Matične ćelije zubne pulpe i njihov potencijalni značaj u regenerativnoj medicini

To date, three types of dental stem cells have been isolated: Dental Pulp Stem Cells (DPSC), Stem Cells From Human Exfoliated Deciduous Teeth (SHED) and Immature Dental Pulp Stem Cells (IDPC). These dental stem cells are considered as mesenchymal stem cells. They reside within the perivascular niche of dental pulp. They are highly proliferative, clonogenic, multipotent and are similar to mesenchymal Bone Marrow Stem Cells (BMSC). Also, they have high plasticity and can be easy isolated. The expressions of the alkaline phosphatase gene, dentin matrix protein 1 and dentinsialophosphoprotein are verified in these cells. Analyses of gene expression patterns indicated several genes which encode extracellular matrix components, cell adhesion molecules, growth factors and transcription regulators, cell signaling, cell communication or cell metabolism. In both conditions, in vivo and in vitro, these cells have the ability to differentiate into odontoblasts, chondrocytes, osteoblasts, adipocytes, neurons, melanocytes, smooth and skeletal muscles and endothelial cells. In vivo, after implantation, they have shown potential to differentiate into dentin but also into tissues like bone, adipose or neural tissue. In general, DPSCs are considered to have antiinflammatory and immunomodulatory abilities. After being grafted into allogenic tissues these cells are ableto induce immunological tolerance. Immunosuppressive effect is shown through the ability to inhibit proliferation of T lymphocytes. Dental pulp stem cells open new perspectives in therapeutic use not only in dentin regeneration, periodontal tissues and skeletoarticular, tissues of craniofacial region but also in treatment of neurotrauma, autoimmune diseases, myocardial infarction, muscular dystrophy and connective tissue damages.Iz zubne pulpe su do danas izolovane tri populacije matičnih ćelija koje su označene kao matične ćelije zubne pulpe (engl. Dental Pulp Stem Cells, DPSC), matične ćelije iz eksfoliranih mlečnih zuba (engl. Stem Cells From Human Exfoliated Decidual Teeth, SHED) i nezrele matične ćelije zubne pulpe (engl. Immature Dental Pulp Stem Cells, IDPC). Sve matične ćelije zubne pulpe su ektomezenhimalnog porekla i lokalizovane su u perivaskularnoj niši. One se lako i efikasno izoluju, visoko su proliferativne, klonogene, multipotentne, ispoljavaju visok stepen plasticiteta i i slične su mezenhimalnim matičnim ćelijama koštane srži (BMSC). U njima je pokazana visoka ekspresija gena alkalne fosfataze, proteina 1 matriksa dentina i dentin-sijalofosfoproteina. Takođe, istaknuta je važnost u ovim ćelijama ekspresije više gena koji kodiraju sintezu komponenti ekstracelularnog matriksa, molekula ćelijske adhezije, faktora rasta, transkripcionih faktora, gena prenosa ćelijskih signala, ćelijske komunikacije i metabolizma. U uslovima in vitro ili in vivo ove ćelije mogu da se diferenciraju, s određenim međusobnim razlikama, u pravcu odontoblasta, hondrocita, osteoblasta, adipocita, neurona/glije, glatkih i skeletnih mišićnih ćelija, endotelnih ćelija i melanocita. U uslovima in vivo, nakon implantacije, pokazuju različit potencijal za formiranje dentina, ali i koštanog, masnog i nervnog tkiva. Generalno se smatra da DPSC imaju anti-inflamatorno dejstvo i ispoljavaju imunom-odulatorni efekat. Takođe, dovode do imunološke tolerancije ukoliko se implantiraju u alogena tkiva. Sposobnost inhibicije proliferacije T limfocita ukazuje na njihovo imunosupresivno dejstvo. Matične ćelije zubne pulpe otvorile su nove perspektive u terapijskoj primeni ovih ćelija ne samo u regeneraciji dentina, tkiva periodoncijuma i koštano-zglobnog tkiva kraniofacijalne regije, već i u lečenju neurotraume, autoimunskih oboljenja, infarkta miokarda, mišićne distrofije i oštećenja vezivnog tkiva

Familial hypercholesterolaemia in children and adolescents from 48 countries: a cross-sectional study

Background: Approximately 450 000 children are born with familial hypercholesterolaemia worldwide every year, yet only 2·1% of adults with familial hypercholesterolaemia were diagnosed before age 18 years via current diagnostic approaches, which are derived from observations in adults. We aimed to characterise children and adolescents with heterozygous familial hypercholesterolaemia (HeFH) and understand current approaches to the identification and management of familial hypercholesterolaemia to inform future public health strategies. Methods: For this cross-sectional study, we assessed children and adolescents younger than 18 years with a clinical or genetic diagnosis of HeFH at the time of entry into the Familial Hypercholesterolaemia Studies Collaboration (FHSC) registry between Oct 1, 2015, and Jan 31, 2021. Data in the registry were collected from 55 regional or national registries in 48 countries. Diagnoses relying on self-reported history of familial hypercholesterolaemia and suspected secondary hypercholesterolaemia were excluded from the registry; people with untreated LDL cholesterol (LDL-C) of at least 13·0 mmol/L were excluded from this study. Data were assessed overall and by WHO region, World Bank country income status, age, diagnostic criteria, and index-case status. The main outcome of this study was to assess current identification and management of children and adolescents with familial hypercholesterolaemia. Findings: Of 63 093 individuals in the FHSC registry, 11 848 (18·8%) were children or adolescents younger than 18 years with HeFH and were included in this study; 5756 (50·2%) of 11 476 included individuals were female and 5720 (49·8%) were male. Sex data were missing for 372 (3·1%) of 11 848 individuals. Median age at registry entry was 9·6 years (IQR 5·8-13·2). 10 099 (89·9%) of 11 235 included individuals had a final genetically confirmed diagnosis of familial hypercholesterolaemia and 1136 (10·1%) had a clinical diagnosis. Genetically confirmed diagnosis data or clinical diagnosis data were missing for 613 (5·2%) of 11 848 individuals. Genetic diagnosis was more common in children and adolescents from high-income countries (9427 [92·4%] of 10 202) than in children and adolescents from non-high-income countries (199 [48·0%] of 415). 3414 (31·6%) of 10 804 children or adolescents were index cases. Familial-hypercholesterolaemia-related physical signs, cardiovascular risk factors, and cardiovascular disease were uncommon, but were more common in non-high-income countries. 7557 (72·4%) of 10 428 included children or adolescents were not taking lipid-lowering medication (LLM) and had a median LDL-C of 5·00 mmol/L (IQR 4·05-6·08). Compared with genetic diagnosis, the use of unadapted clinical criteria intended for use in adults and reliant on more extreme phenotypes could result in 50-75% of children and adolescents with familial hypercholesterolaemia not being identified. Interpretation: Clinical characteristics observed in adults with familial hypercholesterolaemia are uncommon in children and adolescents with familial hypercholesterolaemia, hence detection in this age group relies on measurement of LDL-C and genetic confirmation. Where genetic testing is unavailable, increased availability and use of LDL-C measurements in the first few years of life could help reduce the current gap between prevalence and detection, enabling increased use of combination LLM to reach recommended LDL-C targets early in life

Association of BMI, lipid-lowering medication, and age with prevalence of type 2 diabetes in adults with heterozygous familial hypercholesterolaemia: a worldwide cross-sectional study

Background: Statins are the cornerstone treatment for patients with heterozygous familial hypercholesterolaemia but research suggests it could increase the risk of type 2 diabetes in the general population. A low prevalence of type 2 diabetes was reported in some familial hypercholesterolaemia cohorts, raising the question of whether these patients are protected against type 2 diabetes. Obesity is a well known risk factor for the development of type 2 diabetes. We aimed to investigate the associations of known key determinants of type 2 diabetes with its prevalence in people with heterozygous familial hypercholesterolaemia. Methods: This worldwide cross-sectional study used individual-level data from the EAS FHSC registry and included adults older than 18 years with a clinical or genetic diagnosis of heterozygous familial hypercholesterolaemia who had data available on age, BMI, and diabetes status. Those with known or suspected homozygous familial hypercholesterolaemia and type 1 diabetes were excluded. The main outcome was prevalence of type 2 diabetes overall and by WHO region, and in relation to obesity (BMI ≥30·0 kg/m2) and lipid-lowering medication as predictors. The study population was divided into 12 risk categories based on age (tertiles), obesity, and receiving statins, and the risk of type 2 diabetes was investigated using logistic regression. Findings: Among 46 683 adults with individual-level data in the FHSC registry, 24 784 with heterozygous familial hypercholesterolaemia were included in the analysis from 44 countries. 19 818 (80%) had a genetically confirmed diagnosis of heterozygous familial hypercholesterolaemia. Type 2 diabetes prevalence in the total population was 5·7% (1415 of 24 784), with 4·1% (817 of 19 818) in the genetically diagnosed cohort. Higher prevalence of type 2 diabetes was observed in the Eastern Mediterranean (58 [29·9%] of 194), South-East Asia and Western Pacific (214 [12·0%] of 1785), and the Americas (166 [8·5%] of 1955) than in Europe (excluding the Netherlands; 527 [8·0%] of 6579). Advancing age, a higher BMI category (obesity and overweight), and use of lipid-lowering medication were associated with a higher risk of type 2 diabetes, independent of sex and LDL cholesterol. Among the 12 risk categories, the probability of developing type 2 diabetes was higher in people in the highest risk category (aged 55–98 years, with obesity, and receiving statins; OR 74·42 [95% CI 47·04–117·73]) than in those in the lowest risk category (aged 18–38 years, without obesity, and not receiving statins). Those who did not have obesity, even if they were in the upper age tertile and receiving statins, had lower risk of type 2 diabetes (OR 24·42 [15·57–38·31]). The corresponding results in the genetically diagnosed cohort were OR 65·04 (40·67–104·02) for those with obesity in the highest risk category and OR 20·07 (12·73–31·65) for those without obesity. Interpretation: Adults with heterozygous familial hypercholesterolaemia in most WHO regions have a higher type 2 diabetes prevalence than in Europe. Obesity markedly increases the risk of diabetes associated with age and use of statins in these patients. Our results suggest that heterozygous familial hypercholesterolaemia does not protect against type 2 diabetes, hence managing obesity is essential to reduce type 2 diabetes in this patient population. Funding: Pfizer, Amgen, MSD, Sanofi-Aventis, Daiichi-Sankyo, and Regeneron

Comparative analysis of effectiveness parameters of laparoscopic and open appendectomy in children

Introduction: Acute appendicitis is one of the most common indications for urgent surgical intervention in children. An open appendectomy has been applying since the end of the XIX century, but for the last thirty years, laparoscopic appendectomy became an alternative. There is no consensus among surgeons, which method is suitable for pediatric patients. Aim: The aim of the research is to establish which of these two methods has therapeutic advantages in children, analyzing comparatively their operative and postoperative course in the treatment of acute appendicitis. Material and methods: The study includes 330 children with acute appendicitis. Patients were divided into 2 groups: operated by the method of open appendectomy (OA) and laparoscopic surgery (LA). Analyzed parameters were: methods of operation, age and sex of patients, intraoperative findings, duration of surgery, length of hospital stay, intraoperative and postoperative complications, preoperative levels of leukocytes, duration of antibiotic therapy and pain therapy. Results: In total of 330 children with acute appendicitis, 255 (77%) were operated by OA and 75 (23%) by LA. Catarrhal appendicitis is significantly more frequent intraoperative finding in LA (61%, p < 0.05), while phlegmonous is in OA (49%, p < 0.05). Mean operative time is 61 minutes in LA, compared with 38 minutes in OA. Mean length of hospital stay is 4.2 days after both operative techniques. Antibiotic treatment lasts as the hospital stay. Mean length of pain treatment is 1.3 days after LA and 1.5 days after OA. Conclusion: More children and especially with severe forms of acute appendicitis were treated by OA. Lesser number of patients and less severe forms of acute appendicitis were treated by LA. Total length of hospital stay, antibiotic and pain therapy are shorter in laparoscopic comparing to OA

The influence of obesity on laparoscopic appendectomy as an option choice in children

Introduction: Acute appendicitis is the most common indication for abdominal surgery in children. Despite the increase of obesity in children, little data exist that examine the impact of obesity on surgical outcomes in children. Laparoscopic appendectomy has become more popular and it is used more often in treating obese children. Aim: The aim of this study is to evaluate the impact of obesity on operative and postoperative recovery of children with acute appendicitis, treated with open and laparoscopic method, and to determine which of these techniques is superior. Material and methods: We analyzed medical reports of children treated for acute appendicitis. This study included 330 patients. We used age percentile tables for weight and defined child obesity as weight at or above 90th percentile. Both obese and non-obese patients were divided into two groups: patients who underwent open surgery (OA), and those who underwent laparoscopic surgery (LA). We analyzed gender and age of our patients, their operative time, hospital stay, leukocytes count and complication rate. Results: The study includes 330 patients treated for acute appendicitis. There were 69 obese children (21%). The open technique was more often used in treating both obese (84%) and non-obese patients (76%). Obesity has been shown to increase operative time and hospital stay with no statistical significance, but with significantly higher incidence of complications after OA (67%) than after LA (33%). In the group of non-obese children, there was also significantly higher incidence of complications after OA (91%) than after LA (9%). Conclusion: According to observed parameters, obesity in our patients has no significant impact on the outcome of appendectomy in children. Concerning that there is no significantly high incidence of total number of complications referring to OA, laparoscopic technique should be recommended in treating all stages of acute appendicitis, both in obese and non-obese children

Elongation index derivative as a new mechanobiological parameter in Diabetes mellitus patients

Erythrocyte deformability plays pivotal role in blood circulation in Diabetes Mellitus (DM) patients [1,2]. Comparative examination of erythrocyte mechanical properties in DM patients and healthy donors was conducted. We mainly focused on calculating the Elongation Index (EI) which was obtained from diffraction images, measured by using ektacytometry. No statistically significant difference in EI values on specific physiologically relevant pressures (3Pa or at half maximal value of deformability curve) between DM and control was found, and we introduced the first derivative of deformability curve (dEI/dSS) as a measure for assessing erythrocyte response to deformation (SS denotes shear stress in Pa). Significantly higher values of dEI/dSS at the half maximum value of the deformability curve were noted in healthy people comparing to DM patients, indicating a slower erythrocyte response to shear stress in DM. Scatter plot analysis exhibited a linear decline in the dEI/dSS ratio with increasing shear stress, suggesting reduced erythrocyte responsiveness to higher shear stress, particularly evident in DM. Although preliminary, this investigation indicates that dEI/dSS may offer valuable insights into the hemorheological alterations of DM and enhance comprehension of erythrocyte mechanobiological behavior under different shear stress conditions. Correlations between the proposed measure of erythrocyte mechanical properties and established clinical indicators of DM and its complications such as serum cholesterol and creatinine, will be further explored to gain understanding of the potential application of dEI/dSS for improved diagnosis and/or patient care

Elongation index derivative: a promising hemorheological parameter in Diabetes mellitus erythrocyte analysis

Aim. Diabetes mellitus (DM) is greatly impacted by alterations in the deformability of the erythrocyte membrane [1,2]. Moreover, there is strong evidence suggesting that hemorheological determinants, such as the calculated Elongation Index (EI) from deformability measurements, play a crucial role in the development of diabetic organ complications, notably retinopathy, nephropathy, and neuropathy [3,4]. We measured the deformability of red blood cells in patients with DM by analyzing the erythrocyte EI and its change in response to shear stress (SS) (dEI/dSS) at specific physiologically relevant points, to be used as a new hemorheological parameter. Materials and methods. For all samples tested, DM erythrocytes (n=13) and age- and sex-matched controls (n=11), we used a RheoScan-D300 ektacytometer (RheoMeditech Inc., Korea). Results and discussion. Although statistically insignificant differences were found in EI values between the DM and control groups, the study uncovered specific data points on the deformability curve and highlighted the potential of the first derivative of the deformability curve (dEI/dSS) as an indicator of red blood cell response to deformation. Notably, statistically significant variations in dEI/dSS at the half maximum value of the deformability curve were observed, suggesting a delayed red blood cell response to shear stress in individuals with DM. Moreover, scatter plot analysis revealed a linear decline in the dEI/dSS index with increasing shear stress, indicating a reduced red blood cell responsiveness to higher shear stress levels, particularly prominent in DM patients. Conclusion. Despite being a pilot study, understanding the relationship between dEI/dSS and DM pathology could significantly enhance our knowledge of how red blood cells respond the different shear stress levels, offering crucial insights into the hemorheological implications and contributing to a comprehensive understanding of their mechanobiological behavior [5]

Elongation index derivative as a potential hemorheological parameter in a diffraction analysis of diabetes mellitus patients’ erythrocytes

Comparative analysis of erythrocyte deformability in individuals with Diabetes Mellitus (DM) and healthy individuals (Control) was represented, focusing on the Elongation index (EI) calculation based on diffraction images. While no statistically significant differences in EI values were observed between the groups, we determined specific points along the deformability curve and revealed the first derivative of deformability curve (dEI / dSS) as a potential metric for quantifying erythrocyte response to deformation, where SS represents shear stress in Pa (Pascal). Statistically significant differences in dEI / dSS at the half maximum value of the deformability curve were identified, suggesting a slower erythrocyte response to shear stress in DM patients. Scatter plot analysis demonstrated a linear declining trend in dEI / dSS index with an increase in shear stress, indicating decaying erythrocyte responsiveness to higher shear stress values, particularly pronounced in DM patients. Although pilot, this study suggests that dEI / dSS can provide valuable insights into the hemorheological aspects of DM pathology and contribute to a comprehensive understanding of erythrocyte mechanobiological behavior in response to varying shear stress levels. Correlations between the proposed measure of RBC mechanical properties and established clinical markers of DM and its complications (serum cholesterol, creatinine, and urea level) are obtained to get preliminary insight into dEI / dSS application for better diagnosis and/or patient management