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Background: Autologous fat grafting is a popular technique in plastic surgery. A mechanical processing method is used to facilitate fat injection. No study has investigated whether this process affects cell quality and preservation of biological functionality. This study analyzed the influence of quick mechanical processing through two interconnected small-diameter syringes ("shuffling") on both structure and viability of fat tissue, and on viability, clonogenicity, and differentiation of the freshly isolated stromal vascular fraction., Methods: Lipoaspiration was performed in six healthy donors, followed by shuffling the fat either zero, five, or 30 times between two 10-cc syringes. Thereafter, fat was applied through a 1.5-mm cannula as in a clinical setting for autologous fat grafting. Analysis of different treatment conditions was conducted. Immunofluorescent staining allowed assessment of morphology, viability, composition, and damage of the tissue. The stromal vascular fraction was examined for isolation yield, viability, clonogenicity, and differentiation capacity., Results: The process of shuffling changed the macroscopic but not the microscopic structure of the lipoaspirated fat. No difference in cell number, viability, number of lipid droplets, vascular architecture, or ratio of cell composition was found. Analysis of the stromal vascular fraction, apart from large interdonor variability, did not show a significant change in isolation yield, viability, clonogenicity, or adipogenic differentiation capacity of the expanded cells., Conclusions: The mechanical procedure of shuffling lipoaspirated fat does not alter its tissue viability or its microscopic structure. The absence of impact on the stromal vascular fraction in the assessed parameters suggests that shuffling can be executed according to surgical needs.

Background: Tendon hydrogel is a promising new injectable substance that has been shown to improve repair strength after tendon injury. This study assesses the capacity of platelet-rich plasma to stimulate proliferation and migration of rat adipose-derived stem cells in tendon hydrogel in vitro., Methods: To assess proliferation, adipose-derived stem cells were exposed to plasma, plasma supplemented with growth factors, or platelet-rich plasma in culture medium and tendon hydrogel. To assess migration, adipose-derived stem cells were plated onto tendon hydrogel -coated wells and covered with medium containing plasma, plasma supplemented with growth factors, platelet-rich plasma, or bovine serum albumin. Migration from cell-seeded to cell-free zones was assessed at 12-hour intervals., Results: Platelet-rich plasma augmented proliferation to a greater extent compared with plasma and plasma supplemented with growth factors (10%: optical density, 1.18 versus 0.75 versus 0.98, respectively). Platelet-rich plasma was superior to plasma in tendon hydrogel (10%: optical density, 1.19 versus 0.85) but did not augment proliferation to the extent that plasma supplemented with growth factors did (10%: optical density, 1.19 versus 1.56). Platelet-rich plasma enhanced the migration of adipose-derived stem cells compared with serum-free medium (bovine serum albumin) (36 hours: platelet-rich plasma, 1.88; plasma, 1.51; plasma plus growth factor, 1.80; bovine serum albumin, 1.43)., Conclusions: Tendon healing is mediated by migration of cells to the injured area and cellular proliferation at that site. Tendon hydrogel supplemented with platelet-rich plasma stimulates these processes. Future studies will evaluate this combination's ability to stimulate healing in chronic tendon injuries in vivo.

Background: Recent research indicates that the adipose tissue of nonvascularized grafts is completely remodeled within 3 months, although origins of next-generation cells are unclear., Methods: Inguinal fat pads of green fluorescent protein mice and wild-type mice were cross-transplanted beneath the scalp. At 1, 2, 4, and 12 weeks after transplantation, grafted fat was harvested, weighed, and analyzed through immunohistochemistry, whole-mount staining, and flow cytometry of cell isolates. Bone marrow of green fluorescent protein mice was transplanted to wild-type mice (after irradiation). Eight weeks later, these mice also received fat grafts, which were analyzed as well., Results: The majority of host-derived cells detected during remodeling of grafted fat were macrophages (>90 percent at the early stage; 60 percent at 12 weeks). Cell origins were analyzed at 12 weeks (i.e., when completely regenerated). At this point, mature adipocytes were largely derived from adipose-derived stem/stromal cells of grafts. Although vascular wall constituents were chiefly graft derived, vascular endothelial cells originated equally from graft and host bone marrow. Adipose-derived stem/stromal cells of regenerated fat were an admixture of grafted, host nonbone marrow, and host bone marrow cells., Conclusions: The above findings underscore the importance of adipose stem/stromal cells in the grafted fat for adipocyte regeneration. Host bone marrow and local tissues contributed substantially to capillary networks and provided new adipose-derived stem/stromal cells. An appreciation of mechanisms that are operant in this setting stands to improve clinical outcomes of fat grafting and cell-based therapies.

Calcinated antler cancellous bone (CACB) is useful in repair of bone defects, as its composition and architecture is analogous to natural extracellular bone matrix. The use of CACB scaffolds with adipose-derived stem cells (ADSCs) in repair of rabbit mandibular bone defects was investigated along with the underlying molecular signaling pathways involved. CACB promoted the adhesion, spreading, and viability of ADSCs. Increased extracellular matrix production and expression of osteogenic markers in ADSCs were observed when seeded in CACB. The temporal kinetics of mRNA expression of ADSCs cultured in CACB lagged in comparison with that observed in cells grown in medium with osteogenic supplements. Activation of the extracellular signal-related kinases (ERK) 1/2 and RUNX-2 in CACB-cultured ADSCs was observed, and this activation was attenuated by the MeK inhibitor U0126. Microcomputed tomography scanning analysis and histological evaluations showed that loading the CACB with ADSCs resulted in enhanced new bone formation and angiogenesis when the composites were implanted in rabbit mandibular defects. These results indicated that the osteogenic behavior of ADSCs might be driven by the microenvironment formed by CACB via the ERK signaling pathway. These CACB/ADSCs composites have promising therapeutic potential for large bone defect repairs.

Background: Fat tissue graft has been commonly used for soft tissue augmentation. However, the mechanisms underlying the maintenance of graft volume and weight are still unclear. As morphological features provide direct evidences for cell death and survival, we aimed to investigate the fate of grafted adipocytes and the dynamic changes in the remodeling of adipose tissues by transmission electron microscopy technique., Methods: The unilateral inguinal fat pad of C57BL/6J mice was autografted to the dorsa of the mice. Perilipin expression and morphological changes were investigated by immunohistochemistry staining and transmission electron microscopy, respectively, in grafted tissues collected at posttransplantation days 0, 1, 3, 5, 7, 14, and 30., Results: Transmission electron microscopy analysis revealed that most adipocytes in grafts showed traits of cell death on postgrafting day 3. Multilocular adipocytes with naive nuclei were observed as early as day 5 and a larger number of multilocular adipocytes were found on day 14. Perilipin immunostaining revealed that only some adipocytes located in the margin of grafts survived through the ischemic injury. New adipocytes were visualized at the periphery of the grafts, although the scope of viable adipocyte zonal areas increased from day 5 to day 30., Conclusions: The results provide ultrastructural evidences associated with the remodeling dynamics of adipose tissue grafts. It is suggested that maximized volume of graft should be obtained through promoting regeneration other than improving survival of grafted adipose tissues.

Background: The aim of this study was to evaluate the effects on adipocyte morphology of 2 techniques of fat harvesting and of fat purification in lipofilling, considering that the number of viable healthy adipocytes is important in fat survival in recipient areas of lipofilling., Methods: Fat harvesting was performed in 10 female patients from flanks, on one side with a 2-mm Coleman cannula and on the other side with a 3-mm Mercedes cannula. Thirty milliliter of fat tissue from each side was collected and divided into three 10 mL syringes: A, B, and C. The fat inside syringe A was left untreated, the fat in syringe B underwent simple sedimentation, and the fat inside syringe C underwent centrifugation at 3000 rpm for 3 minutes. Each fat graft specimen was processed for examination under low-vacuum scanning electron microscope. Diameter (μ) and number of adipocytes per square millimeter and number of altered adipocytes per square millimeter were evaluated. Untreated specimens harvested with the 2 different techniques were first compared, then sedimented versus centrifuged specimens harvested with the same technique were compared. Statistical analysis was performed using Wilcoxon signed rank test., Results: The number of adipocytes per square millimeter was statistically higher in specimens harvested with the 3-mm Mercedes cannula (P = 0.0310). The number of altered cells was statistically higher in centrifuged specimens than in sedimented ones using both methods of fat harvesting (P = 0.0080) with a 2-mm Coleman cannula and (P = 0.0050) with a 3-mm Mercedes cannula. Alterations in adipocyte morphology consisted in wrinkling of the membrane, opening of pore with leakage of oily material, reduction of cellular diameter, and total collapse of the cellular membrane., Conclusions: Fat harvesting by a 3-mm cannula results in a higher number of adipocytes and centrifugation of the harvested fat results in a higher number of morphologic altered cells than sedimentation.

Background: The use of cryopreserved adipose tissue for soft tissue augmentation is common, but unpredictability of fat graft viability remains a limitation. Adipose-derived stem cell (ADSC) and stromal vascular fraction (SVF) have been introduced to enhance viability and improve the survival of transplanted fat tissue., Objective: To investigate whether supplementation with ADSC or SVF improved the survival of cryopreserved fat grafts., Methods: The cryopreserved fat grafts were treated with ADSC, SVF, or normal saline in 30 six-week-old male nude mice to test whether ADSC and SVF could improve the survival of the transplanted fat tissue. The authors examined the weight, volume, and histological features of each group (n = 10) at 8 weeks after transplantation to evaluate the survival of the fat tissue., Results: There was no difference between the control and SVF groups with respect to weight, volume, and histological findings. However, the ADSC group showed a significant increase in weight and volume compared with the control and SVF groups. Histological examination showed that the ADSC supplementation improved the quality of the transplanted fat grafts., Conclusion: Taken together, these results suggest a potential clinical utility of ADSC but no advantage of SVF in facilitating cryopreserved fat transfer.

Background: Because of their easy accessibility and versatile biological properties, mesenchymal stem cells taken from fatty tissue (adipose-derived stem cells, ADSC) are attractive for various potential clinical uses. For example, ADSC can be added to fatty tissue before transplantation in the hope of improving the outcome of autologous lipotransfer: the modified procedure is called cell-assisted lipotransfer. The clinical use and commercial promotion of this novel stem-cell treatment (and others) are spreading rapidly, even though there is not yet any clear clinical evidence for its safety and efficacy., Methods: In cooperation with the German Cochrane Center, we systematically searched the literature according to the PRISMA criteria. Eight major medical databases were searched. The retrieved publications were examined by two independent reviewers and assessed using objective criteria., Results: After screening of the 3161 retrieved publications by title, abstract, and (where appropriate) full text, 78 were still considered relevant. 13 of these were reports of clinical studies; only 3 of the 13 met criteria for grade II or III evidence. The studies that were analyzed involved a total of 286 cell-assisted lipotransfer procedures with a longest follow-up time of 42 months. Oncological safety was not demonstrated., Conclusion: The studies published to date have not shown that cell-assisted lipotransfer is generally superior to conventional autologous lipotransfer. They dealt with safety aspects inappropriately or not at all. The case of cell-assisted lipotransfer illustrates the indispensability of high-quality clinical evidence before the introduction of novel stem-cell-based treatments.

"Adipose tissue is a valuable, exploitable, appealing source of regenerative cells that can be used for a variety of clinical challenges. This article reviews the history of the development of adipose-derived cell science, particularly in the context of tissue engineering and regenerative medicine. It describes some of the advancements made in the field, as well as highlighting challenges and obstacles.", (Copyright © 2015 Elsevier Inc. All rights reserved.)

This article discusses adipose-derived stem cell (ASC) biology, describes the current knowledge in the literature for the safety and regulation of ASCs, and provides a brief overview of the regenerative potential of ASCs. It is not an exhaustive listing of all available clinical studies or every study applying ASCs in tissue engineering and regenerative medicine, but is an objective commentary of these topics., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Fat injection empirically started 100 years ago to correct contour deformities mainly on the face and breast. The German surgeon Eugene Hollaender (1867-1932) proposed a cocktail of human and ram fat, to avoid reabsorption. Nowadays, fat injection has evolved, and it ranks among the most popular procedures, for it provides the physician with a range of aesthetic and reconstructive clinical applications with regenerative effects on the surrounding tissues. New research from all over the world has demonstrated the role of adipose-derived stem cells, present in the adipose tissue, in the repair of damaged or missing tissues., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Adipose-derived stromal/stem cells (ASC) are multipotent with abilities to differentiate into multiple lineages including connective tissue and neural cells. Despite unlimited opportunity and needs for human and veterinary regenerative medicine, applications of adipose-derived stromal/stem cells are at present very limited. Furthermore, the fundamental biological factors regulating stemness in ASC and their stable differentiation into other tissue cells are not fully understood. The objective of this review was to provide an update on the current knowledge of the nature and isolation, molecular and epigenetic determinants of the potency, and applications of adipose-derived stromal/stem cells, as well as challenges and future directions. The first quarter of the review focuses on the nature of ASC, namely their definition, origin, isolation and sorting methods and multilineage differentiation potential, often with a comparison to mesenchymal stem cells of bone marrow. Due to the indisputable role of epigenetic regulation on cell identities, epigenetic modifications (DNA methylation, chromatin remodeling and microRNAs) are described broadly in stem cells but with a focus on ASC. The final sections provide insights into the current and potential applications of ASC in human and veterinary regenerative medicine.

Aspirated fat contains unnecessary components such as water, oil, and blood cells. For better outcomes, tissue purification and condensation are useful, especially when injection volume to the recipient site is limited. Because aspirated fat is relatively poor in adipose-derived stem/stromal cells (ASCs), ASC condensation seems important for obtaining better regeneration and retention. Reducing tissue volume by removing some adipocytes or supplementation of stromal vascular fraction or ASCs can increase the ASC/adipocyte ratio in the graft. Clinical results of ASC supplementation remain controversial, but ASC condensation seems to lead to expanding applications of fat grafting into revitalization of stem cell-depleted tissue., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Adipocyte-derived stem cells have emerged as a novel source of stem cell therapy for their autologous and readily accessible and pluripotent potential to differentiate into different lineages such as neural stem cells (NSCs) and endothelial progenitor cells (EPCs). Transplantation of NSCs and EPCs has been promising for the repair of brain injury. We explored using co-transplanted hydrogel scaffold to improve the survival of the transplanted cells and recovery of neurological function. Adult Wistar rats were transplanted with EPC-hydrogel, NSC-hydrogel, NSC-EPC-hydrogel, EPC only, or NSC only 7 days after cortical contusion injury. Behavioral tests were performed to evaluate neurological function before, and 1, 2, 3, and 4 weeks after transplantation. Size of injury, extent of vascularization, as well as the survival and differentiation of the transplanted EPCs and NSCs, were evaluated at week 5. All transplantation groups displayed significantly better neurological function compared with the control groups. Improved neurological function correlated with significantly smaller injury volumes than that of the saline group. Using immunostaining, we have shown that while transplanted NSCs differentiated into both neurons and astrocytes, the EPCs were incorporated into vessel epithelia. The extent of reactive gliosis (based on glial fibrillary acidic protein immunostaining) was significantly reduced in all treatment groups (NSC-EPC-hydrogel, NSC-hydrogel, and EPC-hydrogel) when compared with the saline group, with the highest reduction in the NSC-EPC-hydrogel transplantation group. Thus, co-transplantation of hydrogel scaffold provides a more conducive environment for the survival and differentiation of NSCs and EPCs at the site of brain injury, leading to improved vascularization and better recovery of neurological function.

The abilities of intracavernous injection of autologous stromal vascular fraction (SVF) and adipose-derived stem cells (ADSCs) to facilitate recovery of erectile function in a rat model of cavernous nerve (CN) injury were compared. Forty male Sprague-Dawley rats were randomly divided into four groups: sham and control groups (intracavernous injection of phosphate-buffered saline), SVF group (intracavernous injection of SVF), and ADSC group (intracavernous injection of ADSCs). Rats in the latter three groups underwent bilateral CN injury prior to injection. The evaluation of erectile function and histomorphometric studies were performed 4 weeks after injection. The ratio of maximal intracavernous pressure to mean arterial pressure was significantly lower in the control group than in the sham group (0.18 vs. 0.56, p < .001). Intracavernous injection of SVF (0.36, p = .035) significantly improved erectile function compared with that in the control group, whereas the ADSC group (0.35, p = .052) showed marginally significant improvement. The smooth muscle/collagen ratio, smooth muscle content, number of neuronal nitric-oxide synthase-positive nerve fibers, and expression of von Willebrand factor were significantly higher in the SVF and ADSC groups than in the control group. Expression of endothelial nitric-oxide synthase was significantly increased in the SVF group. The increases in the smooth muscle/collagen ratio and von Willebrand factor expression were larger in the SVF group than in the ADSC group. Intracavernous injection of SVF or ADSCs was equally effective in recovering penile erection in a rat model of CN injury., (©AlphaMed Press.)

Autologous fat grafting has become an important procedure for volumization and revitalization, although clinical outcomes depend greatly on technique. It was revealed recently how grafted fat tissue survives, regenerates, or dies. Experimental results provided the underlying mechanism and clinical implications for therapeutic strategies to maximize the effects of fat grafting, minimize necrosis, and avoid oil cyst formation., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Transplanted adipose tissue has many applications in regenerative medicine. However, fat grafting yields unpredictable results because the fat that is transferred can suffer variable degrees of fat reabsorption. It is necessary to identify methods and maneuvers to minimize reabsorption rates and provide predictable long-term results., Objectives: Our study aimed to identify the optimal method of harvesting, as well as the optimal pressure regime for fat aspiration. The primary objective was to assess the degree of adipocyte and mesenchymal stem cell death that occurred with the various devices and pressure levels used to harvest fat., Methods: This study was a prospective, randomized, comparative study in 15 healthy male and female subjects aged 25 to 60 who were undergoing abdominal cosmetic surgery. Various apparatuses and pressure regimens were used to harvest 8 samples of fat tissue. These samples (R1 = R8) underwent histological analysis in order to verify the integrity and functionality of the adipocytes and mesenchymal stem cells that had been harvested., Results: A total of 14 females and 1 male underwent abdominal cosmetic surgery. Quantitative analysis revealed that the adipocytes in all 8 samples had homogeneous quantitative profiles. The adipose mesenchymal stem cell (AMSC) analysis, according to Friedman ANOVA, revealed no significant variation in the percentage of mesenchymal stem cells (P = .045) between the various samples., Conclusions: The type of device, nozzle diameter tip, and pressure regimen used in this study for harvesting fat tissue did not significantly affect the number of the adipocytes or viable AMSC harvested., Level of Evidence: 3 Therapeutic., (© 2015 The American Society for Aesthetic Plastic Surgery, Inc. Reprints and permission: journals.permissions@oup.com.)

Background: The success of soft-tissue augmentation is offset by the low survival rates of grafted fat tissue. Research shows that adipose-derived stem cells (ASCs) and platelet-rich plasma (PRP) are beneficial to tissue healing., Objectives: To evaluate the long-term effects of different volume fractions of PRP combined with ASCs on fat graft., Methods: ASCs were isolated from human fat tissue, and PRP was obtained from human blood. Cell count kit-8 and real-time polymerase chain reaction (PCR) were used to evaluate the influence of PRP (0%, 10%, 20%, and 30%; volume/volume [v/v]) in medium on ASC proliferation and adipogenic differentiation, respectively. A novel lipoinjection consisting of granular fat, PRP, and ASCs was subcutaneously transplanted into nude mice. The grafts were volumetrically and histologically evaluated 10, 30, 60, and 90 days after transplantation., Results: The addition of PRP improved ASC proliferation. Expression of adipogenic-related genes, peroxisome proliferator-activated receptor-γ, lipoprotein lipase, and adipophilin were up-regulated in PRP-induced ASCs. Compared with other groups, granular fat grafts formed with 20% (v/v) and 30% (v/v) PRP significantly improved residual volumes. More intact adipocytes and capillary formation, but less vacuolization, were observed in the 20% (v/v) and 30% (v/v) PRP groups at 30, 60, and 90 days. However, no significant difference was observed between the 20% (v/v) and 30% (v/v) PRP groups in retaining fat grafts and improving histology., Conclusions: Fat grafting with 20% (v/v) PRP and ASCs constitutes an appropriate transplantation strategy for improving graft survival and provides a potential approach for soft-tissue restoration in plastic and reconstructive surgery., (© 2015 The American Society for Aesthetic Plastic Surgery, Inc. Reprints and permission: journals.permissions@oup.com.)

Although lipotransfer, or fat grafting, is a commonly used procedure in aesthetic and reconstructive surgery, there is still variability in graft survival and neoadipogenesis from one procedure to the next. A better understanding of the sequential molecular events occurring with grafting would allow us to strategize methods to improve the regenerative potency of the grafted tissue. These steps begin with an autophagic process, followed by the inclusion of stromal vascular fraction and matrix components. By tailoring and modifying each of these steps for a particular type of aesthetic or reconstructive procedure, strategic sequencing represents a dynamic approach to lipotransfer with the aim of maximizing adipocyte viability and growth. In the implementation of the strategic sequence, it remains important to consider the clinical viability of each step and its compliance with the US Food and Drug Administration regulations. This review highlights the basic science behind clinically translatable approaches to supplementing various fat grafting procedures.

Inflammatory bowel disease (IBD) is a complex immunological disorder characterized by chronic inflammation caused mainly by unknown factors. The interleukin-10 knockout (IL-10 KO) mouse is a well-established murine model of IBD which develops spontaneous intestinal inflammation that resembles Crohn's disease. In the present study, human adipose-derived mesenchymal stem cells (hAMSCs) were administrated to IL-10 KO mice to evaluate the anti-inflammatory effects of hAMSCs that may attenuate the progress of or treat IBD. After IBD was induced by feeding the IL-10 KO mouse a 125-250 ppm piroxicam mixed diet for 1 week, 2×10(6) hAMSCs were injected into the peritoneum and the mice were switched to a normal diet. After 1 week, the mice were sacrificed and tissue samples were harvested. Tissue scores for inflammation and inflammation-related genes expression were determined. The hAMSC-treated group showed significantly reduced inflammatory changes in histological analysis. Reverse transcription-PCR analysis showed that RANTES, Toll-like receptor 9, and IL-4 expression levels were not significantly different between the groups while IL-12, INF-γ, and TNF-α levels were significantly decreased in the hAMSC treated group. hAMSC attenuated IBD in the IL-10 KO mice by suppressing inflammatory cytokine expression, was mediated by the type 1 helper T cell pathway. Even though only a single injection of hAMSCs was delivered, the effect influenced chronic events associated with inflammatory changes and demonstrated that hAMSCs are a powerful candidate for IBD therapy., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Bone is one of the most dynamic tissues in the human body that can heal following injury without leaving a scar. However, in instances of extensive bone loss, this intrinsic capacity of bone to heal may not be sufficient and external intervention becomes necessary. Several techniques are available to address this problem, including autogenous bone grafts and allografts. However, all these techniques have their own limitations. An alternative method is the technique of distraction osteogenesis, where gradual and controlled distraction of two bony segments after osteotomy leads to induction of new bone formation. Although distraction osteogenesis usually gives satisfactory results, its major limitation is the prolonged duration of time required before the external fixator is removed, which may lead to numerous complications. Numerous methods to accelerate bone formation in the context of distraction osteogenesis have been reported. A viable alternative to autogenous bone grafts for a source of osteogenic cells is mesenchymal stem cells from bone marrow. However, there are certain problems with bone marrow aspirate. Hence, scientists have investigated other sources for mesenchymal stem cells, specifically adipose tissue, which has been shown to be an excellent source of mesenchymal stem cells. In this paper, the potential use of adipose stem cells to stimulate bone formation is discussed.

Background: Clinical prognosis is critically poor in fulminant myocarditis, while it's initiation or progression is fated, in part, by T cell-mediated autoimmunity. Adiponectin (APN) and associated adipokines were shown to be immune tolerance inducers, although the clinically relevant delivery method into target pathologies is under debate. Whether the cell sheet-based delivery system of adipokines might induce immune tolerance and functional recovery in experimental autoimmune myocarditis (EAM) was tested., Methods and Results: Scaffold-free-induced adipocyte cell-sheet (iACS) was generated by differentiating adipose tissue-derived syngeneic stromal vascular-fraction cells into adipocytes on temperature-responsive dishes. Rats with EAM underwent iACS implantation or sham operation. Supernatants of iACS contained a high level of APN and hepatocyte growth factor (HGF), and reduced proliferation of CD4-positive T cells in vitro. Immunohistolabelling showed that the iACS implantation elevated the levels of APN and HGF in the myocardium compared to the sham operation, which attenuated the immunological response by inhibiting CD68-positive macropharges and CD4-positive T-cells and activating Foxp3-positive regulatory T cells. Consequently, left ventricular ejection fraction was significantly greater after the iACS implantation than after the sham operation, in association with less collagen accumulation., Conclusions: The targeted delivery of adipokines using tissue-engineered iACS ameliorated cardiac performance of the EAM rat model via effector T cell suppression and induction of immune tolerance. These findings might suggest a potential of this tissue-engineered drug delivery system in treating fulminant myocarditis in the clinical setting.

Volume stability and growth of tissue engineered adipose tissue equivalents using adipose-derived stem cells (ASCs) rely strongly on angiogenesis and neovascularization to support the maintenance of cells. An attractive cellular approach is based on coimplantation of endothelial cells to create a vascular network. Endothelial progenitor cells (EPCs) are a promising cell population, since they can be easily isolated from autologous human peripheral blood and thus represent a clinically feasible option. We have previously shown in in vitro and semi-in vivo studies that ASCs exert beneficial effects on EPCs in terms of enhanced tube formation and formation of blood vessels, respectively. In this study, we investigated the in vivo effects of coimplantation on endothelial cell-mediated neovascularization and ASC-mediated adipose tissue formation. For this purpose, human ASCs and human EPCs (or HUVECs as direct comparison to EPCs) were suspended alone or in coculture in fibrin and subcutaneously injected into the back of athymic nude mice and explanted after 1, 3 or 6months. Our results show that monocultures of EPCs or HUVECs were not able to perform vasculogenesis and constructs exhibited complete resorption already after 1month. However, a remarkable difference between EPCs and HUVECs was detected when coimplanted with ASCs. While coimplanted HUVECs gave rise to a stable neovasculature which was characterized by perfusion with erythrocytes, coimplanted EPCs showed no ability to form vascular structures. In the case of HUVEC-derived neovasculature, coimplanted ASCs displayed perivascular properties by stabilizing these neovessels. However, formation of human adipose tissue was independent of coimplanted endothelial cells. Our results indicate that HUVECs are superior to EPCs in terms of promoting in vivo neovascularization and recruiting perivascular cells for vessel stabilization when coimplanted with ASCs., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The management of chondral defects has long been a challenge because of the poor self-healing capacity of articular cartilage. Many approaches ranging from symptomatic treatment to structural cartilage regeneration have obtained very limited satisfactory results. Cartilage tissue engineering, which involves an optimized combination of novel scaffolds, cell sources and growth factors, has emerged as a promising strategy for cartilage regeneration and repair. In this study, the cellular morphologies and the adhesion, migration and proliferation capabilities of adipose-derived stem cells (ASCs) and chondrocytes seeded on 3D scaffolds composed of electrospun poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P3HB4HB) were evaluated. Next, TGF-β1/scaffolds with 4:1 co-culture of ASCs and chondrocytes were implanted into the full thickness cartilage defects in rabbit knee for 16 weeks. ASCs and chondrocytes seeded on the scaffolds showed better adhesion, migration and proliferation than that on petri dishes in vitro. Importantly, implantation with TGF-β1/scaffolds with delivery of ASCs and chondrocytes revealed desirable in vivo healing outcomes. These results demonstrate that ASCs have great potential in the field of tissue engineering. It is possible that the improvement in ASC-seeded electrospun 3D P3HB4HB scaffolds may ultimately lead to improved repair of cartilage injuries.

Objective: Unpredictability in graft retention remains a significant drawback of fat grafting. Processing of fat grafts has been the focus of several studies to improve graft survival. The objective of this study was to systematically review the outcomes of different fat graft processing techniques with the goal of (1) deriving clinically oriented insights and (2) identifying gaps in knowledge to stimulate future research., Methods: PubMed, EMBASE, and Cochrane Databases were searched to identify studies that compared different fat graft processing techniques. Outcome measures of interest were any subjective or objective measures of fat graft survival or reports of adverse events., Results: A total of 2056 abstracts were generated from the literature searches; 13 studies met the criteria for data extraction and analysis. Processing methods assessed included decantation, washing, gauze filtration, and centrifugation. Each processing method was found to be better than other methods, depending on the outcome measure used to study graft survival. As well, several studies found statistical equipoise in the outcome measures when analyzing the results of the different techniques. Adverse events were rarely reported and did not correlate with any processing method in particular., Conclusions: No firm concluding recommendation can be made to deem 1 processing technique superior to the others. However, it would seem that techniques, which use a combination of gentle washing and centrifugation, strike the optimal balance of preserving adipocyte viability while removing bulk of the contaminants.

Objective: To investigate the effects of allogeneic adipose-derived stem cells (ADSCs) of rat on the early neovascularization of autologous fat transplantation., Methods: (1) Experiment 1. Adipose tissue was collected from both inguinal regions of two SD rats to isolate, culture, and purify ADSCs through collagen enzyme digestion, density gradient centrifugation, and adherence method. The fourth passage of cells were collected for morphologic observation, detection of expressions of surface markers CD34, CD49d, CD106, and CD45 of ADSCs with flow cytometer, identification of adipogenic and osteogenic differentiation, and determination of the cell proliferation ability with thiazolyl blue method. (2) Experiment 2. Another 30 SD rats were divided into allogeneic adipose granule (AG) group (A, n = 6), autologous AG group (B, n = 8), autologous ADSCs+autologous AG group (C, n = 8), and allogeneic ADSCs+autologous AG group (D, n = 8) according to the random number table. The fourth passage of ADSCs were obtained from adipose tissue from one side of inguinal region of SD rats in group C. Adipose tissue obtained from one side of inguinal region of SD rats of the other 3 groups was abandoned. The AG was prepared from another side of inguinal region of SD rats in the 4 groups. The mixture of 0.6 g AG from one rat and 1 mL DMEM/F12 nutrient solution was injected subcutaneously into the back of another rat in group A, and so on. Autologous AG was injected into its own body of the rats in group B. The mixture of 1 mL autologous ADSCs mixture which contains 3.0 × 10⁶ cells per mililitre autologous ADSCs combined with autologous AG was injected into the rats in group C. The mixture of 1 mL allogeneic ADSCs mixture which contains 3.0 × 10⁶ cells per mililitre ADSCs extractived from the former 2 rats in experiment 1 combined with autologous AG was injected into the rats in group D. At 7 days post transplantation, fat transplants were harvested for gross observation, measurement of wet weight, pathological observation, and assessment of cells with positive expression of CD31 with immunohistochemical method. Data were processed with one-way analysis of variance and SNK test., Results: (1) The fourth passage of cells proliferated well showing fusiform shape similar to fibroblasts. These cells showed positive expression of CD34 and CD49d and weak positive expression of CD106 and CD45. They were able to differentiate into adipocytes and osteoblasts. These cells were identified as ADSCs. The fourth passage of cells grew faster than that of the tenth passage. (2) At 7 days post transplantation, no liquifying necrosis or infection was observed in the fat transplants of the rats in the 4 groups. Wet weight of the fat transplants in groups A and B was respectively (0.25 ± 0.04) and (0.26 ± 0.03) g, which were less than those of groups C and D [(0.36 ± 0.03) and (0.35 ± 0.04) g, with P values below 0.05]. HE staining showed that there were less fat cells and more fibroblasts in the transplants of group A, visible fibrous tissue around uneven shape of fat cells in the transplants of group B, and almost identical size and shape of fat cells and unobvious fibrous tissues were found in the transplants of groups C and D. The cells with positive expression of CD31 were distributed in fibrous tissues in larger number but less around fat cells in the transplants of group A, while more of these cells were observed surrounding fat cells in the transplants of group B. There were more cells with positive expression of CD31 distributed surrounding fat cells in the transplants of groups C and D than that in group B. The cells with positive expression of CD31 observed under 400 times field were more in number in groups C (20.5 ± 1.1) and D (22.1 ± 1.0) than in groups A (8.0 ± 3.6) and B (10.9 ± 1.7), with P values below 0.05., Conclusions: Allogeneic ADSCs combined with autologous AG can significantly improve the early vascularization of fat transplantation as well as autologous ADSCs combined with autologous AG.

Evolving atrophic macular degeneration represents at least 80% of all macular degenerations and is currently without a standardized care. Autologous fat transplantation efficacy was demonstrated by several studies, as these cells are able to produce growth factors. The aim of the work was to demonstrate possible therapeutic effect of the joined suprachoroidal graft of adipocytes, adipose-derived stem cells (ADSCs) in stromal vascular fractions (SVFs) of adipose tissue, and platelet-rich plasma (PRP). Twelve eyes in 12 dry age-related macular degeneration (AMD) patients, aged 71.25 (SD ± 6.8) between 62 and 80 years, were analyzed. A complete ocular evaluation was performed using best corrected visual acuity (BCVA), retinographic analysis, spectral-domain optical coherence tomography, microperimetry, computerized visual field, and standard electroretinogram (ERG). Each eye received a cell in graft between choroid and sclera of mature fat cells and ADSCs in SVF enriched with PRP by means of the variant second Limoli (Limoli retinal restoration technique [LRRT]). In order to test if the differences pre- and post-treatment were significant, the Wilcoxon signed-rank test has been performed. Adverse effects were not reported in the patients. After surgery with LRRT, the most significant increase in the ERG values was recorded by scotopic rod-ERG (answer coming from the rods), from 41.26 to 60.83 μV with an average increase of 47.44% highly significant (P < 0.05). Moderately significant was the one recorded by scotopic maximal ERG (answer coming from the rods and cones), from 112.22 to 129.68 μV with an average increase of 15.56% (P < 0.1). Cell-mediated therapy based on growth factors used appears interesting because it can improve the retinal functionality responses in the short term. The ERG could, therefore, be used to monitor the effect of cell-mediated regenerative therapies.

Mature adipocyte-derived dedifferentiated fat cells (DFAT) have a potential to be useful as new cell-source for cell-based therapy for spinal cord injury (SCI), but the mechanisms remain unclear. The objective of this study was to examine whether DFAT-induced functional recovery is achieved through remyelination and/or glial scar reduction in a mice model of SCI. To accomplish this we subjected adult female mice (n=22) to SCI. On the 8th day post-injury locomotor tests were performed, and the mice were randomly divided into two groups (control and DFAT). The DFAT group received stereotaxic injection of DFAT, while the controls received DMEM medium. Functional tests were conducted at repeated intervals, until the 36th day, and immunohistochemistry or staining was performed on the spinal cord sections. DFAT transplantation significantly improved locomotor function of their hindlimbs, and promoted remyelination and glial scar reduction, when compared to the controls. There were significant and positive correlations between promotion of remyelination or/and reduction of glial scar, and recovery of locomotor function. Furthermore, transplanted DFAT expressed markers for neuron, astrocyte, and oligodendrocyte, along with neurotrophic factors, within the injured spinal cord. In conclusion, DFAT-induced functional recovery in mice after SCI is probably mediated by both cell-autonomous and cell-non-autonomous effects on remyelination of the injured spinal cord., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Traditional adipose tissue transplantation has unpredictable viability and poor absorption rates. Recent studies have reported that treatment with platelet-rich plasma (PRP), adipose-derived stem cells (ASCs), and stromal vascular fraction (SVF) are related to increased survival of grafted adipose tissue. This study was the first simultaneous comparison of graft survival in combination with PRP, ASCs, and SVF. Adipose tissues were mixed with each other, injected subcutaneously into the back of nude mice, and evaluated at 4, 8, and 12 weeks. Human adipocytes were grossly maintained in the ASCs and SVF mixtures. Survival of the adipose tissues with PRP was observed at 4 weeks and with SVF at 8 and 12 weeks. At 12 weeks, volume reduction in the ASCs and SVF mixtures were 36.9% and 32.1%, respectively, which were significantly different from that of the control group without adjuvant treatment, 51.0%. Neovascular structures were rarely observed in any of the groups. Our results suggest that the technique of adding ASCs or SVF to transplanted adipose tissue might be more effective than the conventional grafting method. An autologous adipose tissue graft in combination with ASCs or SVF may potentially contribute to stabilization of engraftment.

Soft tissue augmentation is a process of implanting tissues or materials to treat wrinkles or soft tissue defects in the body. Over the years, various materials have evolved to correct soft tissue defects, including a number of tissues and polymers. Autogenous dermis, autogenous fat, autogenous dermis-fat, allogenic dermis, synthetic implants, and fillers have been widely accepted for soft tissue augmentations. Tissue engineering technology has also been introduced and opened a new venue of opportunities in this field. In particular, a long-lasting filler consisting of hyaluronic acid filler and living human mesenchymal cells called "injectable tissue-engineered soft tissue" has been created and applied clinically, as this strategy has many advantages over conventional methods. Fibroblasts and adipose-derived stromal vascular fraction cells can be clinically used as injectable tissue-engineered soft tissue at present. In this review, information on the soft tissue augmentation method using the injectable tissue-engineered soft tissue is provided.

Background: Fat grafting is a promising modality for soft-tissue augmentation/reconstruction. However, grafted fat tissue is not initially perfused and relies on plasmatic diffusion from the recipient bed until revascularization occurs. The authors evaluated the therapeutic effects of normobaric hyperoxygenation for enhancing fat graft retention., Methods: Aspirated human fat tissue was cultured under tissue hypoxia (1% oxygen), normoxia (6%), and hyperoxia (20%) levels, and evaluated for adipocyte viability. Inguinal fat pads were autografted under mouse scalps (n=36), and mice were housed in either 20% (control) or 60% (normobaric hyperoxygenation) atmospheric oxygen for the first 3 days, and then returned to normoxia. Samples harvested at 0, 1, 2, 4, 8, and 12 weeks were analyzed immunohistochemically for adipocyte viability and regeneration., Results: Organ culture adipocytes died more quickly under lower oxygen tensions; thus, hyperoxygenation of recipient tissues may delay adipocyte death after fat grafting. Autografted mouse adipose tissue underwent dynamic remodeling, from ischemic degeneration to partial regeneration, over 12 weeks. Normobaric hyperoxygenation grafted samples showed significantly larger survival zones and engraftment scores (calculated using sample weight and adipocyte viability) at 1 and 12 weeks, respectively, than control samples. In addition, adipocyte regeneration (number of perilipin-positive preadipocytes), which peaked at 4 weeks, was significantly increased in normobaric hyperoxygenation samples., Conclusion: The normobaric hyperoxygenation protocol using 60% oxygen can be safely applied to enhance adipocyte survival, regeneration, and final engraftment after fat grafting.

Background: Centrifugation is a popular processing method, with an unclear mechanism of action. Hypotheses include fat concentration, reduced inflammatory response by removal of blood, and concentration of adipose-derived stem cells. The authors performed multiple experiments to determine the role of centrifugation and compared it with a different processing method (mesh/gauze technique)., Methods: Lipoaspirate components were quantified after centrifugation at increasing speed to determine concentration efficacy. For comparison, the authors quantified the concentration efficacy of mesh/gauze. They also compared the number of adipose-derived stem cells isolated by either method. To determine the effects of each component, they compared fat alone to fat mixed with various spinoff components in a mouse model. They also compared centrifugation to mesh/gauze., Results: The adipocyte fraction remains constant above 5000 g, whereas 1200 g results in 91 percent concentrated fat. Mesh/gauze also results in 90 percent concentrated fat. The number of adipose-derived stem cells in 1 g of fat was 1603 ± 2020 and 1857 ± 1832 in the centrifuge and mesh/gauze groups, respectively (p = 0.86). Five "add-back" groups were created: fat plus oil, fat plus surgical tumescence, fat plus fresh tumescence, fat plus cell pellets and fresh tumescence, and fat plus cell pellets. The fat-only group had better retention than the groups mixed with tumescence, regardless of whether it was surgical, fresh, or had cell pellets. Oil did not affect grafts. Centrifugation at 1200 g was equivalent to mesh/gauze (0.73 ± 0.12 g and 0.72 ± 0.13 g, respectively)., Conclusions: Centrifugation improves graft retention by concentration of the adipocyte fraction. The concentration efficacy of mesh/gauze is equivalent to centrifugation at 1200 g, with equivalent in vivo outcomes.

Introduction: Stem cells (SCs) show significant benefits in the treatment of postprostatectomy erectile dysfunction (ED). However, the low retention rate of the traditional single-cell strategy at the injection sites limits its therapeutic potential., Aim: This study aims to investigate the feasibility and mechanism of adipose-derived stem cells (ADSCs)-based micro-tissues (MTs) in the treatment of ED in a rat model of bilateral cavernous nerves (CNs) injury., Methods: ADSCs labeled with 5-ethynyl-2-deoxyuridine (EdU) were used to generate MTs with hanging drop method. 10 Sprague-Dawley (SD) rats underwent sham surgery and intracavernous (IC) injection of phosphate buffer solution (PBS) (the sham group). Another 70 rats underwent bilateral CN crush and were then treated with PBS (n = 10, the crush group), dissociated ADSCs (n = 30, the ADSCs group), and MTs (n = 30, the MTs group), respectively. At day 1, 3, 7, 14 (n = 5), and 28 (n = 10) postsurgery, specimens were harvested for histology. At day 28, 10 rats in each group were examined for erectile function before tissue harvest., Main Outcome Measures: Light microscopy of the dynamic aggregation of the MT, immunohistologic examination of the MTs, the retention and distribution of EdU + ADSCs in the corpus cavernosum (CC), and the penis histological analyses of collagen content, Western blot of functional proteins in MTs, intracavernous pressure recording on CN electrostimulation., Results: Three-day-old MTs became stable and expressed nerve growth factor, vascular endothelial growth factor, C-X-C chemokine receptor type 4, Wnt5a, and collagen IV. More EdU + ADSCs retained in the CC in the MTs group than that in the ADSCs group. IC injection of MTs resulted in significant restoration of the erectile function and histopathological changes compared with the ADSCs group., Conclusion: IC-injected MTs resulted in a better restoration of erectile function than traditional single-cell strategy. The underlying mechanisms of recovery appear to involve enhanced cellular retention in the penis and upregulation of some paracrine factors., (© 2014 International Society for Sexual Medicine.)

Background: Postoperative cerebrospinal fluid (CSF) leaks and headaches remain potential complications after retrosigmoid approaches for lesions in the posterior fossa and cerebellopontine angle. The authors describe a simple repair technique with an autologous fat graft-assisted Medpor Titan cranioplasty and investigate the incidence of postoperative CSF leaks and headaches using this technique., Methods: A retrospective chart review was conducted on all cases (n = 60) of retrosigmoid craniectomy from September 2009 to May 2014 in patients who underwent fat graft-assisted cranioplasty. After obtaining a watertight dural closure and sealing off any visible mastoid air cells with bone wax, an autologous fat graft was placed over the dural suture line and up against the waxed-off air cells. The fat graft filled the retrosigmoid cranial defect and was then bolstered with a Medpor Titan (titanium mesh embedded in porous polyethylene) cranioplasty. A postoperative mastoid pressure dressing was applied for 48 h, and prophylactic lumbar drainage was not used. Factors examined in this study included postoperative CSF leak (incisional, rhinorrhea, otorrhea), pseudomeningocele formation, incidence and severity of postoperative headache, length of hospital stay, and length of follow-up., Results: No patients developed postoperative CSF leaks (0 %), pseudomeningoceles (0 %), or new-onset postoperative headaches (0 %) with the described repair technique. There were no cases of graft site morbidity such as hematoma or wound infection. Mean duration of postoperative hospital stay was 3.8 days (range 2-10 days). Mean postoperative follow-up was 12.4 months (range 2.0-41.1 months)., Conclusions: Our multilayer repair technique with a fat graft-assisted Medpor Titan cranioplasty appears effective in preventing postoperative CSF leaks and new-onset postoperative headaches after retrosigmoid approaches. Postoperative lumbar drainage may not be necessary.

Background: Dedifferentiated fat cells, obtained from the ex vivo ceiling culture of mature adipocytes of mammals, have a high proliferative potential and pluripotency. The authors transplanted dedifferentiated fat cells into a nerve defect created in rat facial nerve and evaluated nerve regeneration ability., Methods: The buccal branch of the facial nerve of rats was exposed, and a 7-mm nerve defect was created. Green fluorescent protein-positive dedifferentiated fat cells prepared from adipocytes were mixed with type 1 collagen scaffold and infused into a silicone tube, which was then transplanted into the nerve defect in a green fluorescent protein-negative rat (the dedifferentiated fat group). Regenerated nerves were excised at 13 weeks after transplantation and examined histologically and physiologically. The findings were compared with those of autografts and silicone tubes loaded with collagen gel alone (the control group) transplanted similarly., Results: Axon diameter of regenerated nerve increased significantly in the dedifferentiated fat group compared with the control group, whereas no significant difference was found between the dedifferentiated fat and autograft groups. Myelin thickness was found to be largest in the autograft group, followed by the dedifferentiated fat and the control groups, showing significant differences between all pairs of groups. Evaluation of physiologic function of nerves by compound muscle action potential revealed a significantly better result in the dedifferentiated fat group than in the control group. The regenerated nerves in the dedifferentiated fat group had S100 and green fluorescent protein-double-positive Schwann-like supportive cells., Conclusion: After being transplanted into a facial nerve defect, dedifferentiated fat cells promoted the maturation of the regenerated nerve.

Lipofilling with autologous fat tissue is widely used in plastic and reconstructive surgery to treat soft-tissue deficiency. Unfortunately, implanted cells disappear gradually and make it difficult to predict the resorption rate. Several adjuvants are used to improve the success of fat tissue grafting. In this study, the effect of botulinum toxin (BoNT) on mature adipocytes, as well as adipose-derived stem cells (ASC) and fibroblasts was evaluated. As lidocaine is the most prevalent drug to anesthetize the donor site as well as the area to be treated with autologous fat, this local anesthetic was examined too. Primary ASCs, fibroblasts, and mature adipocytes were exposed to 1, 10, and 20 IU/ml BoNT A and 1% lidocaine. Cells were tested on proliferation, viability, and LDH release. Adipogenic differentiation potential was evaluated by quantitative real-time PCR analyzing the expression of FABP4. BoNT had no significant influence on the proliferation or viability of tested cells. By trend, low concentrations of BoNT improved adipogenic potential of ASCs. Lidocaine had a strong diminishing effect on the proliferation of ASCs and fibroblasts and on the viability of these cells. Mature adipocytes show no significant inferior viability after BoNT or lidocaine treatment. BoNT has no negative effect on ASCs, mature adipocytes, or fibroblasts in vitro. Lidocaine (1%) negatively influences the proliferation and viability of fibroblasts and partly of ASCs but not of mature adipocytes., (Copyright © 2014 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.)

Neovascularization plays an important role in adipose tissue transplantation, because survival of implanted cells strongly relies on sufficient oxygen and nutrient supply. Vascular endothelial growth factor (VEGF) is known as the master regulator of angiogenesis. It is capable of starting the complex cascade of events leading to endothelial cell activation, assembly of new vascular structures, mural cell recruitment, and vessel stabilization. However, consensus is lacking regarding safe and efficient methods for applying VEGF in free fat transplantation in the clinical setting. We investigated whether chitosan nanospheres, a biocompatible high-molecular-weight material, safely improve the efficiency of VEGF application in free fat transplantation. Immunologically compromised nude mice were used as adipose tissue transplantation receptors. Nanospheres loaded with VEGF were mixed with adipocytes and injected subcutaneously to the dorsa of mice. Grafts were harvested at weeks 3, 6, and 12. We found that treated-graft weight and vascularization were significantly higher than controls in a time-dependent manner. We demonstrated that chitosan nanospheres loaded with VEGF significantly promote the fat graft neovascularization and improve adipocyte survival.

Human and canine periodontium are often affected by an inflammatory pathology called periodontitis, which is associated with severe damages across tissues, namely, in the periodontal ligament, cementum, and alveolar bone. However, the therapies used in the routine dental practice, often consisting in a combination of different techniques, do not allow to fully restore the functionality of the periodontium. Tissue Engineering (TE) appears as a valuable alternative approach to regenerate periodontal defects, but for this purpose, it is essential to develop supportive biomaterial and stem cell sourcing/culturing methodologies that address the complexity of the various tissues affected by this condition. The main aim of this work was to study the in vitro functionality of a newly developed double-layer scaffold for periodontal TE. The scaffold design was based on a combination of a three-dimensional (3D) fiber mesh functionalized with silanol groups and a membrane, both made of a blend of starch and poly-ɛ-(caprolactone). Adipose-derived stem cells (canine adipose stem cells [cASCs]) were seeded and cultured onto such scaffolds, and the obtained constructs were evaluated in terms of cellular morphology, metabolic activity, and proliferation. The osteogenic potential of the fiber mesh layer functionalized with silanol groups was further assessed concerning the osteogenic differentiation of the seeded and cultured ASCs. The obtained results showed that the proposed double-layer scaffold supports the proliferation and selectively promotes the osteogenic differentiation of cASCs seeded onto the functionalized mesh. These findings suggest that the 3D structure and asymmetric composition of the scaffold in combination with stem cells may provide the basis for developing alternative therapies to treat periodontal defects more efficiently.

NK cell development and homeostasis require IL-15 produced by both hematopoietic and parenchymal cells. Certain hematopoietic IL-15 sources, such as macrophages and dendritic cells, are known, whereas the source of parenchymal IL-15 remains elusive. Using two types of adipocyte-specific Il15(-/-) mice, we identified adipocytes as a parenchymal IL-15 source that supported NK cell development nonredundantly. Both adipocyte-specific Il15(-/-) mice showed reduced IL-15 production specifically in the adipose tissue but impaired NK cell development in the spleen and liver in addition to the adipose tissue. We also found that the adipose tissue harbored NK progenitors as other niches (e.g. spleen) for NK cell development, and that NK cells derived from transplanted adipose tissue populated the recipient's spleen and liver. These findings suggest that adipocyte IL-15 contributes to systemic NK cell development by supporting NK cell development in the adipose tissue, which serves as a source of NK cells for other organs., (Copyright © 2014 by The American Association of Immunologists, Inc.)

Objective: Perivascular adipose tissue (PVAT) expands during obesity, is highly inflamed, and correlates with coronary plaque burden and increased cardiovascular risk. We tested the hypothesis that PVAT contributes to the vascular response to wire injury and investigated the underlying mechanisms., Approach and Results: We transplanted thoracic aortic PVAT from donor mice fed a high-fat diet to the carotid arteries of recipient high-fat diet-fed low-density lipoprotein receptor knockout mice. Two weeks after transplantation, wire injury was performed, and animals were euthanized 2 weeks later. Immunohistochemistry was performed to quantify adventitial macrophage infiltration and neovascularization and neointimal lesion composition and size. Transplanted PVAT accelerated neointimal hyperplasia, adventitial macrophage infiltration, and adventitial angiogenesis. The majority of neointimal cells in PVAT-transplanted animals expressed α-smooth muscle actin, consistent with smooth muscle phenotype. Deletion of monocyte chemoattractant protein-1 in PVAT substantially attenuated the effects of fat transplantation on neointimal hyperplasia and adventitial angiogenesis, but not adventitial macrophage infiltration. Conditioned medium from perivascular adipocytes induced potent monocyte chemotaxis in vitro and angiogenic responses in cultured endothelial cells., Conclusions: These findings indicate that PVAT contributes to the vascular response to wire injury, in part through monocyte chemoattractant protein-1-dependent mechanisms., (© 2014 American Heart Association, Inc.)

Background: It is important to protect fat viability during grafting. This study aimed to clarify whether physical or chemical factors damage fat viability., Methods: Fat was harvested under high and low tumescence pressure, using syringes and a liposuction machine; this fat was injected through different gauge needles and cultured with lidocaine or epinephrine. Samples were subjected to glucose transport test and observed histologically., Result: The viability and microstructure of fat harvested under high and low tumescence pressure, and by syringe and liposuction machine, were similar (P > 0.05). The viability and intactness of fat cells injected through needles decreased with decreasing needle diameter (P < 0.05). Lidocaine (P < 0.05) and epinephrine (P < 0.05) weakened fat viability., Conclusions: Fat used for grafting can be harvested by tumescent techniques and a liposuction machine. Fat cell viability during injection increases with increasing needle diameter. Fat should be purified to remove drugs.

Aims: Adipose-derived regenerative cells (ADRCs) can be isolated from liposuction aspirates and prepared as fresh cells for immediate administration in cell therapy. We performed the first randomized, placebo-controlled, double-blind trial to examine the safety and feasibility of the transendocardial injections of ADRCs in no-option patients with ischemic cardiomyopathy., Methods and Results: Procedural, postoperative, and follow-up safety end points were monitored up to 36 months. After baseline measurements, efficacy was assessed by echocardiography and single-photon emission computed tomography (6, 12, and 18 months), metabolic equivalents and maximal oxygen consumption (MVO2) (6 and 18 months), and cardiac magnetic resonance imaging (6 months). We enrolled 21 ADRC-treated and 6 control patients. Liposuction was well tolerated, ADRCs were successfully prepared, and transendocardial injections were feasible in all patients. No malignant arrhythmias were seen. Adverse events were similar between groups. Metabolic equivalents and MVO2 values were preserved over time in ADRC-treated patients but declined significantly in the control group. The difference in the change in MVO2 from baseline to 6 and 18 months was significantly better in ADRC-treated patients compared with controls. The ADRC-treated patients showed significant improvements in total left ventricular mass by magnetic resonance imaging and wall motion score index. Single-photon emission computed tomography results suggested a reduction in inducible ischemia in ADRC-treated patients up to 18 months., Conclusion: Isolation and transendocardial injection of autologous ADRCs in no-option patients were safe and feasible. Our results suggest that ADRCs may preserve ventricular function, myocardial perfusion, and exercise capacity in these patients., (Copyright © 2014 Mosby, Inc. All rights reserved.)

The aim of this study was to evaluate the local behavior of intracutaneously injected human mesenchymal stem cells from adipose tissue and to determine the safety of a cell-based cutaneous therapy in an animal model.Human mesenchymal stem cells from adipose tissue were labeled with red fluorochrome and were injected intradermally in the paravertebral area in immunodeficient BalbC/nude mice (n = 21). As a control, cell culturemedium was injected in the same fashion on the contralateral paravertebral side. Four weeks, 6 months, and 12 months after the injection, seven mice were examined. In addition to the injected areas, the lungs, kidneys,spleens, and brains were excised and processed for histological evaluation. Serial sections of all the tissues excised were evaluated for adipose tissue-derived stem cells by means of emerging red fluorescent signals.The injected stem cells could be detected throughout the follow-up period of 1-year at the injection site within the dermal and subcutaneous layers. Bar these areas, adipose tissue-derived stem cells were not found in any otherexamined tissue at any point in time. The adipose tissue-derived stem cells showed a slow transition to deeper subcutaneous adipose tissue layers and, in part, a differentiation into adipocytes. No ulceration, inflammation, ortumor induction could be detected.The present study shows that intracutaneously injected human mesenchymal stem cells from adipose tissue stay at the site of injection, survive in vivo for up to 1-year, and partly differentiate into adipocytes. This is a new andvery important finding needed to safely apply therapies based on such stem cells in fat transplants in regenerative medicine., (Copyright © 2014 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.)

Access to human tissue is critical to medical research, however the laws and regulations surrounding gaining ethical and legal access to tissue are often poorly understood. Recently, there has been a huge increase in the interest surrounding the therapeutic application of adipose tissue, and adipose-derived stem cells. To facilitate our own research interests and possibly assist our local colleagues and collaborators, we established a Research Tissue Bank (RTB) to collect, store and distribute human adipose tissue derived cells with all the appropriate ethical approval for subsequent downstream research. Here we examine the legal, ethical and practical issues relating to the banking of adipose tissue for research in the UK, and discuss relevant international guidelines and policies. We also share our experiences of establishing an RTB including the necessary infrastructure and the submission of an application to a Research Ethics Committee (REC)., (Copyright © 2014 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.)

Since their isolation and characterization nearly a decade ago, adipose-derived stem cells (ASCs) have become one of the most popular adult stem cell populations for soft tissue engineering and regenerative medicine applications. Compared with other stem cell sources, ASCs offer several advantages including abundant autologous source, minor invasive harvesting (liposuction), significant proliferative capacity in culture, and multilineage potential. In this mini review, we focus on some of the more salient published clinical and preclinical data to date regarding ASC treatment for breast and facial soft tissue reconstruction., (© 2014 by the Wound Healing Society.)