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. 2025 Jul 29;38:14520. doi: 10.3389/ti.2025.14520

TABLE 2.

Overview of Interventions, Immunosuppressive Strategies and Outcomes of studies included.

Author and year Interventions Immunosuppresion Outcomes Complications
Barone et al. [23] Bone marrow transplantation Low-dose total body irradiation (100cGy 2 days prior to surgery or 200cGy divided in 2 × 100 cGy doses on preoperative day 2 and 3), T-cell depletion wirh CD3-IT (0.05 mg/kg), CXA (target level 400–800 ng/mL), donor bone marrow cells (7.8 × 108 to 4 × 109 cells/kg of recipient body weight) Bone marrow infusion led to better clinical outcomes; chimerism detected but insufficient for tolerance Mixed chimerism after bone marrow transplantat; VCA appeared insufficient for tolerance induction
Berkane et al. [33] Two study groups: supercooling intervention group and cold storage control group undergoing subsequent normothermic machine perfusion No immunosuppressive therapy used Supercooled VCAs restored vascular flow and had lower resistance during machine perfusion N/A
Blades et al., 2024 [43] Investigation of possible surgical complications No immunosuppressive therapy used All flaps survived initially, with adequate perfusion for 4 days. Flap rejection occurred between POD 5 and POD 9 in all animals Minimal erythema observed post-transplant, no surgery-related deaths or infections
Elgendy et al. [26] Treatment with Co-stimulation blockade and mTOR inhibitor, with or without preceding short-term calcineurin inhibitor therapy mTOR inhibitor (rapamycin [0.02–0.2 mg/kg] or tacrolimus [0.1–0.125 mg/kg]) TAC delayed AR (grade-I AR on POD 30, grade-IV on POD 74); rapid rejection with rapamycin (grade-I AR by POD 2 and 7, grade-IV AR by POD 17–20) Rejection of allograft, erythema, severe necrotizing T cell mediated rejection with deep dermal arterial thrombosis
Fries et al. [35] Tacrolimus eluting hydrogel implants with various concentrations (91 mg, high dose/49 mg, low dose) Graft-implanted enzyme-responsive, TAC eluting hydrogel platform Low-dose TAC prolonged survival; high-dose TAC caused poor tolerance (grade IV AR from POD 56–93) High dose TAC group: one sample excluded due to flap failure on POD 1; four animals showed poor feeding and weight loss, requiring early euthanasia; four animals from high dose TAC group developed pancreatitis
Ibrahim et al. [36] Development of novel tranlational VCA research model Short-term tacrolimus monotherapy (target levels of 10–15 ng/mL) with or without bone marrow infusion Long-term graft survival (>150 days) with viable vascularized bone marrow; successful immune monitoring Venous thrombus in one case resolved by reanastomosis, no graft-versus-host disease
Kim et al. [25] Treatment with tacrolimus for 30 days and ASC therapy (donor-derived ASCs [1.0 × 10^6 cells/kg]) TAC, ASC-therapy Adipose-derived stem cells demonstrated grade IV AR on POD 119 and rejection-free survival over POD 200 as well as upregulated T-regulatory cells The control group reached Banff grade 4 acute rejection by an average of 7.5 days after transplantation. Allografts treated with ASCs demonstrated grade 4 rejection on day 119
Kotsougiani et al. [28] AV-bundle implantation in tibial allotransplant TAC (target levels of 5–30 ng/mL), MMF (target levels of 1–3.5 ng/mL), MPDN (tapered to 0.1 mL) Micro-CT showed bone formation and remodeling at the distal allograft junction; allograft survived without any healing problems or limited hindlimb perfusion during the 4-month follow-up N/A
Kuo et al. [29] Treatment with various dosages of mesenchymal stem cells, CXA, bone marrow transplantation and irradiation Irradiation, bone marrow trnsplantation and CXA Mesenchymal cells extended graft survival, combined CXA and stem cells showed significantly better survival, allografts with CXA exhibited delayed AR, examination of bromodeoxyuridine-labeled mesenchymal stem cells revealed donor mesenchymal stem cells engraftment into the recipient and donor skin Graft-versus-host disease evident in CXA group
Kuo et al. [29] Comparison of rejection in untreated, control and CXA-treatment groups CXA in treatment group, untreated and control: N/A 100% survival rate, CXA treatment delayed flap rejection significantly (POD 38-49), no significant difference in rejection signs in allo-cartilage Swelling for 2 weeks (postoperative saliva gland hypersecretion), control group: progressive rejection by POD 7-28, lymphoid gland tissue and skin were susceptible to early rejection
Kuo et al. [30] Various combinations of mesenchymal stem cells cyclosporine or irradiation Mesenchymal stem cells, CXA, irradiation Mesenchymal stem cells with irradiation and CXA: significantly increased allograft survival compared with other groups (>120 days; p < 0.01); histology showed lowest degree of AR in grafted skin and interstitial muscle layers in mesenchymal stem cell/irradiation/CXA group; significant increase in percentage of CD4+/CD25+ and CD4+/FoxP3+ T in the mesenchymal stem cell/irradiation/CXA group Rejection episodes
Kuo et al. [31] Various dosages of ASCs, tacrolimus or irradiation TAC, irradiation Multiple injections of adipose-derived stem cells, irradiation and TAC increased allograft survival significantly Lymphocyte infiltration in the alloskin and interstitial muscle layers of treatment group
Leonard et al. [24] Stem cell transfusion 100 cGy irradiation, T cell depletion with CD3-IT (50 μg/kg), hematopoietic cell transplantation (15 × 109 cells/kg) Following withdrawal of immunosuppression both VCAs transplanted into stable chimeras
Recipients of hematopoietic cell transplantation displayed no clinical signs of AR up to POD 504
Two animals developed skin graft versus host disease
Mathes et al. [32] Treatment with CXA and bone marrow transplantation (2 × 109 cells/kg) CXA (target levels of 400–800 ng/mL) Donor cell engraftment and multilineage macro chimerism after in utero transplantation of adult bone marrow cells, and chimeric animals were unresponsive to donor antigens in vitro; both control VCAs rejected by POD 21; chimeric animals accepted VCAs (no DSAs or alloreactivity) All grafts demonstrated some mild lymphocytic infiltration at the day 7 biopsy. All of the animals developed a severe dermal perivascular lymphocytic infiltration with scattered eosinophils and went on to reject their donor skin grafts
Park et al. [44] Vascular anastomosis of the carotid artery and jugular vein, fixation of the maxillo-mandibular complex with titanium plates No immunosuppressive therapy used Successful transplant without early arterial or venous insufficiency, acute rejection from POD 7-8 onwards Acute rejection POD 7-8, pink discoloration, edema, erythematous papule with flap necrosis on POD 14–18
Shanmugarajah et al. [38] Hematopoietic stem cell transplantant, irradiation T cell depletion with CD3-IT (50 μg/kg), 100 cGy TBI and 45 days of CXA (target levels of 400–800 ng/mL) HC class II–mismatched chimeras were tolerant of VCAs;
HC class I–mismatched animals rejected VCA skin, (infiltration of CD8+ lymphocytes)
One HC class II mismatched model displayed clinical features of chronic graft versus host disease (euthanized on POD 190)
Tratnig-Frankl et al. [39] Treatment with either saline (control), sodium iodide (NaI), or hydrogen sulfide (H2S) injections No postoperative immunosuppression No effect of H2S or NaI treatment in comparison to NaCl in delaying AR, flap survival and histology revealed no significant differences between the groups One technical failure occurred in the saline MISMATCH subgroup
Wachtman et al. [27] Bone marrow infusion and irradiation Total body (100 cGy) and thymic (700 cGy) irradiation, bone marrow infusion, tacrolimus (0.1 mg/kg/day), CTLA4-Ig (20 mg/kg) Experimental groups rejected allografts (skin and muscle) on POD 5 to 30; skin and muscle histology in all long-term survivors were normal Rejection episodes
Waldner et al. [40] Investigation of VRAM flap applicability in VCA research TAC, rapamycin, CTLA4-Ig POD 5: all grafts demonstrated pale-pink skin color without edema;
follow-up showed improved correlation between clinical appearance and progression of graft rejection in histology
Intraoperative cardiac arrest in one sample (death due to anesthesia); one recipient experienced flap loss due to venous compromise; Banff grade I AR with erythemous and edematous grafts
Wang et al. [45] Treatment with sub-normothermic ex-vivo perfusion using hyper-oxygenated University of Wisconsin (UW) solution No immunosuppressive therapy used Experimental group showed significantly later onset of grade 1 AR at 13.7 days (SD = 0.52, p < 0.05); by POD 15 75% of the flaps showed no evidence of grade 4 AR Rejection episodes
Wu et al. [42] Treatment with various dosages (28 mg/4cc and 49 mg/4cc) of tacrolimus-eluting hydrogel injected into the donor flap TAC-eluting hydrogel (28 mg/4cc and 49 mg/4cc) TAC-eluting hydrogel prolonged graft survival in both groups (grade 4 AR on average by POD 20 and 28) Rejection episodes
Zhang et al. [34] Treatment with various combinations of TGMS and TAC Locally administered TAC-loaded on-demand drug delivery system Repeated intra-graft TGMS-TAC administrations prolong graft survival Grade III-IV rejection

MGH, Massachusetts General Hospital; SLA, Swine leukocyte antigen; HC, histocompatibility complex; MHC, major histocompatibility complex; VCA, vascularized composite allotransplant; SH, single haplotype; PAA, pig allelic antigen; CS, Cold Storage; TAC, tacrolimus; MMF, Mycophenolate Mofetil; MPDN, Methylprednisolone; CXA, cyclosporine A; CD3-IT, CD3-Immunotoxin; (CTLA4-Ig), cytotoxic T-lymphocyte antigen 4 immunoglobulin; POD, postoperative day; AR, acute rejection; DSAs, donor specific antibodies; N/A, not applicable; TGMS, triglycerol monostearate; VRAM, vertical rectus abdominus myocutaneous flap; ASC, adipose-derived stem cell; AV, arteriovenous.