Marek Cegielski

Experimental application of xenogenous antlerogenic cells in replacement of auricular cartilage in rabbits

Abstract:  Background:  Regenerative medicine in the recent years aims at explaining involvement of stem cells in regenerative processes and applying this knowledge in fulfilling human needs to find new, more efficient therapeutic methods. Growing antlers constitute a model organ for examining regeneration processes of tissues because they are the only mammalian appendages capable of regeneration. The rate of growth of deer antlers makes them one of the quickest growing structures in mammals. The cells taking part in this process have a considerable proliferating potential. The aim of the study was to analyze the possibility of using xenogenous antlerogenic cells (AC) in regeneration of cartilaginous tissues in non‐immunosuppressed animals.

The effect of physical training on the N-methyl-N-nitrosourea-induced mammary carcinogenesis of Sprague–Dawley rats

The impact of physical activity on carcinogenesis has been demonstrated in many studies. Taking into account the discrepant results of physical exercise on the cell proliferation and apoptosis of breast cancer, we aimed to examine the impact of physical training on N-methyl-N-nitrosourea-(MNU)-induced mammary carcinogenesis. Fifty female rats were divided into four groups according to the intensity of physical activity they undertook. The number of developed tumors, tumor volume, and histopathological diagnoses were noted. Apoptosis and cell proliferation were studied by the number of TUNEL-positive and Ki-67-expressing cells. We demonstrated a statistically significant decrease in the tumor number between all trained groups and the control group. The results were most pronounced in the group with a moderate intensity of training. Moreover, we showed a decrease in tumor volume as training intensity increased, though the differences were not statistically significant. The mean number of TUNEL-positive cancer cells was significantly higher in the training groups than in the control group. These data suggest that physical training, especially of moderate intensity, may alleviate MNU-induced mammary carcinogenesis. The results could suggest that physical exercise-induced apoptosis may be a protective mechanism.

Antlerogenic stem cells: molecular features and potential in rabbit bone regeneration

ABSTRACT Aim: (i) To assess the expression profiles of stem cell-associated markers including Oct4, Sox2, Klf4, Nanog, C-myc, Stat3 and Cd9, (ii) analyze the nanotopography of the MIC-1 stem cells and (iii) evaluate the efficiency of live stem cell implants and stem cell culture derivatives on the regeneration of bone deficiencies in rabbit mandibles. Materials and methods: The expression profiles of stem cell-associated genes, including Oct4, Sox2, Klf4, Nanog, C-myc, Stat3 and CD9 were assessed using reverse transcription polymerase chain reaction and flow cytometry. Nanotopography of the antlerogenic MIC-1 cell lineage was analyzed using atomic force microscopy. The effect of MIC-1 stem cells, their homogenate and supernatant on the regeneration of bone deficiencies in rabbit mandibles was evaluated using histological analysis. The effect of MIC-1 stem cells and stem cell-based derivatives on the immune responses of the animals was assessed by analyses of acute phase protein levels (haptoglobin and fibrinogen). Results: We found that the MIC-1 cells isolated from the apical regions of growing antlers exhibited molecular features that were characteristics of pluripotent stem cells. Using atomic force microscopy, we determined the details of the cell surface morphologies with a particular emphasis on the patterns of formation of plasma extensions for interlinking adjacent cells. We also demonstrated that not only implanted stem cells but also cell homogenates and cell post-culture supernatants have potential in the regeneration of bone deficiencies in the rabbit mandible. Conclusions: Our findings indicate that the use of both antlerogenic stem cell implants and the preparations derived from the cells offer alternative approaches to those based on autologous stem cells in the biological stimulation of osteogenesis and in bone regeneration.

Characteristics of Antlerogenic Stem Cells and Their Potential Application

Marek Cegielski1,4, Ilona Iżykowska1, Wojciech Dziewiszek2,4, Maciej Zatoński3, Marek Bochnia3,4 and Olga Kalisiak5 1Departments of Histology and Embryology, Medical University of Wroclaw 2 Departments of Pharmacology, Medical University of Wroclaw, 3Department of Otolaryngology, Faculty of Dentistry, Medical University of Wroclaw, 4Stem Cells spin Ltd. Wrocław Technology Park, Wroclaw, 5Department of Clinical Sciences, Faculty of Veterinary Medicine, Warsaw University of life Sciences SGGW2 Poland Corresponding author Marek Cegielski, VD, PhD Medical University of Wroclaw, Dept. of Histology and Embryology ul. Chalubinskiego 6a 50-368 Wroclaw, Poland marcel@hist.am.wroc.pl

Reconstruction of cranial cruciate ligament in rabbits using polyester implants saturated with PRP, antlerogenic stem cells MIC-1 and their homogenate

ABSTRACT Aim of the study: The attempt to limit the negative effects of polyester implants on the articular cavity by using preparations containing growth factors. Materials and Methods: Polyester implants used for the reconstruction of a rabbit’s cranial cruciate ligament (CCL) were saturated with autogenic platelet-rich plasma (PRP), antlerogenic stem cells MIC-1 and their homogenate prior to the surgery. Six months after CCL reconstruction, morphological, and biochemical blood tests were carried out, including proteinogram and acute phase proteins. The knee joints were also examined macro- and microscopically. Results: The results, compared to the control group, showed a favorable effect of the PRP and homogenate of antlerogenic cells on limiting the inflammation caused by the presence of polyester implant in the knee joint. The addition of growth factors caused covering the implant faster with the recipient’s connective tissue, thus contributing to reducing the inflammatory reaction of the articular capsule to the presence of polyester. At the same time, no enhanced local or general reaction of the rabbit organism was observed to the presence of xenogenic antlerogenic stem cells MIC-1 homogenate which, like the PRP, may provide an easily available source of growth factors, increasingly often used in regenerative medicine. Conclusions: Applying antlerogenic stem cells, their homogenate or PRP increases the volume of connective tissue that surrounds and intertwines polyester CCL implant, separating it from synovial cavity environment.