Adipose-derived mesenchymal stem cells cultured in serum-free medium attenuate acute contrast-induced nephropathy by exerting anti-apoptotic effects.

BACKGROUND: Contrast-induced nephropathy (CIN) is a major clinical problem associated with acute kidney injury during hospitalization. However, effective treatments for CIN are currently lacking. Mesenchymal stem cells (MSCs) have protective effects against kidney injury by suppressing inflammation and fibrosis. We previously showed that MSCs cultured in serum-free medium (SF-MSCs) enhance their anti-inflammatory and anti-fibrotic effects. However, whether SF-MSCs potentiate their anti-apoptotic effects is unknown. Here, we investigated the effects of SF-MSCs on a CIN mouse model. METHODS: To create CIN model mice, we removed right kidney at first. One week later, the left renal artery was clamped for 30 min to cause ischemia-reperfusion injury, and mice were injected with iohexol. Then the kidney received 10 Gy of irradiation, and MSCs or SF-MSCs were injected immediately. At 24 h post-injection, mice were sacrificed, and their blood and kidneys were collected to evaluate renal function, DNA damage, and apoptosis. In addition, apoptosis was induced in HEK-293 cells by irradiation and cells were treated with conditioned medium from MSCs or SF-MSCs. RESULTS: Treatment of CIN model mice with SF-MSCs markedly improved renal function compared with MSCs treatment. Cleaved caspase-3 levels and TUNEL-positive cell numbers were strongly suppressed in CIN model mice treated with SF-MSCs compared with the findings in those treated with MSCs. γH2AX levels, a chromosome damage marker, were reduced by MSCs and further reduced by SF-MSCs. In addition, cleaved caspase-3 in irradiated HEK-293 cells was more strongly suppressed by conditioned medium from SF-MSCs than by that from MSCs. Secretion of epidermal growth factor (EGF) was enhanced by culturing MSCs in serum-free medium. Knockdown of EGF by siRNA attenuated the inhibitory effects of SF-MSCs on CIN-induced renal dysfunction and tubular apoptosis. CONCLUSIONS: These findings strongly suggest that SF-MSCs improve CIN in model mice by exerting anti-apoptotic effects in a paracrine manner. Thus, SF-MSCs represent a potential novel therapy for CIN.

Osteocutaneous superficial circumflex iliac perforator flap for the lower extremity bone and soft tissue reconstruction with perforator-to-perforator anastomosis after radical debridement of tibia osteomyelitis: A case report.

Reconstruction of soft tissue and bone defects in tibia chronic osteomyelitis is challenging and often managed by free flap with bone graft. However, the use of osteocutaneous free flap combined with perforator-to-perforator anastomosis has not been reported. We report the case of a 62-year-old man presenting with soft tissue and bone defects with right tibial chronic osteomyelitis, which was successfully treated with an osteocutaneous superficial circumflex iliac perforator (SCIP) flap with perforator-to-perforator anastomosis. After radical debridement and excision of the sequestrum, a 17 × 10-cm skin defect and a 4 × 3-cm bone defect remained. An osteocutaneous SCIP flap, containing a 16 × 9-cm skin paddle and 4 × 2-cm iliac bone, was transferred and anastomosed to the posterior tibial perforator in an end-to-end fashion. An artificial dermis was placed to cover the soft tissue. At 1 week postoperatively, the artificial dermis was partially infected, which required small debridement. Full weight-bearing was permitted 5 weeks postoperatively, and the patient walked independently. No evidence of recurrence of osteomyelitis or skin ulcers was observed at 15 months postoperatively. Therefore, osteocutaneous SCIP flap with perforator-to-perforator anastomosis may be a potential alternative treatment for soft tissue and bone defects after radical debridement of tibia osteomyelitis.

Peripheral T cell profiling reveals downregulated exhaustion marker and increased diversity in lymphedema post-lymphatic venous anastomosis.

Lymphedema is a progressive condition accompanying cellulitis and angiosarcoma, suggesting its association with immune dysfunction. Lymphatic venous anastomosis (LVA) can provide relief from cellulitis and angiosarcoma. However, the immune status of peripheral T cells during lymphedema and post-LVA remains poorly understood. Using peripheral blood T cells from lymphedema, post-LVA, and healthy controls (HCs), we compared the profile of T cell subsets and T cell receptor (TCR) diversity. PD-1+ Tim-3 + expression was downregulated in post-LVA compared with lymphedema. IFN-γ levels in CD4+PD-1+ T cells and IL-17A levels in CD4+ T cells were downregulated in post-LVA compared with lymphedema. TCR diversity was decreased in lymphedema compared with HCs; such TCR skewing was drastically improved in post-LVA. T cells in lymphedema were associated with exhaustion, inflammation, and diminished diversity, which were relieved post-LVA. The results provide insights into the peripheral T cell population in lymphedema and highlight the immune modulatory importance of LVA.

Comparison of therapeutic effects of mesenchymal stem cells derived from superficial and deep subcutaneous adipose tissues.

BACKGROUND: Fibrosis is a common histological feature in the process from chronic organ injury to organ failure. Chronic tissue injury causes inflammatory cell infiltration into the injured tissue. The persistence of this inflammatory cell infiltration leads to fibrosis and organ failure. Adipose-derived mesenchymal stem cells (ASCs) have received much attention as a regenerative therapeutic tool to prevent progression from organ injury to failure. Subcutaneous abdominal adipose tissue is divided into superficial and deep layers by a superficial fascia. Adipose tissue easily collected by liposuction is usually obtained from a deep layer, so ASCs derived from a deep layer are generally used for regenerative medicine. However, no research has been conducted to investigate differences in the therapeutic effects of ASCs from the superficial and deep layers (Sup-ASCs and Deep-ASCs, respectively). Therefore, we compared the therapeutic potencies of Sup-ASCs and Deep-ASCs. METHODS: ASCs were isolated from superficial and deep subcutaneous abdominal adipose tissues collected from patients who underwent breast reconstruction. We first compared cell characteristics, such as morphology, cell proliferation, cell surface markers, adipogenic and osteogenic differentiation, cell senescence markers, and expression of coagulation and anticoagulant factors between Sup-ASCs and Deep-ASCs. Furthermore, we compared their ability to promote polarization of M2 macrophages and to inhibit transforming growth factor (TGF)-ß/Smad signaling using THP-1 cells and TGF-ß1 stimulated HK-2 cells incubated with conditioned media from Sup-ASCs or Deep-ASCs. In in vivo experiments, after renal ischemia-reperfusion injury (IRI) procedure, Sup-ASCs or Deep-ASCs were injected through the abdominal aorta. At 21 days post-injection, the rats were sacrificed and their left kidneys were collected to evaluate fibrosis. Finally, we performed RNA-sequencing analysis of Sup-ASCs and Deep-ASCs. RESULTS: Sup-ASCs had greater proliferation and adipogenic differentiation compared with Deep-ASCs, whereas both ASC types had similar morphology, cell surface markers, senescence markers, and expression of coagulation and anticoagulant factors. Conditioned media from Sup-ASCs and Deep-ASCs equally promoted polarization of M2 macrophages and suppressed TGF-ß/Smad signaling. Moreover, administration of Sup-ASCs and Deep-ASCs equally ameliorated renal fibrosis induced by IRI in rats. RNA-sequencing analysis revealed no significant difference in the expression of genes involved in anti-inflammatory and anti-fibrotic effects between Sup-ASCs and Deep-ASCs. CONCLUSIONS: These results indicate that both Sup-ASCs and Deep-ASCs can be used effectively and safely as an intravascular ASC therapy for organ injury.

Correlation between Lymphatic Surgery Outcome and Lymphatic Image-Staging or Clinical Severity in Patients with Lymphedema.

Lymphoscintigraphy and indocyanine green (ICG) lymphography reveal the severity of extremity lymphedema. Lower extremity lymphedema (LEL) index and NECST classification are related to the clinical severity of lymphedema. We aimed to investigate the correlation between lymphatic surgery, lymphatic imaging, and clinical severity in patients with lymphedema. Thirty-five patients with lower-extremity lymphedema who underwent lymphatic venous anastomosis (LVA) were evaluated. Ten of the thirty-five patients underwent multi-surgery (additional vascularized lymphatic transfer and/or liposuction). We investigated the correlation between the LEL index, NECST classification, lymphoscintigraphy staging, ICG lymphography staging, and rate of improvement (RI: [preoperative LEL index − postoperative LEL index]/[preoperative LEL index] × 100). The LEL index in 35 patients after LVA and all procedures decreased significantly compared to that of preoperative (272.4 vs. 256.2 vs. 243.5, p < 0.05). RI after LVA and all procedures showed positive correlations with the preoperative LEL index; however, there was no correlation with any other lymphatic image or clinical severity. LVA can reduce lymphedema circumference at any stage. Additional surgery improved the circumference. Hence, LVA as the first line of treatment, and vascularized lymphatic transfer and liposuction as additional procedures, should be considered as the standard treatment for lymphedema.

Combined Lymphovenous Anastomosis and Great Saphenous Vein Stripping for Comorbid Lymphedema and Varicose Veins.

Background: Treatment for patients with comorbid lymphedema and varicose veins is controversial. Surgical options for these patients are limited. The study was aimed to investigate the validity of combined lymphovenous anastomosis (LVA) and great saphenous vein stripping (GSVS) for comorbid lymphedema and varicose veins. Methods: Thirteen patients were involved in the study, and the detail was 21 edematous lower limbs (with coexisting varicose veins and lymphedema; the varicose vein and lymphedema [VL] group) who underwent combined GSVS and LVA therapy. Fifteen patients (with 30 edematous lower limbs and lymphedema only; the lymphedema [L] group) who underwent LVA only were included as a control group. GSVS was performed before LVA in the VL group. Results: No significant differences were seen between the groups at baseline. There were no cases indocyanine green (ICG) lymphography pattern deteriorated after GSVS. No significant difference was seen in lymphatic detection rate; 129.71% ± 58.27% (67%-333%) in the VL group and 122.27% ± 39.47% (50%-250%) in the L group (p = 0.59 > 0.05), respective lymphatic diameters 0.66 ± 0.13 (0.45-0.9) mm and 0.75 ± 0.17 (0.45-1.0) mm (p = 0.07 > 0.05), and respective lymphedema improvement rate 12.17% ± 7.35% (0%-27.4%) and 12.65% ± 7.43% (3.7%-22.3%) (p = 0.86 > 0.05). Discussion: In this study, stripping surgery does not cause lymphatic impairment, at least to the extent that would impede the success of an LVA procedure. Comorbid varicose veins and lymphedema can be treated surgically by a combination of LVA and GSVS.

Effect of Postoperative Compression Therapy on the Success of Liposuction in Patients with Advanced Lower Limb Lymphedema.

OBJECTIVE: There is limited information on postoperative care after liposuction for lymphedema limb. The aim of this retrospective study was to identify the threshold compression pressure and other factors that lead liposuction for lower limb lymphedema to success. MATERIALS AND METHODS: Patients were divided according to whether they underwent compression therapy with both stockings and bandaging (SB group), stockings alone (S group), or bandaging alone (B group) for 6 months after liposuction. The postoperative compression pressure and rate of improvement were compared according to the postoperative compression method. We also investigated whether it was possible to decrease the compression pressure after 6 months. Liposuction was considered successful if improvement rate was >15. RESULTS: Mean compression pressure was significantly lower in the S group than in the SB group or B group. The liposuction success rate was significantly higher in the SB group than in the B group or S group. There was not a significant difference between the values at 6 months after liposuction and at 6 months after a decrease in compression pressure in the successful group. CONCLUSION: Our results suggest that stable high-pressure postoperative compression therapy is key to the success of liposuction for lower limb lymphedema and is best achieved by using both stockings and bandages. The postoperative compression pressure required for liposuction to be successful was >40 mmHg on the lower leg and >20 mmHg on the thigh. These pressures could be decreased after 6 months.

Lymphaticovenous Anastomosis for Age-Related Lymphedema.

INTRODUCTION: Primary lymphedema is usually caused by intrinsic disruption or genetic damage to the lymphatics but may also be the result of age-related deterioration of the lymphatics. The aims of this study were to determine the characteristics of age-related lymphedema and to assess the effectiveness of lymphaticovenous anastomosis (LVA) in its treatment. METHODS: Eighty-six patients with primary lymphedema affecting 150 lower limbs were divided into three groups according to whether the age of onset was younger than 35 years, 35-64 years, or 65 years or older. Indocyanine green (ICG) lymphography was performed, followed by LVA surgery. ICG lymphography images were visually classified according to whether the pattern was linear, low enhancement (LE), distal dermal backflow (dDB), or extended dermal backflow (eDB). The lower extremity lymphedema (LEL) index score was calculated before and after LVA. Lymphatic vessel diameter and detection rates were also recorded. RESULTS: In the ≥65 group, the lymphedema was bilateral in 54 patients and unilateral in 1 patient. There was statistically significant deterioration in the LEL index score with progression from the linear, LE, dDB through to the eDB pattern in the ≥65 group. The lymphatic vessel diameter was significantly greater in the ≥65 group. The rate of improvement was highest in the ≥65 group. CONCLUSION: Age-related lymphedema was bilateral and deterioration started distally. The lymphatic vessels in patients with age-related lymphedema tended to be ectatic, which is advantageous for LVA and may increase the improvement rate.

Successful Treatment of Rheumatoid Lymphedema with Lymphatic Venous Anastomosis.

Rheumatoid lymphedema is a rare but severely disabling condition caused by reduced lymphatic drainage. Most treatment methods are conservative and may lead to the exacerbation of lymphedema. Lymphatic venous anastomosis (LVA) is an effective treatment for lymphedema after surgery involving the lymphatic system, such as lymph node dissection for cancer treatment. LVA has not been used to treat rheumatoid lymphedema. We present a case of rheumatoid lymphedema treated with surgical procedures, including LVA. Following LVA, objective and subjective symptom relief was noted, along with decreased swelling and pain in the affected area. The postoperative course was uneventful. LVA for the treatment of rheumatoid lymphedema may provide definitive clinical improvements.

Investigation of flow velocity in recipient perforator artery for a reliable indicator for the flap transfer with perforator to perforator anastomosis.

INTRODUCTION: In flap transfer with perforator to perforator anastomosis (FTPPA), encountering poor pulsation and blood flow in a recipient perforator intraoperatively often makes FTPPA impossible. This study sought to identify color and spectral Doppler ultrasonography (CSDUS) parameters that can aid reliable preoperative selection of a recipient perforator artery. PATIENTS AND METHODS: The study enrolled 38 patients with lower extremity lymphedema who underwent vascularized lymphatic tissue transfer with perforator to perforator anastomosis for physiological lymphatic flow reconstruction. In all cases, three candidate recipient perforators were searched in each lower extremities, and vessel diameter and peak systolic flow velocity (PSFV) were measured. The inclusion criteria for candidates were a vessel diameter of >0.5 mm and a PSFV of >10 cm/s. These measures were compared with intraoperative findings, diameters and if there was pulsation and visible spurting evident. RESULTS: A total of 114 candidates were selected, and 52 of the candidates were dissected until suitable perforators were found. PSFV (cm/s) on CSDUS was ≥20.0 in 32 perforators (84.2%) and was 15.0-19.9 in 6 perforators (15.8%) in the group with pulsation and visible spurting evident, and 15.0-19.9 in one perforator (7.1%) and ≤ 14.9 in 13 perforators (92.9%) in the group without pulsation and visible spurting evident. There was a statistically significant correlation between preoperative PSFV and intraoperative pulsation and visible spurting evident after dissection (P = 0.021 × 10-3 ). The flap survival rate was 92.1%. CONCLUSION: PSFV is an important preoperative determinant of the suitability of a recipient perforator artery for FTPPA.

Modified intraoperative distal compression method for lymphaticovenous anastomosis with high success and a low venous reflux rates.

INTRODUCTION: For successful lymphaticovenous anastomosis (LVA), it is important to create anastomoses with high flow to maintain patency. To ensure that this can be achieved, we compared the efficacy of a modified intraoperative distal compression (IDC) technique with the conventional no compression (NC) method for lower limb lymphedema. PATIENTS AND METHODS: In the IDC group, compression was applied to an area of the foot distal to the first LVA site. After completion of the first LVA, the distal compression was extended over the first LVA site to the distal end of the second LVA site. RESULTS: There was no significant difference between the IDC (n = 25) and NC (n = 25) groups in detection rate. However, significant differences were observed in lymphatic vessel diameter and LVA success rate. No intraoperative anastomotic obstruction was seen at the conclusion of surgery. Intraoperative congestion with blood was detected in lymphatic vessels in 8 of 79 anastomoses (10.1%) in the NC group, but not in any cases in the IDC group (p = 0.002). There was a significant between-group difference in the rate of improvement in lymphedema between the IDC (16.1±3.6) and NC groups (14.0±3.4; p = 0.03). DISCUSSION: IDC during LVA is thought to increase lymph flow in larger caliber lymphatics, leading to a high success rate and a low rate of venous reflux. IDC is beneficial when performing LVA.

Lymphovenous Anastomosis for Morbidly Obese Patients with Lymphedema.

Although patients with obesity-induced lymphedema can be treated by weight loss therapy, they find it difficult to lose the required amount of weight. The aims of this study were to clarify the characteristics of the lymphatic vessels in patients with obesity-induced lymphedema and to determine the feasibility and efficacy of lymphovenous anastomosis (LVA) in these patients. METHODS: Twenty-two patients (44 edematous lower limbs) with a body mass index (BMI) >35 kg/m2 (obese group) and 91 patients with lymphedema (141 edematous lower limbs) and BMI <25 kg/m2 were enrolled as a control group (nonobese group) and underwent LVA. The diameter and depth of lymphatics and the effect of LVA were compared. RESULTS: Lymphatics were detectable within 10-mm depth in the nonobese group and the obese group (3.0 ± 1.4 mm versus 3.5 ± 2.1 mm; P < 0.01). The lymphatic diameter was significantly greater in the obese group than in the nonobese group (0.79 ± 0.30 mm versus 0.54 ± 0.22 mm; P < 0.01). There was no significant difference in the rate of improvement in lymphedema after LVA between the nonobese group (9.1% ± 9.2%) and the obese group (8.9% ± 7.3%; P = 0.84). There was no correlation between the improvement rate of lymphedema and that of BMI in the obese group (P = 0.57). CONCLUSIONS: LVA is a feasible procedure even in morbidly obese patients. Considering that substantial weight loss is a difficult and time-consuming task for patients, LVA combined with not gaining weight is a good option for these patients.

A Simple, Versatile Device for Multiplanar Photography.

BACKGROUND: Taking pictures of protruding body parts (such as fingertips, toes, nipples, auricles, hands, and feet) from multiple directions is difficult. To solve this problem, we developed a simple and easy-to-use device, using mirrors. METHODS: The device is composed of 4 trapezoidal plane plastic mirrors firmly connected to each other. It is possible to photograph the object from 5 directions at once, including the real image of the object at the center, and 4 images that are reflected in the mirrors around the sides. RESULTS: The device allowed photographing various body parts from multiple directions at once. CONCLUSION: This simple and easy-to-use device helps us to take additional photographs in a single shot, which previously took more time and effort.

Gluteal-fold flap repair of rectovaginal fistula caused by aluminum potassium sulfate hydrate-tannic acid injection for internal hemorrhoids: a case report.

BACKGROUND: Rectovaginal fistula (RVF) following aluminum potassium sulfate hydrate-tannic acid (ALTA) injection therapy for hemorrhoids is a rare complication. We report the first case of RVF after ALTA injection therapy successfully treated by gluteal-fold flap. CASE PRESENTATION: A 49-year-old female suffered from a fever and rectal ulcer after undergoing internal hemorrhoid treatment with a submucosal ALTA injection at a previous clinic. One week after ALTA therapy, she noted obvious passage of flatus and stool through the vagina, and was diagnosed with RVF by anoscope at another clinic. She was referred to our hospital 3 weeks after ALTA therapy. Sigmoid colostomy was performed for fecal diversion as a preliminary step for fistula repair. However, the fistula was scarred and the defect between the rectum and vagina did not improve at all. Ten months after ALTA therapy, we performed fistula repair by gluteal-fold flap. Seven months later, sigmoid-colostomy reversal was performed. The patient has experienced no RVF in the 3 years since sigmoid-colostomy reversal. CONCLUSIONS: The gluteal-fold flap strategy is a useful option for severe RVF management.