Presentation number: MG 44
ARE BIOLOGICAL TREATMENTS FOR KNEE OSTEOARTHRITIS EFFECTIVE? KOOS, WOMAC AND VAS SCORE ANALYSIS
Zrinka Djukic Koroljevic1, Petar Brlek1, Vilim Molnar1, Damir Hudetz1,2,3, Zeljko Jelec1,4, Eduard Rod1, Dinko Vidovic1,5,6, Trpimir Vrdoljak1,2, Igor Boric1,7,8,9, Tadija Petrovic1,5, Fabijan Cukelj1,5,7, Vid Matisic1, Eduard Pavelic1, David Karli10, Dragan Primorac1,3,7,8,9, 11,12,13,14,15,16
1St. Catherine Specialty Hospital, Zagreb, Croatia, 2Clinical Hospital Sveti Duh, Zagreb, Croatia, 3School of Medicine, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia, 4Department of Nursing, University North, Varaždin, Croatia, 5Clinic for Traumatology, University Hospital Sestre Milosrdnice, Zagreb, Croatia, 6School of Dental Medicine, University of Zagreb, Zagreb, Croatia, 7Medical School, University of Split, Split, Croatia, 8Medical School, University of Rijeka, Rijeka, Croatia, 9Medical School, University of Mostar, Mostar, Bosnia and Herzegovina, 11Eberly College of Science, The Pennsylvania State University, University Park, State College, PA, USA, 10Steadman Clinic, Vail, CO USA, 12The Henry C. Lee College of Criminal Justice and Forensic Sciences, University of New Haven, West Haven, CT, USA, 13Medical School REGIOMED, Coburg, Germany, 14School of Medicine, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia, 15Faculty of Dental Medicine and Health, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia, 16National Forensic Sciences University, Gujarat, India
Osteoarthritis is the most common progressive musculoskeletal condition. It affects not only cartilage but all the joint tissues, causing irreversible morphological changes and decreased joint function. Changes are mediated by numerous cytokines, chemokines, adipokines, growth factors, and new, biological treatments are commonly used to slow down the natural course of disease but also to reduce patient’s symptoms. In this study, we included 8 women and 8 men with grade 2 and 3 knee osteoarthritis. Patients were treated with the intraarticular application of 2 mL of autologous micro-fragmented adipose tissue (MFAT) containing stromal vascular fraction (SVF) (Arthrex ACP, Double-Syringe System, Arthrex, Munich, Germany) in combination with 5 mL of leukocyte-poor platelet-rich plasma (PRP) (The Angel System, Arthrex, Munich, Germany).. After the initial assessment, patients were followed up on their Knee Injury and Osteoarthritis Outcome Score (KOOS) score, Visual analog scale score and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score 3 and 6 months after intraarticular application. Statistical analysis revealed a difference in the total KOOS score, total WOMAC score and the VAS score, both for resting and movement, over time, and the Wilcoxon Signed Ranks Test showed a statistically significant improvement three months (p = 0.001) and six months (p = 0.005) after the application of SVF and PRP. However, we did not observe changes in the glycosaminoglycans level (GAG) by using delayed gadolinium (Gd)-enhanced magnetic resonance imaging of cartilage (dGEMRIC). In conclusion, biological treatments are effective in reducing signs and symptoms of knee osteoarthritis, measured three months after intraarticular application, with the effect still consistent six months after application.
Key words: knee osteoarthritis, stromal vascular fraction, KOOS, WOMAC, VAS
Presentation number: MG 45
THE POTENTIAL USE OF MESENCHYMAL STEM CELLS IN GYNECOLOGY
Matea Hodonj1, Petar Brlek2, Ivana Erceg Ivkošić2,3, Dragan Primorac2,3,4,5,6,7,8,9,10,11
1Private family practice “dr. Ivica Cvetković”, Samobor, Croatia, 2St. Catherine Specialty Hospital, Zagreb, Croatia, 3Faculty of Dental Medicine and Health, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia, 4Medical School, University of Split, Split, Croatia, 5Department of Biochemistry & Molecular Biology, The Pennsylvania State University, State College, PA, USA, 6The Henry C. Lee College of Criminal Justice and Forensic Sciences, University of New Haven, West Haven, CT, USA, 7Medical School REGIOMED, Coburg, Germany, 8School of Medicine, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia, 9Medical School, University of Rijeka, Rijeka, Croatia, 10University of Split, University Department of Forensic Sciences, Split, Croatia, 11National Forensic Sciences University, Gujarat, India
Mesenchymal stem cells (MSCs) are our own body’s mechanism for healing and regeneration, all due to the secretion of bioactive factors which are having an immunomodulatory and regenerative ability. However, in the past decade, there is a rising number of studies that indicated their exceptional role in regenerative medicine. Their potential is very promising and can be used as a treatment for diseases in rheumatology, orthopedics, neurosurgery, endocrinology and many other fields. Many clinical trials, as well as published scientific articles, provided evidence showing the beneficial effect of their application, due to their analgesic, anti-inflammatory, antiapoptotic, angiogenic and immunomodulatory effects. In gynecology, they are being used both in animal and human studies for treating various innate and acquired diseases and especially in conditions that until now, haven’t been treated effectively. There are published case series evaluating the long-term (3 years) safety and effectiveness of MSC for urogenital atrophy which is according to literature and clinical practice, possibly a very effective new treatment for clinical problem that affects several millions of patients. Lichen sclerosus treatment is also very demanding and there have been published articles about successful results with MSC. As one of the world’s most common disorders, infertility and its possible causes are very challenging and often with poor therapeutic results. Studies have shown that endometrial, menstrual, umbilical, bone marrow and adipose-derived MSCs are efficient as a treatment and can restore fertility in treated individuals. In many cases, the use of MSCs in gynecology has proved efficient, safe and with the possibility of a wide application. In Asherman syndrome, this therapy enabled restoring the endometrium thickness – the patients had improvement of menstrual cycles and had successful pregnancies. So far, in St. Catherine’s specialty hospital, we have experience and great results with MSC in orthopedics – they have been successfully used in our hospital for more than six years. We are now preparing a clinical trial with MSC application in gynecology, hopefully with the same accomplishment. Yet more clinical and scientific studies are needed on the use of MSC in gynecology, but so far, they are very encouraging.
Key words: mesenchymal stem cell, gynecology, regenerative medicine, infertility, Asherman syndrome
Presentation number: MG 46
CLINICAL APPLICATION OF CULTURED KERATINOCYTES AS ADVANCED THERAPY MEDICINAL PRODUCTS: A TWENTY-YEAR EXPERIENCE IN CROATIA
Marija Zekušić1, Marina Bujić Mihica1, Tamara Dolenec1, Marija Skoko1, Anamarija Jularić1, Dominik Puljić1, Ivana Vrgoč Zimić1, Milivoje Boranić2, Hrvoje Tomičić3, Antun Kljenak4, Ivanka Batarilo5, Dinko Vidović6, Tiha Vučemilo1
1University Hospital Center “Sestre milosrdnice”, Department of Transfusion and Regenerative Medicine, Zagreb, Croatia, 2Ruđer Bošković Institute, Zagreb, Croatia, 3University Hospital Center “Sestre milosrdnice”, Department of Burns, Zagreb, Croatia, 4Children’s Hospital Zagreb, Zagreb, Croatia, 5Croatian Institute of Transfusion Medicine, Department of Microbiology, Zagreb, Croatia, 6University Hospital Center “Sestre milosrdnice”, Clinic for Traumatology, Zagreb, Croatia
The aim of this study is to present development of tissue engineering and clinical application of cultured keratinocytes. Advanced therapy medicinal products (ATMPs) are medicines for humans based on gene therapy, somatic cell therapy and tissue-engineered products. Cultured keratinocytes regenerate the epithelium and belong to the category of ATMP as tissue-engineered products. The development of ATMPs in Croatia began during 2002 in collaboration with the Ruđer Bošković Institute, the Clinic of Traumatology and the Children’s Hospital Zagreb on the project “Production of skin grafts in vitro”. Tissue Engineering Laboratory was built in May 2005 in accordance with Good Manufacturing Practice and clean room technology. Tissue and Cell Bank (TCB) was established during 2007. The procedure includes isolation of keratinocytes from the skin biopsy (about 4-6 cm2/ 0.3 mm thick) after which they are seeded onto a feeder layer of 3T3 cells and incubated at 37 °C, 5% CO2. Preparation of the optimal number of grafts is accomplished within 3-4 weeks depending on the area of the injury. Quality control involves potency (yield, viability, CFE), purity (p63, CK14, CK19), impurities (rest of 3T3 cells) and safety (sterility, mycoplasma, bacterial endotoxins). The first successful production of epidermal grafts began in the Clinic of Traumatology in September 2002 with 10 epidermal grafts of 700 cm2. This retrospective analysis covers period from February 2002 to October 2003. The project included donors (n=15), from 2 to 66 years old with 92 cultured grafts. Microbiological control has proven the sterility of all keratinocytes, cell media as well as epidermal transplants. From July 2007 to March 2022 in TCB, donors (n=62) were from 2 to 74 years old with 2175 cultured grafts from which 88,9% were transplanted and 11,1% discarded. The most common reasons for discardment were patient’s death, initial microbiological contamination, and technical reasons. Keratinocytes prepared as epidermal grafts or suspension with fibrin glue contributed to the survival of severely burned patients. Twenty years of successful cooperation between TCB employees and clinicians have resulted in successful application of cultured autografts in severely burnt patients.
Key words: tissue engineering, keratinocytes, ATMP
Presentation number: MG 47
INJECTION OF AUTOLOGOUS PLATELET-RICH PLASMA FOR TREATING ANDROGENIC ALOPECIA: PILOT STUDY OF A NOVEL TREATMENT PROTOCOL
Ana Maletic1, Ivo Dumic-Cule2, Petar Brlek3, Rado Zic4, Dragan Primorac3,5,6,7,8,9,10,11,12,13
1Polyclinic Maletic, Daruvar, Croatia, 2University North, Varazdin, Croatia, 3St. Catharine Special Hospital, Zagreb, Croatia, 4University Hospital Dubrava, Department of Plastic Surgery, Zagreb, Croatia, 5University of Split, School of Medicine, Split, Croatia, 6University of Split, University Department of Forensic Sciences, Split, Croatia, 7 University of Osijek, Faculty of Medicine, Osijek, Croatia, 8University of Osijek, Faculty of Dental Medicine and Health, Osijek, Croatia, 9University of Rijeka, School of Medicine, Rijeka, Croatia, 10Pennsylvania State University, Eberly College of Science, University Park, PA, USA, 11University of New Haven, Henry C. Lee College of Criminal Justice and Forensic Sciences, West Haven, CT, USA, 12Medical School REGIOMED, Coburg, Germany, 13The National Forensic Sciences University, Gandhinagar, Gujarat, India
Autologous platelet-rich plasma (PRP) treatment has emerged as a valuable, effective, and affordable treatment for androgenetic alopecia in recent years. Androgenetic alopecia is the most common type of alopecia, affecting both men and women, characterized by diminished hair follicles mainly pronounced in the frontal region and vertex. A huge variety of PRP treatment regimens were described but so far there is no consensus for standardization of PRP preparation or administration protocol. Our study enrolled two patients (ages 56 and 33 years old) with the aim to test the efficacy of a new PRP application protocol of only 2 treatments by using a combination of a PRP collecting device and a conventional kit. Efficacy of treatment was assessed after a 6-month follow-up by AI-driven software on microscopic images of treated regions. An average number of hairs, cumulative hair thickness, and the number of follicular units increased in the vertex region of both patients by 30/59%, 35/53%, and 14/48% respectively (P<0.05). A novel PRP treatment regimen with a decreased number of treatments was shown significantly effective in only 6 months of follow-up.
Key words: hair loss, vertex, autologous platelet-rich plasma
Presentation number: MG 48
NOVEL CELL-BASED THERAPIES IN CROHN’S DISEASE
Ana Dimova1, Petar Brlek1, Branko Bogdanić1,2, Mislav Rakić1,3, Tihomir Grgić1,4, Ivica Grgurević1,3, Tomislav Bokun1,3, Ivana Kovačić1,5, Dragan Primorac1,6,7,8,9,10,11,12,13,14
1St. Catherine Specialty Hospital, Zagreb, Croatia, 2University Hospital Centre Zagreb, Zagreb, Croatia, 3University Hospital Dubrava, Zagreb, Croatia, 4University hospital “Sveti Duh”, Zagreb, Croatia, 5General Hospital Karlovac, Karlovac, 6Medical School, University of Mostar, Mostar, Bosnia and Herzegovina, Croatia 7Faculty of Dental Medicine and Health, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia, 8Medical School, University of Rijeka, Rijeka, Croatia, 9Medical School, University of Split, Split, Croatia, 10Department of Biochemistry & Molecular Biology, The Pennsylvania State University, State College, PA, USA, 11University of New Haven, Henry C. Lee College of Criminal Justice and Forensic Sciences, West Haven, CT, USA, 12Medical School REGIOMED, Coburg, Germany, 13School of Medicine, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia, 14National Forensic Sciences University, Gujarat, India
Stem cells (SCs) are undifferentiated or partially differentiated cells with the potential to specialize in more mature cells and, at the same time, self-replicate in daughter stem cells. In addition to the high proliferative potential for regeneration, SCs have many other effects on tissues, including growth support, immunomodulatory effect, and effect on paracrine signaling. Mesenchymal stem cells (MSCs) are multipotent stem cells that can differentiate into a limited number of cell types of the same lineage. MSCs are considered new therapeutic agents for immune-mediated diseases, including Crohn’s disease (CD) thanks to their differentiation potential into gut cells and their pro-angiogenic and immunomodulatory characteristics. There are currently two methods for MSC use in patients with CD: systemic (intravenous) use for systemic control of intestinal inflammation in luminal CD and local administration as a therapeutic approach for patients with perianal fistulizing CD. Current therapy for CD involves immunosuppressive drugs that promote remission of intestinal inflammation and related symptoms. Recent research shows that depending on the intercellular environment in which stem cells are located, they have an increased secretion of anti-inflammatory cytokines (IL-2, IL-4, IL-10, and TGF-β) and a decrease in pro-inflammatory cytokines (IL-1, IL-6, IL-17, and TNF-α). MSCs in a pro-inflammatory environment with high concentrations of TNF-α and IFN-γ secrete anti-inflammatory mediators and become so-called MSC-2, which can inhibit the activation of dendritic cells, T and B lymphocytes, and NK cells. The results obtained in a large number of clinical trials suggest that topical application of autologous as well as allogeneic adipose-derived stem cells is a safe and useful therapeutic approach for the healing of perianal fistulas in patients with CD. The safety of MSC-based therapies, after systemic administration of MSCs, remains to be investigated to be safely used as new therapeutic agents for the treatment of CDs due to their differentiating potential as well as their proangiogenic and immunomodulatory properties.
Key words: Stem cell, Crohn’s disease, mesenchymal stem cell, TNF-α, adipose-derived stem cell