ABSTRACT
Hydrogel-based bioinks are the main formulations used for Articular Cartilage (AC) regeneration due to their similarity to chondral tissue in terms of morphological and mechanical properties. However, the main challenge is to design and formulate bioinks able to allow reproducible additive manufacturing and fulfil the biological needs for the required tissue. In our work, we investigated an innovative Manuka honey (MH)-loaded photocurable gellan gum methacrylated (GGMA) bioink, encapsulating mesenchymal stem cells differentiated in chondrocytes (MSCs-C), to generate 3D bioprinted construct for AC studies. We demonstrated the beneficial effect of MH incorporation on the bioink printability, leading to the obtainment of a more homogenous filament extrusion and therefore a better printing resolution. Also, GGMA-MH formulation showed higher viscoelastic properties, presenting complex modulus G∗ values of â¼1042 âPa, compared to â¼730 âPa of GGMA. Finally, MH-enriched bioink induced a higher expression of chondrogenic markers col2a1 (14-fold), sox9 (3-fold) and acan (4-fold) and AC ECM main element production (proteoglycans and collagen).
ABSTRACT
Ageing is among the main risk factors for human disease onset and the identification of the hallmarks of senescence remains a challenge for the development of appropriate therapeutic target in the elderly. Here, we compare senescence-related changes in two cell populations of mesenchymal stromal cells by analysing their miRNA profiling: Human Dental Pulp Stromal Cells (hDPSCs) and human Periosteum-Derived Progenitor Cells (hPDPCs). After these cells were harvested, total RNA extraction and whole genome miRNA profiling was performed, and DIANA-miRPath analysis was applied to find the target/pathways. Only 69 microRNAs showed a significant differential expression between dental pulp and periosteum progenitor cells. Among these, 24 were up regulated, and 45 were downregulated in hDPSCs compared to hPDPCs. Our attention was centered on miRNAs (22 upregulated and 34 downregulated) involved in common pathways for cell senescence (i.e. p53, mTOR pathways), autophagy (i.e. mTOR and MAPK pathways) and cell cycle (i.e. MAPK pathway). The p53, mTOR and MAPK signaling pathways comprised 43, 37 and 112 genes targeted by all selected miRNAs, respectively. Our finding is consistent with the idea that the embryological origin influences cell behavior and the ageing process. Our study strengthens the hypothesis that ageing is driven by numerous mediators interacting through an intricate molecular network, which affects adult stem cells self-renewal capability. Graphical abstract.
Subject(s)
Mesenchymal Stem Cells , Aged , Aging/genetics , Humans , MicroRNAs/genetics , TOR Serine-Threonine Kinases , Tumor Suppressor Protein p53/geneticsABSTRACT
The cervical fasciae have always represented a matter of debate. Indeed, in the literature, it is quite impossible to find two authors reporting the same description of the neck fascia. In the present review, a historical background was outlined, confirming that the Malgaigne's definition of the cervical fascia as an anatomical Proteus is widely justified. In an attempt to provide an essential and a more comprehensive classification, a fixed pattern of description of cervical fasciae is proposed. Based on the morphogenetic criteria, two fascial groups have been recognized: (1) fasciae which derive from primitive fibro-muscular laminae (muscular fasciae or myofasciae); (2) fasciae which derive from connective thickening (visceral fasciae). Topographic and comparative approaches allowed to distinguish three different types of fasciae in the neck: the superficial, the deep and the visceral fasciae. The first is most connected to the skin, the second to the muscles and the third to the viscera. The muscular fascia could be further divided into three layers according to the relationship with the different muscles.
Subject(s)
Fascia , Neck , HumansABSTRACT
Human adipose derived stem cells have shown chondrogenic differentiation potential in cartilage tissue engineering in combination with biomimetic materials. In this study, the chondrogenic potential of a porous gelatin based scaffold genipin (GNP) crosslinked was investigated in human mesenchymal stem cells obtained from adipose tissue. Cells were cultured up to 4 weeks on the scaffold and on monolayer, MTT assay was performed to evaluate cell viability, light, and transmission electron microscopy were carried out to demonstrate cell proliferation, scaffold adhesion, and cell colonization inside the porous architecture of the biomaterial. The expression of chondrogenic markers such as SOX9, collagen type II, aggregan, and versican was investigated by Real Time PCR. Results showed an high cell viability, adhesion, and colonization of the scaffold. Real Time PCR data demonstrated an upregulation of all the chondrogenic markers analyzed. In conclusion, 3D gelatin GNP crosslinked porous scaffold provides an improved environment for chondrogenic differentiation of stem cells compared with cell monolayer culture system.
Subject(s)
Adipose Tissue/cytology , Biomimetic Materials , Cell Differentiation/physiology , Chondrogenesis/physiology , Iridoids , Mesenchymal Stem Cells/physiology , Tissue Scaffolds , Adipose Tissue/metabolism , Adipose Tissue/ultrastructure , Aggrecans/biosynthesis , Collagen Type II/biosynthesis , Gelatin , Humans , Mesenchymal Stem Cells/metabolism , Mesenchymal Stem Cells/ultrastructure , Microscopy , Microscopy, Electron, Transmission , Real-Time Polymerase Chain Reaction , SOX9 Transcription Factor/biosynthesis , Versicans/biosynthesisABSTRACT
The purpose of the study was to investigate the biological risk of asbestiform antigorite, which is a fibrous variety of antigorite, one of the natural mineral fibres of the serpentine group to which asbestos chrysotile belongs. Asbestiform antigorite is very abundant and commonly found associated with asbestos chrysotile in serpentinites, a kind of rock outcropping present in many geographical locations worldwide. In this study we evaluated the morphological, immunohistochemical and functional effects of antigorite fibres in alveolar epithelial cancer cells (A549), a standardized human cell line currently used as a model to study cytotoxicity induced by pharmacological agents. The antigorite fibres were identified and characterized morphologically and chemically by X-ray powder diffractometry, transmission and scanning electron microscopy, both with annexed energy dispersive spectrometry. The effects of 50 microg/ml of antigorite in A549 lung cells treated at 24 and 48 h resulted in increased synthesis of VEGF, Cdc42 and beta-catenin that represent potential risks for cancer development. Phalloidin labelling showed an irregular distribution of filamentous actin resulting from antigorite contact. Our studies indicate potential cellular toxicity of antigorite in vivo, providing the opportunity to elucidate the effect of asbestos on cancer induction and possible modes of therapy.