Protective Topical Dual-Sided Nanofibrous Hemostatic Dressing Using Mussel and Silk Proteins with Multifunctionality of Hemostasis and Anti-Bacterial Infiltration.

Topical hemostatic agents are preferred for application to sensitive bleeding sites because of their immediate locoregional effects with less tissue damage. However, the majority of commercial hemostatic agents fail to provide stable tissue adhesion to bleeding wounds or act as physical barriers against contaminants. Hence, it has become necessary to investigate biologically favorable materials that can be applied and left within the body post-surgery. In this study, a dual-sided nanofibrous dressing for topical hemostasis is electrospun using a combination of two protein materials: bioengineered mussel adhesive protein (MAP) and silk fibroin (SF). The wound-adhesive inner layer is fabricated using dihydroxyphenylalanine (DOPA)-containing MAP, which promotes blood clotting by aggregation of hemocytes and activation of platelets. The anti-adhesive outer layer is composed of alcohol-treated hydrophobic SF, which has excellent spinnability and mechanical strength for fabrication. Because both proteins are fully biodegradable in vivo and biocompatible, the dressing would be suitable to be left in the body. Through in vivo evaluation using a rat liver damage model, significantly reduced clotting time and blood loss are confirmed, successfully demonstrating that the proposed dual-sided nanofibrous dressing has the right properties and characteristics as a topical hemostatic agent having dual functionality of hemostasis and physical protection.

Vessel-Derived Decellularized Extracellular Matrices (VdECM): Novel Bio-Engineered Materials for the Wound Healing.

BACKGROUND: Decellularized extracellular matrix (dECM) is a non-cellular scaffold with various functions in tissue engineering and regenerative medicine. Elastin is related to tissue elasticity and scarless wound healing, abundantly found in lung and blood vessel tissues. We studied the characteristics of blood vessel-derived dECM (VdECM) and its effect in wound healing. METHODS: VdECM was prepared from porcine blood vessel tissue. Weight percentages of elastin of VdECM and atelocollagen were analyzed. Migratory potential of VdECM was tested by scratch assay. VdECM in hydrogel form was microscopically examined, tested for fibroblast proliferation, and examined for L/D staining. Cytokine array of various growth factors in adipocyte-derived mesenchymal stem cell (ASC) media with VdECM was done. Animal wound model showed the wound healing effect of VdECM hydrogel in comparison to other topical agents. RESULTS: VdECM contained 6.7 times more elastin than atelocollagen per unit weight. Microscopic view of 0.35% VdECM hydrogel showed consistent distribution. Compared to 3% atelocollagen, 0.35% VdECM showed superior results in fibroblast migration. Fluorescent microscopic findings of L/D assay had highest percentage of cell survival in 1% VdECM compared to atelocollagen. Growth factor expression was drastically amplified when VdECM was added to ASC media. In the animal study model, epithelialization rate in the VdECM group was higher than that of control, oxytetracycline, and epidermal growth factor ointments. CONCLUSION: VdECM contains a high ratio of elastin to collagen and amplifies expressions of many growth factors. It promotes fibroblast migration, proliferation, and survival, and epithelialization comparable to other topical agents.

Bone Regeneration with 3D-Printed Hybrid Bone Scaffolds in a Canine Radial Bone Defect Model.

BACKGROUND: The repair of large bone defects remains a significant challenge in clinical practice and requires bone grafts or substitute materials. In this study, we developed a unique hybrid bone scaffold comprising a three dimensional (3D)-printed metal plate for weight bearing and a biodegradable polymer tube serving as bone conduit. We assessed the long-term effect of the hybrid bone scaffold in repairing radial bone defects in a beagle model. METHODS: Bone defects were created surgically on the radial bone of three beagle dogs and individually-tailored scaffolds were used for reconstruction with or without injection of autologous bone and decellularized extracellular matrix (dECM). The repaired tissue was evaluated by X-ray, micro-computed tomography, and histological observation 6 months after surgery. The functional integrity of hybrid bone scaffold-mediated reconstructions was assessed by gait analysis. RESULTS: In vivo analysis showed that the hybrid bone scaffolds maintained the physical space and bone conductivity around the defect. New bone was formed adjacent to the scaffolds. Addition of autologous bone and dECM in the polymer tube improved healing by enhancing bone induction and osteoconduction. Furthermore, the beagles' gait appeared normal by 4 months. CONCLUSION: The future of bone healing and regeneration is closely related to advances in tissue engineering. Bone production using autologous bone and dECM loaded on 3D-printed hybrid bone scaffolds can successfully induce osteogenesis and provide mechanical force for functional bone regeneration, even in large bone defects.

Two-team-approached free flap reconstruction for plantar malignant melanoma: An observational (STROBE-compliant) trial.

Reconstructive treatments of heel defects usually involve regional flap techniques such as medial plantar flap procedures due to the limited availability of adjacent soft tissues. Although free flaps have advantages in terms of function and aesthetics, they remain challenging due to the longer operation time required than for regional flaps. Thus, we introduce an appropriate 2-team surgical protocol to reconstruct plantar defects after wide excision of malignant melanoma using free flap coverage. From 2015 to 2020, a retrospective study was performed including 21 patients who underwent free flap surgeries to reconstruct defects due to plantar malignant melanoma. Lymphoscintigraphy was performed to localize sentinel lymph nodes, and the procedure was carried out by 2 teams working together, a tumor-ablative team and a reconstructive team. The present study is adhered to the STROBE guidelines for cohort studies. The average operation time was 241.4 minutes and was not significantly different even in cases with inguinal dissection (P value: 0.641). All flaps survived after 2 cases of venous insufficiency and 1 case of hematoma were resolved by immediate revision surgery. The 2-team approach to surgically reconstruct heel defects after wide excision of malignant melanoma using free flap coverage offers favorable results and lower morbidity than regional flap approaches.

Quantitative Assessment of the Restoration of Original Anatomy after 3D Virtual Reduction of Long Bone Fractures.

Background: The purpose of this study was to demonstrate the usefulness of 3D image-based virtual reduction by validating the evaluation criteria according to guidelines suggested by the AO Surgery Reference. Methods: For this experiment, 19 intact radial ORTHObones (ORTHObones radius, 3B Scientific, Germany, Hamburg) without any fractures were prepared. All ORTHObones with six cortical marking holes (three points on the distal part and three points on the proximal part) were scanned using a CT scanner twice (before/after intentional fracture of the ORTHObone). After the virtual reduction of all 19 ORTHObones, accuracy evaluations using the four criteria (length variation, apposition variation, alignment variation, Rotation Variation) suggested in the AO Surgery Reference were performed. Results: The mean (M) length variation was 0.42 mm, with 0.01 mm standard deviation (SD). The M apposition variation was 0.48 mm, with 0.40 mm SD. The M AP angulation variation (for alignment variation) was 3.24°, with 2.95° SD. The M lateral angulation variation (for alignment variation) was 0.09°, with 0.13° SD. The M angle of axial rotation was 1.27° with SD: 1.19°. Conclusions: The method of accuracy evaluation used in this study can be helpful in establishing a reliable plan.

Perineal perforator switch flap for three-dimensional vulvovaginal reconstruction.

BACKGROUND: Vulvovaginal reconstruction is challenging. In this study, we evaluated the outcomes of vulvovaginal reconstruction with "perineal perforator switch flap" (PPSF) and compared it with other conventional flaps. In addition, the long-term esthetic results were compared with the perineal perforator propeller flap (PPPF), which we previously used. METHODS: We retrospectively reviewed the clinical data of 16 patients (27 flaps) who underwent vulvovaginal reconstruction with PPSF. After tumor resection, perineal perforators close to the genitofemoral sulcus were identified. The flap was designed as an island with the perforator at the center. The flap was elevated while preserving the soft tissue surrounding the pedicle, and transferred to the defect via the subcutaneous tunnel without pedicle skeletonization. RESULTS: All flaps survived and no major surgical complications were observed. The total follow-up period was 16.13±3.38 months. The mean operation time was 79.38±19.65 min, and the initiation of walking and the length of hospitalization were 1.69 ± 0.79 and 5.69 ± 0.79 days, respectively. Perineal function was well preserved. Comparison of esthetic results with PPPF showed that PPSF showed better results in symmetrical and labial shape (2.29 ± 0.73 vs. 3.13 ± 0.81; p=0.015, 2.43 ± 1.02 vs. 3.25 ± 0.68; p=0.031, respectively), and in total score. (10.29 ± 2.16 vs. 12.31 ± 1.82; p=0.017). CONCLUSION: PPSF was technically simple and significantly reduced the duration of operation and the overall recovery time. PPSF also prevented delay in radiation. Therefore, PPSF is a promising method for vulvovaginal reconstruction.

Mechanical and Biocompatibility Properties of Sintered Titanium Powder for Mimetic 3D-Printed Bone Scaffolds.

A composite comprising Ti and NaCl powders was sintered similar to a three-dimensional (3D)-printed patient-customized artificial bone scaffold. Additionally, a proper microstructure of the mimetic scaffold and the optimum processing parameters for its development were analyzed. The mechanical properties of the metal-based porous-structured framework used as an artificial bone scaffold were an optimum replacement for the human bone. Thus, it was confirmed that patient-customized scaffolds could be manufactured via 3D printing. The 3D-printed mimetic specimens were fabricated by a powder-sintering method using Ti for the metal parts, NaCl as the pore former, and polylactic acid as the biodegradable binder. Scanning electron microscopy (SEM) images showed that pores were formed homogeneously, while X-ray computed tomography confirmed that open pores were generated. The porosity and pore size distribution were measured using a mercury porosimeter, while the flexural strength and flexural elastic modulus were calculated using the three-point bending test. Based on these measurements, a pore-former content of 15 vol % optimized the density and flexural strength to 2.52 g cm-2 and 283 MPa, respectively, similar to those of the actual iliac bone. According to the 3D-printing production method, a selective laser-sintering process was applied for the fabrication of the mimetic specimen, and it was determined that the microstructure and properties similar to those of previous metal specimens could be achieved in the as-prepared specimen. Additionally, a decellularized extracellular matrix (dECM) was used to coat the surfaces and interiors of the specimens for evaluating their biocompatibilities. SEM image analysis indicated that the adipose-derived stem cells grew evenly inside the pores of the coated specimens, as compared with the bulky Ti specimens without the dECM coating. The doubling time at 65% was measured at 72, 75, and 83 h for specimens with pore-former contents of 5, 10, and 15 vol %, respectively. The doubling time without the pore former was 116 h. As compared with the specimens without the pore former (73 h), 15% of the dECM-coated specimens showed a doubling time of 64%, measured at 47 h.

A Novel Hypothesis and Characterization to Isolate Microvascular Endothelial Cells Simultaneously with Adipose-Derived Stem Cells from the Human Adipose-Derived Stromal Vascular Fraction.

BACKGROUND: Angiogenesis and vasculogenesis are essential processes for successful tissue regeneration in tissue engineering and regenerative medicine. The adipose-derived stromal vascular fraction (SVF) is not only a source of adipose stem cells (ASC) but also a suitable source of microvascular endothelial cells because it is a rich capillary network. So, we propose a new hypothesis for isolating adipose-derived human microvascular endothelial cells (HMVEC-A) from the SVF and developed a dual isolation system that isolates two cell types from one tissue. METHOD: To isolate HMVEC-A, we analyzed the supernatant discarded when ASC is isolated from the adipose-derived SVF. Based on this analysis, we assumed that the SVF adherent to the bottom of the culture plate was divided into two fractions: the stromal fraction as the ASC-rich fraction, and the vascular fraction (VF) as the endothelial cells-rich fraction floating in the culture supernatant. VF isolation was optimized and the efficiency was compared, and the endothelial cells characteristics of HMVEC-A were confirmed by flow cytometric analysis, immunocytochemistry (ICC), a DiI-acetylated low-density lipoprotein (DiI-Ac-LDL) uptake, and in vitro tube formation assay. RESULTS: Consistent with the hypothesis, we found a large population of HMVEC-A in the VF and isolated these HMVEC-A by our isolation method. Additionally, this method had higher yields and shorter doubling times than other endothelial cells isolation methods and showed typical morphological and phenotypic characteristics of endothelial cells. CONCLUSION: Cells obtained by the method according to our hypothesis can be applied as a useful source for studies such as tissue-to-tissue networks, angiogenesis and tissue regeneration, patient-specific cell therapy, and organoid chips.

Verapamil-containing silicone gel reduces scar hypertrophy.

A hypertrophic scar is a common dermal fibroproliferative lesion usually treated with topical silicone. Verapamil, a type of calcium channel blocker, is considered a candidate drug for the treatment of hypertrophic scars. Here, we report that the addition of verapamil to topical silicone gel enhances treatment outcomes of hypertrophic scars. Upon creation of hypertrophic scars with the rabbit ear model, varying concentrations of verapamil-added silicone gel (0.1, 1, and 10 mg/g) were applied daily for 28 days. After the animals were euthanised, microscopic measurement was performed for (a) scar elevation index (SEI), (b) fibroblast count, and (c) capillary count. On gross analysis, features of hypertrophic scars were significantly alleviated in the verapamil-added groups. On histologic examination, verapamil-added groups showed (a) reduced SEI (1.93 (1.79-2.67) for control vs 1.34 (1.21-1.51) for silicone only and 1.13 (1.01-1.65) for verapamil-added silicone), (b) fibroblast count 700.5 (599.5-838.5) for control, 613.25 (461-762.5) for silicone only, and 347.33 (182.5-527) for verapamil-added silicone), and (c) capillary formation (52 (35.5-96.5) for control, 46 (28-64.5) for silicone only, and 39.83(24-70) for verapamil-added silicone) (Kruskal-Wallis test, P < .05). On western blot, expression levels of collagen I protein was lower in the 1 mg/g and 10 mg/g verapamil-added silicone compared with control. Therefore, we suggest a therapeutic concentration of verapamil-added silicone gel of at least over 1 mg/g. Further study regarding maximally effective concentration and deeper insight into the mechanism of action should follow.

Nitric Oxide Produced by the Antioxidant Activity of Verapamil Improves the Acute Wound Healing Process.

BACKGROUND: Verapamil is used in the treatment of hypertension, angina pectoris, cardiac arrhythmia, hypertrophic scars, and keloids to block transmembrane calcium ion flux. Verapamil has antioxidant activity, which enhances the production of nitric oxide (NO). NO promotes the proliferation of fibroblasts, keratinocytes, endothelial cells, and epithelial cells during wound healing. In this study, we investigated the effect of verapamil and its antioxidant properties on the enhancement of acute wound healing via NO. METHODS: A full-thickness wound healing model was created on the rat dorsal with a silicone ring. The wound closure rate was estimated every 2 days for 14 days. A histological study was performed to evaluate wound healing. Immunofluorescence staining was analyzed for angiogenesis. The expressions of collagen type I (COL I), collagen type III (COL III), and vascular endothelial growth factor (VEGF) were assessed by Western blot. Real-time polymerase chain reaction (qRT-PCR) was performed to examine the expression of endothelial NO synthase and inducible NO synthase, which are related to antioxidant activity in the process of wound healing. RESULTS: The wound closure rate was faster in the verapamil group compared to the control and silicone groups. Histologic analysis revealed capillaries and stratum basale in the verapamil group. Immunofluorescence staining was shown vessel formation in the verapamil group. Western blot and qRT-PCR analysis revealed high expression levels of COL I, VEGF, eNOS, and FGF in the verapamil. CONCLUSION: Verapamil's antioxidant activity enhances NO production in acute wound healing. We suggest that verapamil can be used to promote acute wound healing.

Flexible Adipose-Vascular Tissue Assembly Using Combinational 3D Printing for Volume-Stable Soft Tissue Reconstruction.

A new concept, assembling cell-laden tissue modules, is for the first time proposed for soft tissue engineering. Adipose-vascular tissue modules composed of a synthetic polymer-based substructure and customized bioinks using planar 3D cell printing are engineered. Such tissue modules are systematically assembled into a synthetic polymer-based module holder fabricated with rotational 3D printing, resulting in the development of a flexible and volumetric tissue assembly. Whereas most of the previous studies about the construction of adipose tissue are limited to hypoxia, poor vascularization, rapid resorption, and mismatch in mechanical properties, it is aimed to realize the construction of nonhypoxic, flexible, and volume-stable tissue assembly in this study. The significance of engineered tissue assembly is proven through various in vitro and in vivo evaluations. In particular, stable volume and remarkable neovascularization/adipogenesis are observed in the implanted assembly over four weeks. Interestingly, the size of newly formed lipid droplets and the remodeled morphology in the assembly are comparable to those in native adipose tissue. As far as it is known, this work is a first report suggesting a cell printing-based tissue assembly for functional reconstruction of soft tissue.

Clinical Efficacy and Safety of Injection of Stromal Vascular Fraction Derived from Autologous Adipose Tissues in Systemic Sclerosis Patients with Hand Disability: A Proof-Of-Concept Trial.

BACKGROUND: Stromal vascular fraction (SVF) has recently emerged as a potential therapeutic modality, due to its multipotent cellular components in tissue regeneration. Systemic sclerosis (SSc) is a progressive autoimmune disease that results in hand disability by skin fibrosis and microangiopathies. We performed an open-label study to investigate the efficacy and safety of SVF injection in SSc patients (Clinical Trial number: NCT03060551). METHODS: We gathered 20 SSc patients with hand disability, planning for a 24-week follow-up period. SVF was extracted from autologous adipose tissues, processed by the closed system kit, and injected into each finger of SSc patients. We observed various efficacy and safety profiles at each follow-up visit. RESULTS: Among the 20 initially enrolled patients, eighteen received SVF injection, and were completely followed-up for the whole study period. Patients received 3.61 × 106 mesenchymal stem cells into each finger on average. Skin fibrosis, hand edema, and quality of life were significantly improved, and 31.6% of active ulcers were healed at 24 weeks after injections. Semiquantitative results of nailfold capillary microscopy were ameliorated. There was no single serious adverse event related to the procedure. CONCLUSIONS: Injection of SVF derived from autologous adipose tissues is tolerable, and shows clinical efficacy in SSc patients.

Diametric Comparison between the Thoracodorsal Vessel and Deep Inferior Epigastric Vessel in Breast Reconstruction.

BACKGROUND: In microvascular anastomosis, size discrepancy is common and can increase thrombotic complications. If size differences can be predicted, then vessels of the appropriate size can be selected. This study documented the difference in diameter between the thoracodorsal (TD) vessel and deep inferior epigastric perforator (DIEP) pedicle in each patient who underwent breast reconstruction using free tissue transfer. Patients and Methods. This retrospective study included 32 anastomoses (27 breasts including five cases of supercharged anastomosis) of breast reconstruction with the free DIEP flap and TD recipient between August 2018 and June 2019. In the microscopic view, the caliber of the TD vessel, the largest branch to the serratus anterior muscle, the descending branch, the largest and the second largest branches to the latissimus dorsi muscle, and the DIEP pedicle were measured. RESULTS: The diameter of the deep inferior epigastric artery was similar to that of the descending branch, and their anastomosing rate was 56.3%. The diameter of the deep inferior epigastric vein was similar to the branch to the serratus anterior muscle and the descending branch, and their anastomosing rates were 29.3% and 29.3%, respectively. All flaps were survived; however, in one case, a reoperation was needed to remove the hematoma, in which case fat necrosis occurred as the only complication. CONCLUSION: TD branches of similar size to the DIEP pedicle were prioritized in anastomosis. The descending branch and the branch to the serratus anterior muscle are expected to be good candidates as recipients in breast reconstruction with DIEP free flap. Moreover, supercharged anastomosis of DIEP pedicles can be achieved within TD branches.

Unilateral autologous breast reconstruction with unipedicled and bipedicled deep inferior epigastric artery perforator flap: A review of 168 cases over 3 years.

BACKGROUND: Harvesting a DIEP flap based on bipedicled perforators can enhance vascular perfusion, which can reduce complication rates, minimize fat necrosis, and flap failure. This study summarizes our experience with using unipedicled and bipedicled DIEP flaps for breast reconstruction. PATIENTS AND METHODS: A total of 168 consecutive patients undergoing unilateral breast reconstruction with DIEP flaps over a 3-year period were retrospectively reviewed. Primary microvascular anastomoses were performed to the thoracodorsal vessels in both unipedicled and bipedicled DIEP groups. In bipedicled DIEP flap cases, additional secondary microvascular anastomoses were performed either by extraflap or intraflap options. Clinical characteristics and outcomes were recorded. RESULTS: Unipedicled (n = 89; 53%) and bipedicled flaps were used. Both groups were comparable for mean age, diabetes mellitus, hypertension, smoking, and chemotherapy. BMI was 24.9 ± 3.6 in the unipedicled group and 22.8 ± 2.9 in the bipedicled group (p < .001). The surgical duration was longer in bipedicled group (367 ± 86.5 minu vs 403.7 ± 65.6 min, p < .05) but incidence of fat necrosis decreased in the bipedicle group (24 patients [27%] vs. 7 patients [8.9%] p < .05). There was no flap loss or instance of abdominal hernia in any group. CONCLUSIONS: The bipedicled DIEP flaps may be a feasible option for large breast reconstruction in thin patients. However, the additional microsurgical technical complexity and longer operative time must be considered.

Cryopreservation of lipoaspirates: in vitro measurement of the viability of adipose-derived stem cell and lipid peroxidation.

As the storage time of the fat tissue passes by, lipid peroxidation and creation of by-products may take place. The objective of this study was to evaluate the cell viability and functional changes of adipose-derived stem cells (ADSCs) in the cryopreserved lipoaspirates at different temperatures in accordance with lipid peroxidation. Lipoaspirates acquired from liposuction were divided into four different temperature groups and stored at 4°C, -20°C, -80°C, and -196°C. After isolating ADSC from each sample, gross cell morphology and cell viability were compared with doubling time and colony-forming unit (CFU) formation ability. Acid value, that is, thiobarbituric acid value was measured to assess lipid peroxidation. No viable ADSC was observed in -20°C and -196°C samples for past 1 week and a superior number of the live cells were detected in the 4°C group compared with the -80°C group. However, the persistence of cell division and CFU formation after 1 week was only observed in adipocytes stored at -80°C. Lipid peroxidation mainly occurred at 4°C and -20°C storage samples. If the lipoaspirates were planned to be cryopreserved, it is advised to store at -80°C. However, the number of actually functional ADSCs is very low. Furthermore, even in the cryopreserved status, continuous lipid peroxidation and by-product creation took place, suggesting shorter preservation period as possible in the clinics.

Efficacy of Altered Two-Point Fixation in Zygomaticomaxillary Complex Fracture.

PURPOSE: To reconstruct a zygomaticomaxillary complex (ZMC) fracture, zygomaticofrontal (ZF) suture is the most reliable site to assess anatomical alignment and to secure rigidity. It has been chosen primary site to be fixed, but approach through the lateral eyebrow incision may leave a visible scar. This study suggests altered two-point fixation of ZMC fracture without accessing the ZF suture. METHODS: In the retrospective study, a total of 40 patients with ZMC fracture were divided into two groups (group 1, two-point fixation and group 2, three-point fixation). Patient demographics and follow-up were evaluated, and degree of reduction including cortical gaps of ZF and inferior orbital (IO) area, protruding difference of zygoma, and malar difference using asymmetry index were measured through preoperative and postoperative CT. RESULTS: Preoperatively, the means of ZF displacement, IO displacement, protruding difference of zygoma, and facial asymmetry index between the groups were not statistically different. The result was the same after the operation. However, all variables were significantly different before and after surgery within each group. Moreover, mean operation time was significantly different between groups (P value = 0.026). CONCLUSION: Altered two-point fixation in ZMC fracture excluding incision approaching the ZF provides surgical efficacy and similar surgical outcomes to three-point fixation but offers reduced operation time and fewer complications.

Characterization of cell signaling, morphology, and differentiation potential of human mesenchymal stem cells based on cell adhesion mechanism.

It is essential to characterize the cellular properties of mesenchymal stem cell populations to maintain quality specifications and control in regenerative medicine. Biofunctional materials have been designed as artificial matrices for the stimulation of cell adhesion and specific cellular functions. We have developed recombinant maltose-binding protein (MBP)-fused proteins as artificial adhesion matrices to control human mesenchymal stem cell (hMSC) fate by using an integrin-independent heparin sulfate proteoglycans-mediated cell adhesion. In this study, we characterize cell adhesion-dependent cellular behaviors of human adipose-derived stem cells (hASCs) and human bone marrow stem cells (hBMSCs). We used an MBP-fused basic fibroblast growth factor (MF)-coated surface and fibronectin (FN)-coated surface to restrict and support, respectively, integrin-mediated adhesion. The cells adhered to MF exhibited restricted actin cytoskeleton organization and focal adhesion kinase phosphorylation. The hASCs and hBMSCs exhibited different cytoplasmic projection morphologies on MF. Both hASCs and hBMSCs differentiated more dominantly into osteogenic cells on FN than on MF. In contrast, hASCs differentiated more dominantly into adipogenic cells on MF than on FN, whereas hBMSCs differentiated predominantly into adipogenic cells on FN. The results indicate that hASCs exhibit a competitive differentiation potential (osteogenesis vs. adipogenesis) that depends on the cell adhesion matrix, whereas hBMSCs exhibit both adipogenesis and osteogenesis in integrin-mediated adhesion and thus hBMSCs have noncompetitive differentiation potential. We suggest that comparing differentiation behaviors of hMSCs with the diversity of cell adhesion is an important way to characterize hMSCs for regenerative medicine.

Optimal Condition of Isolation from an Adipose Tissue-Derived Stromal Vascular Fraction for the Development of Automated Systems.

BACKGROUND: The stromal vascular fraction (SVF) isolated from adipose tissue, which contains stem cells as well as other cell types, has been applied in various research fields. Although different enzymatic concentrations and treatment durations have been applied to isolate the SVF, optimal conditions have not been established. Thus, we aimed to establish the optimal conditions for isolation of the SVF from adipose tissue by automated systems. METHODS: The SVF was collected from removed adipose tissues of five donors during surgery. The SVF was treated with 0.1% or 0.2% collagenase type I for 20, 40, or 60 min. Then, colony forming unit (CFU) assays and flow cytometry were performed to characterize the adipose stem cells (ASCs). A cytokine array was used to investigate the correlation between colony-formation ability and the secretion of isolated ASCs. RESULTS: Treatment with 0.1% collagenase type I for 60 min resulted in a higher SVF yield, whereas treatment with 0.1% collagenase for 40 min resulted in higher CFU values. In addition, expression of interleukin (IL)-6, IL-8, and monocyte chemoattractant protein-1 in the SVF was higher in the high-CFU group than in the low-CFU group. CONCLUSION: The optimal conditions for isolation of the SVF from adipose tissue were treatment with 0.1% collagenase type I for 40 min. We identified the conditions required for efficient SVF isolation based on high CFU values, and our results will facilitate the development of automated systems.

One-stage nipple and breast reconstruction using a deep inferior epigastric perforator flap after a skin-sparing mastectomy.

BACKGROUND: Nipple reconstruction is usually performed as a delayed procedure in patients with breast cancer who undergo skin-sparing mastectomy and breast reconstruction surgery using a deep inferior epigastric perforator (DIEP) flap. The authors designed this study to evaluate the utility of breast reconstruction based on a DIEP flap and immediate nipple reconstruction. METHODS: A retrospective review was conducted of all patients who underwent breast reconstruction performed by a single plastic surgeon from October 2016 to June 2018. Through a questionnaire and chart review, we compared surgical. RESULTS: and complications in cases of single-stage nipple reconstruction after skin-sparing mastectomy (n=17) with patients who underwent delayed nipple reconstruction after skin-sparing mastectomy, modified radical mastectomy, or simple mastectomy (n=7). RESULTS: In a subjective analysis using clinical photos, the immediate nipple reconstruction group had higher scores than their counterparts in an evaluation of the nipple-areolar complex (NAC) (NAC placement, 3.34 vs. 3.04; nipple projection, 3.05 vs. 3.03; nipple size, 3.30 vs. 3.29). No significant differences between the groups were found in terms of complications. CONCLUSIONS: Simultaneous nipple reconstruction is a reliable surgical method with economic advantages. No differences were found in terms of outcomes and complications in comparison to delayed reconstruction. Therefore, surgeons can consider simultaneous nipple reconstruction without particular concerns about asymmetry or necrosis.

Trochanteric area reconstruction with free flap using perforators as recipients: An alternative and effective option.

BACKGROUND: Reconstructing soft tissue defect on the trochanteric area is challenging. Due to the significant complications associated with regional flap, free tissue transfer is an appropriate option. However, this area has poor recipient vessels. Therefore, we present perforators as suitable recipient vessels to facilitate the use of free flap coverage for the successful reconstruction of defects in the trochanteric area. PATIENTS AND METHODS: From 2013 to 2017, 10 patients underwent free flap reconstruction for soft tissue defects of the trochanteric area. After preoperative computed tomography or magnetic resonance imaging images confirmed the enhanced perforating artery, the skin site was identified by Doppler. If the vessel was confirmed as reliable, the operation was performed in the same manner as for other free flaps. RESULTS: All of the flaps survived, and the perforators selected for surgery included four superficial circumflex iliac artery perforators, four tensor fasciae latae artery perforators, and two inferior gluteal artery perforators. The average diameter of the recipient artery was 0.97 mm and that of the vein was 0.94 mm. One case exhibited arterial insufficiency caused by compression of hematoma; however, complete flap survival was achieved in this case with revised surgery. CONCLUSION: Reconstructing soft tissue defects in the trochanteric area is limited in recipient vessels. However, using a perforator vessel as a recipient facilitates the reconstruction by free flap coverage. This method leads to acceptable flap survival and sufficient padding, with reduced morbidity and collateral injury.