Тканевой и клеточный банк в обеспечении здоровья населения

Şcoala de Management în Sănătate Publică, USMF Nicolae Testemiţanu, Laborator Inginerie tisulară şi Culturi celulare, Banca de Ţesuturi Umane, IMSP SCTOThis article represents an overview of the modern activities in tissues and cells banking activity for transplantation, including in the Republic of Moldova. It describes the types of tissues banks and grafts, history of tissue and cells harvesting, processing and preservation. It provides an analysis of the European and national legislation in this area and discusses the needs of modern medicine in such medical structures.Эта статья представляет собой обзор актуальных проблем, связанных с банковской деятельностью в области консервации тканей и клеток для трансплантации, в том числе в Молдове, видов банков и трансплантатов. Также проанализированы история консервации тканей, европейское и национальное законодательства в этой области, современные потребности медицины в таких медицинских структурах

The tissue engineered grafts for bone defects recovering

Laborator Inginerie tisulară şi culturi celulare, catedra Chirurgie operatorie şi anatomie topografică USMF “Nicolae Testemiţanu”Biological methods for bone regeneration are needed to reverse bone loss caused by diverse agents. Regenerative therapies could use traditional autogenous, allogenous bone grafting or composite grafts including resorbable 3D scaffolds and differentiated or no differentiated cells. As cells grafts could be: autogenous bone marrow cells or allogenous cord blood stem cells. Also gene expression systems will be combined with cells seeded in precise three-dimensional configurations on synthetic scaffolds to control both temporal and spatial distribution of regenerative factors. The natural processes of bone formation and repair require the coordinated expression of many molecules; including growth factors, bone morphogenetic proteins, and specific transcription factors. Current knowledge related to bone and fracture repair describes how tissue engineering, may mimic critical aspects of these natural processes

Surgical treatment of unicameral calcaneus cyst by introducing demineralized allogeneic bone paste: a case report

Introducere. Osul calcaneu nu este un loc obișnuit pentru chisturile osoase unicamerale. Nu este cunoscută etiologia acestor leziuni. Astfel de chisturi sunt simptomatice, din cauza concentrării forțelor pe călcâi și necesită tratament chirurgical în majoritatea cazurilor. Scopul lucrării. Această lucrare constituie un caz clinic al unui pacient căruia i s-a efectuat plastia defectului osos cu pastă osoasă demineralizată. Material și metode. Scopul acestei lucrări este de a prezenta cazul unui sportiv de 20 de ani care a fost internat la Secția de Microchirurgie și Chirurgie Reconstructivă cu dureri în regiunea calcaneului stâng în timpul mersului, apărute în urmă cu aproximativ 4 săptămâni. Pacientul a fost supus unei radiografii toracice, analize generale de sânge și urină, toate având parametri in limitele normei, cu excepția radiografiei piciorului stâng care a evidențiat o leziune chistică de 4,0x3,0x3,0 cm a calcaneului drept. Pacientul a fost supusă unui tratament chirurgical, care a constat în chiuretaj al chistului urmat de umplerea defectului osos cu 12 cm3 de pastă osoasă alogenă demineralizată amestecată cu 10 ml de sânge al pacientului. Pasta osoasă alogenică demineralizată a fost preparată de Banca de țesuturi umane. A fost efectuată o radiografie intraoperatorie care a arătat repararea defectului osos. Nu au fost raportate complicații postoperatorii precoce sau tardive. Pacientul a fost externat a doua zi după operație. Concluzii. Datele imagistice sunt necesare atunci când se suspectează o masă chistică. Tratamentul chirurgical în combinație cu utilizare de pasta osoasă demineralizată ar trebui să fie considerat un ca fiind un tratament eficient pentru chisturile osoase.Background. The calcaneus is not a commonplace for unicameral bone cysts. Not much is known about the etiology of these lesions. Such kind of cysts are symptomatic, because of the concentration of forces through the heel and require surgical treatment in most of the cases. Objective of the study. This paper is a case report of bone defect substitution using demineralized bone paste. Material and methods. The purpose of this work is to report a case of a 20-year-old athlete who was admitted to the Department of Microsurgery and Reconstructive Surgery with pain in the region of the left calcaneus during walking, which appeared about 4 weeks ago. The patient underwent plain chest X-ray, general blood and urine test, all of them came normal beside the X-ray of the left foot which showed a 4.0x3.0x3.0 cm cystic lesion of the right calcaneus. The patient underwent surgical treatment, which consisted of extended curettage of the cyst followed by filling of the bone defect with 12 cm3 demineralized allogeneic bone paste mixed with 10 ml of patients’ blood. The Human Tissue Bank prepared the demineralized allogeneic bone paste. An intraoperative X-ray was performed and showed the bone defect repair. There were no early nor late postoperative complications reported. The patient was discharged from the hospital the second day after the operation. Conclusions. Imaging data are required when a cystic mass is suspected. Surgical treatment in combination with demineralized bone paste should strongly be considered as an effective treatment for bone cysts

Izolarea colagenului din complexul ombilico-placentar pentru utilizarea în ingineria tisulară

Background. Collagen-based bone substitutes are widely used in bone tissue regeneration in orthopedic, traumatic and oromaxillofacial surgery. The predominant sources of collagen extraction are from animals, but they can transmit zoonoses from animal to human. Thus, collagen from the umbilicalplacental complex is a particular interest in the use as a graft for bone regeneration. Objective of the study. Obtaining collagen from the umbilical-placental complex for use in tissue engineering. Material and Methods. Collagen was isolated from umbilical-placental complexes (n = 3) with a mass of 66,5± 0,06 g received from the Tissue Bank. Preliminarily from the material, the non-collagenous proteins were removed with 0.05 M Na2HPO4, pH 8.7-9.1. The extraction was performed with pepsin and СН3СООН 0,5М and 5 mМ EDTA. Statistical processing was performed in Excel 2003. Results. The collagen extracts that were obtained after precipitation with 0.9% NaCl initially settled to the bottom of the container in the form of collagen fibers. After centrifugation and decantation of the supernatant, white mucilaginous substances were obtained which were purified by dialysis. The collagen concentrations that were obtained constituted 5,86± 0,04 mg/ml and were determined on the basis of dry mass. Conclusion. The amount of collagen obtained from the placental umbilical complex is significant and indicates that the source of production is a safe one and the method of production is efficient. Introducere. Substituenții osoși pe bază de colagen sunt utilizați pe scară largă în regenerarea țesutului osos în chirurgia ortopedică, traumatică și oromaxilo-facială. Sursele de extragere a colagenului predominant sunt de la animale, însă acestea pot transmite de la animal la om zoonoze. Astfel, colagenul din complexul ombilico-placentar prezintă un interes deosebit în utilizare ca grefă pentru regenerarea osoasă. Scopul lucrării. Obținerea colagenului din complexul ombilico-placentar pentru utilizarea în ingineria tisulară. Material și Metode. Colagenul a fost izolat din complexele ombilico-placentare (n=3) cu masa 66,5± 0,06 g primite de la Banca de Țesuturi. Preliminar, din material au fost îndepărtate proteinele necolagenice cu 0,05М Na2HPО4, pH 8,7-9,1. Extracția a fost efectuată cu pepsină și СН3СООН 0,5М și 5 mМ EDTA. Prelucrarea statistică a fost efectuată în Excel 2003. Rezultate. Extractele colagenice care au fost obținute după precipitarea cu NaCl 0,9 % inițial s-au sedimentat la fundul recipientului având aspect de fibre de colagen. După centrifugare și decantarea supernatantului au fost obținute substanțe mucilaginoase de culoare albă care au fost purificate prin dializă. Concentrațiile de colagen care au fost obținute au constituit 5,86± 0,04 mg/ml și au fost determinate în baza masei uscate. Concluzii. Cantitatea de colagen obținut din complexul ombilico-placentar este semnificativă și denotă că sursa de obținere este una sigură, iar metoda de obținere este eficientă

Angiogenesis stimulation modalities in experimental ischemia of the lower limb

Catedra Anatomie Topografică şi Chirurgie Operatorie, USMF „Nicolae Testemiţanu” Laboratorul Inginerie tisulară şi culturi celulareObstructive arterial diseases of different etiologies remain a significant cause of surgical and medical hospital treatment. The number of performed classic surgery is a continuous falling. Due to the development of efficient techniques involving gene or stem cell therapy could be the most promising treatment methods in severe lower limb ischemia. Arteriopatiile obstructive de toate etiologiile rămân procentual la un număr important de cazuri internate şi tratate în clinicele chirurgicale şi medicale. Numărul de intervenţii chirurgicale clasice este în continuă scădere, datorită dezvoltării a unor tehnici relativ eficiente, cum ar fi: terapia genică şi terapia cu celule progenitoare, care este una din cele mai promiţătoare metode de tratament al ischemiei critice ale membrelor pelviene

Stem cells therapy in infertility

Foreword. Worldwide, infertility affects 8-12% of couples of reproductive ages. In recent years, stem-cell therapy raised new hope in the field of reproductive disability management. Infertility is a major health problem, and the data presented in this review suggest that the treatment of infertility with stem cells seems to be plausible, because some types of treatments have already been tested in humans, achieving live births, while others show great potential only in animal studies. Material and methods. Was provided scientific research of the preclinical and clinical studies for the last 15 years in order to determine the effects of stem cell-based therapy in the treatment of infertility. Articles were selected from the PubMed, Scopus, and ScienceDirect databases. Results. Studies using the experimental model have shown that stem cell therapy for infertility treatment gives positive results. The main types of used stem cells are embryonic stem cells (ESCs), mesenchymal stem cells (MSCs), spermatogonia stem cells (SSCs), and induced-pluripotent stem cells (iPSCs). ESCs can produce germ cells and thereby treat infertility. ESC-derived PGCs are susceptible to ethical controversy because the process involves the destruction of human embryo. Sperm/oocytelike cells can now be produced from embryonic stem cells. SSC transplantation is presented as a novel and promising strategy, based on the premise of spermatogenesis and stem cell self-renewal. SSCs are responsible for the continuous production of male sperm. This technology can also be used to treat human infertility and does not cause ethical or immune problems. These results strongly suggest that SSC transplantation can be a successful treatment for male infertility caused by premature chemotherapy. Some studies have recently reported that both human iPSCs can differentiate into male germ cells. Functional tests showed that sperm produced by iPSCs were able to fertilize oocytes after intra-cytoplasmic injection and generate fertile offspring after embryo transfer. Ovarian failure is inevitable with age. In addition, injecting BM stem cells can stimulate ovarian function, restore normal ovaries and hormone levels, and possibly allow pregnancy. Conclusion. Several aspects of stem cell therapy remain unexplored. Thus, vast untapped potential still exists regarding applications in treating diseases such as infertility. We are confident that science will be able to cure infertility once the right approach is found. The reason for great promise is that stem cell treatment can be utilized on many different levels, from direct transplantation of stem cells, or their paracrine factors into reproductive organs, to in vitro differentiation into germ cells or gametes

The dental pulp chamber evaluation by using cone-beam computed tomography

Laboratory of Tissue Engineering and Cells Culture, Nicolae Testemitanu State University of Medicine and Pharmacy, Chisinau, the Republic of MoldovaAbstract. Background: Cone-beam computed tomographic (CBCT) imaging is a valuable tool in dental practice. It is widely used in endodontic treatment for the root canal morphology examination. Therefore, the purpose of this study was to use CBCT to calculate the volume of the pulp chamber at different tooth groups. Material and methods: This study conforms to protocols approved and in accordance with the ethics committee’s requirements, informed consent was obtained from each patient. Morphologic measurements of 120 maxillary and 120 mandibular molars (from 40 patients, aged 18–45 years) were included in this study. CBCT images were taken using a Kodak 9500 (Dental Systems, Carestream Health) operated at 90 kVp with a voxel size of 300 mm and a field of view of 90-50 mm. All scans were taken following the manufacturer’s recommendation protocol. According to the examination requirements, C-shaped roots, single-rooted molars, crowned teeth, and teeth with caries and/or restorations violating the pulp chamber were excluded. All measurements were taken on the coronal plane view. Results: In the present study, we used CBCT imaging to gather information regarding pulp chamber volume. With the scanned 3-dimensional images, we were able to clinically determine the pulp chamber parameters using a standardized and defined spatial approach. Conclusions: The data we collected here serve as a proof of principle for the analysis of dental landmarks before collecting stem cells. In this particular study, existing CBCT scans were used to provide useful information that can be used as a guide to determine the volume of the pulp chamber

Colagenul în medicicna regenerativă – present şi perspective

Laboratory of Tissue Engineering and Cells Cultures, SMPhU „Nicolae Testemiţanu”In the new paradigms of regenerative medicine, the use of materials in contact with biological materials (cells, tissues/organs, physiological fluids and biomolecules) is a current illustration of the need of interdisciplinary scientific approaches that combine the most recent advances in materials science and technology, basic sciences and life sciences. In tissue engineering, matrices are developed for cells support, promoting their differentiation and proliferation towards the formation of a new tissue. Such strategies allow for producing hybrid constructs that can be implanted in patients to induce the regeneration of tissues or replace failing or malfunctioning organs. Different materials have been proposed to be used in the processing of scaffolds, namely biodegradable polymers. In this review, the natural-based materials of collagen that have been proposed to be used in tissue engineering strategies and isolation methods of collagen will be overviewed. Colagenul în medicicna regenerativă – present şi perspective În paradigma nouă a medicinei regenerative utilizarea materialelor care vin în contact cu materialele biologice (celule, ţesututi, organe, lichide fiziologice şi biomolecule) necesită studii ştiinţifice interdisciplinare, care combină cele mai recente performanţe în ştiinţa şi tehnologia materialelor, ştiinţele de bază şi a vieţii. În ingineria tisulară sunt dezvoltate matrice pentru suportul celulelor, cu proprietăţi de a induce diferenţierea şi prolifirarea în direcţia formării noului ţesut. Se necesită obţinerea construcţiilor hibride care pot fi implantantate la pacienţi pentru a induce regenerarea ţesuturilor sau înlocuirea oreganelor nefuncţionale sau cu funcţia redusă. În acest articol de sinteză sunt descrise şi analizate materiale de origine naturală, anume a colagenului, metodele cunoscute de izolare şi purificare a colagenului ce sunt propuse pentru a fi utilizate în perspectiva ingineriei tisulare. Cuvinte cheie: Ingineria tisulară, colagen tip I, metode de izolare, matrice 3D

Management of the cornea grafts in the tissue and human cells bank of the Republic of Moldova during the years 2013-2019

Nicolae Testemitanu State University of Medicine and Pharmacy of the Republic of Moldova, Laboratory of Tissue Engineering and Cells Cultures, Congresul consacrat aniversării a 75-a de la fondarea Universității de Stat de Medicină și Farmacie „Nicolae Testemițanu” din Republica Moldova, Ziua internațională a științei pentru pace și dezvoltareIntroduction: Tissue Bank serves with corneal grafts a population of 4 million people, which offers over 40 corneas per year for transplant in the Republic of Moldova. The high quality of the cornea due to international standards (based on both the standards of the American Eye Bank Association and the European Eye Bank Association) described in all areas of operation, including donor selection and screening, corneal collection, storage, testing, evaluation and transport to transplant centers. A standard protocol is the National Corneal Transplant Protocol of 2019, which includes maintaining a comprehensive database, supported by ophthalmic surgeons in the Republic of Moldova, in which potential data are collected on all aspects of corneal donation and transplantation. The authors previously published data on the indications for the corneal transplantation and in this study we analyzed this database to evaluate the source and management of donor corneal tissue in the Republic of Moldova. • Purpose: Evaluation of the cornea transplant application rate in the Republic of Moldova the period of years 2013 - 2020. • Material and methods: Prospective study, electronic database of Tissue Bank for the period of 7 years 2013 - 2019 were analyzed for each year in terms of the donor number, the indications for the sampling, cause of death, interval from death to corneal preservation, storage methods, endothelial evaluation, bacteriological contamination and distribution. Results: During the study period, 306 corneas were taken from 153 donors (69,8% male, 30,2% female), with a mean age of donors 59,4 years (18,3 years SD) and between 18 and 91 years old. Donors were from forensic medicine (23,5%), public hospitals (67,6%) and multi-organ donors (7.1%). The most common causes of the donor deaths were the cardiovascular disease, trauma and the cerebrovascular diseases. The average storage time increased from 3,5 to 11,8 days, from when the culture medium replaced hypothermic storage. Invalidation of the corneas was in 22,8% of cases, of which were determined by serological infections ( HBsAg - positive, HCV positive, HIV / AIDS) - 15%, and biological contamination occurred in 7,8% of all donor corneas. The most common bacterial and fungal isolates were coagulase-negative staphylococci and Candida spp., respectively. A significant decrease in the contamination rate was identified during the study years. Overall, 77,2% of the corneal tissue taken was used for corneal transplantation (74,8% for penetrating keratoplasty, 2,1% for lamellar keratoplasty and 1.3% for unspecified transplants) and 22,8% were destroyed. The most common reasons for tissue settlement were biological contamination, serology, and endothelial cell integrity assessment. Conclusions: 1. The analysis of the Tisue Bank database provides valuable information about corneal transplantation in the Republic of Moldova. 2. The rate of corneal utilization increased during each year of the study, reflecting improvements in all areas of Tissue Bank operation, in particular, corneal storage and decreased microbiological contamination. 3. The Human Tissue and Cell Bank, founded is the main supplier of the corneas graft for the transplantation in the Republic of Moldova