Improving dermal fibroblast-to-epidermis communications and aging wound repair through extracellular vesicle-mediated delivery of Gstm2 mRNA.

Skin aging is characterized by the disruption of skin homeostasis and impaired skin injury repair. Treatment of aging skin has long been limited by the unclear intervention targets and delivery techniques. Engineering extracellular vesicles (EVs) as an upgraded version of natural EVs holds great potential in regenerative medicine. In this study, we found that the expression of the critical antioxidant and detoxification gene Gstm2 was significantly reduced in aging skin. Thus, we constructed the skin primary fibroblasts-derived EVs encapsulating Gstm2 mRNA (EVsGstm2), and found that EVsGstm2 could significantly improve skin homeostasis and accelerate wound healing in aged mice. Mechanistically, we found that EVsGstm2 alleviated oxidative stress damage of aging dermal fibroblasts by modulating mitochondrial oxidative phosphorylation, and promoted dermal fibroblasts to regulate skin epidermal cell function by paracrine secretion of Nascent Polypeptide-Associated Complex Alpha subunit (NACA). Furthermore, we confirmed that NACA is a novel skin epidermal cell protective molecule that regulates skin epidermal cell turnover through the ROS-ERK-ETS-Cyclin D pathway. Our findings demonstrate the feasibility and efficacy of EVs-mediated delivery of Gstm2 for aged skin treatment and unveil novel roles of GSTM2 and NACA for improving aging skin.

Knockdown of ZEB1 Inhibits Hypertrophic Scarring through Suppressing the Wnt/ß-catenin Signaling Pathway in a Mouse Model.

BACKGROUND: Hypertrophic scars (HS) cause functional impairment and cosmetic deformities following surgeries or burns (30% to 94%). There is no target therapy yet because the pathogenesis of HS progression is not well-known. In tissue fibrosis, Zinc finger E-box binding homeobox 1 (ZEB1) abnormal upregulation is an important cause for extracellular matrix (ECM) overexpression, which is the main molecular change in HS. Therefore, we hypothesized that ZEB1-knockdown inhibits HS formation. METHODS: ZEB1 expression in human HS and TGF-ß1-induced fibroblasts were identified by PCR and western blotting. ZEB1 was knockdown by siRNA in HS fibroblasts (HSFs) and mouse HS model (C57/BL6, male, 8-12 weeks). After 8-hour-transfection, HSFs were subjected to PCR, western blotting and CCK-8, apoptosis, migration and contraction assays. Mice HS were analyzed by HE staining, PCR and western blotting after 56 days. RESULTS: ZEB1 was upregulated in HS tissue (2.0-fold; p < 0.001). ZEB1 knockdown inhibited HSFs activity (0.6 to 0.7-fold; p < 0.001), the expression of fibrotic markers (0.4 to 0.6-fold; p < 0.001) and ß-catenin, cyclinD1 and c-Myc expression (0.5-fold; p < 0.001). In mouse HS models, HS skin thickness was thinner (1.60 ± 0.40 mm vs. 4.04 ± 0.36 mm; p < 0.001) after ZEB1 knockdown. CONCLUSIONS: Knockdown of ZEB1 inhibits HS formation both in vitro and in vivo. However, this is an in vitro/mouse model and more validation is needed. CLINICAL RELEVANCE STATEMENT: The discovery of ZEB1 as a mediator of HS formation might be a potential therapeutic target in HS treatment.

Conjoint fascial sheath suspension with levator muscle advancement for severe blepharoptosis.

In patients with severe blepharoptosis, the function of the levator muscle is usually weak. Even if a large amount of levator is resected, under-correction and recurrence often occur postoperatively. Frontalis suspension is the first choice for severe ptosis; however, the external orbital lifting force of the frontalis causes non-physiological eyelid movement. Conjoint fascial sheath (CFS) is a fibrous tissue which can provide dynamic movement of upper eyelids and has been applied for the treatment of mild and moderate blepharoptosis in recent years. This study aims to assess the efficacy and safety of CFS suspension combined with levator muscle advancement for treating severe blepharoptosis. A retrospective study included 44 patients (60 eyelids) with severe ptosis who underwent the modified technique. Preoperatively, levator muscle function and margin reflex distance 1 (MRD1) were measured. Surgical outcomes, symmetry results and complications were evaluated postoperatively. At the 12-18 months follow-up, adequate or normal correction was achieved in 56 eyelids (93.3%), and 37 patients (84.1%) presented good or fair symmetry results. The most common complication was conjunctival prolapse, which was observed in six eyelids (10.0%), followed by lid fold deformity and under-correction. No exposure keratitis was recorded. In conclusion, the modified technique can physically elevate the eyelid with limited tissue injury and is effective for the correction of severe ptosis. Both satisfactory functional and esthetic results were achieved, and severe complications (such as exposure keratitis) were not observed.

Correction of Mild-to-Moderate Sunken Upper Eyelids of Asians with Stromal Vascular Fraction Gel.

INTRODUCTION: Fat grafting is an efficient and safe procedure for the correction of upper eyelid sulcus deepening. Stromal vascular fraction (SVF) gel has been proven to be an ideal fat derivative and can be widely used for facial augmentation. We aimed to determine the efficacy of SVF gel for the correction of a mild-to-moderate sunken superior sulcus among Asian patients. METHODS: Patients with a mild-to-moderate sunken superior sulcus underwent SVF gel grafting of the sunken upper periorbital area. The primary result was the quantitative volume difference in the superior sulcus region before and after grafting. This was evaluated through three-dimensional VECTRA® imaging. The secondary results included the aesthetic quantitative evaluation results (upper lid area and pretarsal space ratio), global aesthetic improvement scale (GAIS) score, and complications. RESULTS: Thirty-one patients with mild-to-moderate sunken upper eyelids were included in this study. The average unilateral injected amount was 1.235 mL (± 0.171 mL). The 1-year delta volume was 0.801 ± 0.086 mL, and the effective survival volume was 65.3% (± 6.1%). The median preoperative pretarsal space and upper lid area ratio was 1.010 (± 0.150). The median postoperative pretarsal space and upper lid area ratio at 1 year was 0.159 (± 0.031) (n = 62; P < 0.0001), indicating a significantly reduced sunken appearance. The average GAIS score was 2.174 (± 0.391). All patients were satisfied with their surgical outcomes. The reoperation rate was 12.9%. CONCLUSIONS: SVF gel is safe and effective for the treatment of a mild-to-moderate sunken superior sulcus and is associated with satisfactory clinical outcomes and short recovery times.

Angiogenesis modulation-mediated inhibitory effects of tacrolimus on hypertrophic scar formation.

Hypertrophic scar (HS) is a fibroproliferative disorder that causes cosmetic as well as functional problems; however, to our knowledge, there is no satisfactory treatment for HS to date. Previous studies have indicated that angiogenesis plays a crucial role in HS formation; therefore, anti-angiogenetic therapies are considered effective in improving HS. Although tacrolimus (TAC) has been proven effective in preventing HS formation in vivo and in vitro, its underlying mechanism remains controversial and ambiguous. Because of its anti-angiogenic effects in other diseases, we aimed to determine whether TAC reduces HS by suppressing angiogenesis. Using a rabbit ear HS model that we developed, HS was treated once a week with normal saline, dimethyl sulfoxide, or TAC for 3 weeks. Histological evaluation indicated that TAC significantly reduced collagen deposition and microvessel density in scar tissues. Moreover, immunofluorescence staining for CD31 and vascular endothelial growth factor (VEGF)-A revealed that TAC significantly inhibited HS angiogenesis. In vitro analysis showed that TAC inhibited endothelial cell migration and tubulogenesis as well as the viability and proliferation of human umbilical vascular endothelial cells (HUVECs) and HS fibroblasts (HSFBs). Furthermore, TAC significantly downregulated the expression of the human angiogenetic factors VEGF-A, FGF-2, PDGF-ß, and TGF-ß1 in HUVECs and HSFBs. Additionally, TAC-mediated inhibition of angiogenesis decreased the gene expression of crucial fibrotic markers, including α- smooth muscle actin and collagens 1 and 3, in HSFBs. This is the first study to demonstrate the inhibitory effects of TAC on HS formation mediated by a mechanism involving the suppression of scar angiogenesis.

Management of Complications in 257 Cases of Breast Augmentation with Polyacrylamide Hydrogel, using Two Different Strategies: A Retrospective Study.

BACKGROUND: Polyacrylamide hydrogel (PAAG) has been used globally for breast augmentation, leading to long-term clinical complications. However, whether the infiltrated fibrotic capsule should be removed with PAAG to alleviate the complications remains unclear. This study aimed to ascertain different causes of complications and proper management strategies for PAAG removal in augmented breasts. METHODS: From July 2015 to December 2019, patients who underwent breast augmentation with PAAG and in whom surgical intervention was undertaken for PAAG-associated adverse events at Shanghai Ninth People's Hospital were retrospectively reviewed. Patients were categorized into two groups according to whether the fibrotic capsule was removed (RFC) or not (NRFC). Aesthetic outcomes, PAAG residues, and adverse events were evaluated post-operatively to assess whether important issues pertaining to these arose following fibrotic capsule removal. Tissue histology and PAAG degradation analysis were implemented to investigate immune response, degradability, and toxicity of PAAG. RESULTS: Altogether, 257 patients (88 RFC and 169 NRFC patients) were enrolled. 73.4% and 79.5% of the RFC and NRFC groups showed fairly good outcomes, with no significant difference, respectively. (X2 = 0.0804, p = 0.79) Significant differences were found between two surgical techniques upon patient satisfaction, respectively. (X2 = 3.529; p = 0.0301). Predictor of poor outcomes identified scar (OR, 4.555, p = 0.0019) and PAAG residue (OR, 5.379, p = 0.0003). Predictor of patient satisfaction identified post-operative outcomes (OR, 3.797; 95% CI, 1.860-8.923; p = 0.0002) and surgical technique (NRFC) (OR, 2.519; 95% CI, 1.449-4.434; p = 0.0008). CONCLUSIONS: Both treatment strategies showed good results in our study. Removal of the fibrotic capsule from infiltration of PAAG largely depends on the individual psychological condition, aesthetic expectations, complications, and magnetic resonance imaging results. While PAAG does not degrade in the host's body over time, it may elicit immune reactions and chronic inflammation in the long term. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine Ratings, please refer to Table of Contents or online Instructions to Authors www.springer.com/00266.

Aesthetic Correction of Mild-to-Moderate Blepharoptosis Among Asians: The Bridge Technique.

INTRODUCTION: Although several surgical methods have been introduced to treat mild-to-moderate blepharoptosis, including levator-based techniques such as Müller muscle-conjunctival resection, few complications and better functional and aesthetic outcomes remain elusive for plastic surgeons. Hence, this study aimed to provide a new technique (bridge technique) using the levator aponeurosis-Müller's muscle flap to achieve optimal blepharoptosis correction for function and aesthetics among Asians. METHODS: From January 2019 to May 2020, this new technique was performed on 157 consecutive patients with mild-to-moderate blepharoptosis. Our technique was based on the anchor of the levator complex to the tarsus using mattress stitches and three-layer fixation, which provided a reliable motion transmitter for elevating the upper eyelid. The patients' medical records and photographs were reviewed 12 months postoperatively to assess the margin reflex distance 1 (MRD1), incidence of complications, and aesthetic outcomes. RESULTS: The average preoperative and postoperative MRD1 measured 1.2 ± 0.31 mm and 3.7 ± 0.34 mm, respectively. A significant difference was observed between the preoperative and postoperative distance values (p < 0.001). In the aesthetic evaluation, the grading was good, fair, and poor for 142 (90.4%) patients, 10 (6.4%) patients, and 5 (3.2%) patients, respectively. Complications included undercorrection in 12 (5.2%) cases and overcorrection in nine (3.9%) cases, and no cases of residual lagophthalmos were recorded. CONCLUSION: Mild-to-moderate blepharoptosis among Asians can be corrected effectively using this new technique. However, the long-term outcomes of this procedure should be explored.

Comparison of the Rheological Properties and Structure of Fat Derivatives Generated via Different Mechanical Processing Techniques: Coleman Fat, Nanofat, and Stromal Vascular Fraction-Gel.

Importance: Coleman fat, nanofat, and stromal vascular fraction-gel (SVF-gel) are three widely used fat derivatives. However, their rheological properties and structure remain unknown. Objectives: To disclose the rheological properties and structure of three different fat derivatives. Design, Settings, and Participants: Fat tissues obtained from eight different donors were processed into three separate groups: Coleman fat, nanofat, and SVF-gel (n = 8); their viscoelastic properties and structure were determined. Intervention: Oscillation measurements were performed in the context of serrated 25-mm parallel-plate geometry with a 1.2-mm gap at 25°C. In addition, fat samples were fixed using a patented protocol and observed under scanning electron microscopy. Main Outcomes and Measures: Comparison of the viscoelastic properties, microstructure, and particle size. Results: At 0.77 Hz, the elastic modulus of SVF-gel, Coleman fat, and nanofat was 201.6 ± 0.74, 69.94 ± 15.61, and 34.89 ± 3.484 Pa, respectively; their viscosity was 44.06 ± 3.038, 15.37 ± 2.0380, and 7.516 ± 0.7250 mPa, respectively. The particle size of SVF-gel, Coleman fat, and nanofat was 106.0 ± 4.796, 86.93 ± 3.597, and 12.61 ± 7.603 µm, respectively. Conclusion and Relevance: Mechanical processing may impact graft efficacy. The characterization of the rheological properties and structure of different fat derivatives in this study may help surgeons select the better type of tissue for a given intervention; however, further studies are still required.

Impact of human adipose tissue-derived stem cells on dermatofibrosarcoma protuberans cells in an indirect co-culture: an in vitro study.

BACKGROUND: Autologous adipose tissue transfer may be performed for aesthetic needs following the resection of dermatofibrosarcoma protuberans (DFSP), the most common cutaneous soft tissue sarcoma, excluding Kaposi sarcoma. The regenerative effectiveness of cell-assisted lipotransfer is dependent on the presence of adipose tissue-derived stem cells (ADSCs). This is the first study to evaluate the potential oncological risks as ADSCs could unintentionally be sited within the proximity of the tumor microenvironment of DFSP cells. METHODS: Primary DFSP cells were indirectly co-cultured with ADSCs in a conditioned medium or in a Transwell system. The impact was analyzed by assessing proliferation, migration, invasion, angiogenesis, and tumor-associated genes and proteins. Results of these assays were compared between co-culture and mono-culture conditions. RESULTS: Our experimental results showed that ADSCs were able to promote proliferation, migration, invasion, and angiogenesis of DFSP cells; this was accompanied by a significant increase in the expression levels of beta-type platelet-derived growth factor receptor, collagen type I alpha 1 chain, vascular endothelial growth factor, hepatocyte growth factor, and basic fibroblast growth factor. CONCLUSIONS: The current report clearly demonstrates that ADSCs can enhance different malignant properties of DFSP cells in vitro, which should not be neglected when considering the clinical use of human ADSCs and its related derivatives in skin regenerative therapies.

Experience of Two Different Techniques of Medial Epicanthoplasty with Four-Point Homologous Design: A Cohort Study.

BACKGROUND: Various methods have been introduced to eliminate the epicanthus; however, there are no clear guidelines to determine the optimal technique for epicanthus. We aimed to investigate the clinical effect of modified Z-plasty and modified Y-V flap on epicanthus and report our experience. METHODS: Medical records, including photographic information and scar score, of 81 eyes of 43 patients with congenital epicanthus were collected. All patients underwent epicanthus correction surgery by the modified Z-plasty or modified Y-V flap methods with a four-point design from January 2018 to December 2019 in Shanghai Ninth People's hospital and completed a follow-up of at least 6 months. The cosmetic outcome was evaluated in terms of by photographic evaluation and scar score, and the structural outcome was assessed by intercanthal distance (ICD) and palpebral fissure length, inner canthal angle, the canthal tilt angle. The results of two surgical techniques were compared and analyzed. RESULTS: The cosmetic outcome of both group is good in both groups. ICD is smaller and inner canthal angle is larger in modified Z-plasty group, compared to modified Y-V flap group, respectively (P < 0.05). Patients with severe epicanthus in the modified Z-plasty group showed greater improvement in structural outcomes than in the modified Y-V flap group (P < 0.05), while those with mild epicanthus in the modified Y-V flap group showed more improvement in ICD than those in the modified Z-plasty group (P < 0.05). By 6 months, scarring in the modified Z-plasty group was more hypertrophic and obvious compared to the modified Y-V flap group (P < 0.05). CONCLUSION: Modified Y-V flap technique caused less scarring. Conversely, the modified Z-plasty technique was more efficient in shortening the ICD and increasing inner canthal angle. While modified Z-plasty reconstruction is more suitable for patients with severe epicanthus, the modified Y-V flap is more suitable for patients with mild epicanthus. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Combination of Extended Paramedian Forehead Flap and Laser Hair Removal in the Reconstruction of Distal Nasal Defect.

AIM: The study sought to determine whether extended forehead flap by using part of the scalp combined with laser hair removal is a practical approach when repairing distal nasal defect. METHODS: A retrospective study was conducted with 6 patients who underwent reconstruction of nasal defects with extended forehead flaps (including scalp combined with subsequent laser hair removal) at Shanghai Ninth People's Hospital between June 2016 and December 2018. Surgical data collected included age, causes of defects, sizes of defects, invaded nasal subunits, implantation of tissue expander, time elapsed between the formation of defects and surgical treatments, sessions of laser hair removal, and follow-up results. All patients had nasal defects after removal of a benign nevus (diameter, >1.5 cm), which invaded distal nasal subunits (including the nasal tip, soft triangle, columella, and nasal ala) and could not be repaired using adjacent tissue. Preoperative and postoperative assessments of esthetic outcomes were analyzed, including skin color and texture match, appearance deformity, and scar appearance. RESULTS: All 6 female patients (ages 4 to 24 years) accepted the extended forehead flap technique and subsequent laser hair removal treatments. Among the 6 patients, 2 had nasal tip defects, 1 had a columella nasi defect, one had an alar defect, and 2 had multiunit defects. All transferred forehead flaps survived with no necrosis and underwent 2 to 5 subsequent laser hair removal treatments. The color and texture of the transferred flaps was similar to the adjacent skin in all patients. All patients were satisfied with the final outcome of the reconstructed nose with no obvious scarring during follow-up. CONCLUSIONS: Extending the forehead flap using the scalp is a practical and safe approach for the repair of large and distal nasal defects. Subsequent laser hair removal can eliminate the hair follicles of the scalp and improve the appearance and color of the flap.

Human adipose-derived stem cells enriched with VEGF-modified mRNA promote angiogenesis and long-term graft survival in a fat graft transplantation model.

BACKGROUND: Fat grafting, as a standard treatment for numerous soft tissue defects, remains unpredictable and technique-dependent. Human adipose-derived stem cells (hADSCs) are promising candidates for cell-assisted therapy to improve graft survival. As free-living fat requires nutritional and respiratory sources to thrive, insufficient and unstable vascularization still impedes hADSC-assisted therapy. Recently, cytotherapy combined with modified mRNA (modRNA) encoding vascular endothelial growth factor (VEGF) has been applied for the treatment of ischemia-related diseases. Herein, we hypothesized that VEGF modRNA (modVEGF)-engineered hADSCs could robustly enhance fat survival in a fat graft transplantation model. METHODS: hADSCs were acquired from lipoaspiration and transfected with modRNAs. Transfection efficiency and expression kinetics of modRNAs in hADSCs were first evaluated in vitro. Next, we applied an in vivo Matrigel plug assay to assess the viability and angiogenic potential of modVEGF-engineered hADSCs at 1 week post-implantation. Finally, modVEGF-engineered hADSCs were co-transplanted with human fat in a murine model to analyze the survival rate, re-vascularization, proliferation, fibrosis, apoptosis, and necrosis of fat grafts over long-term follow-up. RESULTS: Transfections of modVEGF in hADSCs were highly tolerable as the modVEGF-engineered hADSCs facilitated burst-like protein production of VEGF in both our in vitro and in vivo models. modVEGF-engineered hADSCs induced increased levels of cellular proliferation and proangiogenesis when compared to untreated hADSCs in both ex vivo and in vivo assays. In a fat graft transplantation model, we provided evidence that modVEGF-engineered hADSCs promote the optimal potency to preserve adipocytes, especially in the long-term post-transplantation phase. Detailed histological analysis of fat grafts harvested at 15, 30, and 90 days following in vivo grafting suggested the release of VEGF protein from modVEGF-engineered hADSCs significantly improved neo-angiogenesis, vascular maturity, and cell proliferation. The modVEGF-engineered hADSCs also significantly mitigated the presence of fibrosis, apoptosis, and necrosis of grafts when compared to the control groups. Moreover, modVEGF-engineered hADSCs promoted graft survival and cell differentiation abilities, which also induced an increase in vessel formation and the number of surviving adipocytes after transplantation. CONCLUSION: This current study demonstrates the employment of modVEGF-engineered hADSCs as an advanced alternative to the clinical treatment involving soft-tissue reconstruction and rejuvenation.

Tarsal-Fixation With Aponeurotic Flap Linkage in Blepharoplasty: Bridge Technique.

BACKGROUND: Double eyelid blepharoplasty is the most popular cosmetic surgery among Asian populations. Although various surgical procedures have been reported, certain complications still arise, including asymmetry or the disappearance of the double eyelid crease, an unnatural double eyelid crease curve, and distinct scars. OBJECTIVES: The aim of this study was to describe a new blepharoplasty technique intended to reduce the various complications and create dynamic, natural-looking double eyelids. METHODS: From September 2018 to June 2019, the new technique was performed on 92 Chinese patients. The core of our technique was to construct a reliable motion transmitter, the orbital septum-levator aponeurosis flap, which was sutured to the tarsus. The outcomes were assessed 6 months after the surgery by reviewing medical records and photographs to assess the aesthetic outcomes, the relapse rate, and the incidence of complications. RESULTS: In the aesthetic outcome assessment, 86 patients were graded as good and 4 were graded as fair; of the latter, 2 had uneven scars and 2 showed showed a slight asymmetry of the double eyelid curve in length or width. Two patients were graded as poor because of the disappearance of the double eyelid crease. The relapse rate was 2.2%. Two patients had temporary hematoma. Six patients had lagophthalmos that subsided within 30 days. No cases of infection or ptosis were observed. CONCLUSIONS: The bridge technique is an effective method for creating dynamic, natural-looking double eyelids. However, the long-term outcome of this new surgical approach requires further study.