Simple and Rapid Non-Enzymatic Procedure Allows the Isolation of Structurally Preserved Connective Tissue Micro-Fragments Enriched with SVF

Busato, Alice; Francesco, Francesco De; Biswas, Reetuparna; Mannucci, Silvia; Conti, Giamaica; Fracasso, Giulio; Conti, Anita; Riccio, Vittorio; Riccio, Michele; Sbarbati, Andrea

doi:10.3390/cells10010036

Cited by 28 publications

(29 citation statements)

References 46 publications

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“…In non‐enzymatic separation processes, Victor [24] reported that the number of cells in the SVF aspirates obtained using the ultrasonic cavitation method was 3.93–12.8 × 10 6 and the cell viability rate was 86.5–96.2%. Conversely, Busato et al [7] reported the average number of non‐enzymatic yet free‐nucleated cells in SVF, obtained by the tumescent anaesthesia method, was 4.1 × 10 4 . In the literature, no study has reported the methods of obtaining SVF using peripheral local anaesthesia.…”

Section: Discussionmentioning

confidence: 99%

Midterm results of intra‐articular stromal vascular fraction injection for the treatment of knee osteoarthritis

Çimen,

Irgıt,

Bekmezci

et al. 2023

Knee surg. sports traumatol. arthrosc.

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Purpose This study aimed to evaluate the safety and eicacy of intra-knee stromal vascular fraction (SVF) injection in patients with symptomatic knee osteoarthritis at the midterm (3-year) follow-up. Methods SVF injection was applied to 25 knees of 20 patients. Eighteen patients (90%) were female, and the means ± standard deviations of age was 61.9 ± 7.8 (range, 50-76) years. Patients who received conservative treatment for at least 6 months and had radiographic Kellgren-Lawrence (K-L) grades 2 and 3 varus gonarthrosis were included in the study. SVF was obtained from the umbilical region by liposuction using local anaesthesia. Patients were followed-up for 36 months. Their visual analogue scale (VAS), Western Ontario and McMaster University Osteoarthritis Index (WOMAC) and Lysholm scores were evaluated before and at 6, 12, 24 and 36 months post-SVF injection. Results A statistically signiicant improvement (p < 0.05) was observed in VAS, WOMAC and Lysholm scores at the irst 2-year follow-up compared to baseline. However, no statistically signiicant diference (n.s.) was observed in VAS, WOMAC and Lysholm scores at the 3-year follow-up compared with baseline. Conclusion Intra-articular SVF injection decreased pain and signiicantly improved the functional outcomes in the irst 2 years in knees with grade 2-3 osteoarthritis; however, these positive efects of the injection disappeared in the 3rd year. Although short-term successful results of SVF have been reported in the literature, prospective studies are needed for medium-and long-term results.

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Section: Discussionmentioning

confidence: 99%

Midterm results of intra‐articular stromal vascular fraction injection for the treatment of knee osteoarthritis

Çimen,

Irgıt,

Bekmezci

et al. 2023

Knee surg. sports traumatol. arthrosc.

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“…In addition, some authors reported a reduced level of cell contamination after mechanical isolation when compared to enzymatic treatment [32]. The main pitfall is represented by the higher content of blood mononuclear cells, which appeared to be related to the location of ASCs precursors and pericytes in the perivascular niches, requiring releases [23,[33][34][35][36]. In the authors' opinion, the only real advantage of enzymatic digestion using collagenase to isolate and extract the SVF from the adipose tissue is that eventually, it can offer the possibility of obtaining the tissue breakdown to the single cell.…”

Section: Methods Of Isolationmentioning

confidence: 99%

New perspectives in regenerative medicine and surgery: the bioactive composite therapies (BACTs)

et al. 2021

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Regenerative medicine and surgery is a rapidly expanding branch of translational research in tissue engineering, cellular and molecular biology.To date, the methods to improve cell intake, survival, and isolation need to comply with a complex and still unclear regulatory frame, becoming everyday more restrictive and often limiting the effectiveness and outcome of the therapeutic choices. Thus, the authors developed a novel 360° regenerative strategy based on the synergic action of several new components called the bioactive composite therapies (BACTs) to improve grafted cells intake, and survival in total compliance with the legal and ethical limits of the current regulatory frame.The rationale at the origin of this new technology is based on the evidence that cells need supportive substrate to survive in vitro and this observation, applying the concept of translational medicine, is true also in vivo. Bioactive composite mixtures (BACMs) are tailor-made bioactive mixtures containing several bioactive components that support cells’ survival and induce a regenerative response in vivo by stimulating the recipient site to act as an in situ real bioreactor. Many different tissues have been used in the past for the isolation of cells, molecules, and growth factors, but the adipose tissue and its stromal vascular fraction (SVF) remains the most valuable, abundant, safe, and reliable source of regenerative components and particularly of adipose-derived stems cells (ADSCs). The role of plastic surgeons as the historical experts in all the most advanced techniques for harvesting, manipulating, and grafting adipose tissue is fundamental in this constant process of expansion of regenerative procedures. In this article, we analyze the main causes of cell death and the strategies for preventing it, and we present all the technical steps for preparing the main components of BACMs and the different mixing modalities to obtain the most efficient regenerative action on different clinical and pathological conditions. The second section of this work is dedicated to the logical and sequential evolution from simple bioactive composite grafts (BACGs) that distinguished our initial approach to regenerative medicine, to BACTs where many other fundamental technical steps are analyzed and integrated for supporting and enhancing the most efficient regenerative activity. Level of Evidence: Not gradable

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“…ASCs were isolated from human lipoaspirate of healthy donors (women of ages between 35 and 45 years) after informed consent, following the protocol described by Peroni et al [22] and Busato et al [23], using an enzymatic method [18,19]. flask with complete growth medium.…”

Section: Isolation and Seeding Of Adipose-derived Stem Cellsmentioning

confidence: 99%

Tissue-Material Integration and Biostimulation Study of Collagen Acellular Matrices

Sierra

Busato

Zingaretti

et al. 2022

Tissue Eng Regen Med

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Background: Breast reconstruction after mastectomy using silicone implants is a surgical procedure that occasionally leads to capsular contracture formation. This phenomenon constitutes an important and persistent cause of morbidity, and no successful therapies are available to date. Recently, the use of acellular membranes as a protective material for silicone prostheses has been gaining attention due to their ability to prevent this adverse outcome. For this reason, the evaluation of the tissue-material integration and the induced biostimulation by acellular membranes results crucial. Evaluation of in vivo tissue integration and biostimulation induced by three different natural acellular collagen membranes. Methods: Scanning electron microscopy was performed to analyse the membrane porosity and cells-biomaterial interaction in vitro, both in dry and wet conditions. Adipose-derived stem cells were cultured in the presence of membranes, and the colonisation capacity and differentiation potential of cells were assessed. In vivo tests and ex vivo analyses have been performed to evaluate dermal integration, absorption degree and biostimulation induced by the evaluated membrane. Results: Analysis performed in vitro on the three different acellular dermal matrices evidenced that porosity and the morphological structure of membranes influence the liquid swelling ratio, affecting the cell mobility and the colonisation capacity. Moreover, the evaluated membranes influenced in different manner the adipose derived stem cells differentiation and their survival. In vivo investigation indicated that the absorption degree and the fluid accumulation surrounding the implant were membrane-dependent. Finally, ex vivo analysis confirmed the membrane-dependent behavior revealing different degree of tissue integration and biostimulation, such as adipogenic stimulation. Conclusion: The physico-chemical characteristics of the membranes play a key role in the biostimulation of the cellular environment inducing the development of well-organized adipose tissue.

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Simple and Rapid Non-Enzymatic Procedure Allows the Isolation of Structurally Preserved Connective Tissue Micro-Fragments Enriched with SVF

Cited by 28 publications

References 46 publications

Midterm results of intra‐articular stromal vascular fraction injection for the treatment of knee osteoarthritis

Midterm results of intra‐articular stromal vascular fraction injection for the treatment of knee osteoarthritis

New perspectives in regenerative medicine and surgery: the bioactive composite therapies (BACTs)

Tissue-Material Integration and Biostimulation Study of Collagen Acellular Matrices

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