Pharmacokinetics and Biodistribution of Tacrolimus after Topical Administration: Implications for Vascularized Composite Allotransplantation.

AIM: The high doses of oral tacrolimus (TAC) (1,2) necessary to prevent acute rejection (AR) after vascularized composite allotransplantation (VCA) are associated with systemic adverse effects. The skin is the most antigenic tissue in VCA and the primary target of AR. However, the short-term use of topical TAC (Protopic®), as an off-label adjunct to oral TAC, to treat AR episodes pro re nata (PRN), has yielded inconsistent results. There is lack of data on the pharmacokinetics and tissue distribution of topical TAC in VCA, that hampers our understanding of the reasons for unreliable efficacy. Toward this goal, we evaluated the ability of topical TAC to achieve high local tissue concentrations at the site of application with low systemic concentrations. MATERIALS AND METHODS: We assessed the pharmacokinetics and tissue distribution of topical TAC (Protopic®, 0.03%) after single or repeated topical application in comparison to those after systemic delivery in rats. Animals received a single topical application of TAC ointment (Group 1) or an intravenous (IV) injection of TAC (Group 2) at a dose of 0.5 mg/kg. In another experiment, animals received daily topical application of TAC ointment (Group 3), or daily intraperitoneal (IP) injection of TAC (Group 4) at a dose of 0.5 mg/kg for 7 days. TAC concentrations in blood and tissues were analyzed by Liquid Chromatography-Mass Spectrometry (LC/MS-MS). RESULTS: Following single topical administration, TAC was absorbed slowly with a Tmax of 4 h and an absolute bioavailability of 11%. The concentrations of TAC in skin and muscle were several folds higher than whole blood concentrations. Systemic levels remained subtherapeutic (< 3 ng/ml) with repeated once daily applications. CONCLUSION: Topical application of TAC ointment (Protopic®, 0.03%) at a dose of 0.5 mg/kg/day provided high concentrations in the local tissues with low systemic exposure. Repeated topical administration of TAC is well tolerated with no local or systemic adverse effects. This study confirms the feasibility of topical application of TAC for site specific graft immunosuppression and enables future applications in VCA.

Effect of Systemic Adipose-derived Stem Cell Therapy on Functional Nerve Regeneration in a Rodent Model.

Regardless of etiology, peripheral nerve injuries (PNI) result in disruption/loss of neuromuscular junctions, target muscle denervation, and poor sensorimotor outcomes with associated pain and disability. Adipose-derived stem cells (ASCs) have shown promise in neuroregeneration. However, there is a paucity of objective assessments reflective of functional neuroregeneration in experimental PNI. Here, we use a multimodal, static, and dynamic approach to evaluate functional outcomes after ASC therapy in a rodent PNI model. METHODS: Lewis rats were divided into 3 groups: 10 mm sciatic nerve resection ("CUT" group; n = 10), transection and repair ("REP" group; n = 10), transection and repair plus single-dose ASCs ("ASC" group; n = 12). Allogeneic (Brown Norway rat) ASCs (1 × 106) were administered intravenously on postoperative day 1. Functional outcome was assessed by static sciatic index, toe spread factor, and a dynamic swim test on a weekly basis for 6 weeks. Sciatic nerves and gastrocnemius muscles were harvested at endpoint (6 weeks) for histological analysis. RESULTS: The ASC group showed accelerated functional recovery on the swim test at 2 weeks postoperatively, with continued improvement over 4 weeks, culminating in superior overall outcomes at 6 weeks compared with the REP group. The CUT group showed no significant improvement from baseline. Nerve histomorphometry correlated well with the swim test results in the ASC group. Gastrocnemius muscle weights showed no difference between the REP and the ASC groups. CONCLUSION: Our study confirms that early, single dose, systemic administration of ASC after PNI accelerates and enhances overall motor recovery on static and dynamic functional tests as evidenced by improvements in voluntary as well as involuntary motions.

High-volume hydrodissection for abdominally based free flaps: Preliminary results.

INTRODUCTION: A novel method of high-volume hydrodissection that provides both subfascial and intramuscular perforator dissection of deep inferior epigastric perforators has been safely demonstrated in animals. This manuscript demonstrates our preliminary results when translating this technique to humans. METHODS: A retrospective review was performed of all free-flap breast reconstructions utilizing the high-volume hydrodissection technique performed by the senior author (DS) at the University of Florida Medical Center from January 2014 to June 2015. RESULTS: Twenty-one patients underwent 31 free-flap breast reconstructions during the study period utilizing the high-volume hydrodissection technique. No patients were lost to follow-up which averaged 8.2 months. Complications included 1 partial flap necrosis (<10%), 1 breast hematoma, 1 abdominal bulge, 2 flaps with minimal fat necrosis (<5%), and 1 abdominal wall cellulitis. CONCLUSION: High-volume hydrodissection is a safe technique to facilitate dissection of abdominally based free flaps for breast reconstruction. © 2015 Wiley Periodicals, Inc. Microsurgery 37:307-311, 2017.

Adipose- and Bone Marrow-Derived Mesenchymal Stem Cells Prolong Graft Survival in Vascularized Composite Allotransplantation.

BACKGROUND: Strategies aiming at minimization or elimination of systemic immunosuppression are key immediate goals for clinical expansion of vascularized composite allotransplantation (VCA). We compared the in vitro and in vivo immunomodulatory efficacy of adipose-derived mesenchymal stem cells (AD-MSCs) and bone marrow (BM)-derived MSCs in a rat VCA model. METHODS: Both cell types were tested in vitro for suppressor function using mixed lymphocyte reactivity assays. AD-MSCs or BM-MSCs were administered intravenously (1 × 10 or 5 × 10 cells/animal) to Lewis rat recipients of mismatched Brown Norway hindlimb transplants. Short course tacrolimus (FK-506) monotherapy was withdrawn at postoperative day 21. In vivo regulatory T-cell induction, peripheral blood chimerism, and microchimerism in lymphatic organs were analyzed. RESULTS: AD-MSCs and BM-MSCs exhibited strong dose-dependent suppressor function in vitro, which was significantly more pronounced for AD cells. In vivo, all animals revealed peripheral multi-lineage chimerism at four weeks (P < 0.01) independent of cell type and dosage. Regulatory T-cell levels were increased with both cell types, the most in AD-MSC groups. These immunomodulatory effects were only transient. MSC treatment resulted in long-term (>120 day) allograft survival in 47% of the animals, which correlated with durable microchimerism in BM and spleen. CONCLUSIONS: AD-MSCs and BM-MSCs exert immunomodulatory effects that prolong survival of immunogenic skin-bearing VCA grafts with short course (21 day) tacrolimus induction therapy. The in vivo findings in terms of allograft survival did not reflect superior immunomodulatory characteristics of AD-MSCs found in vitro.

High-volume hydrodissection: increasing the safety and efficiency of perforator dissection.

BACKGROUND: Although perforator flaps have advanced the field of reconstructive microsurgery, these flaps increase operative time and difficulty of dissection. A prospective experimental animal study was performed to study the use of high-volume hydrodissection to simultaneously decrease the operative time while increasing the safety of perforator dissection. METHODS: Sixteen Sprague-Dawley rats underwent bilateral "deep inferior epigastric perforator" flap dissections with hydrodissection used on the study side and a traditional dissection performed on the control side. Primary outcome measurements included dissection time and dissection difficulty score (1-5 in order of increasing difficulty). RESULTS: The mean (SD) dissection time for the hydrodissected perforators was 9.29 (2.50) minutes versus 13.22 (2.44) minutes for the control perforators (P < 0.001). The mean (SD) dissection difficulty score was 4.44 (0.73) for the dissection of the control side compared with 1.69 (0.87) for the hydrodissected side (P < 0.001). CONCLUSION: The mechanical benefits of hydrodissection of perforators were evident in reduction of perforator dissection time and difficulty.

Aesthetic treatment of pectus excavatum: a new endoscopic technique using a porous polyethylene implant.

BACKGROUND: Pectus excavatum is the most frequent malformation of the rib cage. Functional aspects associated with this malformation often are absent even in adults not involved in competitive sports activities. Overall, these patients often live with extreme psychological discomfort when the malformations are minor. Traditionally, the correction of these malformations has been geared toward interventions that modify the architecture of the rib cage. However, all these interventions, even the most recent, involve considerably invasive major surgery. In fact, optimal results are not always achieved with corrective surgery using the insertion of silicone prosthesis, and patients often experience complications. METHODS: To correct intermediate and modest pectus excavatum in a stable manner and with the least amount of invasiveness, the authors developed a camouflage technique that uses porous prostheses made from high-density linear polyethylene. This material is generally used for reconstruction of the brain case. Between February 2001 and March 2006, in the I Unit of Plastic Surgery of the authors' Institute, 11 adult pectus excavatum patients with no previous cardiorespiratory symptoms underwent the authors' surgical technique. The average patient age was 29 years. RESULTS: Surgical repair was successful in all cases, and the average hospital stay was short. There were no complications during the follow-up period. The described approach repairs nonfunctional pectus excavatum in the adult with satisfying aesthetic and stable results, short hospital stay, and high patient popularity ratings. CONCLUSIONS: The best therapeutic option for pectus excavatum, especially with intermediate or moderate severity, is still controversial: thoracic surgery or camouflage surgery with implant? Trying to address those issues we propose a new technique by a multidisciplinary, not aggressive approach using a high density linear polyethylene implant and Omentus flap and the early analysis of our data.