Adipose tissue stromal vascular fraction:redrawing the lines

The research performed in this thesis aimed to investigate the difference in clinical effect of stromal vascular fraction on decreased skin quality due to a pathological cause (scarring/wounds) or physiological cause (ageing). Stromal vascular fraction of adipose tissue consist of all cell types including adipose derived stromal cells within adipose tissue except for the adipocytes itself. In the past years, the stromal vascular fraction has shown to harbour a high regenerative potential and can be isolated by enzymatic isolation. However, enzymatic isolation is time-consuming and expensive. In this thesis, we have developed an intra-operative mechanical isolation procedure to isolate the stromal vascular fraction, without the use of enzymes. In this way, our mechanical isolation procedure was able to isolate a highly viable stromal vascular fraction containing unaffected adipose derived stromal cells within 20 min. Using this mechanical isolation procedure, the effect of mechanically isolated stromal vascular fraction has been investigated to improve skin quality in patients. Research in this thesis has shown that the stromal vascular fraction was not able to improve decreased skin quality due to ageing in female patients; no improvement in skin texture or elasticity was shown. On the other hand, the stromal vascular fraction was able to improve wound healing after a bilateral mamma-reduction model. After 6 months, a fast healing occurred in the scars of the breasts that received stromal vascular fraction compared to the other control breast. However, after 12 months the increased wound healing rate was not visible in a difference in scar appearance. In vitro studies in this research showed that the clinical effect of a faster wound healing is probably ascribed to adipose derived stromal cells in the stromal vascular fraction which stimulate angiogenesis, decrease fibrosis and act anti-inflammatory. Additionally, it also contains extracellular matrix which is able to bind and release secreted factors by adipose derived stromal cells and thus functions as a natural scaffold to guide tissue regeneration

Potentials and Limitations of the Use of Platelet-Rich Plasma (PRP) in Combination with Lipofilling:An Evidence-Based Approach

The transplantation of adipose tissue or lipofilling can be used to restore volume loss or to regenerate damaged tissue. The regenerative capacity of adipose tissue is most likely to be ascribed to the stromal vascular fraction (SVF) containing all non-adipocyte cell types, e.g., fibroblasts, immune cells, endothelial cells, and adipose derived stromal cells (ASCs). ASCs are believed to be the most important cell type in SVF based on their ability to differentiate into multiple cell lineages such as endodermal, ectodermal, and mesenchymal cell types and to secrete a variety of paracrine factors. In this way, ASCs influence important processes related to healing of damaged tissue such as angiogenesis, extracellular matrix remodeling, and immunomodulation. To enhance the clinical effect of lipofilling, platelet-rich plasma (PRP) has been used to stimulate angiogenesis capacity as well as proliferation and differentiation of ASCs in lipograft by synthesizing large numbers of growth factors, e.g., vascular endothelial growth factor (VEGF) and platelet derived growth factor (PDGF). Due to the variety of growth factors in platelets, the addition of PRP to lipofilling might be even more effective to regenerate damaged tissue. To date, several case series in literature suggest a beneficial effect of the addition of PRP to lipofilling for skin rejuvenation, wound healing purposes, dermal fibrosis, and alopecia androgenetic although well-designed prospective clinical trials are lacking. The aim of this chapter is to highlight the currently available clinical evidence of PRP-enriched lipofilling for the aforementioned indications.</p

Platelet Rich STROMA, the Combination of PRP and tSVF and Its Potential Effect on Osteoarthritis of the Knee

(1) Background: osteoarthritis (OA) of the knee is a degenerative disease accompanied by pain, reduced mobility and subsequent decrease in quality of life. Many studies on OA of the knee have reported that using an intercellular acting-derivate like platelet-rich plasma (PRP) results in a limited effect or none at all. Authors hypothesized that adding tissue-Stromal Vascular Fraction (tSVF) to PRP (Platelet Rich Stroma (PRS)) would reduce pain and improve functionality in osteoarthritis of the knee. (2) Methods: a consecutive case series of fifteen patients (aged 43-75 years) suffering from OA of the knee (Kellgren-Lawrence stage two to three) were treated with a single injection of autologous PRS. tSVF was mechanically isolated by means of the fractionation of adipose tissue (FAT) procedure. Clinical evaluation was done using a visual analogue score (VAS) score, an adapted Western Ontario and McMaster Universities Osteoarthritis index (WOMAC) and Lysholm score at fixed time points: pre-injection as well as three, six and twelve months post injection. (3) Results: VAS and WOMAC scores improved significantly after twelve months (p 0.05) in comparison to pre-injection measurements. No complications were seen in any of the patients. One patient was excluded due to a total knee arthroplasty. (4) Conclusions: a single injection with PRS for OA of the knee seems to lead to an improvement of function and simultaneous reduction of pain and joint stiffness for a period of twelve months. Further controlled trials are required to determine the optimal treatment regimen and evaluate long-term results

Adipose Tissue-Derived Components:From Cells to Tissue Glue to Treat Dermal Damage

In recent decades, adipose tissue transplantation has become an essential treatment modality for tissue (volume) restoration and regeneration. The regenerative application of adipose tissue has only recently proven its usefulness; for example, the method is useful in reducing dermal scarring and accelerating skin-wound healing. The therapeutic effect is ascribed to the tissue stromal vascular fraction (tSVF) in adipose tissue. This consists of stromal cells, the trophic factors they secrete and the extracellular matrix (ECM), which have immune-modulating, pro-angiogenic and anti-fibrotic properties. This concise review focused on dermal regeneration using the following adipose-tissue components: adipose-tissue-derived stromal cells (ASCs), their secreted trophic factors (ASCs secretome), and the ECM. The opportunities of using a therapeutically functional scaffold, composed of a decellularized ECM hydrogel loaded with trophic factors of ASCs, to enhance wound healing are explored as well. An ECM-based hydrogel loaded with trophic factors combines all regenerative components of adipose tissue, while averting the possible disadvantages of the therapeutic use of adipose tissue, e.g., the necessity of liposuction procedures with a (small) risk of complications, the impossibility of interpatient use, and the limited storage options.</p

Mechanically Derived Tissue Stromal Vascular Fraction Acts Anti-inflammatory on TNF Alpha-Stimulated Chondrocytes In Vitro

Enzymatically isolated stromal vascular fraction (SVF) has already shown to be effective as a treatment for osteoarthritis (OA). Yet, the use of enzymes for clinical purpose is highly regulated in many countries. Mechanical preparation of SVF results in a tissue-like SVF (tSVF) containing intact cell–cell connections including extracellular matrix (ECM) and is therefore less regulated. The purpose of this study was to investigate the immunomodulatory and pro-regenerative effect of tSVF on TNFα-stimulated chondrocytes in vitro. tSVF was mechanically derived using the Fractionation of Adipose Tissue (FAT) procedure. Characterization of tSVF was performed, e.g., cellular composition based on CD marker expression, colony forming unit and differentiation capacity after enzymatic dissociation (from heron referred to as tSVF-derived cells). Different co-cultures of tSVF-derived cells and TNFα-stimulated chondrocytes were analysed based on the production of sulphated glycosaminoglycans and the anti-inflammatory response of chondrocytes. Characterization of tSVF-derived cells mainly contained ASCs, endothelial cells, leukocytes and supra-adventitial cells. tSVF-derived cells were able to form colonies and differentiate into multiple cell lineages. Co-cultures with chondrocytes resulted in a shift of the ratio between tSVF cells: chondrocytes, in favor of chondrocytes alone (p < 0.05), and IL-1β and COX2 gene expression was upregulated in TNFα-treated chondrocytes. After treatment with (a conditioned medium of) tSVF-derived cells, IL-1β and COX2 gene expression was significantly reduced (p < 0.01). These results suggest mechanically derived tSVF stimulates chondrocyte proliferation while preserving the function of chondrocytes. Moreover, tSVF suppresses TNFα-stimulated chondrocyte inflammation in vitro. This pro-regenerative and anti-inflammatory effect shows the potential of tSVF as a treatment for osteoarthritis

Lifting the Mouth Corner:A Systematic Review of Techniques, Clinical Outcomes, and Patient Satisfaction

BACKGROUND: Mouth corners are an essential part of the centro-facial area for perception of attractiveness and emotions. Downturned mouth corners are a result of aging or have a congenital origin. Different mouth corner lifting techniques are described in the literature. OBJECTIVES: This review was performed to systematically assess and compare invasive and non-invasive mouth corner lifting techniques and their effectiveness, patient satisfaction and adverse effects. METHODS: MEDLINE (via PubMed), EMBASE and The Cochrane Central Register of controlled trials databases were searched for clinical and observational studies published in peer-reviewed academic journals with abstracts available (searched from May 18, 2019, to December 18, 2021). Outcomes of interest were aesthetic mouth corner lifting techniques, the degree of lift as well as the longevity of the lifting effect, patient satisfaction and adverse effects. Techniques were subdivided in invasive techniques and non-invasive techniques. RESULTS: Eleven out of 968 studies were included in the qualitative analysis. In general, surgical techniques seem to have a better mouth corner lifting effect in comparison to non-surgical techniques; however, objective evidence is weak, and surgery inevitably results in a scar. Reported patient satisfaction was good for both surgical and non-surgical techniques and no severe complications have been described. CONCLUSIONS: Surgical techniques seem to have a better lifting effect on mouth corners than non-surgical techniques. Nevertheless, objective evidence is weak, and a scar is inevitable

Matrix Metalloproteases from Adipose Tissue-Derived Stromal Cells Are Spatiotemporally Regulated by Hydrogel Mechanics in a 3D Microenvironment

Adipose tissue-derived stromal cells (ASCs) are of interest in tissue engineering and regenerative medicine (TERM) due to their easy acquisition, multipotency, and secretion of a host of factors that promote regeneration. Retention of ASCs in or around lesions is poor following direct administration. Therefore, for TERM applications, ASCs can be ‘immobilized’ via their incorporation into hydrogels such as gelatine methacryloyl (GelMA). Tweaking GelMA concentration is a common approach to approximate the mechanical properties found in organs or tissues that need repair. Distinct hydrogel mechanics influence the ability of a cell to spread, migrate, proliferate, and secrete trophic factors. Mesenchymal cells such as ASCs are potent remodellers of the extracellular matrix (ECM). Not only do ASCs deposit components, they also secrete matrix metalloproteases (MMPs) which degrade ECM. In this work, we investigated if GelMA polymer concentration influenced the expression of active MMPs by ASCs. In addition, MMPs’ presence was interrogated with regard to ASCs morphology and changes in hydrogel ultrastructure. For this, immortalised ASCs were embedded in 5%, 10%, and 15% (w/v) GelMA hydrogels, photopolymerised and cultured for 14 d. Zymography in situ indicated that MMPs had a variable, hydrogel concentration-dependent influence on ASCs-secreted MMPs. In 5% GelMA, ASCs showed a high and sustained expression of MMPs, while, in 10% and 15% GelMA, such expression was almost null. ASCs morphology based on F-actin staining showed that increasing GelMA concentrations inhibit their spreading. Scanning electron microscopy (SEM) showed that hydrogel ultrastructure in terms of pore density, pore size, and percentage porosity were not consistently influenced by cells. Interestingly, changes in ultrastructural parameters were detected also in cell-free materials, albeit without a clear trend. We conclude that hydrogel concentration and its underlying mechanics influenced MMP expression by ASCs. The exact MMPs that respond to these mechanical cues should be defined in follow-up experiments

Skin measurement devices to assess skin quality:A systematic review on reliability and validity

Background Many treatments aim to slow down or reverse the visible signs of skin aging and thereby improve skin quality. Measurement devices are frequently employed to measure the effects of these treatments to improve skin quality, for example, skin elasticity, color, and texture. However, it remains unknown which of these devices is most reliable and valid. Materials and methods MEDLINE, Embase, Cochrane Central, Web of Science, and Google Scholar databases were searched. Instruments were scored on reporting construct validity by means of convergent validity, interobserver, intraobserver, and interinstrument reliability. Results For the evaluation of skin color, 11 studies were included describing 16 measurement devices, analyzing 3172 subjects. The most reliable device for skin color assessment is the Minolta Chromameter CR-300 due to good interobserver, intraobserver, and interinstrument reliability. For skin elasticity, seven studies assessed nine types of devices analyzing 290 subjects in total. No intra and interobserver reliability was reported. Skin texture was assessed in two studies evaluating 72 subjects using three different types of measurement devices. The PRIMOS device reported excellent intra and interobserver reliability. None of the included reviewed devices could be determined to be valid based on construct validity. Conclusion The most reliable devices to evaluate skin color and texture in ordinary skin were, respectively, the Minolta Chromameter and PRIMOS. No reliable device is available to measure skin elasticity in ordinary skin and none of the included devices could be determined to be designated as valid