scholarly journals Developing a Glyoxal-Crosslinked Chitosan/Gelatin Hydrogel for Sustained Release of Human Platelet Lysate to Promote Tissue Regeneration

2021 ◽  
Vol 22 (12) ◽  
pp. 6451
Author(s):  
Ching-Cheng Tsai ◽  
Tai-Horng Young ◽  
Guang-Shih Chen ◽  
Nai-Chen Cheng
Keyword(s):  
Growth Factor ◽  
Sustained Release ◽  
Tissue Regeneration ◽  
Transforming Growth Factor ◽  
Human Platelet ◽  
Umbilical Vein ◽  
Platelet Lysate ◽  
Release Pattern ◽  
Human Platelet Lysate

The clinical application of human platelet lysate (HPL) holds promise for tissue regeneration, and the development of an efficient vehicle for its delivery is desired. Chitosan-based hydrogels are potential candidates, but they often exhibit weak mechanical properties. In this study, a chitosan/gelatin (CS-GE) hydrogel crosslinked by glyoxal was fabricated for sustained release of HPL. The influence of HPL on Hs68 fibroblast and human umbilical vein endothelial cell (HUVEC) culture was evaluated, and we found that supplementing 5% HPL in the medium could significantly improve cell proliferation relative to supplementing 10% fetal bovine serum (FBS). Moreover, HPL accelerated the in vitro wound closure of Hs68 cells and facilitated the tube formation of HUVECs. Subsequently, we fabricated CS-GE hydrogels crosslinked with different concentrations of glyoxal, and the release pattern of FITC-dextrans (4, 40 and 500 kDa) from the hydrogels was assessed. After an ideal glyoxal concentration was determined, we further characterized the crosslinked CS-GE hydrogels encapsulated with different amounts of HPL. The HPL-incorporated hydrogel was shown to significantly promote the proliferation of Hs68 cells and the migration of HUVECs. Moreover, the release pattern of transforming growth factor-β1 (TGF-β1) and platelet-derived growth factor-BB (PDGF-BB) from hydrogel was examined in vitro, demonstrating a sustained release profile of the growth factors. Finally, the chick chorioallantoic membrane assay revealed that HPL encapsulation in the hydrogel significantly stimulated angiogenesis in ovo. These results demonstrate the great potential of the crosslinked CS-GE hydrogel to serve as an effective delivery system for HPL to promote tissue regeneration.

scholarly journals Human Platelet Lysate Acts Synergistically With Laminin to Improve the Neurotrophic Effect of Human Adipose-Derived Stem Cells on Primary Neurons in vitro

2021 ◽  
Vol 9 ◽  
Author(s):  
Martino Guiotto ◽  
Wassim Raffoul ◽  
Andrew M. Hart ◽  
Mathis O. Riehle ◽  
Pietro G. di Summa
Keyword(s):  
Stem Cells ◽  
Neurite Outgrowth ◽  
Animal Studies ◽  
Human Platelet ◽  
Platelet Lysate ◽  
Neural Cells ◽  
Adipose Derived Stem Cells ◽  
Peripheral Nerve Lesions ◽  
Human Platelet Lysate

BackgroundDespite the advancements in microsurgical techniques and noteworthy research in the last decade, peripheral nerve lesions have still weak functional outcomes in current clinical practice. However, cell transplantation of human adipose-derived stem cells (hADSC) in a bioengineered conduit has shown promising results in animal studies. Human platelet lysate (hPL) has been adopted to avoid fetal bovine serum (FBS) in consideration of the biosafety concerns inherent with the use of animal-derived products in tissue processing and cell culture steps for translational purposes. In this work, we investigate how the interplay between hPL-expanded hADSC (hADSChPL) and extracellular matrix (ECM) proteins influences key elements of nerve regeneration.MethodshADSC were seeded on different ECM coatings (laminin, LN; fibronectin, FN) in hPL (or FBS)-supplemented medium and co-cultured with primary dorsal root ganglion (DRG) to establish the intrinsic effects of cell–ECM contact on neural outgrowth. Co-cultures were performed “direct,” where neural cells were seeded in contact with hADSC expanded on ECM-coated substrates (contact effect), or “indirect,” where DRG was treated with their conditioned medium (secretome effect). Brain-derived nerve factor (BDNF) levels were quantified. Tissue culture plastic (TCPS) was used as the control substrate in all the experiments.ResultshPL as supplement alone did not promote higher neurite elongation than FBS when combined with DRG on ECM substrates. However, in the presence of hADSC, hPL could dramatically enhance the stem cell effect with increased DRG neurite outgrowth when compared with FBS conditions, regardless of the ECM coating (in both indirect and direct co-cultures). The role of ECM substrates in influencing neurite outgrowth was less evident in the FBS conditions, while it was significantly amplified in the presence of hPL, showing better neural elongation in LN conditions when compared with FN and TCPS. Concerning hADSC growth factor secretion, ELISA showed significantly higher concentrations of BDNF when cells were expanded in hPL compared with FBS-added medium, without significant differences between cells cultured on the different ECM substrates.ConclusionThe data suggest how hADSC grown on LN and supplemented with hPL could be active and prone to support neuron–matrix interactions. hPL enhanced hADSC effects by increasing both proliferation and neurotrophic properties, including BDNF release.

scholarly journals Anti-Fibrosis Effect of Relaxin and Spironolactone Combined on Isoprenaline-Induced Myocardial Fibrosis in Rats via Inhibition of Endothelial–Mesenchymal Transition

2017 ◽  
Vol 41 (3) ◽  
pp. 1167-1178 ◽  
Author(s):  
Jiejie Cai ◽  
Xiao Chen ◽  
Xingxing Chen ◽  
Lingzhi Chen ◽  
Gaoshu Zheng ◽  
...  
Keyword(s):  
Growth Factor ◽  
Myocardial Fibrosis ◽  
Transforming Growth Factor ◽  
Umbilical Vein ◽  
Combined Therapy ◽  
Fibrous Tissue ◽  
Type I ◽  
Mesenchymal Transition ◽  
Endothelial Mesenchymal Transition

Background: The effect of relaxin and spironolactone combined on myocardial fibrosis has not been reported. Thus, we investigated the effect of the combined therapy on isoprenaline-induced myocardial fibrosis and the mechanism. Methods: Rats were injected subcutaneously with isoprenaline to induce myocardial fibrosis and underwent subcutaneous injection with relaxin (2 µg·kg-1·d-1) and given a gavage of spironolactone (30 mg·kg-1·d-1) alone or combined for 14 days. In vitro, the endothelial–mesenchymal transition was induced with transforming growth factor β (TGF-β) in human umbilical vein endothelial cells (HUVECs) pretreated with relaxin, 200 ng/ml, and/or spironolactone, 1uM. Results: Relaxin and spironolactone used alone or combined improved cardiac function and decreased cardiac weight indices; reduced fibrous tissue proliferation; reduced levels of type I and III collagen; decreased the expression of α–smooth muscle actin (α-SMA) and transforming growth factor-β1 (TGF-β1), and increased the expression of cluster of differentiation-31 (CD31) in rats with isoprenaline-induced myocardial fibrosis. In vitro, compared with TGF-β treatment, relaxin and spironolactone used alone or combined with TGF-β decreased cell mobility, α-SMA and vimentin levels but increased vascular endothelial cadherin (VE-cadherin) and endothelial CD31levels. Especially, combined therapy had more remarkable effect than relaxin and spironolactone used alone both in vitro and in vivo. Conclusion: Relaxin and spironolactone combined affected isoprenaline-induced myocardial fibrosis in rats that the mechanism might be inhibition of the cardiac endothelial–mesenchymal transition.

scholarly journals Effects of plant-derived immunomodulators and lizak preparation in vitro taken separately or in combination on the immune factors of palatine tonsils of children with chronic tonsillitis

2020 ◽  
pp. 13-18
Author(s):  
Alexander Yu. Bredun ◽  
Oleg F. Melnikov ◽  
Мarina D. Timchenko
Keyword(s):  
Growth Factor ◽  
Cell Viability ◽  
Tissue Regeneration ◽  
Transforming Growth Factor ◽  
Interferon Γ ◽  
Interleukin 1Β ◽  
Combined Use ◽  
Antibacterial Preparation ◽  
Regeneration Factor

Introduction: Chronic inflammatory diseases of the upper respiratory tract of both microbial and viral origin occur due to the immune deficiency of systemic and local nature. Among a large number of immunomodulation agents, a special role belongs to plant preparations, which are characterized by relative harmlessness and high efficiency, both in case of systemic and local use and they are often combined with antimicrobial agents, therefore the aim of the study was to investigate their effect on the reactions and condition of lymphoid cells of tonsilswith separate and combined use in vitro. Methods: Cell suspensions were prepared mechanically and adjusted to a concentration of 2 million/ml in medium 199 with additives. Then, Lizak preparation was added to the cells and starch solution was used in control. Imupret and Esberitox preparations were added to the culture with or without Lizak preparation. After cultivation, the levels of proinflammatory cytokine – Interleukin-1β, pro-allergic factor – Interleukin-4, Th-1 – derivative of cytokine – interferon-γ were studied in the supernatant using the «Tsitokin LLC» reagent kits (RF), as well as tissue regeneration factor – transforming growth factor –TGF-1β (Austria). The preparation was prepared from the cell pellet, in which the relative nonviable cell count was determined in a sample with trypan blue using a light microscope (Olympus CX21FS1).The statistical analysis was performed using Student's t-distribution. Results: Plant-derived preparations did not have any effect on cell viability in culture, did not reduce the level of the pro-inflammatory cytokine interleukin-1β and did not stimulate the production of the regeneration factor TGF-1β. At the same time, the combined use of plant-derived preparations and the antibacterial preparation Lizak in cell culture of tonsils was accompanied by an increase in cell viability compared to the use of Lizak preparation separately, the stimulation of the regulatory antiviral factor interferon-γ and an increase in the production of tissue regeneration factor. Conclusion: The immunomodulating properties of the antibacterial preparation Lizak and plant-derived immunomodulator Esberitox significantly differ in the points of application. Both plant-derived preparations stimulated the production of γ-interferon by tonsil cells in the presence of Lizak preparation, the transforming growth factor TGF-1β, the viability of tonsil cells was the highest with the combination of Lizak and Esberitox.

scholarly journals Combined Effects of Methyldopa and Flavonoids on the Expression of Selected Factors Related to Inflammatory Processes and Vascular Diseases in Human Placenta Cells—An In Vitro Study

Molecules ◽  
2021 ◽  
Vol 26 (5) ◽  
pp. 1259
Author(s):  
Anna Bogacz ◽  
Przemysław Ł. Mikołajczak ◽  
Marlena Wolek ◽  
Aleksandra Górska ◽  
Michał Szulc ◽  
...  
Keyword(s):  
Growth Factor ◽  
Mrna Expression ◽  
Cell Line ◽  
Transforming Growth Factor ◽  
Umbilical Vein ◽  
Combined Effects ◽  
Vegf Mrna ◽  
Protein Levels ◽  
Tnf Α

The aim of the study was to investigate combined effects of flavonoids (apigenin, baicalein, chrysin, quercetin, and scutellarin) and methyldopa on the expression of selected proinflammatory and vascular factors in vitro for prediction of their action in pregnancy-induced hypertension. The research was conducted on a trophoblast-derived human choriocarcinoma cell line and a primary human umbilical vein endothelial cell line. Cytotoxicity of compounds in selected concentrations (20, 40, and 100 µmol) was measured using the MTT test and the concentration of 40 µmol was selected for further analysis. Subsequently, their effects with methyldopa on the expression of selected markers responsible for inflammation (TNF-α; IL-1β; IL-6) and vascular effects (hypoxia-inducible factor 1α—HIF-1α; placental growth factor—PIGF; transforming growth factor β—TGF-β; vascular endothelial growth factor—VEGF) at the mRNA and protein levels were assessed. It was found that every combined administration of a flavonoid and methyldopa in these cells induced a down-regulating effect on all tested factors, except PIGF, especially at the mRNA expression level. As hypertension generally raises TNF-α, IL-1β, IL-6, HIF-1α, TGF-β, and VEGF mRNA expression and/or protein levels, the results obtained in the studied model may provide a positive prognostic factor for such activity in vivo.

Exosomes derived from human platelet lysate affect MSC functions in vitro

2014 ◽  
Author(s):  
Elena Torreggiani ◽  
Francesca Perut ◽  
Laura Roncuzzi ◽  
Nicola Baldini
Keyword(s):  
Human Platelet ◽  
Platelet Lysate ◽  
Human Platelet Lysate

The effect of human platelet lysate on corneal nerve regeneration

2019 ◽  
pp. bjophthalmol-2019-314408 ◽  
Author(s):  
Chin-Te Huang ◽  
Hsiao-Sang Chu ◽  
Kuo-Chi Hung ◽  
Lily Wei Chen ◽  
Mei-Yun Chen ◽  
...  
Keyword(s):  
Wound Healing ◽  
Nerve Regeneration ◽  
Neurotrophic Factors ◽  
Ganglion Cells ◽  
Human Platelet ◽  
Platelet Lysate ◽  
Human Platelet Lysate ◽  
Corneal Nerve

AimThis study aimed to test whether human platelet lysate (HPL) has neurotrophic ability for corneal nerve regeneration.MethodsWe measured the neurotrophic factors in human peripheral serum (HPS) and two commercially available HPLs, UltraGRO and PLTMax. In vitro, we compared the growth rates, neuronal differentiation and immunostaining of neuron markers in mouse neuroblastoma cell line (Neuro-2a) and primary culture of mouse trigeminal ganglion cells that were cultivated in different concentrations of fetal bovine serum, HPS and HPL. In vivo, we created corneal wounds on Sprague Dawley rats with a rotating burr and evaluated the effects of topical HPL on wound healing and corneal nerve regeneration by in vivo confocal microscopy and corneal aesthesiometry.ResultsHPLs had significantly higher concentrations of various neurotrophic factors compared with HPS (p<0.05). In Neuro-2a cells, 3% HPL was better at promoting neuronal growth and differentiation compared with HPS at the same concentration. HPL was also found to have superior neurotrophic effects compared with HPS in primary cultures of mouse trigeminal ganglion cells. In vivo, HPL-treated eyes had better corneal epithelial wound healing rate, nerve regeneration length and corneal touch threshold compared with eyes treated with artificial tears (p<0.05).ConclusionHPL has significantly higher concentrations of neurotrophic factors compared with HPS. It showed not only in vitro but also in vivo corneal neurotrophic abilities. Our results suggest that HPL may have a potential role in the treatment of diseases related to corneal nerve damage or degeneration.

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