Bensidhoum M, Mouiseddine M. , et al. Local irradiation not only induces homing of human mesenchymal stem cells at exposed sites but promotes their widespread engraftment to multiple organs: a study of their quantitative distribution after irradiation damage. Stem Cells 2006; 24 (04) 1020-1029
    CrossrefPubMedGoogle Scholar
  • 35 Eweida A, Fathi I, Eltawila AM. , et al. Pattern of bone generation after irradiation in vascularized tissue engineered constructs. J Reconstr Microsurg 2018; 34 (02) 130-137
    Artikel in Thieme ConnectPubMedGoogle Scholar