Revealing the Mechanism ofIn VitroWound Healing Properties ofCitrus tamuranaExtract

In the present investigation, we examined the effect of Hyuganatsu (Citrus tamurana) extract (HE) on skin fibroblast (TIG-119) proliferation and migration duringin vitrowound healing. HE selectively inhibited proliferation of TIG-119 cells at higher concentration (>1.0 mg/mL); at lower concentrations (0.1, 0.25, 0.5, and 0.75 mg/mL), it exhibited linear and time-dependent cell proliferation.In vitroscratch wound healing studies showed that the HE also accelerated the migration of cells towards the wounded region. Cytometric analysis demonstrated that HE extract did not alter G1/0 and S phases of cell cycle in any concentration studied; however, G2/M phases of cell cycle were significantly (P<0.05) accelerated at 0.75 mg/mL dose. RT-PCR and Western blotting analysis indicated that HE markedly overexpressed levels of Rac-1, Rho-A, and Cdc-42 mRNA and the respective proteins. Cyclin-dependent kinases (Cdk-1 and -2) gene expression activity was significantly (P<0.05) increased, but protein content decreased during treatment with HE. The induction of Cdk-1 and -2 by HE was abolished by inhibitors, transcription (DRB), and translation (CHX), implying transcriptional regulation that requiredde novoprotein synthesis.

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Migration and Proliferation Effects of Thymoquinone-Loaded Nanostructured Lipid Carrier (TQ-NLC) and Thymoquinone (TQ) on In Vitro Wound Healing Models

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2019/9725738 ◽

2019 ◽

Vol 2019 ◽

pp. 1-14 ◽

Cited By ~ 1

Author(s):

Henna Roshini Alexander ◽

Sharifah Sakinah Syed Alwi ◽

Latifah Saiful Yazan ◽

Fatin Hanani Zakarial Ansar ◽

Yong Sze Ong

Keyword(s):

Wound Healing ◽

Nigella Sativa ◽

Drug Carrier ◽

Healing Process ◽

Diabetic Patients ◽

Nanostructured Lipid Carrier ◽

Impaired Wound Healing ◽

And Migration ◽

Proliferation And Migration

Wound healing is a regulated biological event that involves several processes including infiltrating leukocyte subtypes and resident cells. Impaired wound healing is one of the major problems in diabetic patients due to the abnormal physiological changes of tissues and cells in major processes. Thymoquinone, a bioactive compound found in Nigella sativa has been demonstrated to possess antidiabetic, anti-inflammatory, and antioxidant effects. Today, the rapidly progressing nanotechnology sets a new alternative carrier to enhance and favour the speed of healing process. In order to overcome its low bioavailability, TQ is loaded into a colloidal drug carrier known as a nanostructured lipid carrier (NLC). This study aimed to determine the effect of TQ-NLC and TQ on cell proliferation and migration, mode of cell death, and the antioxidant levels in normal and diabetic cell models, 3T3 and 3T3-L1. Cytotoxicity of TQ-NLC and TQ was determined by MTT assay. The IC10 values obtained for 3T3-L1 treated with TQ-NLC and TQ for 24 hours were 4.7 ± 3.3 and 5.3 ± 0.6 μM, respectively. As for 3T3, the IC10 values obtained for TQ-NLC and TQ at 24 hours were 4.3 ± 0.17 and 3.9 ± 2.05 μM, respectively. TQ-NLC was observed to increase the number of 3T3 and 3T3-L1 healthy cells (87–95%) and gradually decrease early apoptotic cells in time- and dose-dependant manner compared with TQ. In the proliferation and migration assay, 3T3-L1 treated with TQ-NLC showed higher proliferation and migration rate (p<0.05) compared with TQ. TQ-NLC also acted as an antioxidant by reducing the ROS levels in both cells after injury at concentration as low as 3 μM. Thus, this study demonstrated that TQ-NLC has better proliferation and migration as well as antioxidant effect compared with TQ especially on 3T3-L1 which confirms its ability as a good antidiabetic and antioxidant agent.

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Effects of a New Bioceramic Material on Human Apical Papilla Cells

Journal of Functional Biomaterials ◽

10.3390/jfb9040074 ◽

2018 ◽

Vol 9 (4) ◽

pp. 74 ◽

Cited By ~ 16

Author(s):

Diana Sequeira ◽

Catarina Seabra ◽

Paulo Palma ◽

Ana Cardoso ◽

João Peça ◽

...

Keyword(s):

Wound Healing ◽

Mineral Trioxide Aggregate ◽

Cellular Adhesion ◽

Cellular Migration ◽

Cellular Viability ◽

Migration Rates ◽

Apical Papilla ◽

And Migration ◽

Proliferation And Migration

Background: The development of materials with bioregenerative properties is critically important for vital pulp therapies and regenerative endodontic procedures. The aim of this study was to evaluate the cytocompatibility and cytotoxicity of a new endodontic biomaterial, PulpGuard, in comparison with two other biomaterials widely used in endodontic procedures, ProRoot Mineral Trioxide Aggregate (MTA) and Biodentine. Methods: Apical papilla cells (APCs) were isolated from third molars with incomplete rhizogenesis from patients with orthodontic indication for dental extraction. Cultured APCs were incubated for 24, 48, or 72 h with different dilutions of eluates prepared from the three materials. Cellular viability, mobility, and proliferation were assessed in vitro using the Alamar Blue assay and a wound-healing test. The cells were also cultured in direct contact with the surface of each material. These were then analyzed via Scanning Electron Microscopy (SEM), and the surface chemical composition was determined by Energy-Dispersive Spectroscopy (EDS). Results: Cells incubated in the presence of eluates extracted from ProRoot MTA and PulpGuard presented rates of viability comparable to those of control cells; in contrast, undiluted Biodentine eluates induced a significant reduction of cellular viability. The wound-healing assay revealed that eluates from ProRoot MTA and PulpGuard allowed for unhindered cellular migration and proliferation. Cellular adhesion was observed on the surface of all materials tested. Consistent with their disclosed composition, EDS analysis found high relative abundance of calcium in Biodentine and ProRoot MTA and high abundance of silicon in PulpGuard. Significant amounts of zinc and calcium were also present in PulpGuard discs. Concerning solubility, Biodentine and ProRoot MTA presented mild weight loss after eluate extraction, while PulpGuard discs showed significant water uptake. Conclusions: PulpGuard displayed a good in vitro cytocompatibility profile and did not significantly affect the proliferation and migration rates of APCs. Cells cultured in the presence of PulpGuard eluates displayed a similar profile to those cultured with eluates from the widely used endodontic cement ProRoot MTA.

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Extracellular matrix derived peptides and mesenchymal stem cell motility

10.26686/wgtn.17152088.v1 ◽

2021 ◽

Author(s):

◽

Sandi Grainne Dempsey

Keyword(s):

Extracellular Matrix ◽

Wound Healing ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Cell Motility ◽

Cell Attachment ◽

Mass Spectrometry Analysis ◽

And Migration ◽

Proliferation And Migration

<p>Biomaterials derived from decellularised extracellular matrices have shown promise as tools in tissue regeneration and wound healing. Such materials display biocompatibility as well as inherent bioactivity, promoting constructive remodelling in healing tissues. In this study, the bioactivity of ovine forestomach matrix (a decellularised extracellular matrix biomaterial) is assessed based on its ability to affect the proliferation and migration of wound healing cells. This material supported cell attachment and proliferation, but did not allow cell infiltration in vitro. Enzymatic digestion of the material rendered soluble components that were able to induce proliferation and migration of some cell types. Cell-mediated processing of the material generated a protein or proteins with chemotactic activity for mesenchymal stem cells in vitro. Mass spectrometry analysis indicated the bioactive component consisted of the proteoglycan decorin, or fragments thereof. Decorin has not previously been shown to induce mesenchymal stem cell motility, and these findings may add to what is known about decorin and its role in constructive remodelling. Furthermore, this cell-mediated approach for ECM breakdown could lead to the discovery of other bioactive peptides involved in ECM remodelling and wound healing.</p>

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Achyrocline satureioides (Lam.) DC (Asteraceae) Extract-Loaded Nanoemulsions as a Promising Topical Wound Healing Delivery System: In Vitro Assessments in Human Keratinocytes (HaCaT) and HET-CAM Irritant Potential

Pharmaceutics ◽

10.3390/pharmaceutics13081241 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1241

Author(s):

Lucélia Albarello Balestrin ◽

Tainá Kreutz ◽

Flávia Nathiely Silveira Fachel ◽

Juliana Bidone ◽

Nicolly Espindola Gelsleichter ◽

...

Keyword(s):

Wound Healing ◽

Hacat Cell ◽

Chorioallantoic Membrane ◽

Human Keratinocytes ◽

Scratch Assay ◽

Hen’S Egg ◽

And Migration ◽

Achyrocline Satureioides ◽

Proliferation And Migration

Achyrocline satureioides (Lam.) DC Asteraceae extracts (ASEs) have been investigated for the treatment of various skin disorders. This study reports the effects of ASE-loaded nanoemulsions (NEASE) on the cellular viability, death by necrosis, and migration of immortalized human keratinocytes (HaCaT cell line), as well as the irritant potential through the hen’s egg chorioallantoic membrane test (HET-CAM). NEASE exhibited a polydispersity index above 0.12, with a droplet size of 300 nm, ζ-potential of −40 mV, and content of flavonoids close to 1 mg/mL. No cytotoxicity of the ASE was observed on HaCaT by MTT assay (up to 10 µg/mL). A significant increase of HaCaT viability was observed to NEASE (up to 5 μg/mL of flavonoids), compared to treatment with the ASE. The necrosis death evaluation demonstrated that only NEASE did not lead to cell death at all the tested concentrations. The scratch assay demonstrated that NEASE was able to increase the cell migration at low flavonoid concentrations. Finally, the HET-CAM test proved the non-irritative potential of NEASE. Overall, the results indicate the potential of the proposed formulations for topical use in wound healing, in view of their promising effects on proliferation and migration in keratinocytes, combined with an indication of the absence of cytotoxicity and non-irritating potential.

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The Activity of Combination of Hydrolyzed Virgin Coconut Oil and Chitosan Toward Wound Healing Parameters on NIH 3T3 Cells Using in Vitro Methods

Asian Journal of Pharmaceutical Research and Development ◽

10.22270/ajprd.v7i3.534 ◽

1970 ◽

Vol 7 (3) ◽

pp. 14-19 ◽

Cited By ~ 1

Author(s):

Hekdin Marsius Sipayung ◽

Jansen Silalahi ◽

Yuandani Y

Keyword(s):

Cell Proliferation ◽

Wound Healing ◽

Protein Expression ◽

Scratch Wound ◽

Cell Proliferation Activity ◽

Proliferation Activity ◽

Cox 2 ◽

Nih 3T3 ◽

Vegf Protein

Objectives: The objective of this study was to investigate the activity of combination of hydrolyzed VCO (HVCO) and chitosan on NIH 3T3 cell proliferation activity, NIH 3T3 cell migration, COX-2 and VEGF protein expression. Design: In vitro cytotoxic assay was determined by MTT (MicrocultureTetrazoliumTehnique) assay, cell proliferation activity was measured by calculating cell viability incubated 24 hours, 48 hours and 72 hours, wound closure percentage was tested by scratch wound healing method, expression of COX-2 protein and VEGF protein were measured by immunocytochemical method. Interventions: The variable that was intervened in this study was the concentration of HVCO and chitosan. Main Outcome Measures: The main measurements carried out in this study were the absorbance value of HVCO and chitosan which was converted into viability cell, proliferation activity, percentage of wound closure, and percentage of COX-2 and VEGF protein expression. Results: Cytotoxic activity of HVCO and chitosan resulted the best concentration at 31.25 μg/ml, scratch wound healing assay from a combination HVCO and chitosan resulted the best migration of fibroblast cells at a ratio of 1:1 with HVCO 62.5 μg/ml and chitosan 62.5 μg/ml, combination of HVCO 62.5 μg/ml and chitosan 62.5 μg/ml (1:1) increased expression of COX-2 and VEGF. Conclusion: Combination of HVCO and chitosan could increase NIH 3T3 cell migration, COX-2 and VEGF protein expression. Combination of HVCO and chitosan had better wound healing activity in vitro than single use. Keywords: Rhizomucor miehei, viability, proliferation, migration, expression

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Positive feedback between lncRNA FLVCR1-AS1 and KLF10 may inhibit pancreatic cancer progression via the PTEN/AKT pathway

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-021-02097-0 ◽

2021 ◽

Vol 40 (1) ◽

Author(s):

Jiewei Lin ◽

Shuyu Zhai ◽

Siyi Zou ◽

Zhiwei Xu ◽

Jun Zhang ◽

...

Keyword(s):

Cell Cycle ◽

Pancreatic Cancer ◽

Cell Proliferation ◽

Cancer Progression ◽

Positive Feedback ◽

Molecular Mechanisms ◽

Tumor Tissues ◽

And Migration

Abstract Background FLVCR1-AS1 is a key regulator of cancer progression. However, the biological functions and underlying molecular mechanisms of pancreatic cancer (PC) remain unknown. Methods FLVCR1-AS1 expression levels in 77 PC tissues and matched non-tumor tissues were analyzed by qRT-PCR. Moreover, the role of FLVCR1-AS1 in PC cell proliferation, cell cycle, and migration was verified via functional in vitro and in vivo experiments. Further, the potential competitive endogenous RNA (ceRNA) network between FLVCR1-AS1 and KLF10, as well as FLVCR1-AS1 transcription levels, were investigated. Results FLVCR1-AS1 expression was low in both PC tissues and PC cell lines, and FLVCR1-AS1 downregulation was associated with a worse prognosis in patients with PC. Functional experiments demonstrated that FLVCR1-AS1 overexpression significantly suppressed PC cell proliferation, cell cycle, and migration both in vitro and in vivo. Mechanistic investigations revealed that FLVCR1-AS1 acts as a ceRNA to sequester miR-513c-5p or miR-514b-5p from the sponging KLF10 mRNA, thereby relieving their suppressive effects on KLF10 expression. Additionally, FLVCR1-AS1 was shown to be a direct transcriptional target of KLF10. Conclusions Our research suggests that FLVCR1-AS1 plays a tumor-suppressive role in PC by inhibiting proliferation, cell cycle, and migration through a positive feedback loop with KLF10, thereby providing a novel therapeutic strategy for PC treatment.

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Growth Differentiation Factor-9 Promotes Fibroblast Proliferation and Migration in Keloids through the Smad2/3 Pathway

Cellular Physiology and Biochemistry ◽

10.1159/000452538 ◽

2016 ◽

Vol 40 (1-2) ◽

pp. 207-218 ◽

Cited By ~ 8

Author(s):

Zhaohua Jiang ◽

Qingxiong Yu ◽

Lingling Xia ◽

Yi Zhang ◽

Xiuxia Wang ◽

...

Keyword(s):

Cell Cycle ◽

Wound Healing ◽

Growth Factor ◽

Fibroblast Proliferation ◽

Differentiation Factor ◽

Cox 2 ◽

And Migration ◽

Tgf Β1 ◽

Keloid Fibroblasts ◽

Proliferation And Migration

Background: Keloids are fibroproliferative scars that develop as a result of a dysregulated wound healing process; however, the molecular mechanisms of keloid pathogenesis remain unclear. Keloids are characterized by the ability to spread beyond the original boundary of the wound, and they represent a significant clinical challenge. Previous work from our group suggested that growth differentiation factor (GDF)-9 plays a role in the invasive behavior of keloids. Here, we examined the involvement of GDF-9 in keloid formation and spread and elucidated a potential underlying mechanism. Methods: The expression of GDF-9, cyclooxygenase (COX)-2, vascular epidermal growth factor (VEGF)-C, matrix metalloprotease (MMP)-2, MMP-9, transforming growth factor (TGF)-β1, and the related signaling pathway components in human keloid tissues or keloid fibroblasts (kFBs) was monitored by qRT-PCR and western blot. A series of overexpression and silencing experiments in normal and keloid fibroblasts were used to modify the expression of GDF-9. The effects of GDF-9 on kFB proliferation and migration were assessed using the CCK-8, cell cycle and scratch wound healing assays. Results: GDF-9 promotes fibroblast proliferation and migration. GDF-9 silencing in kFBs decreased cell proliferation, blocked cell cycle progression, downregulated the angiogenic markers COX-2 and VEGF-C, and downregulated MMP-2 and MMP-9 expression, whereas it had no effect on the levels of TGF-β1. GDF-9 silencing significantly inhibited Smad2 and Smad3 phosphorylation in kFBs. Conclusions: GDF-9 promotes the proliferation and migration of kFBs via a mechanism involving the Smad2/3 pathway.

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A Novel Gold Calreticulin Nanocomposite Based on Chitosan for Wound Healing in a Diabetic Mice Model

Nanomaterials ◽

10.3390/nano9010075 ◽

2019 ◽

Vol 9 (1) ◽

pp. 75 ◽

Cited By ~ 10

Author(s):

Sara Paola Hernández Martínez ◽

Teodoro Iván Rivera González ◽

Moisés Armides Franco Molina ◽

Juan José Bollain y Goytia ◽

Juan José Martínez Sanmiguel ◽

...

Keyword(s):

Wound Healing ◽

Topical Administration ◽

Nuclear Antigen ◽

Histological Evaluation ◽

Mice Model ◽

Fibroblast Migration ◽

And Migration ◽

Proliferating Cell

The development of new nanomaterials to promote wound healing is rising, because of their topical administration and easy functionalization with molecules that can improve and accelerate the process of healing. A nanocomposite of gold nanoparticles (AuNPs) functionalized with calreticulin was synthetized and evaluated. The ability of the nanocomposite to promote proliferation and migration was determined in vitro, and in vivo wound healing was evaluated using a mice model of diabetes established with streptozotocin (STZ). In vitro, the nanocomposite not affect the cell viability and the expression of proliferating cell nuclear antigen (PCNA). Moreover, the nanocomposite promotes the clonogenicity of keratinocytes, endothelial cells, and fibroblasts, and accelerates fibroblast migration. In vivo, mice treated with the nanocomposite presented significantly faster wound healing. The histological evaluation showed re-epithelization and the formation of granular tissue, as well as an increase of collagen deposition. Therefore, these results confirm the utility of AuNPs–calreticulin nanocomposites as potential treatment for wound healing of diabetic ulcers.

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Elacestrant (RAD1901) exhibits anti-tumor activity in multiple ER+ breast cancer models resistant to CDK4/6 inhibitors

Breast Cancer Research ◽

10.1186/s13058-019-1230-0 ◽

2019 ◽

Vol 21 (1) ◽

Cited By ~ 9

Author(s):

Hitisha K. Patel ◽

Nianjun Tao ◽

Kyung-Min Lee ◽

Mariela Huerta ◽

Heike Arlt ◽

...

Keyword(s):

Breast Cancer ◽

Cell Cycle ◽

Estrogen Receptor ◽

De Novo ◽

Single Agent ◽

Metastatic Breast ◽

Cancer Resistance ◽

Treatment Paradigm ◽

Anti Tumor Activity

Abstract Background Addition of CDK4/6 inhibitors (CDK4/6i) to endocrine therapy significantly increased progression-free survival, leading to their approval and incorporation into the metastatic breast cancer treatment paradigm. With these inhibitors being routinely used for patients with advanced estrogen receptor-positive (ER+) breast cancer, resistance to these agents and its impact on subsequent therapy needs to be understood. Considering the central role of ER in driving the growth of ER+ breast cancers, and thus endocrine agents being a mainstay in the treatment paradigm, the effects of prior CDK4/6i exposure on ER signaling and the relevance of ER-targeted therapy are important to investigate. The objective of this study was to evaluate the anti-tumor activity of elacestrant, a novel oral selective estrogen receptor degrader (SERD), in preclinical models of CDK4/6i resistance. Methods Elacestrant was evaluated as a single agent, and in combination with alpelisib or everolimus, in multiple in vitro models and patient-derived xenografts that represent acquired and “de novo” CDK4/6i resistance. Results Elacestrant demonstrated growth inhibition in cells resistant to all three approved CDK4/6i (palbociclib, abemaciclib, ribociclib) in both ESR1 wild-type and mutant backgrounds. Furthermore, we demonstrated that elacestrant, as a single agent and in combination, inhibited growth of patient-derived xenografts that have been derived from a patient previously treated with a CDK4/6i or exhibit de novo resistance to CDK4/6i. While the resistant lines demonstrate distinct alterations in cell cycle modulators, this did not affect elacestrant’s anti-tumor activity. In fact, we observe that elacestrant downregulates several key cell cycle players and halts cell cycle progression in vitro and in vivo. Conclusions We demonstrate that breast cancer tumor cells continue to rely on ER signaling to drive tumor growth despite exposure to CDK4/6i inhibitors. Importantly, elacestrant can inhibit this ER-dependent growth despite previously reported mechanisms of CDK4/6i resistance observed such as Rb loss, CDK6 overexpression, upregulated cyclinE1 and E2F1, among others. These data provide a scientific rationale for the evaluation of elacestrant in a post-CDK4/6i patient population. Additionally, elacestrant may also serve as an endocrine backbone for rational combinations to combat resistance.

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Pharmacological Inhibition of Cyclin-Dependent Kinases Triggers Anti-Fibrotic Effects in Hepatic Stellate Cells In Vitro

International Journal of Molecular Sciences ◽

10.3390/ijms21093267 ◽

2020 ◽

Vol 21 (9) ◽

pp. 3267

Author(s):

Anna Hübbers ◽

Julia Hennings ◽

Daniela Lambertz ◽

Ute Haas ◽

Christian Trautwein ◽

...

Keyword(s):

Cell Cycle ◽

Liver Fibrosis ◽

Hepatic Stellate Cells ◽

Healing Process ◽

Stellate Cells ◽

Wound Healing Process ◽

Cdk Inhibitor ◽

Cyclin E1 ◽

Cyclin Dependent Kinases

Liver fibrosis is a wound healing process in response to chronic liver injury, which is characterized by the accumulation of extracellular collagen produced by Hepatic Stellate Cells (HSCs). This process involves cell cycle re-entry and proliferation of normally quiescent HSCs controlled by cyclins and associated cyclin-dependent kinases (Cdks). Cdk2 mediates the entry and progression through S-phase in complex with E-and A-type cyclins. We have demonstrated that cyclin E1 is essential for liver fibrogenesis in mice, but it is not known if this is dependent on Cdk2 or related Cdks. Here, we aimed to evaluate the benefit of the pan-Cdk inhibitor CR8 for treatment of liver fibrosis in vitro. CR8-treatment reduced proliferation and survival in immortalized HSC lines and in addition attenuated pro-fibrotic properties in primary murine HSCs. Importantly, primary murine hepatocytes were much more tolerant against the cytotoxic and anti-proliferative effects of CR8. We identified CR8 dosages mediating anti-fibrotic effects in primary HSCs without affecting cell cycle activity and survival in primary hepatocytes. In conclusion, the pharmacological pan-Cdk inhibitor CR8 restricts the pro-fibrotic properties of HSCs, while preserving proliferation and viability of hepatocytes at least in vitro. Therefore, CR8 and related drugs might be beneficial for the treatment of liver fibrosis.

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