Improvement in autologous human fat transplant survival with SVF plus VEGF-PLA nano-sustained release microspheres.

Early neovascularization is important for autologous fat transplant survival. SVF cells are ideal seed cells. Both vascular endothelial growth factor (VEGF) and SVF cells can promote neovascularization. However, the half-life (about 50 min) of VEGF is too short to sustain an adequate local concentration. We have investigated whether VEGF-polylactic acid (PLA) nano-sustained release microspheres plus SVF cells can improve neovascularization and survival of transplanted fat tissues. SVF cells were harvested and constructed VEGF-PLA nano-sustained release microspheres in vitro. Human fat tissues was mixed with SVF cells plus VEGF-PLA, SVF cells alone or Dulbecco's modified Eagle's medium as the control. These three mixtures were injected into random sites in 18 nude mice. Two months later, the transplants were weighed and examined histologically; and capillaries were counted to quantify neovascularization. Hematoxylin-eosin (HE) and anti-VEGF stains were applied to reveal cell infiltration. The mean wet weight of fat in the SVF plus VEGF-PLA, SVF alone, and control transplants were 0.18 ± 0.013 g, 0.16 ± 0.015 g, and 0.071 ± 0.12 g, respectively; the differences between groups were statistically significant. More vessels were present in the SVF plus VEGF-PLA transplants than in the other two types. Transplants mixed with SVF cells also had an acceptable density of capillaries. Histological analysis revealed that both the SVF plus VEGF-PLA and SVF alone transplants, but not the control transplants, were composed of adipose tissue, and had less fat necrosis and less fibrosis than control specimens. SVF plus VEGF-PLA transplants had significantly greater capillary density and VEGF expression than the other two transplant groups. Thus transplanted fat tissue survival and quality can be enhanced by the addition of VEGF-PLA nano-sustained release microspheres plus SVF cells.

Three-dimensional spheroid formation of adipose-derived stem cells improves the survival of fat transplantation by enhance their therapeutic effect.

Adipose-derived stem cells (ADSCs) have important applications in basic research, especially in fat transplantation. Some studies have found that three-dimensional (3D) spheroids formed by mesenchymal stem cells have enhanced therapeutic potential. However, the fundamental basics of this effect are still being discussed. ADSCs were harvested from subcutaneous adipose tissues and 3D spheroids were formed by the automatic aggregation of ADSCs in a non-adhesive 6-well plate. Oxygen glucose deprivation (OGD) was used to simulate the transplantation microenvironment. We found that 3D culture of ADSCs triggered cell autophagy. After inhibiting autophagy by Chloroquine, the rates of apoptosis were increased. When the 3D ADSC-spheroids were re-planked, the number of senescent ADSCs decreased, and the proliferation ability was promoted. In addition, there were more cytokines secreted by 3D ADSC-spheroids including VEGF, IGF-1, and TGF-ß. After adding the conditioned medium with human umbilical vein endothelial cells (HUVECs), 3D ADSC-spheroids were more likely to promote migration, and tube formation, stimulating the formation of new blood vessels. Fat grafting experiments in nude mice also showed that 3D ADSC-spheroids enhanced survival and neovascularization of fat grafts. These results suggested that 3D spheroids culturing of ADSCs can increase the therapeutic potential in fat transplantation.

Effectiveness and safety of the improved mini-incision surgery for osmidrosis treatment.

BACKGROUND: As the mainstream treatment of axillary osmidrosis, surgical treatment is still limited by various complications, such as paresthesia, scars, local infection, hematoma, flap necrosis, and long recovery time. In this study, we tried to adopt the improved mini-incision surgery for osmidrosis treatment. OBJECTIVES: The paper aims to evaluate the clinical effectiveness and safety of the improved mini-incision surgery for axillary osmidrosis treatment. PATIENTS/METHODS: Clinical series of patients underwent improved mini-incision surgery were retrospectively reviewed. Dates of complications, including paresthesia, scars, infection, hematoma, skin necrosis, and recurrence were analyzed. RESULTS: Among 61 cases, 58 cases had a preoperative osmidrosis score of 3 and 3 cases had a preoperative score of 2; while 13 cases had a postoperative osmidrosis score of 0, 43 cases had a postoperative score of 1 and 5 cases had a postoperative score of 2, significantly lower than that before (p < 0.001). A total of 12 axillae complications occurred, 2 axillae (1.6%) had paresthesia; 5 axillae (4.1%) had hematoma; 2 axillae (1.6%) had local flap necrosis due to hematoma; and 3 axillae (2.4%) had hypertrophic scars. CONCLUSIONS: The results showed that the improved mini-incision surgery was safe and effective for osmidrosis treatment.

Anti-aging effects of fetal dermal mesenchymal stem cells in a D-galactose-induced aging model of adult dermal fibroblasts.

The main characteristic of skin aging is the change in the composition of the dermis, mainly resulting from fibroblast senescence. Mesenchymal stem cells derived from fetal dermis are defined as fetal dermal mesenchymal stem cells; they reportedly exert wound healing effects on the skin and regulate keloid fibroblast proliferation. D-Galactose is widely used in animal aging models. In this study, we confirmed that D-galactose inhibits adult dermal fibroblast proliferation, and the inhibitory effect gradually increased with increasing concentration. Finally, we chose a concentration of 40 g/L D-galactose to induce adult dermal fibroblast senescence. D-Galactose increased the intensity of senescence-associated ß-galactosidase staining and the levels of reactive oxygen species in adult dermal fibroblasts. Furthermore, D-galactose increased the mRNA expression of p16, p21, and p53. The fetal dermal mesenchymal stem cell-conditioned medium improved the above-mentioned effects. Overall, fetal dermal mesenchymal stem cells exerted anti-aging effects against adult dermal fibroblasts induced by D-galactose via paracrine functions.

Creating natural double eyelids with continuous buried suture and mini-incision technique using subcutaneous absorbable suture for patients with puffy eyelids.

IMPORTANCE Incision and buried suture are 2 primary techniques for creating double eyelids. The incision method is suitable for all kinds of eyelids, but operational trauma and prolonged recovery time limit its application. The non-incision approach can shape a natural and vivid crease with a relatively short recovery time. However, it is not suitable for patients with puffy eyelids, and in those patients the duration of the supratarsal crease is not long.We propose a method of combining the techniques of continuous buried suture and mini-incision. OBJECTIVE To explore a new kind of double eyelid plasty, a corrective surgical procedure for patients with puffy eyelids. DESIGN, SETTING, AND PARTICIPANTS Observational study of 221 patients with puffy single eyelids who underwent this new blepharoplasty from May 2007 to and March 2012. INTERVENTIONS Combined continuous buried-suture and mini-incision surgery using subcutaneous absorbable suture to create natural double eyelids under local anesthesia. All procedures were performed by the same surgeon. MAIN OUTCOMES AND MEASURES All patients were observed after surgery for a period ranging from 1 to 28 months (mean follow-up, 16 months). Data collection included operative time, postoperative recovery and complications. RESULTS All double eyelids appeared natural after short operative time and rapid postoperative recovery, leaving an invisible scar and long-lasting supratarsal crease. No corneal damage or infection occurred.CONCLUSIONS AND RELEVANCE Combined continuous buried-suture and mini-incision surgery using subcutaneous absorbable suture to create natural double eyelids is a reliable,durable, and less-invasive technique for patients with puffy eyelids.

[Experimental study on fluorescent labeling and optimization method of purifying human stromal vascular fraction cells].

OBJECTIVE: To find a kind of simple and effective method for purifying and labeling stromal vascular fraction cells (SVFs) so as to provide a theoretical basis for clinical application of SVFs. METHODS: The subcutaneous adipose tissue were harvested form volunteers. The adipose tissue was digested with 0.065%, 0.125%, and 0.185% type I collagenase, respectively. SVFs were harvested after digestion and counted. After trypan blue staining, the rate of viable cells was observed. SVFs was labeled by 1, l'-dioctadecyl-3, 3, 3', 3'-2-tetramethy-lindocyanine perchlorate (DiI). The fluorescent labeling and growth was observed under an inverted fluorescence microscope. MTT assay was used to detect cell proliferation. RESULTS: The number of SVFs was (138.68 +/- 11.64) x 10(4), (183.80 +/- 10.16) x 10(4), and (293.07 +/- 8.31) x 10(4) in 0.065% group, 0.125% group, and 0.185% group, respectively, showing significant differences among 3 groups (P < 0.01). The rates of viable cells were 91% +/- 2%, 90% +/- 2%, and 81% +/- 2% in 0.065% group, 0.125% group, and 0.185% group, respectively, and it was significantly higher in 0.065% group and 0.125% group than in 0.185% group (P < 0.01), but no significant difference was found between 0.065% group and 0.125% group (P=0.881). Inverted fluorescence microscope showed that the cell membranes could be labeled by DiI with intact cell membrane, abundant cytoplasm, and good shape, but nucleus could not labeled. SVFs labeled by DiI could be cultured successfully and maintained a normal form. MTT assay showed that similar curves of the cell growth were observed before and after DiI labeled to SVFs. CONCLUSION: The optimal collagenase concentration for purifying SVFs is 0.125%. DiI is a kind of ideal fluorescent dye for SVFs.

[Experimental study of the effect of adipose stromal vascular fraction cells with VEGF on the neovascularization of free fat transplantation].

OBJECTIVE: To investigate the effect of adipose stromal vascular fraction cells (SVFs) with VEGF on the neovascularization of free fat transplantation. METHODS: SVFs were obtained from subcutaneous fat and labelled with DiI. 0.3 ml autologous fat tissue was mixed with 0.2 ml cells: 1) autologous SVFs with VEGF (Group A); 2) autologous SVFs (Group B); 3) complete DMEM (Group C) And then the mixture was injected randomly under the back skin of 12 nude mice. The transplanted fat tissue in three groups was harvested at 2 months after implantation. Wet weight and diameter of fat grafts was measured. After HE and CD31 staining,blood vessel density, viable adipocytes and fibrous proliferation were observed. RESULTS: Trace of SVFs labeled by DiI in vivo could be detected by fluorescent microscope. The wet weight of fat grafts was (191.90 +/- 9.81) mg in group A, (177.01 +/- 10.50) mg in group B, and (92.05 +/- 8.30) mg in group C (P<0.01). The diameter of fat grafts was (0.49 +/- 0.24) cm in group A, (0.40 +/- 0.26) cm in group B, and (0.32 +/- 0.28) cm in group C (P<0.01). Histological analysis showed the blood vessel density was (14.58 +/- 2.06)/HPL in group A, (11.55 +/- 2.18)/HPL in group B, (7.87 +/- 1.55)/HPL in group C. Compared with group B and group C, group A had more adipose tissue with less fat necrosis and fibrosis and had significantly higher capillary density. CONCLUSIONS: The autologous adipose stromal vascular fraction cells with VEGF could improve the neovascularization of free fat significantly. It indicates a wide clinical application in the future.