Sub-zero non-freezing of vascularized composite allografts in a rodent partial hindlimb model

I Filz von Reiterdank; P Tawa; Y Berkane; E de Clermont-Tonnerre; A T Dinicu; C Pendexter; M Goutard; A G Lellouch; A B Mink van der Molen; J H Coert; C L Cetrulo Jr; K Uygun

doi:10.1016/j.cryobiol.2024.104950

Sub-zero non-freezing of vascularized composite allografts in a rodent partial hindlimb model

Cryobiology. 2024 Sep:116:104950. doi: 10.1016/j.cryobiol.2024.104950. Epub 2024 Aug 24.

Authors

I Filz von Reiterdank¹, P Tawa², Y Berkane³, E de Clermont-Tonnerre², A T Dinicu⁴, C Pendexter⁵, M Goutard², A G Lellouch⁶, A B Mink van der Molen⁷, J H Coert⁷, C L Cetrulo Jr⁸, K Uygun⁹

Affiliations

¹ Center for Engineering for Medicine and Surgery, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA; Shriners Children's Boston, Boston, MA, USA; Department of Plastic, Reconstructive and Hand Surgery, University Medical Center Utrecht, Utrecht, the Netherlands; Vascularized Composite Allotransplantation Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
² Shriners Children's Boston, Boston, MA, USA; Vascularized Composite Allotransplantation Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA; Department of Plastic, Reconstructive et Aesthetic Surgery, Hôpital Paris Saint-Joseph, Paris, France.
³ Center for Engineering for Medicine and Surgery, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA; Shriners Children's Boston, Boston, MA, USA; Vascularized Composite Allotransplantation Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA; Department of Plastic, Reconstructive and Aesthetic Surgery, Hôpital Sud, CHU Rennes, University of Rennes, Rennes, France.
⁴ Center for Engineering for Medicine and Surgery, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA; Shriners Children's Boston, Boston, MA, USA.
⁵ Center for Engineering for Medicine and Surgery, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
⁶ Shriners Children's Boston, Boston, MA, USA; Vascularized Composite Allotransplantation Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA; Innovative Therapies in Haemostasis, INSERM UMR-S 1140, University of Paris, F-75006, Paris, France.
⁷ Department of Plastic, Reconstructive and Hand Surgery, University Medical Center Utrecht, Utrecht, the Netherlands.
⁸ Shriners Children's Boston, Boston, MA, USA; Vascularized Composite Allotransplantation Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
⁹ Center for Engineering for Medicine and Surgery, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA; Shriners Children's Boston, Boston, MA, USA. Electronic address: [email protected].

PMID: 39134131
PMCID: PMC11404353 (available on 2025-09-01)
DOI: 10.1016/j.cryobiol.2024.104950

Abstract

Ischemia is a major limiting factor in Vascularized Composite Allotransplantation (VCA) as irreversible muscular injury can occur after as early as 4-6 h of static cold storage (SCS). Organ preservation technologies have led to the development of storage protocols extending rat liver ex vivo preservation up to 4 days. Development of such a protocol for VCAs has the added challenge of inherent ice nucleating factors of the graft, therefore, this study focused on developing a robust protocol for VCA supercooling. Rodent partial hindlimbs underwent subnormothermic machine perfusion (SNMP) with several loading solutions, followed by a storage solution with cryoprotective agents (CPA) developed for VCAs. Storage occurred in suspended animation for 24h and VCAs were recovered using SNMP with modified Steen. This study shows a robust VCA supercooling preservation protocol in a rodent model. Further optimization is expected to allow for its application in a transplantation model, which would be a breakthrough in the field of VCA preservation.

Keywords: Preservation; Subzero non-freezing; Supercooling; Vascularized composite allografts.

MeSH terms

Animals
Composite Tissue Allografts
Cryopreservation* / methods
Cryoprotective Agents* / pharmacology
Hindlimb* / blood supply
Male
Organ Preservation Solutions / pharmacology
Organ Preservation* / methods
Perfusion / methods
Rats
Vascularized Composite Allotransplantation* / methods

Substances

Cryoprotective Agents
Organ Preservation Solutions

Abstract

MeSH terms

Substances

Grants and funding