In Vitro Characterization of Adipose Stem Cells Non-Enzymatically Extracted from the Thigh and Abdomen.

Autologous fat grafting is a surgical technique in which adipose tissue is transferred from one area of the body to another, in order to reconstruct or regenerate damaged or injured tissues. Before reinjection, adipose tissue needs to be purified from blood and cellular debris to avoid inflammation and preserve the graft viability. To perform this purification, different enzymatic and mechanical methods can be used. In this study, we characterized in vitro the product of a closed automatic device based on mechanical disaggregation, named Rigenera®, focusing on two sites of adipose tissue harvesting. At first, we optimized the Rigenera® operating timing, demonstrating that 60 s of treatment allows a higher cellular yield, in terms of the cell number and growth rate. This result optimizes the mechanical disaggregation and it can increase the clinical efficiency of the final product. When comparing the extracted adipose samples from the thigh and abdomen, our results showed that the thigh provides a higher number of mesenchymal-like cells, with a faster replication rate and a higher ability to form colonies. We can conclude that by collecting adipose tissue from the thigh and treating it with the Rigenera® device for 60 s, it is possible to obtain the most efficient product.

A Non-Enzymatic Method to Obtain a Fat Tissue Derivative Highly Enriched in Adipose Stem Cells (ASCs) from Human Lipoaspirates: Preliminary Results.

Adipose tissue possesses phenotypic gene expression characteristics that are similar to human mesenchymal stem cells (hMSCs). Nevertheless, the multilineage potential may be inhibited, and cells may not expand adequately to satisfy the requirements of Good Manufacturing Practice (cGMP). An autologous hMSC-enriched fat product would fulfil the void from a biomedical and clinical perspective. In this study, we suggest a novel mechanism using a closed system without enzymes, additives or other modifications, which will produce non-expanded, accessible material. This decentralized fat product, unlike unprocessed lipoaspirates, adequately encloses the vascular stroma with adipocytes and stromal stalks along with their vascular channels and lumina. This fat product contained hASCs and fewer hematopoietic elements such as lipoaspirates, which were digested enzymatically according to flow cytometric investigations, and molecular analysis also showed significant hASC uniformity within the cells of the stromal vascular tissue. Moreover, the fat product produced a higher quantity of hASCs similar to hMSCs in isolation with the typical characteristics of an osteogenic, chondrogenic and adipogenic lineage. Interestingly, these properties were evident in the non-enzymatic derived adipose tissue, as opposed to hASCs in isolation from the enzymatically digested lipoaspirates, suggesting that the aforementioned procedure may be an adequate alternative to regenerate and engineer tissue for the treatment of various medical conditions and promote efficient patient recovery.

Management of Fournier's gangrene non-healing wounds by autologous skin micrograft biotechnology: a new technique.

Fournier's gangrene is an acute bacterial infection producing necrosis of the perineum and external genitalia that generally affects elderly men. Although skin grafts and flaps are the standard procedure for reconstruction, sometimes wounds can become chronic. Rigenera Protocol is a new technique based on autologous skin micrografts that reactivates and supports wound healing. A 40-year-old male with Fournier's gangrene, due to a rectal microperforation following diarrhoea, was treated with surgical debridement, negative pressure wound therapy and subsequently coverage with skin grafts. He developed non-healing wounds treated by Rigenera protocol after two months of advanced wound dressings. This technique is based on skin micrografts obtained by mechanical dermal disgregation to provide mesenchymal stem cells and extracellular matrix to the wound. The suspension injected into the wound triggers reactivation of healing without significant residual scarring on both donor site and treated area. Non-healing wounds were reduced by 15% at day 7 and by 50% after 30 days. Wounds completely healed after seventy days. The regenerated tissue appeared closer to skin graft than to scar tissue. This report shows how the use of skin micrografts through Rigenera protocol can be a useful method to reactivate wound healing resulting from Fournier's gangrene, with no discomfort for patient in a practical, safe and easy way.

In Vitro and In Vivo Studies of Alar-Nasal Cartilage Using Autologous Micro-Grafts: The Use of the Rigenera® Protocol in the Treatment of an Osteochondral Lesion of the Nose.

Cartilage defects represent a serious problem due to the poor regenerative properties of this tissue. Regarding the nose, nasal valve collapse is associated with nasal blockage and persistent airway obstruction associated with a significant drop in the quality of life for patients. In addition to surgical techniques, several cell-based tissue-engineering strategies are studied to improve cartilage support in the nasal wall, that is, to ameliorate wall insufficiency. Nevertheless, there are no congruent data available on the benefit for patients during the follow-up time. In this manuscript, we propose an innovative approach in the treatment of cartilage defects in the nose (nasal valve collapse) based on autologous micro-grafts obtained by mechanical disaggregation of a small portion of cartilage tissue (Rigenera® protocol). In particular, we first analyzed in vitro murine and human cartilage micro-grafts; secondly, we analyzed the clinical results of a patient with pinched nose deformity treated with autologous micro-grafts of chondrocytes obtained by Rigenera® protocol. The use of autologous micro-graft produced promising results in surgery treatment of cartilage injuries and could be safely and easily administrated to patients with cartilage tissue defects.

Treatment of Oncological Post-surgical Wound Dehiscence with Autologous Skin Micrografts.

BACKGROUND/AIM: The closure of postoperative wounds is essential in order to prevent surgical site infections or wound dehiscence, mainly in oncological patients. We aimed to demonstrate the efficacy of autologous micrografts in the management of wound dehiscence in an oncology patient undergoing decompressive spinal laminectomy. CASE REPORT: A 57-year-old man with IgG multiple myeloma and medullary plasmocytoma C7-T3, was to undergo decompressive spinal laminectomy and vertebral fixation leading to a wound dehiscence with exposed instrumentation. Autologous micrografts were obtained by Rigenera protocol and directly applied to the dehisced wound. After 60 days of negative pressure wound therapy, we observed reduction of the diameter and depth of wound dehiscence, with a coverage of instrumentation, without complete re-epithelialization, that instead was reached by application of autologous micrografts after 70 days. CONCLUSION: The Rigenera protocol may be the solution for complex wounds in oncological and immune-compromised patients where other treatments are contraindicated.