The effects of low level laser and electromagnetic field on morphology, proliferation, differentiation and aging of human adipose tissue-derived mesenchymal stem cells

Poslednjih nekoliko godina otkriveno je da laser niske snage (engl. Low Level Laser, LLL) i elektromagnetno polje (engl. Electromagnetic Field, EMF) utiču na brojne biološke procese, rast i proliferaciju ćelija, a naročito na matične ćelije. Cilj ove studije bio je ispitivanje efekata LLL-a i EMF-a na karakteristike i potencijal humanih mezenhimalnih matičnih ćelija iz masnog tkiva (engl. human Adipose TissueDerived Mesenchymal Stem Cells, hAT-MSCs) i na njihovo prihvatanje nakon eventualne terapijske aplikacije. hAT-MSCs su izolovane iz masnog tkiva 12 osoba starosti 18- 65 godina. LLL tretman se primenjivao tokom 7 dana, jednom dnevno u različitom trajanju, u nekoliko različitih doza od 1 J/cm2, 3 J/cm2, 5 J/cm2 and 10 J/cm2, sa talasnom dužinom od 808 nm, snagom od 200 mW i gustinom energije od 0.2 W/cm2. EMF tretman se, takođe, primenjivao u trajanju od 7 dana, jednom dnevno u različitom trajanju, preko magnetnih jastučića, frekvencije od 50 Hz i različitim intenzitetom od 1 mT, 3 mT, 5 mT i 10 mT. Kontrolna grupa ćelija kultivisana je u istim uslovima, ali bez tretmana fizikalnim agensima. Sedam dana nakon tretmana utvrđene su pojedinačne optimalne doze LLL-a i EMF-a i ispitana je vijabilnost, morfologija, proliferacija, diferencijacija, starenje i oksidativni stres tretiranih ćelija. Rezultati su pokazali da je nakon 7 dana broj hAT-MSCs tretiranih EMF-om bio znatno veći od broja kontrolnih, netretiranih ćelija, hAT-MSCs tretirane LLL-om su bile brojnije od onih tretiranih EMF-om, a da su hAT-MSCs koje su stimulisane kombinovano LLL-om i EMF-om bile najbrojnije. LLL i/ili EMF tretman nije značajno uticao na vijabilnost tretiranih ćelija. Isti pozitivni efekti su registrovani i kada se ispitala diferencijacija. LLL i/ili EMF tretman nije uticao na starenje, niti je povećao oksidativni stres ćelija. Takođe su primećene promene u morfologiji ćelija u smislu povećanja površine ćelija i fraktalne dimenzije u grupama koje su tretirane EMF-om i kombinacijom EMF-a i LLL-a. U zaključku, LLL i/ili EMF tretman je ubrzao proliferaciju i poboljšao diferencijaciju hAT-MSCs, bez ugrožavanja njihove održivosti, bez uticaja na starenje i oksidativni stres, te predstavlja terapijski potencijal u ćelijskom inženjeringu.In recent years, electromagnetic field (EMF) and low-level laser (LLL) have been found to affect various biological processes, the growth and proliferation of cells, and especially that of stem cells. The aim of this study was to investigate the effects of EMF and LLL on the characteristics and abilities of human adipose tissue-derived mesenchymal stem cells (hATMSCs) and thus to examine the impact of these therapeutic physical modalities on stem cell engraftment. hAT-MSCs were isolated from subcutaneous adipose tissue of 12 persons ranging in age from 18 to 65 years. LLL was applied also for 7 days, for varying periods of time, at different radiation energies of 1 J/cm2, 3 J/cm2, 5 J/cm2 and 10 J/cm2, with a wavelength of 808 nm, power output of 200 mW, and power density of 0.2 W/cm2. EMF was applied for a period of 7 days, once a day for varying periods of time, via a magnetic cushion surface at a frequency of 50 Hz and an different intensity of 1 mT, 3 mT, 5 mT and 10 mT. Nonexposed cells (control) were cultivated under the same culture conditions. Seven days after treatment, optimum doses of LLL and EMF are determined, and the cells were examined for cell viability, morphology, proliferation, differentiation, aging and oxidative stress. We found that after 7 days, the number of EMF-treated hATMSCs was significantly higher than the number of the untreated cells, LLL-treated hAT-MSCs were more numerous than EMFtreated cells, and hAT-MSCs that were treated with the combination of EMF and LLL were the most numerous. EMF and/or LLL treatment did not significantly affect hAT-MSC viability by itself. The same positive effects were registered also when differentiation was examined. LLL and/or EMF treatment did not affect aging and oxidative stress of hAT-MSCs. Changes in cell morphology were also observed, in terms of an increase in cell surface area and fractal dimension in hAT-MSCs treated with EMF and the combination of EMF and LLL. In conclusion, LLL and/or EMF treatment accelerated the proliferation and enhance differentiation of hAT-MSCs without compromising their viability, without affecting aging and oxidative stress, and therefore, they may be used in stem cell tissue engineering

Corneal Stem Cells as a Source of Regenerative Cell-Based Therapy

© 2020 Jasmin S. Nurković et al. In the past few years, intensive research has focused on corneal stem cells as an unlimited source for cell-based therapy in regenerative ophthalmology. Today, it is known that the cornea has at least two types of stem cells: limbal epithelial stem cells (LESCs) and corneal stromal stem cells (CSSCs). LESCs are used for regeneration of corneal surface, while CSSCs are used for regeneration of corneal stroma. Until now, various approaches and methods for isolation of LESCs and CSSCs and their successful transplantation have been described and tested in several preclinical studies and clinical trials. This review describes in detail phenotypic characteristics of LESCs and CSSCs and discusses their therapeutic potential in corneal regeneration. Since efficient and safe corneal stem cell-based therapy is still a challenging issue that requires continuous cooperation between researchers, clinicians, and patients, this review addresses the important limitations and suggests possible strategies for improvement of corneal stem cell-based therapy

Measurement of Bone Mineral Density in Children with Cerebral Palsy from an Ethical Issue to a Diagnostic Necessity

© 2020 Jasmin S. Nurković et al. Introduction. Due to concerns about cumulative radiation exposure in the pediatric population, it is not standard practice to perform dual-energy X-ray absorptiometry (DXA) analysis in the diagnostic process of musculoskeletal disorders, such as cerebral palsy (CP). This study aimed to evaluate the bone mineral density (BMD) in children with CP and the ethical justification of applying DXA analysis in these children. Material and Methods. In this monocentric retrospective analysis, data were collected from children and adolescents with CP who were treated for a primary illness for three years. A clinical examination, which included a DXA analysis, recommended by the multidisciplinary team, was performed. After applying inclusion and exclusion criteria, 60 scans remained for statistical analysis. BMD and Z-scores for the lumbar spine (LS), and hip right and left femoral neck (RFN and LFN, respectively), and total hip (TH) were recorded. Results. The average age of children with CP when DXA analysis was first performed was about 7 years. The BMD (mean±SD) at LS (LS-BMD) of all patients was 0.612±0.12, at RFN 0.555±0.11, at LFN 0.572±0.1, and at TH (TH-BMD) 0.581±0.13. The values of the Z-score (mean±SD) at LS of all patients were-2.5±0.22, at RFN-2.2±0.21, at LFN-2.25 (SD=0.2), and at TH-2.3 (SD=0.23). There was no statistical significance between age and gender; however, BMI, walking ability, fracture history, and pattern of CP had a significant impact on BMD and Z-score values of these children. Conclusion. The results of our study clearly indicate that children with CP have a higher risk of low BMD, osteoporosis, and bone fractures, which makes it ethically justifiable to perform the DXA analysis in these children

Mezenhimalne matične ćelije iz masnog tkiva - izolacija, kultivacija i ciljana diferencijacija

Stem cells are non-specialized body cells at a very early stage of development, which under normal conditions in a given tissue can differentiate into different types of functionally specialized mature cells. Mesenchymal stem cells (MSCs) are attractive candidates for clinical use in the reconstruction of damaged tissue, especially as it can be isolated from various sources and reproduced, and their use does not carry any ethical problems. Isolation methods of MSCs from adipose tissue are based on enzymatic degradation of the obtained materials. Terms for cultivation of mesenchymal stem cells are temperature of 37 °C and the partial pressure of CO2 5%. MSCs are cultured in the medium, often in a-MEM medium with 10% or 20% of fetal calf serum. Under these conditions of cultivation adherent cells form colonies in 7-14 days. MSCs are multipotent and able to differentiate in vitro conditions into Mesodermal differentiation, forming osteoblasts, chondrocytes and adipocytes. However, they can be differentiated into cells of ectodermal origin (such as neurons) and cells of endodermal origin (eg, ß-cells of the pancreatic islets of Langerhans and hepatocytes). The Laboratory for stem cell of the Department of Biomedical Sciences at the State University of Novi Pazar conducts research of MSCs originating from human adipose tissue. In collaboration with surgeons of Health Center of Novi Pazar, and respecting the norms of the Ethics Committee of both institutions in the period from 01.07.2011. to 01.07.2012. were obtained 22 samples of human subcutaneous adipose tissue of patients aged 18 to 65 years. 15 samples successfully completed the process of isolation and cultivation, and 8 induces Mesodermal differentiation.Matične ćelije su nespecijalizovane ćelije organizma u vrlo ranom stadijumu razvića, koje u normalnim uslovima u datom tkivu mogu da se diferenciraju u različite tipove funkcionalno specijalizovanih zrelih ćelija. Mezenhimalne matične ćelije (MSCs) su atraktivni kandidati za kliničku primenu u obnavljanju oštećenih tkiva, pogotovu što se mogu izolovati iz više izvora i umnožavati, a njihova primena ne nosi nikakve etičke probleme. Metode izolacije MSCs iz masnog tkiva zasnivaju se na enzimatskom razlaganju dobijenog materijala. Uslovi kultivacije mezenhimalnih matičnih ćelija su temperatura od 37°C i parcijalni pritisak CO2 5%. MSCs se gaje u medijumu, najčešće u a-MEM medijumu sa 10% ili 20% fetalnim telećim serumom. Pod tim uslovima kultivisanja za 7-14 dana adherisane ćelije formiraju kolonije. MSCs su multipotentne i sposobne da se diferenciraju u uslovima in vitro u mezodermalnom pravcu, stvarajući osteoblaste, hondrocite i adipocite. Međutim, one mogu da se diferenciraju i u ćelije ektodermalnog (npr. neurone) i ćelije endodermalnog porekla (npr. ß-ćelije Langerhansovih ostrvaca pankreasa i hepatocite). U Laboratoriji za matične ćelije Departmana za biomedicinske nauke Državnog univerziteta u Novom Pazaru sprovodi se istraživanje MSCs poreklom iz humanog masnog tkiva. U saradnji sa hirurzima Zdravstvenog centra Novi Pazar, a uz poštovanje normi Etičkog komiteta obe ustanove u periodu od 01.07.2011. do 01.07.2012. godine dobijeno je 22 uzorka potkožnog humanog masnog tkiva od ispitanika starosti od 18 do 65 godina. Od 15 uzoraka uspešno je završen proces izolacije i kultivacije, a od 8 i ciljane mezodermalne diferencijacije

COVID-19 DEATH in NOVI PAZAR - sERBIAN BERGAMO

Serbia was one of the countries in Europe and the world that were most affected by the coronavirus disease (COVID-19) pandemic. City Novi Pazar was the greatest coronavirus hotspot in Europe on July 1st, 2020, due to several hundred infected people. Even though united data was published at the state level, there is no data by region or city, so the interpretation of the COVID-19 epidemic in Serbia at the regional level is difficult. Different levels of health care and health education of citizens and the degree of respect for the proposed epidemiological measures have led to significant differences in the number of tests, a large number of infected, and several deaths by regions and cities. Insufficiently precise and up-to-date keeping of records and statistical data on COVID-19 at the state and local level also complicates the pandemic's scientific and epidemiological analysis. Novi Pazar is a city in southwestern Serbia with a population of 100 000. It is similar in population to the city of Bergamo, in northern Italy in the Lombardy region. As of 1st of July, 2020, Novi Pazar had 300% higher mortality per 100 000 population compared to the same month last year, and almost ten times higher mortality than the rest of Serbia

The effects of low level laser and electromagnetic field on morphology, proliferation, differentiation and aging of human adipose tissue-derived mesenchymal stem cells

Poslednjih nekoliko godina otkriveno je da laser niske snage (engl. Low Level Laser, LLL) i elektromagnetno polje (engl. Electromagnetic Field, EMF) utiču na brojne biološke procese, rast i proliferaciju ćelija, a naročito na matične ćelije. Cilj ove studije bio je ispitivanje efekata LLL-a i EMF-a na karakteristike i potencijal humanih mezenhimalnih matičnih ćelija iz masnog tkiva (engl. human Adipose TissueDerived Mesenchymal Stem Cells, hAT-MSCs) i na njihovo prihvatanje nakon eventualne terapijske aplikacije. hAT-MSCs su izolovane iz masnog tkiva 12 osoba starosti 18- 65 godina. LLL tretman se primenjivao tokom 7 dana, jednom dnevno u različitom trajanju, u nekoliko različitih doza od 1 J/cm2, 3 J/cm2, 5 J/cm2 and 10 J/cm2, sa talasnom dužinom od 808 nm, snagom od 200 mW i gustinom energije od 0.2 W/cm2. EMF tretman se, takođe, primenjivao u trajanju od 7 dana, jednom dnevno u različitom trajanju, preko magnetnih jastučića, frekvencije od 50 Hz i različitim intenzitetom od 1 mT, 3 mT, 5 mT i 10 mT. Kontrolna grupa ćelija kultivisana je u istim uslovima, ali bez tretmana fizikalnim agensima. Sedam dana nakon tretmana utvrđene su pojedinačne optimalne doze LLL-a i EMF-a i ispitana je vijabilnost, morfologija, proliferacija, diferencijacija, starenje i oksidativni stres tretiranih ćelija. Rezultati su pokazali da je nakon 7 dana broj hAT-MSCs tretiranih EMF-om bio znatno veći od broja kontrolnih, netretiranih ćelija, hAT-MSCs tretirane LLL-om su bile brojnije od onih tretiranih EMF-om, a da su hAT-MSCs koje su stimulisane kombinovano LLL-om i EMF-om bile najbrojnije. LLL i/ili EMF tretman nije značajno uticao na vijabilnost tretiranih ćelija. Isti pozitivni efekti su registrovani i kada se ispitala diferencijacija. LLL i/ili EMF tretman nije uticao na starenje, niti je povećao oksidativni stres ćelija. Takođe su primećene promene u morfologiji ćelija u smislu povećanja površine ćelija i fraktalne dimenzije u grupama koje su tretirane EMF-om i kombinacijom EMF-a i LLL-a. U zaključku, LLL i/ili EMF tretman je ubrzao proliferaciju i poboljšao diferencijaciju hAT-MSCs, bez ugrožavanja njihove održivosti, bez uticaja na starenje i oksidativni stres, te predstavlja terapijski potencijal u ćelijskom inženjeringu.In recent years, electromagnetic field (EMF) and low-level laser (LLL) have been found to affect various biological processes, the growth and proliferation of cells, and especially that of stem cells. The aim of this study was to investigate the effects of EMF and LLL on the characteristics and abilities of human adipose tissue-derived mesenchymal stem cells (hATMSCs) and thus to examine the impact of these therapeutic physical modalities on stem cell engraftment. hAT-MSCs were isolated from subcutaneous adipose tissue of 12 persons ranging in age from 18 to 65 years. LLL was applied also for 7 days, for varying periods of time, at different radiation energies of 1 J/cm2, 3 J/cm2, 5 J/cm2 and 10 J/cm2, with a wavelength of 808 nm, power output of 200 mW, and power density of 0.2 W/cm2. EMF was applied for a period of 7 days, once a day for varying periods of time, via a magnetic cushion surface at a frequency of 50 Hz and an different intensity of 1 mT, 3 mT, 5 mT and 10 mT. Nonexposed cells (control) were cultivated under the same culture conditions. Seven days after treatment, optimum doses of LLL and EMF are determined, and the cells were examined for cell viability, morphology, proliferation, differentiation, aging and oxidative stress. We found that after 7 days, the number of EMF-treated hATMSCs was significantly higher than the number of the untreated cells, LLL-treated hAT-MSCs were more numerous than EMFtreated cells, and hAT-MSCs that were treated with the combination of EMF and LLL were the most numerous. EMF and/or LLL treatment did not significantly affect hAT-MSC viability by itself. The same positive effects were registered also when differentiation was examined. LLL and/or EMF treatment did not affect aging and oxidative stress of hAT-MSCs. Changes in cell morphology were also observed, in terms of an increase in cell surface area and fractal dimension in hAT-MSCs treated with EMF and the combination of EMF and LLL. In conclusion, LLL and/or EMF treatment accelerated the proliferation and enhance differentiation of hAT-MSCs without compromising their viability, without affecting aging and oxidative stress, and therefore, they may be used in stem cell tissue engineering

Corneal Stem Cells as a Source of Regenerative Cell-Based Therapy

In the past few years, intensive research has focused on corneal stem cells as an unlimited source for cell-based therapy in regenerative ophthalmology. Today, it is known that the cornea has at least two types of stem cells: limbal epithelial stem cells (LESCs) and corneal stromal stem cells (CSSCs). LESCs are used for regeneration of corneal surface, while CSSCs are used for regeneration of corneal stroma. Until now, various approaches and methods for isolation of LESCs and CSSCs and their successful transplantation have been described and tested in several preclinical studies and clinical trials. This review describes in detail phenotypic characteristics of LESCs and CSSCs and discusses their therapeutic potential in corneal regeneration. Since efficient and safe corneal stem cell-based therapy is still a challenging issue that requires continuous cooperation between researchers, clinicians, and patients, this review addresses the important limitations and suggests possible strategies for improvement of corneal stem cell-based therapy

Concise review: Therapeutic potential of mesenchymal stem cells for the treatment of acute liver failure and cirrhosis

© 2014 AlphaMed Press. Currently, the most effective therapy for acute liver failure and advanced cirrhosis is liver transplantation. However, this procedure has several limitations, including lack of donors, surgical complications, immunological suppression, and high medical costs. The alternative approaches that circumvent the use of a whole liver, such as stem cell transplantation, have been suggested as an effective alternate therapy for hepatic diseases. Mesenchymal stem cells (MSCs), also known as multipotent mesenchymal stromal cells, are self-renewing cells that can be found in almost all postnatal organs and tissues, including liver. During the past decade, great progress has been made in the field of MSC-dependent liver regeneration and immunomodulation. Because of their potential for differentiation into hepatocytes as well as their immunomodulatory characteristics, MSCs are considered as promising therapeutic agents for the therapy of acute liver failure and cirrhosis. In this concise review, we have summarized therapeutic potential of MSCs in the treatment of acute liver failure and cirrhosis, emphasizing their regenerative and immunomodulatory characteristics after engraftment in the liver. We have also presented several outstanding problems including conflicting data regarding MSCs engraftment in the liver and unwanted mesenchymal lineage differentiation in vivo which limits MSC therapy as a mainstream treatment approach for liver regeneration. It can be concluded that efficient and safe MSC-based therapy for acute and chronic liver failure remains a challenging issue that requires more investigation and continuous cooperation between clinicians, researchers, and patients

Measurement of Bone Mineral Density in Children with Cerebral Palsy from an Ethical Issue to a Diagnostic Necessity

Introduction. Due to concerns about cumulative radiation exposure in the pediatric population, it is not standard practice to perform dual-energy X-ray absorptiometry (DXA) analysis in the diagnostic process of musculoskeletal disorders, such as cerebral palsy (CP). This study aimed to evaluate the bone mineral density (BMD) in children with CP and the ethical justification of applying DXA analysis in these children. Material and Methods. In this monocentric retrospective analysis, data were collected from children and adolescents with CP who were treated for a primary illness for three years. A clinical examination, which included a DXA analysis, recommended by the multidisciplinary team, was performed. After applying inclusion and exclusion criteria, 60 scans remained for statistical analysis. BMD and Z-scores for the lumbar spine (LS), and hip right and left femoral neck (RFN and LFN, respectively), and total hip (TH) were recorded. Results. The average age of children with CP when DXA analysis was first performed was about 7 years. The BMD (mean±SD) at LS (LS-BMD) of all patients was 0.612±0.12, at RFN 0.555±0.11, at LFN 0.572±0.1, and at TH (TH-BMD) 0.581±0.13. The values of the Z-score (mean±SD) at LS of all patients were −2.5±0.22, at RFN −2.2±0.21, at LFN -2.25 (SD=0.2), and at TH -2.3 (SD=0.23). There was no statistical significance between age and gender; however, BMI, walking ability, fracture history, and pattern of CP had a significant impact on BMD and Z-score values of these children. Conclusion. The results of our study clearly indicate that children with CP have a higher risk of low BMD, osteoporosis, and bone fractures, which makes it ethically justifiable to perform the DXA analysis in these children

Combined effects of electromagnetic field and low-level laser increase proliferation and alter the morphology of human adipose tissue-derived mesenchymal stem cells

In recent years, electromagnetic field (EMF) and low-level laser (LLL) have been found to affect various biological processes, the growth and proliferation of cells, and especially that of stem cells. The aim of this study was to investigate the effects of EMF and LLL on proliferation of human adipose tissue-derived mesenchymal stem cells (hAT-MSCs) and thus to examine the impact of these therapeutic physical modalities on stem cell engraftment. hAT-MSCs were isolated from subcutaneous adipose tissue of six persons ranging in age from 21 to 56 years. EMF was applied for a period of 7 days, once a day for 30 min, via a magnetic cushion surface at a frequency of 50 Hz and an intensity of 3 mT. LLL was applied also for 7 days, once a day for 5 min, at radiation energies of 3 J/cm(2), with a wavelength of 808 nm, power output of 200 mW, and power density of 0.2 W/cm(2). Nonexposed cells (control) were cultivated under the same culture conditions. Seven days after treatment, the cells were examined for cell viability, proliferation, and morphology. We found that after 7 days, the number of EMF-treated hAT-MSCs was significantly higher than the number of the untreated cells, LLL-treated hAT-MSCs were more numerous than EMF-treated cells, and hAT-MSCs that were treated with the combination of EMF and LLL were the most numerous. EMF and/or LLL treatment did not significantly affect hAT-MSC viability by itself. Changes in cell morphology were also observed, in terms of an increase in cell surface area and fractal dimension in hAT-MSCs treated with EMF and the combination of EMF and LLL. In conclusion, EMF and/or LLL treatment accelerated the proliferation of hAT-MSCs without compromising their viability, and therefore, they may be used in stem cell tissue engineering