Different titanium surfaces modulate the bone phenotype of SaOS-2 osteoblast-like cells.

Commercially pure titanium implants presenting a relatively smooth, machined surface or a roughned endosseous surface show a large percentage of clinical successes. Surface properties of dental implants seem to be important with respect to bone cells response. Implant topography appears to modulate cell growth and differentiation of osteoblasts affecting the bone healing around the titanium implant. The aim of the present study was to examine the effects of three different titanium surfaces on cell morphology, adhesion and bone phenotypic expression of human osteoblast-like cells, SaOS-2. SaOS-2 cells were cultured on commercially pure titanium disks of 1 cm in diameter with three different surface roughness: smooth (S), sandblasted (SB) and titanium plasma sprayed (TPS). Differences in the cell morphology were found on the three surfaces showing an uniform monolayer of shaped cells on the S surfaces, and clusters of multilayered cells with an irregular shape on the rough surfaces. The adhesion of SaOS-2 cells, as measured after 3h of culture, was not affected by surface roughness. ECM components such as collagen I (CoI), fibronectin (FN), vitronectin (VN) and tenascin (TN) were secreted and organized only on SB and TPS surfaces while on S surfaces they remained in the cytoplasm. Osteopontin and BSP-II were largely detected on SB and TPS surfaces, while only minimal production was observed on S surfaces. These data show that titanium surface roughness affects bone differentiation of osteoblast like-cells, SaOS-2, indicating that surface properties may be able to modulate the osteoblast phenotype. These observations also suggest that the bone healing response around dental implants can be affected by surface topography

Effects of physical exercise on adiponectin, leptin, and inflammatory markers in childhood obesity: systematic review and meta-analysis

Background: New findings on adipose tissue physiology and obesity-Associated inflammation status suggest that modification of the adipokine level can be relevant for the long-Term prevention of obesity-Associated chronic disease. Objectives: The scope of the present study was to investigate the effectiveness of physical exercise in reducing the systemic inflammation related to obesity in children. Methods: We conducted a systematic review with meta-Analysis of controlled randomized trials, identified through electronic database search, which investigated the effect of physical exercise, without concomitant dietary intervention, on adiponectin, leptin, and/or other inflammatory markers in children up to age 18 years with a body mass index greater than the 95th percentile for age and sex. Results: Seven trials were included in the meta-Analysis, with a total of 250 participants. Compared with the control group without any lifestyle modification, the physical exercise resulted in a reduction in leptin [standardized mean difference (SMD)-1.13; 95% confidence interval (95%CI):-1.89 to-0.37; I2 = 79.9%] and interleukin-6 (SMD-0.84; 95%CI:-1.45 to-0.23, I2 = 0.9%) and an increase in adiponectin plasma concentration (SMD 0.69; 95%CI: 0.02-1.35; I2 = 74.3%). Conclusions: These results indicate that physical exercise improved the inflammatory state in children with obesity. It is unclear whether this effect can reduce the risk of cardiovascular and metabolic disease in adulthood. Clinical trials with a uniform intervention protocol and outcome measurements are required to put our knowledge on adipose tissue biology into a clinical perspective

Non-modified RNA-Based Reprogramming of Human Dermal Fibroblasts into Induced Pluripotent Stem Cells

The generation of pluripotent stem cells from adult somatic cells by cell reprogramming has put a whole new perspective on stem cell biology and stem cell-based regenerative medicine. Cell reprogramming acts through the introduction of key genes that regulate and maintain the pluripotent cell state. In this chapter, we describe the optimized protocol for the efficient isolation of fibroblasts from a skin punch biopsy and the subsequent easy and effective generation of integration-free induced pluripotent stem cell (iPSC) colonies forcing the expression of specific factors by non-modified RNAs. © 2021, Springer Science+Business Media, LLC

Direct cell reprogramming as a new emerging strategy in cardiac regeneration

Myocardial infarction (MI) is the current leading cause of mortality in the industrialised world. It is due to the irreversible death of billions of cardiomyocytes, secondary to a condition of ischemia. This leads to the formation of a stiff fibrotic tissue, mainly populated by cardiac fibroblasts (CFs). Currently, the only available therapy addressing the irreversible loss of functional cardiomyocytes is heart transplantation. Different tissue engineering approaches and cell therapies are under investigation, aimed at recovering myocardial contractility. Main issues in these strategies are the poor grafting and survival ability of implanted cells as well as the limited endogenous regenerative potential of adult heart. A new strategy is now emerging based on direct reprogramming of CFs into induced cardiomyocytes (iCMs) using transcriptional factors and/ or microRNAs (miRNAs) (miR-combo) [2-4]. Proof of concepts results of in vitro and in vivo conversion of mouse CFs into iCMs have been published and in vitro direct reprogramming of human CFs has also been reported [1-3]. However, such strategy is still an immature approach: reprogramming efficiency is low and partially reprogrammed non-beating cardiomyocytes have been generally obtained. Recently, in vitro direct reprogramming efficiency of mouse CFs cultured in 3D fibrin hydrogels using miR-combo has resulted significantly increased compared to 2D culture systems [4]. Based on these preliminary results, in this work we studied the miR-combo mediated reprogramming efficiency of human dermal and cardiac fibroblasts cultured on hydrogel matrices, including fibrin, fibrin/laminin, fibrin/fibronectin and fibrin/cardiac biomatrix [5], by analysing cell morphology, cell viability, change in gene expression (PCR analysis) and presence of markers of trans-differentiation by immunohistochemistry. The 3D biomimetic hydrogels were able to increase reprogramming efficiency respect to 2D culture environment, both at a genetic and protein level, with an enhancement in the expression of cardiac genes and cardiac proteins such as cardiac troponin I and alpha sarcomeric actinin. [1] J.A. Batty et al. Eur. J. Heart Failure 2016; 18: 145 [2] T.M. Jayawardena et al. Circ. Res. 2012; 110: 1465-1473. [3] T.M. Jayawardena et al. Circ. Res. 2015; 116:418-24. [4] Y. Li et al. Scientific Reports 2016; 6: 38815. [5] C. Castaldo et al. Biomed Res Int. 2013; 2013: 352370. ERC-CoG 2017 BIORECAR project is acknowledge

In vitro cultured progenitors and precursors of cardiac cell lineages from human normal and post-ischemic hearts.

The demonstration of the presence of dividing primitive cells in damaged hearts has sparked increased interest about myocardium regenerative processes. We examined the rate and the differentiation of in vitro cultured resident cardiac primitive cells obtained from pathological and normal human hearts in order to evaluate the activation of progenitors and precursors of cardiac cell lineages in post-ischemic human hearts. The precursors and progenitors of cardiomyocyte, smooth muscle and endothelial lineage were identified by immunocytochemistry and the expression of characteristic markers was studied by western blot and RT-PCR.The amount of proteins characteristic for cardiac cells (alpha-SA and MHC, VEGFR-2 and FVIII, SMA for the precursors of cardiomyocytes, endothelial and smooth muscle cells, respectively) inclines toward an increase in both alpha-SA and MHC. The increased levels of FVIII and VEGFR2 are statistically significant, suggesting an important re-activation of neoangiogenesis. At the same time, the augmented expression of mRNA for Nkx 2.5, the trascriptional factor for cardiomyocyte differentiation, confirms the persistence of differentiative processes in terminally injured hearts. Our study would appear to confirm the activation of human heart regeneration potential in pathological conditions and the ability of its primitive cells to maintain their proliferative capability in vitro. The cardiac cell isolation method we used could be useful in the future for studying modifications to the microenvironment that positively influence cardiac primitive cell differentiation or inhibit, or retard, the pathological remodeling and functional degradation of the heart

Non-integrating Methods to Produce Induced Pluripotent Stem Cells for Regenerative Medicine: An Overview

Induced Pluripotent Stem cells (iPSC) are adult somatic cells genetically reprogrammed to an embryonic stem cell-like state. Due to their autologous origin from adult somatic cells, iPSCs are considered a tremendously valuable tool for regenerative medicine, disease modeling, drug discovery and testing. iPSCs were first obtained by introducing specific transcription factors through retroviral transfection. However, cell reprogramming obtained by integrating methods prevent clinical application of iPSC because of potential risk for infection, teratomas and genomic instability. Therefore, several integration-free alternate methods have been developed and tested thus far to overcome safety issues. The present chapter provides an overview and a critical analysis of advantages and disadvantages of non-integrating methods used to generate iPSCs

Normal versus pathological cardiac fibroblast-derived extracellular matrix differentially modulates cardiosphere-derived cell paracrine properties and commitment

Human resident cardiac progenitor cells (CPCs) isolated as cardiosphere-derived cells (CDCs) are under clinical evaluation as a therapeutic product for cardiac regenerative medicine. Unfortunately, limited engraftment and differentiation potential of transplanted cells significantly hamper therapeutic success. Moreover, maladaptive remodelling of the extracellular matrix (ECM) during heart failure progression provides impaired biological and mechanical signals to cardiac cells, including CPCs. In this study, we aimed at investigating the differential effect on the phenotype of human CDCs of cardiac fibroblast-derived ECM substrates from healthy or diseased hearts, named, respectively, normal or pathological cardiogel (CG-N/P). After 7 days of culture, results show increased levels of cardiogenic gene expression (NKX2.5, CX43) on both decellularized cardiogels compared to control, while the proportion and staining patterns of GATA4, OCT4, NKX2.5, ACTA1, VIM, and CD90-positive CPCs were not affected, as assessed by immunofluorescence microscopy and flow cytometry analyses. Nonetheless, CDCs cultured on CG-N secreted significantly higher levels of osteopontin, FGF6, FGF7, NT-3, IGFBP4, and TIMP-2 compared to those cultured on CG-P, suggesting overall a reduced trophic and antiremodelling paracrine profile of CDCs when in contact with ECM from pathological cardiac fibroblasts. These results provide novel insights into the bidirectional interplay between cardiac ECM and CPCs, potentially affecting CPC biology and regenerative potential

Flatfoot in children: anatomy of decision making

Concern about a child’s foot posture is a common reason for frequent consultations for an array of health care professionals; sports medicine specialists are often the first to recognize and advise on foot pathology. In the decision making process, it is essential to distinguish between the different types of flatfoot deformity: paediatric or adult, congenital or acquired, flexible or rigid. Although flatfoot in children is a common finding, evidence for the techniques of the reliable and reproducible assessment of the foot posture is scant. This general review presents the factors involved in the forming and supporting of the foot arches, discusses the protocols useful in the evaluation of the foot posture, and indicates how to differentiate between flatfoot cases needing treatment and cases that need only reassurance