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-1ß and COX2 gene expression was upregulated in TNFα-treated chondrocytes. After treatment with (a conditioned medium of) tSVF-derived cells, IL-1ß 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.

The minimal access cranial suspension (MACS) lift: A systematic review of literature 18 years after its introduction.

BACKGROUND: One of the most popular short-scar rhytidectomy techniques used nowadays is the minimal access cranial suspension (MACS) lift developed by Tonnard and Vaerpele. The technique uses purse-string sutures in combination with limited skin undermining to obtain a clear vertical tissue repositioning. Since its introduction, the technique has extensively been described and used around the world by facial plastic surgeons. Now, 18 years after its introduction, a systematic review concerning the results and complications of the MACS lift is presented to establish its current position in facial rejuvenation. METHODS: The MEDLINE, Embase, Cochrane Central, and Google Scholar databases were searched for studies evaluating the MACS lift (June 10, 2020). Outcomes of interest were long-term effect, satisfaction, and complications of the MACS lifting as determined by the patient and/or surgeon. RESULTS: Six studies were included, with 739 patients treated with the MACS lift in total. No major complications were reported, four studies did report on the occurrence of minor complications. All of the six studies reported relatively high levels of satisfaction. Three studies reported a shorter procedural duration for the MACS lift compared with the conventional facelift. In three of the six included studies, the level of evidence was low. The effect on neck rejuvenation is limited. CONCLUSION: The MACS lift can be considered a minimally invasive facelift procedure with a relatively low complication rate. The procedure, often combined with additional procedures, results in evident patient and/or surgeon satisfaction. To obtain a better desired result on an aged neck area, additional procedures are warranted.

Tissue Stromal Vascular Fraction Improves Early Scar Healing: A Prospective Randomized Multicenter Clinical Trial.

BACKGROUND: Wound healing and scar formation depends on a plethora of factors. Given the impact of abnormal scar formation, interventions aimed to improve scar formation would be most advantageous. The tissue stromal vascular fraction (tSVF) of adipose tissue is composed of a heterogenous mixture of cells embedded in extracellular matrix. It contains growth factors and cytokines involved in wound-healing processes, eg, parenchymal proliferation, inflammation, angiogenesis, and matrix remodeling. OBJECTIVES: The aim of this study was to investigate the hypothesis that tSVF reduces postsurgical scar formation. METHODS: This prospective, double-blind, placebo-controlled, randomized trial was conducted between 2016 and 2020. Forty mammoplasty patients were enrolled and followed for 1 year. At the end of the mammoplasty procedure, all patients received tSVF in the lateral 5 cm of the horizontal scar of 1 breast and a placebo injection in the contralateral breast to serve as an intrapatient control. Primary outcome was scar quality measure by the Patient and Observer Scar Assessment Scale (POSAS). Secondary outcomes were obtained from photographic evaluation and histologic analysis of scar tissue samples. RESULTS: Thirty-four of 40 patients completed follow-up. At 6 months postoperation, injection of tSVF had significantly improved postoperative scar appearance as assessed by the POSAS questionnaire. No difference was observed at 12 months postoperation. No improvement was seen based on the evaluation of photographs and histologic analysis of postoperative scars between both groups. CONCLUSIONS: Injection of tSVF resulted in improved wound healing and reduced scar formation at 6 months postoperation, without any noticeable advantageous effects seen at 12 months.

The Addition of Tissue Stromal Vascular Fraction to Platelet-Rich Plasma Supplemented Lipofilling Does Not Improve Facial Skin Quality: A Prospective Randomized Clinical Trial.

BACKGROUND: Lipofilling has become popular as a treatment to improve aging-related skin characteristics (eg, wrinkles, pigmentation spots, pores, or rosacea). Different additives such as platelet-rich plasma (PRP) or stromal vascular fraction (SVF) have been combined with lipofilling to increase the therapeutic effect of adipose-derived stromal cells (ASCs). OBJECTIVES: The aim of this study was to examine the hypothesis that mechanically isolated SVF augments the therapeutic effect of PRP-supplemented lipofilling to improve facial skin quality. METHODS: This prospective, double-blind, placebo-controlled, randomized trial was conducted between 2016 and 2019. In total, 28 female subjects were enrolled; 25 completed the follow-up. All patients received PRP-supplemented lipofilling with either mechanically isolated SVF or saline. SVF was isolated by fractionation of adipose tissue (tSVF). Results were evaluated by changes in skin elasticity and transepidermal water loss, changes in skin-aging-related features, ie, superficial spots, wrinkles, skin texture, pores, vascularity, and pigmentation, as well as patient satisfaction (FACE-Q), recovery, and number of complications up to 1 year postoperative. RESULTS: The addition of tSVF to PRP-supplemented lipofilling did not improve skin elasticity, transepidermal water loss, or skin-aging-related features. No improvement in patient satisfaction with overall facial appearance or facial skin quality was seen when tSVF was added to PRP-supplemented lipofilling. CONCLUSIONS: In comparison to PRP-supplemented lipofilling, PRP-supplemented lipofilling combined with tSVF does not improve facial skin quality or patient satisfaction in a healthy population. PRP-supplemented lipofilling combined with tSVF can be considered a safe procedure.

The Rainbow Scale for the Assessment of the Cervicomental Angle: A Validated Scale.

BACKGROUND: Aging of the neck results in an increased cervicomental angle, which can be treated by various surgical and nonsurgical procedures. To measure the success of these procedures, standardized validated objective photographic measurement tools are needed. However, no online standardized photographic measurement tools exist for the assessment of the cervicomental angle. OBJECTIVES: The purpose of this study was to establish a validated and reliable measurement tool for the assessment of the cervicomental angle based on the Rainbow Scale. METHODS: A 5-point photographic rating scale was developed and created from 1 photograph with Adobe Photoshop. Fifteen reference photographs of women, 3 photographs per grade, were included for validation. Seven panelists (ie, plastic and maxillofacial surgeons) assessed the reference photographs 3 times with a minimal interval of 3 days in an online survey. Intra- and inter-observer agreements were calculated utilizing the weighted kappa coefficient. RESULTS: Mean intra-observer agreement was 0.93 (0.78-1.00). Mean interobserver agreement was 0.796 (0.574-0.961) for survey 1, 0.868 (0.690-0.960) for survey 2, and 0.820 (0.676-0.959) for survey 3. CONCLUSIONS: The Rainbow Scale for the assessment of the cervicomental angle has been validated in an online fashion. The scale is reproducible and reliable and requires no learning curve. Potential applications include objective assessment of neck treatment planning and surgical outcome.

Fractionation of Adipose Tissue Procedure With a Disposable One-Hole Fractionator.

BACKGROUND: Adipose tissue has been widely used in regenerative surgery for its therapeutic potential. This potential is often ascribed to the stromal vascular fraction (SVF), which can be mechanically isolated. Mechanical isolation results in an SVF that retains intact cell-cell communication including extracellular matrix and is therefore named tissue-SVF (tSVF). OBJECTIVES: The aim of this study was to evaluate a new disposable 1-hole fractionator for fractionation of adipose tissue (FAT), and compare this new device with the existing reusable 3-hole fractionator. METHODS: The composition of tSVF obtained via the 1-hole fractionator was histologically and histochemically compared to unprocessed adipose tissue. The number of viable nuclear cells in tSVF obtained by the 1-hole and 3-hole fractionators as well as unprocessed adipose tissue were compared after enzymatic isolation and tested for colony-forming capacity. Flow cytometry was used to compare different cell compositions based on surface marker expression between tSVF isolated by the two types of fractionators. RESULTS: Fractionation of adipose tissue with the 1-hole fractionator condenses vasculature and extracellular matrix by disrupting adipocytes. The number of viable nuclear cells in tSVF obtained with the two fractionators was comparable and significantly higher than unprocessed lipoaspirate. Furthermore, tSVF isolated by both fractionators showed similar cell compositions and comparable colony-forming capacities. CONCLUSIONS: FAT with a disposable 1-hole fractionator effectively isolates tSVF with a cell count and cell composition comparable to the fraction obtained with the 3-hole reusable fractionator. The disposable 1-hole fractionator, however, is safer and more user friendly.

The Effects of Facial Lipografting on Skin Quality: A Systematic Review.

BACKGROUND: Autologous lipografting for improvement of facial skin quality was first described by Coleman in 2006. The current dogma dictates that adipose tissue-derived stromal cells that reside in the stromal vascular fraction of lipograft contribute to skin rejuvenation (e.g., increased skin elasticity), a more homogenous skin color, and softening of skin texture. Nowadays, many studies have been reported on this "skin rejuvenation" effect of autologous fat grafting. This systematic review was undertaken to assess the efficacy of autologous lipografting on skin quality. METHODS: The MEDLINE, Embase, Cochrane Central, Web of Science, and Google Scholar databases were searched for studies evaluating the effect of autologous lipografting on facial skin quality (May 11, 2018). Outcomes of interest were skin texture, color, and elasticity in addition to histologic outcomes and number of complications. RESULTS: Nine studies were included, with 301 patients treated in total. No meta-analysis could be performed because of heterogeneity of the metrics and outcomes. Eight studies reported increased skin elasticity; improvement in skin texture; and a more homogeneous skin color after treatment with lipografting, cellular stromal vascular fraction, or nanofat. One study reported no increased skin elasticity after lipografting. Histologic improvement was seen after lipografting and adipose tissue-derived stromal cell injections. However, in general, the level of evidence of the included studies was low. No serious complications were reported. CONCLUSION: Autologous facial lipografting and cellular stromal vascular fraction and adipose tissue-derived stromal cell injections hardly seem to improve facial skin quality but can be considered a safe procedure.

The Development of Facial Lipofilling from a Historical Point of View.

Lipofilling, the transplantation of adipose tissue, has already been used since the end of the 19th century. For decades, lipofilling was used to restore loss of volume due to aging, trauma, or congenital defects. Later on, the indications for the use of lipofilling expanded by treating aged skin, scars, and improving wound healing. The expansion was caused by the discovery of adipose derived stromal cells (ASCs) in adipose tissue and the development of very fine harvesting and injection cannulas which made it possible to inject small adipose tissue particles in small volume areas, such as the face. ASCs are multipotent stromal cells which reside in the stromal vascular fraction (SVF) of adipose tissue and are able to differentiate in multiple cell lineages and secrete a plurality of growth factors with regenerative potentials. The discovery of ASCs led toward more experimental cell-based therapies, that is, ASCs or SVF isolated by means of enzymatic isolation procedures. Later on, enzymatic isolation procedures were forbidden in many countries by legislation and were replaced by mechanical isolation procedures, such as the Nanofat and Fractionation of Adipose Tissue (FAT) procedures. The Nanofat procedure has been extensively investigated, especially as treatment for skin rejuvenation in the face. Though, substantial evidence is lacking for using facial lipofilling or any therapeutic component, that is, ASCs or SVF for skin rejuvenation to date. In contrast, facial lipofilling to restore loss of volume seems to be promising.

Isolation of Stromal Vascular Fraction by Fractionation of Adipose Tissue.

Adipose tissue-derived stromal cells (ASCs) are a promising candidates for cellular therapy in the field of regenerative medicine. ASCs are multipotent mesenchymal stem cell-like and reside in the stromal vascular fraction (SVF) of adipose tissue with the capacity to secrete a plethora of pro-regenerative growth factors. Future applications of ASCs may be restricted through (trans)national governmental policies that do not allow for use of nonhuman-derived (non-autologous) enzymes to isolate ASC. Besides, enzymatic isolation procedures are also time consuming. To overcome this issue, nonenzymatic isolation procedures to isolate ASCs or the SVF are being developed, such as the fractionation of adipose tissue procedure (FAT). This standardized procedure to isolate the stromal vascular fraction can be performed within 10-12 min. The short procedure time allows for intraoperative isolation of 1 mL of stromal vascular fraction derived from 10 mL of centrifuged adipose tissue. The stromal vascular fraction mostly contains blood vessels, extracellular matrix, and ASCs. However, based on the histological stainings an interdonor variation exists which might result in different therapeutic effects. The existing interdonor variations can be addressed by histological stainings and flow cytometry.

Adipose tissue-derived extracellular matrix hydrogels as a release platform for secreted paracrine factors.

Fat grafting is an established clinical intervention to promote tissue repair. The role of the fat's extracellular matrix (ECM) in regeneration is largely neglected. We investigated in vitro the use of human adipose tissue-derived ECM hydrogels as release platform for factors secreted by adipose-derived stromal cells (ASCs). Lipoaspirates from nondiabetic and diabetic donors were decellularized. Finely powdered acellular ECM was evaluated for cell remainders and DNA content. Acellular ECM was digested, and hydrogels were formed at 37°C and their viscoelastic relaxation properties investigated. Release of ASC-released factors from hydrogels was immune assessed, and bio-activity was determined by fibroblast proliferation and migration and endothelial angiogenesis. Acellular ECM contained no detectable cell remainders and negligible DNA contents. Viscoelastic relaxation measurements yielded no data for diabetic-derived hydrogels due to gel instability. Hydrogels released several ASC-released factors concurrently in a sustained fashion. Functionally, released factors stimulated fibroblast proliferation and migration as well as angiogenesis. No difference between nondiabetic and diabetic hydrogels in release of factors was measured. Adipose ECM hydrogels incubated with released factors by ASC are a promising new therapeutic modality to promote several important wound healing-related processes by releasing factors in a controlled way.

Augmentation of Dermal Wound Healing by Adipose Tissue-Derived Stromal Cells (ASC).

The skin is the largest organ of the human body and is the first line of defense against physical and biological damage. Thus, the skin is equipped to self-repair and regenerates after trauma. Skin regeneration after damage comprises a tightly spatial-temporally regulated process of wound healing that involves virtually all cell types in the skin. Wound healing features five partially overlapping stages: homeostasis, inflammation, proliferation, re-epithelization, and finally resolution or fibrosis. Dysreguled wound healing may resolve in dermal scarring. Adipose tissue is long known for its suppressive influence on dermal scarring. Cultured adipose tissue-derived stromal cells (ASCs) secrete a plethora of regenerative growth factors and immune mediators that influence processes during wound healing e.g., angiogenesis, modulation of inflammation and extracellular matrix remodeling. In clinical practice, ASCs are usually administered as part of fractionated adipose tissue i.e., as part of enzymatically isolated SVF (cellular SVF), mechanically isolated SVF (tissue SVF), or as lipograft. Enzymatic isolation of SVF obtained adipose tissue results in suspension of adipocyte-free cells (cSVF) that lack intact intercellular adhesions or connections to extracellular matrix (ECM). Mechanical isolation of SVF from adipose tissue destructs the parenchyma (adipocytes), which results in a tissue SVF (tSVF) with intact connections between cells, as well as matrix. To date, due to a lack of well-designed prospective randomized clinical trials, neither cSVF, tSVF, whole adipose tissue, or cultured ASCs can be indicated as the preferred preparation procedure prior to therapeutic administration. In this review, we present and discuss current literature regarding the different administration options to apply ASCs (i.e., cultured ASCs, cSVF, tSVF, and lipografting) to augment dermal wound healing, as well as the available indications for clinical efficacy.

Introducing Platelet-Rich Stroma: Platelet-Rich Plasma (PRP) and Stromal Vascular Fraction (SVF) Combined for the Treatment of Androgenetic Alopecia.

BACKGROUND: Androgenetic alopecia (AGA) is characterized by miniaturization of the hair follicles gradually causing conversion of terminal hairs into vellus hairs, leading to progressive reduction of the density of hair on the scalp. Approved therapeutic options are limited and show side effects. OBJECTIVES: To evaluate injections of stromal vascular fraction (SVF), which is rich in adipose-derived stromal cells (ASCs) in combination with platelet-rich plasma (PRP) in the upper scalp as a new autologous treatment option for AGA. METHODS: Ten male patients (age range, 25-72 years), suffering from AGA at stage II to III according to the Norwood-Hamilton scale, have been treated with a single injection of autologous PRS (ACPSVF: combination of PRP and SVF) in the upper scalp. Preinjection and 6 and 12 weeks postinjection changes in hair density were assessed using ultra high-resolution photography (Fotofinder). RESULTS: Hair density was significantly increased after 6 weeks and 12 weeks postinjection (P = 0.013 and P < 0.001). In hair-to-hair matching analyses, new hair grew from active follicles. Furhtermore nonfunctioning hair follicles filled with hyperkeartotic plugs, up to today assumed incapable of forming new hair, proved to grow new hair. No side effects were noted after treatment. CONCLUSIONS: A single treatment of platelet-rich stroma injected in the scalp of patients with AGA significantly increased hair density within 6 to 12 weeks. Further research is required to determine the optimal treatment regimen. Preferred options to our opinion include the repetition of PRS or additional treatments with PRP.