Vladimir Petrović

Craniofacial bone tissue engineering

There are numerous conditions, such as trauma, cancer, congenital malformations, and progressive deforming skeletal diseases, that can compromise the function and architectonics of bones of craniofacial region. The need to develop new approaches for treatment of these disorders arises from the fact that conventional therapeutic strategies face many obstacles and limitations. The use of tissue engineering in regeneration of craniofacial bone structures is a very promising possibility and a great challenge for researchers and practitioners. Developments in stem cell biology and engineering have led to the discovery of different stem cell populations and biodegradable materials with suitable properties. This review summarizes the current achievements in tissue engineering of craniofacial bone, temporomandibular joint, and periodontal ligament.

Dental Tissue — New Source for Stem Cells

Stem cells have been isolated from many tissues and organs, including dental tissue. Five types of dental stem cells have been established: dental pulp stem cells, stem cells from exfoliated deciduous teeth, stem cells from apical papilla, periodontal ligament stem cells, and dental follicle progenitor cells. The main characteristics of dental stem cells are their potential for multilineage differentiation and self-renewal capacity. Dental stem cells can differentiate into odontoblasts, adipocytes, neuronal-like cells, glial cells, osteoblasts, chondrocytes, melanocytes, myotubes, and endothelial cells. Possible application of these cells in various fields of medicine makes them good candidates for future research as a new, powerful tool for therapy. Although the possible use of these cells in therapeutic purposes and tooth tissue engineering is still in the beginning stages, the results are promising. The efforts made in the research of dental stem cells have clarified many mechanisms underlying the biological processes in which these cells are involved. This review will focus on the new findings in the field of dental stem cell research and on their potential use in the therapy of various disorders.

Tissue Engineering of the Urinary Bladder: Current Concepts and Future Perspectives

There are many conditions that can affect the normal structure of the urinary bladder wall and lead to the inadequate evacuation of urine or even disable urine excretion. In these cases, the essential task is to restore the function of the urinary bladder, most often through surgical intervention. Some of the disorders, such as bladder acontractility, bladder cancer, and inflammatory disease, represent a great challenge in practice due to the number of complications that can occur after the intervention and due to frequent relapses. The use of tissue engineering strategies that include the use of stem cells and artificially created scaffolds could give solutions for treatment of many disorders of the urinary bladder and transplantation therapies in the future. Although the research in this field is still in its infancy, there are some promising results that raise hope that the tissue engineering approach could offer long-term solutions for many issues in regenerative urology. This review summarizes the current achievements and perspectives in the use of stem cells and tissue engineering techniques in the field of urinary bladder regeneration.

Morphometric Analysis of Connective Tissue Sheaths of Sural Nerve in Diabetic and Nondiabetic Patients

One of the most common complications of diabetes mellitus is diabetic neuropathy. It may be provoked by metabolic and/or vascular factors, and depending on duration of disease, various layers of nerve may be affected. Our aim was to investigate influence of diabetes on the epineurial, perineurial, and endoneurial connective tissue sheaths. The study included 15 samples of sural nerve divided into three groups: diabetic group, peripheral vascular disease group, and control group. After morphological analysis, morphometric parameters were determined for each case using ImageJ software. Compared to the control group, the diabetic cases had significantly higher perineurial index (P < 0.05) and endoneurial connective tissue percentage (P < 0.01). The diabetic group showed significantly higher epineurial area (P < 0.01), as well as percentage of endoneurial connective tissue (P < 0.01), in relation to the peripheral vascular disease group. It is obvious that hyperglycemia and ischemia present in diabetes lead to substantial changes in connective tissue sheaths of nerve, particularly in peri- and endoneurium. Perineurial thickening and significant endoneurial fibrosis may impair the balance of endoneurial homeostasis and regenerative ability of the nerve fibers. Future investigations should focus on studying the components of extracellular matrix of connective tissue sheaths in diabetic nerves.

Stem Cell-Based Dental Tissue Engineering

The development of biological and biomaterial sciences profiled tissue engineering as a new and powerful tool for biological replacement of organs. The combination of stem cells and suitable scaffolds is widely used in experiments today, in order to achieve partial or whole organ regeneration. This review focuses on the use of tissue engineering strategies in tooth regeneration, using stem cells and stem cells/scaffold constructs. Although whole tooth regeneration is still not possible, there are promising results. However, to achieve this goal, it is important to understand and further explore the mechanisms underlying tooth development. Only then will we be able to mimic the natural processes with the use of stem cells and tissue engineering techniques.

Analysis of Fascicular Structure and Connective Tissue Sheaths in Sural Nerve during Aging

Summary The aim of our study was to analyze the changes of connective tissue sheaths of epi-, peri- and endoneurium of sural nerve during aging. The study was conducted on sural nerve samples of 10 cases aged 9-80 years. The specimens were embedded in paraffin using standard procedures, after which 5-μm-thick cross-sections of nerve trunks were made and stained using Masson’s trichrome staining. After morphological analysis of fascicular structure and connective sheaths of the nerve, morphometric analysis was conducted using the software for digital image analysis “ImageJ”. Each investigated case was analyzed for total neural, epineurial and fascicular cross-section area, mean values of perineurial index, volume density of myelinated axons and of endoneurial content. To test the difference in mean values for statistical significance we used the Student’s T-test for small independent sample. The number of fascicles was 5-13, while the majority of the nerves had less than 10 fascicles. Fascicular structure, which included the number of fascicles and epifascicular/fascicular area ratio, did not show significant changes during aging. Perineurial thickness /fascicle size ratio statistically significantly increased in the older investigated group (p<0.05). Myelinated fibres were of smaller diameter, with more irregular form and markedly less frequent in older cases. Quantitative analysis showed statistically significant decrease in volume density of myelinated fibres in the older group. As results of applied investigation methods we found thickening of perineurial sheath of sural nerve during aging, as well as endoneurial fibrosis. Future investigations of age-related changes should focus on analysis of the components of extracellular matrix within perineurium and endoneurium.

Role of stem cells in kidney repair

End-stage renal disease and acute renal failure are the most important issues of practical and clinical nephrology, bearing in mind their high mortality rate, solely symptomatic treatment, and overall economic impact on society. The advances in stem cell biology opened the door for the new era in treatment of many disorders, including renal, offering new therapeutical solutions. Findings suggesting that the adult kidney contains stem cells and that stem cells from bone marrow have potential to differentiate into renal cells focused research on the possible application of these cells in therapy of kidney disorders. The other promising candidates for stem cell therapy for the kidney are embryonic stem cells and amniotic fluid-derived stem cells. This article focuses on the characteristics and possible application of these types of stem cells.

Performance of the new clinical case definitions of pertussis in pertussis suspected infection and other diagnoses similar to pertussis

Background In an effort to improve the pertussis diagnosis, the Global Pertussis Initiative (GPI) proposed an algorithm of the signs/symptoms of pertussis for three age groups: 0–3 months, 4 months to 9 years, and ≥10 years of age. Methods We evaluated the accuracy of the clinical case definitions for pertussis proposed by the GPI using laboratory-confirmed pertussis as a reference standard for four groups: clinically suspected pertussis without comorbidity; asthma exacerbation; allergic constitution, and other diagnoses (bronchitis, bronchiolitis, laryngitis, and tracheitis). We included only patients who fulfilled one or more criteria of clinical case definitions for the age groups (0–3 months, 4 months–9 years, and ≥10 years of age). The data for this prospective epidemiological study were collected between 1st January 2013–31st December 2016 at the outpatients and inpatients health care settings in the South Bačka District of Autonomous Province of Vojvodina, Serbia. We evaluated accuracy of the certain sign and symptom combinations of GPI case definitions based on their sensitivity, specificity, and likelihood ratios. Results A total of 1043 participants were included, with 306 (29.3%) laboratory-confirmed pertussis cases. In patients aged 0–3 months, whoop and apnoea associated with laboratory confirmation of pertussis. In patients aged 4 months-9 years with a pertussis suspicion infection or with one of the other diagnoses, the highest accuracy was found for whoop combined with apnoea or post-tussive emesis. In patients aged 10 years and older, several different sign and symptom combinations were associated with an increased risk of pertussis among all enrolment diagnoses. There were fewer hospitalizations among the fully vaccinated children than in partly or unvaccinated children aged 4 months to 6 years (20.7% vs. 60.0%, p = 0.017). Conclusions The numerous sign and symptom combinations in the observed case definitions were good predictors for laboratory-confirmed pertussis among all enrolment diagnoses, therefore suggesting the necessity for increased awareness of possibility for pertussis in patients with certain pertussis-like medical conditions.

Distribution of Collagen I, III, and IV and Laminin in the Human Liver during Prenatal Development

In the absence of systematized data on the extracellular matrix components during prenatal liver development, the present study aimed to investigate the time of appearance and distribution of collagen types I, III, and IV and laminin. The study material included embryonic and fetal livers, aged 7–37 weeks, categorized into 3 trimesters. The material was stained using hematoxylin-eosin and immunohistochemistry methods for the identification of collagen I, III, and IV and laminin. Collagen I was detected near the end of the first trimester in the capsules and walls of interlobular veins. As the liver matures, collagen I is increasingly abundant in the capsules, portal area connective tissues, arterial walls, interlobular veins, sinusoids, and central veins. Collagen III and collagen IV appear in the middle of the first trimester in the capsules, portal areas, and walls of central veins, as well as the sinusoids particularly. In trimesters 2 and 3, these collagens are increasingly present in all the structures, but collagen IV is also present in nerve fibers. Laminin is sporadically present adjacent to the sinusoids in trimester 1, while in trimesters 2 and 3 this protein commonly appears in the walls of arteries and interlobular veins, in the basal membrane of bile ducts, and in nerve fibers. The contents of collagen I, III, and IV increase during prenatal development in the liver capsule, arterial and vein walls, sinusoids, and portal area. Laminin expression is consistent with that of the collagens with the exception that, within lobules, laminin disappears with liver maturation.

Immunohistochemical Heterogeneity of the Endothelium of Blood and Lymphatic Vessels in the Developing Human Liver and in Adulthood.

The endothelium of liver sinusoids in relation to the endothelium of other blood vessels has specific antigen expression similar to the endothelium of lymphatic vessels. Bearing in mind that there is no consensus as to the period or intensity of the expression of certain antigens in the endothelium of blood and lymphatic vessels in the liver, the aim of our study was to immunohistochemically investigate the dynamic patterns of the expression of CD31, CD34, D2-40, and LYVE-1 antigens during liver development and in adulthood on paraffin tissue sections of human livers of 4 embryos, 38 fetuses, 6 neonates, and 6 adults. The results show that, in a histologically immature liver at the end of the embryonic period, CD34 molecules are expressed only on vein endothelium localized in developing portal areas, whereby the difference between portal venous branches and CD34-negative central veins belongs to the collecting venous system. In the fetal period, with aging, expression of CD34 and CD31 molecules on the endothelium of central veins and blood vessels of the portal areas increases. Sinusoidal endothelium shows light and sporadic CD34 immunoreactivity in the late embryonic and fetal periods, and is lost in the neonatal and adult periods, unlike CD31 immunoreactivity, which is poorly expressed in the fetal and neonatal periods but is present in adults. The endothelium of sinusoids and lymphatic vessels express LYVE-1, and the endothelium of lymphatic vessels express LYVE-1 and D2-40 but not CD34. Similarity between the sinusoidal and lymphatic endothelium includes the fact that both types are LYVE-1 positive and CD34 negative.