scholarly journals The combination of ADSCs and 10% PRP increases Rb protein expression on senescent human dermal fibroblasts

F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 516
Author(s):  
Sinta Murlistyarini ◽  
Lulus Putri Aninda ◽  
Ufida Aini Afridafaz ◽  
Sri Widyarti ◽  
Agustina Tri Endharti ◽  
...  

Background: The senescence process in human dermal fibroblasts (HDFs) is caused by cell cycle withdrawal processes, one of which is the result of the retinoblastoma (Rb) protein being in a hypo-phosphorylated state. Since adipose-derived stem cells (ADSCs) have a paracrine effect, ADSCs were utilized to improve the senescence process of HDFs. The use of non-autologous cell culture media to grow ADSCs can be legally problematic; therefore, platelet-rich plasma (PRP) can be considered as an alternative medium. PRP contains various growth factors that can be used to process the reversal of senescent HDFs. The combination of ADSCs and PRP is expected to increase the expression of Rb protein in HDFs that have undergone the senescence process. Methods: This study was performed in vitro with a randomized sample, and non-blinded pre-and post-test control group. The primary culture of senescent HDFs was transfected with a combination of ADSCs and 10% PRP. The effect on migration was observed through the scratch test, while the effect of PRP on reversal senescence was observed through Sa-β-gal analysis and the expression of protein Rb with ELISA. Results: The senescent HDFs that received a combined transfection of ADSCs and 10% PRP proliferated rapidly in the scratch test. Based on the Sa-β-gal assay, they showed fewer senescent HDFs cells. The combination of ADSCs and 10% PRP elevated the expression of Rb protein significantly (P < 0.001). Conclusions: The combination of ADSCs and 10% PRP was shown to have a reversal effect on the senescence process of HDFs in vitro.

Nanomaterials ◽  
2019 ◽  
Vol 9 (8) ◽  
pp. 1131
Author(s):  
Nouf N. Mahmoud ◽  
Lubna M. Al-Kharabsheh ◽  
Enam A. Khalil ◽  
Rana Abu-Dahab

Herein, the cytotoxicity, cellular uptake and wound healing of human dermal fibroblasts were investigated upon treatment with gold nanorods (GNR) decorated with different ligands. Neutral and cationic poly ethylene glycol (PEG)-decorated GNR demonstrated the least cytotoxicity and cellular internalization, while anionic- and bovine serum albumin (BSA)-coated GNR revealed significant cytotoxicity and cellular uptake into human dermal fibroblasts. The cell scratch test demonstrated that neutral, cationic PEGylated GNR and anionic-decorated GNR have accelerated the wound healing rate in vitro after 24 h of incubation with scratched human dermal fibroblasts compared to control, while there was a drastic retardation of wound healing rate of scratched fibroblasts upon exposure to BSA-GNR accompanied with a significant release of the inflammatory cytokine; interlukin-1β (IL-1β). The cytotoxicity of GNR against the dermal cells and their ability to enhance the wound healing in vitro are greatly linked to their surface modifications.


2021 ◽  
Vol 9 (A) ◽  
pp. 422-427
Author(s):  
Sinta Murlistyarini ◽  
Lulus Putri Aninda ◽  
Sri Widyarti ◽  
Agustina Tri Endharti ◽  
Teguh Wahju Sardjono

BACKGROUND: In the senescence process, the retinoblastoma (Rb) protein binds to E2F in hypophosphorylated conditions, preventing the cell to enter the S-phase in the cell cycle. Human Forkhead Box M1 (FOXM1) protein, key regulator G1/S and G2/M phases, decreases in the senescence process. Many studies have been carried out to reverse this system, one of which used exosomes of adipose-derived stem c ells conditioned media (ADSC-CM). These exosomes contain a variety of specific proteins which have pro-proliferation properties, however, little is known on the role of these exosomes toward the change of phosphorylated Rb and FOXM1. AIM: This study aims to find out the involvement of exosomes of ADSC-CM on these two proteins on senescence human dermal fibroblasts (HDFs). METHODS: In vitro experiment was undergone randomization sample and non-blinded pre-/post-test control group. The primary culture of senescent HDFs was transfected with exosomes of ADSC-CM; then, its effect on migration and senescence reversal was observed through analyzing Sa-β-gal, Rb, and FOXM1 protein expression. RESULTS: The expression of Sa-β-gal was higher in the control group. Our result demonstrated the exosome of ADSC-CM significantly induced the expression of Rb and FOXM1 protein in senescent HDFs (p < 0.05). CONCLUSION: It proved that exosomes of ADSC-CM could shift the senescent fibroblast into metabolically active cells.


2021 ◽  
pp. 036354652110186
Author(s):  
Min Ji Lee ◽  
Kang Sup Yoon ◽  
Sohee Oh ◽  
Sue Shin ◽  
Chris Hyunchul Jo

Background: While platelet-rich plasma (PRP) has been widely studied for musculoskeletal disorders, few studies to date have reported its use for adhesive capsulitis (AC). Fully characterized and standardized allogenic PRP may provide clues to solve the underlying mechanism of PRP with respect to synovial inflammation and thus may clarify its clinical indications. Purpose: To clinically evaluate the safety and efficacy of a fully characterized pure PRP injection in patients with AC and to assess the effects of pure PRP on synoviocytes with or without inflammation in vitro. Study Design: Controlled laboratory study and cohort study; Level of evidence, 3. Methods: For the clinical analysis, a total of 15 patients with AC received an ultrasonography-guided intra-articular PRP injection and were observed for 6 months. Pain, range of motion (ROM), muscle strength, shoulder function, and overall satisfaction in the patients were evaluated using questionnaires at 1 week as well as at 1, 3, and 6 months after the PRP injection and results were compared with the results of a propensity score−matched control group that received a corticosteroid injection (40 mg triamcinolone acetonide). For the in vitro analysis, synoviocytes were cultured with or without interleukin-1β (IL-1β) and PRP. The gene expression of proinflammatory and anti-inflammatory cytokines as well as matrix enzymes and their inhibitors was evaluated. Results: At 6-month follow-up, pure PRP significantly decreased pain and improved ROM, muscle strength, and shoulder function to levels comparable with those after a corticosteroid injection. All pain values, strength measurements, and functional scores significantly improved up to 6 months in the PRP group, but these measures improved up to 3 months and then were decreased at 6 months in the corticosteroid group. ROM was significantly improved in the 2 groups at 6 months compared with baseline. Allogenic PRP did not cause adverse events. For the in vitro findings, PRP induced inflammation but significantly improved the IL 1β−induced synovial inflammatory condition by decreasing proinflammatory cytokines such as IL-1β, tumor necrosis factor−α, IL-6, cyclooxygenase-2, and microsomal prostaglandin E synthase−1 and decreased matrix enzymes (matrix metalloproteinase−1, −3, and −13 as well as a disintegrin and metalloproteinase with thrombospondin motifs−4 and −5) and further increasing anti-inflammatory cytokines such as vasoactive intestinal peptide. Conclusion: This study showed that PRP decreased pain and improved shoulder ROM and function to an extent comparable with that of a corticosteroid in patients with AC. Allogenic pure PRP acted in a pleiotropic manner and decreased proinflammatory cytokines only in the inflammatory condition. Clinical Relevance: Allogenic PRP could be a treatment option for the inflammatory stage of AC.


2009 ◽  
Vol 4 (2) ◽  
pp. 025008 ◽  
Author(s):  
Ikuko Machida-Sano ◽  
Yasushi Matsuda ◽  
Hideo Namiki

Marine Drugs ◽  
2018 ◽  
Vol 16 (7) ◽  
pp. 239 ◽  
Author(s):  
Lei Wang ◽  
WonWoo Lee ◽  
Jae Oh ◽  
Yong Cui ◽  
BoMi Ryu ◽  
...  

Our previous study evaluated the antioxidant activities of sulfated polysaccharides from Celluclast-assisted extract of Hizikia fusiforme (HFPS) in vitro in Vero cells and in vivo in zebrafish. The results showed that HFPS possesses strong antioxidant activity and suggested the potential photo-protective activities of HFPS. Hence, in the present study, we investigated the protective effects of HFPS against ultraviolet (UV) B-induced skin damage in vitro in human dermal fibroblasts (HDF cells). The results indicate that HFPS significantly reduced intracellular reactive oxygen species (ROS) level and improved the viability of UVB-irradiated HDF cells in a dose-dependent manner. Furthermore, HFPS significantly inhibited intracellular collagenase and elastase activities, remarkably protected collagen synthesis, and reduced matrix metalloproteinases (MMPs) expression by regulating nuclear factor kappa B (NF-κB), activator protein 1 (AP-1), and mitogen-activated protein kinases (MAPKs) signaling pathways in UVB-irradiated HDF cells. These results suggest that HFPS possesses strong UV protective effect, and can be a potential ingredient in the pharmaceutical and cosmetic industries.


2021 ◽  
Vol 95 ◽  
Author(s):  
E.S. El-Wakil ◽  
H.F. Abdelmaksoud ◽  
T.S. AbouShousha ◽  
M.M.I. Ghallab

Abstract Our work aimed to evaluate the possible effect of Annona muricata (Graviola) leaf extract on Trichinella spiralis in in vitro and in vivo studies. Trichinella spiralis worms were isolated from infected mice and transferred to three culture media – group I (with no drugs), group II (contained Graviola) and group III (contained albendazole) – then they were examined using the electron microscope. In the in vivo study, mice were divided into five groups: GI (infected untreated), GII (prophylactically treated with Graviola for seven days before infection), GIII (infected and treated with Graviola), GIV (infected and treated with albendazole) and GV (infected and treated with a combination of Graviola plus albendazole in half doses). Drug effects were assessed by adults and larvae load beside the histopathological small intestinal and muscular changes. A significant reduction of adult and larval counts occurred in treated groups in comparison to the control group. Histopathologically, marked improvement in the small intestinal and muscular changes was observed in treated groups. Also, massive destruction of the cultured adults’ cuticle was detected in both drugs. This study revealed that Graviola leaves have potential activity against trichinellosis, especially in combination with albendazole, and could serve as an adjuvant to anti-trichinellosis drug therapy.


1987 ◽  
Author(s):  
J De Crée ◽  
H Geukens ◽  
H Demoen ◽  
H Verhaegen

Red blood cell (RBC) filtration in platelet rich plasma (PRP) and platelet poor plasma (PPP) was equally decreased (p < 0.0001) in 120 patients with acute myocardial infarction (AMI) as compared to a control group. In a double-blind experiment 2 groups of 30 patients with AMI received an acute oral dose of 60 mg of ketanserin, a serotonin (5-HT) antagonist at 5-HT2-receptors, or placebo. Ketanserin treatment improved RBC filtration in PRP with an average increase of 30%. A similar experiment using PPP showed a significant increase of 10%. Filtration of plasma improved after ketanserin treatment in PRP, but not in PPP. Cross-exchange experiments showed the ketanserin-induced improvement of RBC filtration in PRP and PPP to be also plasmadependent. 5-HT in vitro at 10−9M deteriorated RBC filtration in PPP (p < 0.05), and ketanserin in vitro at 10−7M counteracted this phenomenon (p < 0.001). Finally we found that the effect of a subacute treatment with ketanserin on the filtration of RBC Suspensions, enriched with a constant amount of white blood cells (WBC), was more pronounced than on control RBC suspensions of patients with AMI.These results indicate that the impaired RBC filtration, reported in vascular diseases may be dependent on a subtle interaction between platelets, WBC, RBC and plasma. Treatment with ketanserin is capable to interrupt this vicious circle of rheological disturbances at different levels, first of all, by improving RBC deformability, but also by counteracting the platelet mediated effects on RBC and by favourably influencing the physical properties of WBC and so preventing clogging phenomena. Serotonin probably plays a pivotal role in these cascade of events and therapy with ketanserin might be of clinical value in diseases where microcirculatory flow is compromised.


Cartilage ◽  
2019 ◽  
pp. 194760351985873 ◽  
Author(s):  
Alexander M. Satin ◽  
Jolanta B. Norelli ◽  
Nicholas A. Sgaglione ◽  
Daniel A. Grande

ObjectiveGiven the potential applications of combined biologics, the authors sought to evaluate the in vitro effect of combined platelet-rich plasma (PRP) and hyaluronic acid (HA) on cellular metabolism.DesignBone marrow–derived mesenchymal stem cells (BMSCs) and chondrocytes were obtained from the femurs of Sprague-Dawley rats. An inflammatory model was created by adding 10 ng/mL interleukin-1-beta to culture media. Non-crosslinked high-molecular-weight HA, activated-PRP (aPRP), and unactivated-PRP (uPRP) were tested. Cellular proliferation and gene expression were measured at 1 week. Genes of interest included aggrecan, matrix metalloproteinase (MMP)-9, and MMP-13.ResultsCombined uPRP-HA was associated with a significant increase in chondrocyte and BMSC proliferation at numerous preparations. There was a trend of increased chondrocyte aggrecan expression with combined PRP-HA. The greatest and only significant decrease in BMSC MMP-9 expression was observed with combined PRP-HA. While a significant reduction of BMSC MMP-13 expression was seen with PRP and HA-alone, a greater reduction was observed with PRP-HA. MMP-9 chondrocyte expression was significantly reduced in cells treated with PRP-HA. PRP-alone and HA-alone at identical concentrations did not result in a significant reduction. The greatest reduction of MMP-13 chondrocyte expression was observed in chondrocytes plus combined PRP-HA.ConclusionsWe demonstrated a statistically significant increase in BMSC and chondrocyte proliferation and decreased expression of catabolic enzymes with combined PRP-HA. These results demonstrate the additive in vitro effect of combined PRP-HA to stimulate cellular growth, restore components of the articular extracellular matrix, and reduce inflammation.


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