PDGF Gene Research Articles

The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study.

Metallic nanoparticles (NPs) exhibit interesting radiosensitizing effects, and finding a way to accurately deliver them appears to be crucial. Due to their tumor tropism, mesenchymal stem cells (MSCs) represent a strategic approach. Therefore, we aimed to evaluate the impact of core-shell Fe3O4@Au NPs on the functionality of human pulmonary MSCs (HPMSCs). The results showed that 100 µg/mL Fe3O4@Au NPs, accumulated in HPMSCs (revealed by Prussian blue staining), did not alter cell viability as assessed by cell counting, MTT, and LDH assays. However, caspase 9 and Bcl2 gene expression, evaluated by RT-qPCR, was regulated 72 h after exposure to the NPs. Moreover, the NPs also decreased proinflammatory cytokine/chemokine secretions, except for CXCL8 (ELISA). These modulations were associated with the downregulation of AMPK gene expression at 24 h. In contrast, the NPs did not modulate VEGF, PI3K, or PDGF gene expression. Nevertheless, a decrease in VEGF secretion was observed after 24 h of exposure to the NPs. Interestingly, the Fe3O4@Au NPs did not modulate Nrf2 gene expression, but they did regulate the expression of the genes encoding Nox4 and HMOX-1. Additionally, the NPs increased ROS production, suggesting a redox imbalance. Finally, the Fe3O4@Au NPs did not affect the HPMSCs' viability or proangiogenic/tumorigenic markers. These findings are encouraging for investigating the effects of Fe3O4@Au NPs delivered by HPMSCs to tumor sites in combination with radiation.

Alpha terpineol preconditioning enhances regenerative potential of mesenchymal stem cells in full thickness acid burn wounds

Regeneration of full thickness burn wounds is a significant clinical challenge. Direct stem cell transplantation at the wound site has a promising effect on wound regeneration. However, stem cell survival within the harsh wound environment is critically compromised. In this regard, preconditioning of stem cells with cytoprotective compounds can improve the efficiency of transplanted cells. This study evaluated the possible effect of alpha terpineol (αT) preconditioned mesenchymal stem cells (αT-MSCs) in full thickness acid burn wound. An optimized concentration of 10 μM αT was used for MSC preconditioning, followed by scratch assay analysis. A novel rat model of full thickness acid burn wound was developed and characterized via macroscopic and histological examinations. Treatment (normal and αT-MSCs) was given after 48 h of burn wound induction, and the healing pattern was examined till day 40. Skin tissues were harvested at the early (day 10) and late (day 40) wound healing phases and examined by histological grading, neovascularization, and gene expression profiling of healing mediators. In scratch assay, αT-MSCs exhibited enhanced cell migration and wound closure (scratch gap) compared to normal MSCs. In vivo findings revealed enhanced regeneration in the wound treated with αT-MSCs compared to normal MSCs and untreated control. Histology revealed enhanced collagen deposition with regenerated skin layers in normal MSC- and αT-MSC treated groups compared to the untreated control. These findings were correlated with enhanced expression of α-SMA as shown by immunohistochemistry. Additionally, αT-MSC group showed reduced inflammation and oxidative stress, and enhanced regeneration, as witnessed by a decrease in IL-1β, IL-6, TNF-α, and Bax and an increase in BCL-2, PRDX-4, GPX-7,SOD-1, VEGF, EGF, FGF, MMP-9, PDGF, and TGF-β gene expression levels at early and late phases, respectively. Overall findings demonstrated that αT exerts its therapeutic effect by mitigating excessive inflammation and oxidative stress while concurrently enhancing neovascularization. Thus, this study offers new perspectives on managing full thickness acid burn wounds in future clinical settings.

Therapeutic Potential of Secretome from Hypoxic-Mesenchymal Stem Cell (SH-MSC) in Regulating PDGF and IL-1β Gene Expression in Fluconazole-Related Alopecia

Therapeutic Potential of Secretome from Hypoxic-Mesenchymal Stem Cell (SH-MSC) in Regulating PDGF and IL-1β Gene Expression in Fluconazole-Related Alopecia

1H-NMR metabolomics analysis identifies hypoxanthine as a novel metastasis-associated metabolite in breast cancer

Breast cancer is one of the leading causes of death in females, mainly because of metastasis. Oncometabolites, produced via metabolic reprogramming, can influence metastatic signaling cascades. Accordingly, and based on our previous results, we propose that metabolites from highly metastatic breast cancer cells behave differently from less-metastatic cells and may play a significant role in metastasis. For instance, we aim to identify these metabolites and their role in breast cancer metastasis. Less metastatic cells (MCF-7) were treated with metabolites secreted from highly metastatic cells (MDA-MB-231) and the gene expression of three epithelial-to-mesenchymal transition (EMT) markers including E-cadherin, N-cadherin and vimentin were examined. Some metabolites secreted from MDA-MB-231 cells significantly induced EMT activity. Specifically, hypoxanthine demonstrated a significant EMT effect and increased the migration and invasion effects of MCF-7 cells through a hypoxia-associated mechanism. Hypoxanthine exhibited pro-angiogenic effects via increasing the VEGF and PDGF gene expression and affected lipid metabolism by increasing the gene expression of PCSK-9. Notably, knockdown of purine nucleoside phosphorylase, a gene encoding for an important enzyme in the biosynthesis of hypoxanthine, and inhibition of hypoxanthine uptake caused a significant decrease in hypoxanthine-associated EMT effects. Collectively for the first time, hypoxanthine was identified as a novel metastasis-associated metabolite in breast cancer cells and represents a promising target for diagnosis and therapy.

The Effect of Secretome Hypoxia Mesenchymal Stem Cells on PDGF and IL-1b Gene Expression (Experimental Study on Wistar Rats Hyperglycemic Wound Models)

Hyperglycemic wounds take longer to heal, and high glucose results in an increase in inflammatory cells, a decrease in the angiogenesis process, and the production of growth factors. Antibiotic treatment, surgery, or debridement can increase the risk of amputation. This study aimed to determine the effect of administering gel secretome hypoxia mesenchymal stem cells (SH-MSCs) on the expression of PDGF and IL-1b in Wistar rats with a hyperglycemic wound model. In vivo laboratory experimental research with a Post Test Only Control Group Design. The samples consisted of 30 Wistar rats, divided into 5 groups, namely healthy rats (P1), negative control (P2), positive control with gentamicin (P3), gel secretome dose of 20 μL/rat (P4), and gel secretome dose of 40 μL/rat (P5). MSC secretome hypoxia gel treatment for 10 days, then skin tissue samples were examined using the RTq-PCR method to analyze PDGF and IL-1b gene expression. Analysis of PDGF gene expression showed significant differences between treatment groups using the Kruskal-Wallis test with a result of 0.018 (p<0.05), but there were no significant differences between groups using the Mann-Whitney test (p<0.05). Analysis of IL-1b gene expression showed significant differences between treatment groups using the Kruskal Wallis test with a result of 0.001 (p<0.05), various doses of secretome gel affected reducing IL-1b gene expression using the Mann Whitney test with a result of 0.004 (p<0, 05). The most significant decrease was at a secretome dose of 40 μL/mouse. MSC secretome hypoxia gel at a dose of 40 μL/rat effectively reduces IL-1b gene expression in Wistar rats with a hyperglycemic wound model. However, various doses of MSC secretome hypoxia gel do not significantly increase PDGF gene expression.

Abstract 14924: SM22-Cre-Mediated SOCS3 Deficient Mice Exhibit Pericardial Fibrosis, Diastolic Dysfunction, and Increased Mortality Rates Due to Augmented Activation of STAT3 in Fibroblasts

Background: Suppressor of cytokine signaling-3 (SOCS3) is a cytokine-inducible potent negative regulator of STAT3 signaling pathway. Here, we aimed to determine whether SOCS3 in non-cardiomyocytes would play a role in cardiovascular pathophysiology. Methods and Results: To investigate the role of SOCS3 in non-cardiomyocytes, we generated conditional knockout mice by crossing SOCS3-flox mice with SM22-α-Cre mice (SM22-SOCS3-KO mice). Mortality rates in the SM22-SOCS3-KO mice was significantly higher than that in the wild-type mice (WT). Heart weight to body weight ratio was significantly increased in SM22-SOCS3-KO mice compared with WT at 12 months of age. Echocardiographic analyses revealed that SM22-SOCS3-KO mice showed significantly increased left ventricular diastolic dysfunction compared with WT from 12 months of age. Sirius-red staining revealed that thickness of pericardium and cardiac interstitial fibrosis in SM22-SOCS3-KO mice were markedly greater compared with WT mice at 12 months of age. Western blot analyses showed that phosphorylated STAT3 was significantly increased in SM22-SOCS3-KO hearts compared with WT mice at 12 months of age. PCR array analysis revealed that the expression of pro-fibrotic ctgf , pdgf , and TGF-beta family genes including tgf-beta1 , tgf-beta2 , and tgf-beta3 , were significantly higher in SM22-SOCS3-KO hearts than those in WT at 6 months of age. We also found that circulating interleukin-6 concentration was significantly higher in SM22-SOCS3-KO mice compared with WT mice at 12 months of age. After interleukin-6 administration, in the heart tissue from SM22-SOCS3-KO mice, coimmunostaining of phosphorylated STAT3 and ER-TR7 revealed that STAT3 activation occurred in fibroblasts in the coronary arterial adventitia and pericardium. Conclusion: Thus, SM22-Cre-mediated SOCS3 deletion induces higher mortality rates, increased pericardial fibrosis, cardiac interstitial fibrosis, and increased diastolic dysfunction in aging mice, possibly through the increased activation of STAT3 in fibroblasts.

The Effect of Secretome Hypoxia Mesenchymal Stem Cell (Sh-MSC) Therapy on PDGF and IFN-Gama Gene Expression in Male Wistar Rat Excision Models

Wounds are damage to normal anatomical structures and functions due to pathological processes that originate internally or externally and affect certain organs. The wound healing process consists of four highly integrated and overlapping hemostasis phases, inflammation, proliferation, and remodeling or network resolution. Therapy using Mesenchymal Stem Cells (MSC) is considered effective in improving skin wound healing because it has the potential for differentiation and immunoregulation and there is no potential for post-treatment rejection. This research aims to prove the influence of secretome hypoxia mesenchymal stem cells (SH-MSC) on PDGF and IF-Gamma gene expression in Wistar rats using the excision wound model. This research is in-vivo experimental research with Post-test Only Control Group Design. The research subjects were 18 male Wistar rats with excision wound models divided into three groups there are placebo gel (negative control), Clobetasol at a dose of 0.25g/kg (positive control), and the P1 group SH-MSC gel at a dose of 400 μl/kg BW, were given treatment for 5 days. On the 6th day, the skin tissue was examined using the RT-PCR method to see the expression of the PDGF and IF-gamma genes. Statistical analysis using One-way ANOVA and Post Hoc LSD test. The average PDGF gene expression in the P1 group was the highest, followed by the positive control. The lowest average expression of the PDGF gene was found in the negative control. The average IFN-γ gene expression in the negative control was the highest, followed by the positive control group. The lowest average expression of the IFN-γ gene was found in the P1 group. The administration of SH-MSCs topical gel 400μl/kg BW increased PDGF expression in male Wistar rats with excision wound models, whereas the administration of gel secretome SH-MSC at a dose of 400μl/kg BW can reduce IFN-γ levels in male Wistar rats with the excision wound model.

Genetic Influence of Fracture Nonunion (FNU): A Systematic Review.

Nonunion of fractures occurs in about 15% of all fractures causing repeated surgical interference and prolonged morbidity. We performed this systematic review to assess genes and polymorphisms influencing fractures' nonunion (FNU). We searched between 2000 and July 2022 in PubMed, EMBASE, the Cochrane Central Register of Controlled Trials and Cochrane Database of Systematic Reviews, Genome Wide Association Studies (GWAS) Catalog, and the Science Citation Index, with the keywords nonunion of fractures, genetic influence, and GWAS. The exclusion criteria were review articles and correspondence. The data were retrieved to determine the number of studies, genes, and polymorphisms and the total number of subjects screened. A total of 79 studies were reported on nonunion of fractures and genetic influence. After the inclusion and exclusion criteria, ten studies with 4402 patients' data were analyzed. Nine studies were case-controlled, and 1 GWAS. It was identified that patients with polymorphisms in the genes ANXA3, BMP2, CALY, CYR61, FGFR1, IL1β, NOG, NOS2, PDGF gene, and TACR1 are prone to develop a nonunion of fractures. We believe that for patients who develop an early nonunion of fractures, a genetic study should be conducted for single nucleotide polymorphism (SNP) and genes so that alternative and more aggressive treatment can be performed to heal fractures without prolonged morbidity.

Gene Expression Behavior of a Set of Genes in Platelet and Tissue Samples from Patients with Breast Cancer.

The tumor microenvironment (TME) is constituted by a great diversity of highly dynamic cell populations, each of which contributes ligands, receptors, soluble proteins, mRNAs, and miRNAs, in order to regulate cellular activities within the TME and even promote processes such as angiogenesis or metastasis. Intravasated platelets (PLT) undergo changes in the TME that convert them into tumor-educated platelets (TEP), which supports the development of cancer, angiogenesis, and metastasis through the degranulation and release of biomolecules. Several authors have reported that the deregulation of PF4, VEGF, PDGF, ANG-1, WASF3, LAPTM4B, TPM3, and TAC1 genes participates in breast cancer progression, angiogenesis, and metastasis. The present work aimed to analyze the expression levels of this set of genes in tumor tissues and platelets derived from breast cancer patients by reverse transcription-quantitative polymerase chain reaction (RTqPCR) assays, in order to determine if there was an expression correlation between these sources and to take advantage of the new information to be used in possible diagnosis by liquid biopsy. Data from these assays showed that platelets and breast cancer tumors present similar expression levels of a subset of these genes' mRNAs, depending on the molecular subtype, comorbidities, and metastasis presence.

Insertion/Deletion (InDel) Variants within the Sheep Fat-Deposition-Related PDGFD Gene Strongly Affect Morphological Traits.

Platelet-derived growth factor D (PDGFD) is a member of the PDGF gene family, and it plays an important role in the regulation of adipocyte development in mammals. Furthermore, genome-wide association studies (GWAS) have previously identified it as a candidate gene associated with fleece fiber variation, body size, and the fat-tail phenotype in domestic Chinese sheep. In this study, a total of 1919 indigenous Chinese sheep were genotyped to examine the association between nucleotide sequence variations in PDGFD and body morphology. Our results detected both a 14 bp insertion in intron 2 and a 13 bp deletion in intron 4 of PDGFD. Moreover, these two InDel loci had low to moderate polymorphism. Notably, the 13 bp deletion mutation of PDGFD was found to significantly affect sheep body size. Yearling rams in the Luxi black-headed sheep (LXBH) containing a heterozygous genotype (insertion/deletion, ID) were found to have larger body length, chest depth, and body weight than those with wild genotypes. Furthermore, adult ewes in the Guiqian semi-fine wool sheep (GSFW) containing a homozygous mutation (deletion/deletion, DD) were found to have smaller chest width than their peers. Moreover, yearling ewes in this group with the same homozygous mutation were found to have lower body weight, chest width, and cannon circumference compared to those of other individuals. This study demonstrates that PDGFD InDel polymorphisms have the potential to be effective molecular markers to improve morphological traits in domestic Chinese sheep.

Precision medicine in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune disease that has diverse clinical manifestations, ranging from restricted cutaneous involvement to life-threatening systemic organ involvement. The heterogeneity of pathomechanisms that lead to SLE contributes to between-patient variation in clinical phenotype and treatment response. Ongoing efforts to dissect cellular and molecular heterogeneity in SLE could facilitate the future development of stratified treatment recommendations and precision medicine, which is a considerable challenge for SLE. In particular, some genes involved in the clinical heterogeneity of SLE and some phenotype-related loci (STAT4, IRF5, PDGF genes, HAS2, ITGAM and SLC5A11) have an association with clinical features of the disease. An important part is also played by epigenetic varation (in DNA methylation, histone modifications and microRNAs) that influences gene expression and affects cell function without modifying the genome sequence. Immune profiling can help to identify an individual's specific response to a therapy and can potentially predict outcomes, using techniques such as flow cytometry, mass cytometry, transcriptomics, microarray analysis and single-cell RNA sequencing. Furthermore, the identification of novel serum and urinary biomarkers would enable the stratification of patients according to predictions of long-term outcomes and assessments of potential response to therapy.

Isolation of oleanolic acid from Lavandula stoechas and its potent anticancer properties against MCF-7 cancer cells via induced apoptosis

Medicinal plants are natural resources of diversified therapeutic metabolites. Lavandula stoechas has been used traditionally for the management of different metabolic disorders. However, studies on the anticancer characteristics of certain bioactive compounds present in this plant are very limited. In view of this, the aim of the present study was to isolate oleanolic acid (a triterpenoid) from L. stoechas and assess its anticancer trait against breast carcinoma cells (MCF-7) via induced apoptosis. Initially, oleanolic acid was isolated from the ethyl acetate fraction of L. stoechas using standard extraction protocols. Furthermore, the structure of compound was established using mass spectrometry, FT-IR, 1H NMR, and 2D NMR (HSQC) techniques. In vitro antiproliferative effect of oleanolic acid was assessed against breast carcinoma cells (MCF-7 and MDA-MB-231) and non-cancerous cells (BHK-21) using the MTT assay. Oleanolic acid showed prominent cytotoxicity of 72.44 ± 3.33 and 78.55 ± 2.67 % against MCF-7 and MDA-MB-231 cells with IC50 values of 13.09 and 160.22 µg/mL, respectively. Moreover, the compound showed negligible toxicity against the BHK-21 cell line. Furthermore, the apoptotic features in oleanolic acid-treated MCF-7 cells were determined using fluorescent microscopy, which showed typical nuclear condensation, increased ROS generation, and the loss of outer mitochondrial membrane potentials. The levels of activated caspases in MCF-7 cancer cells were estimated using a fluorescent kit, showing a significant increment in caspase-9 and caspase-7 for oleanolic acid-treated MCF-7 cells. Moreover, oleanolic acid showed down-regulation of the Bcl2 and PDGF genes. Our findings demonstrated the pivotal role of oleanolic acid as pronounced anticancer agent against MCF-7 cell lines by inducing apoptotic mechanism via intrinsic pathway.

PDGF-AA promotes gap junction intercellular communication in chondrocytes via the PI3K/Akt pathway

ABSTRACT Background Gap junction intercellular communication (GJIC) plays an important role in cell growth, development and homeostasis. Connexin 43 (Cx43) is an important half-channel protein responsible for gap junction formation. Platelet-derived growth factor AA (PDGF-AA) regulates the proliferation, migration, metabolism, apoptosis and cell cycle of chondrocytes. However, the role of PDGF-AA in gap junction intercellular communication in chondrocytes is not fully understood. In the current study, we performed experiments to explore the effect of PDGF-AA on GJIC and its underlying biomechanical mechanism. Methods qPCR was performed to determine the expression of PDGF, PDGFR and connexin family genes in chondrocytes and/or cartilage. A scrape loading/dye transfer assay was used to determine GJIC. Western blot analysis was applied to detect the expression of Cx43 and PI3K/Akt signaling pathway proteins. Immunofluorescence staining was utilized to examine protein distribution. Scanning electron microscopy was used to delineate the morphology of chondrocytes. Results Expression of PDGF-A mRNA was highest among the PDGF family in chondrocytes and cartilage tissues. PDGF-AA promoted functional GJIC formation in chondrocytes by upregulating the expression of Cx43. Enhanced functional GJIC formation in chondrocytes induced by PDGF-AA occurred through the activation of PI3K/Akt signaling and its nuclear accumulation. Conclusion For the first time, this study provides evidence demonstrating the role of PDGF-AA in cell-to-cell communication in chondrocytes through mediating Cx43 expression.

The effect of Chaihu-shugan-san on cytotoxicity induction and PDGF gene expression in cervical cancer cell line HeLa in the presence of paclitaxel +cisplatin.

Chaihu-shugan-san, as a traditional Chinese herbal formula, is composed of seven different herbs. This medicine can treat cancer due to its antioxidant compounds. In this study, the effect of Chaihu-shugan-san was considered on cytotoxicity induction and PDGF gene expression in cervical cancer cell line HeLa at different concentrations and at different times, by the MTT method. Paclitaxel + cisplatin were used as a control in this study. The expression of the PDGF gene was quantitatively evaluated in treated cells by real-time PCR, and a generalized linear model was used to evaluate the effect of the medicine, and Duncan's multiple range tests were used to evaluate the data. The results of the MTT test showed that Chaihu-shugan-san had antitumor properties in different concentrations, but there was a significant difference between this medicine and paclitaxel +cisplatin. Also, examination of gene expression showed that this medicine reduced the expression of the PDGF gene in the HeLa cancer cell line (P ? 0.04). Therefore, Chaihu-shugan-san could be suggested as an effective factor in preventing the growth of cervical cancer cells and controlling angiogenic factors that play an important role in the metastasis of cancerous tumors.

Abstract 3062: Proton irradiation sensitizes cancer cell lines with acquired radioresistance by exploiting differentially dysregulated DNA damage response (DDR) pathways

Abstract Radiotherapy (RT) is a primary treatment modality that is used in 50% of cancer patients. Clinically, tumor radioresistance poses a challenge to achieve complete anti-tumor response following conventional photon-RT. Diverse mechanisms have been proposed to underpin radioresistance. However, targeting these aberrant pathways to overcome tumor radioresistance remains a clinical challenge. Here, we investigate if proton beam therapy (PBT) invokes differential cellular responses compared to photon-RT in a broad panel of radioresistant (RR) cancer cell lines. We generated isogenic RR human cancer cell lines: 22Rv1 prostate cancer, FaDu hypopharyngeal cancer and C666-1 nasopharyngeal cancer, by exposure of respective wildtype (wt) to 90 Gy photon-RT (2 Gy x 45 fr). RR was confirmed by clonogenic survival, with surviving fraction ratio (SFRR/SFwt) at 1-4 Gy (range: 1.2-1.8). Whole exome sequencing (100x; Illumina NovaSeq) was performed to profile RR-associated mutational drivers. We performed transcriptomic and proteome profiling of cellular response such as non-homologous end-joining (NHEJ) and homologous recombination (HR) repair at 1-24 h after 4 Gy PBT (2.5 MeV, 2 Gy/min) and photon-RT (0.66 MeV, 0.716 Gy/min). At baseline, we found common mutations and differential activation of several pro-survival (Akt, mTOR), epithelial-mesenchymal transition (EMT) (PDGF, TGFb) and DNA repair pathways (ATM, BRCA1, NHEJ- and HR-associated) genes in RR- relative to wt-22Rv1. Post-photon-RT, we observed residual persistent DNA repair in wt-cell lines up to 48 h. In contrast, repair in RR-cell lines was more proficient, as shown by recovery of γH2AX, ATM-Chk2 within 6 h, with significant NHEJ activation (DNA-PKcs). Next, we observed that RR-cell lines were more sensitive to PBT than photon-RT (mean SF4Gy PBT/SF4Gy photon-RT = 0.61), with delayed DNA repair (γH2AX) observed up to 48 h, which may be attributed to the difficulty in repairing PBT-induced DNA damage. This phenomenon, seems to be primarily orchestrated by decreased NHEJ activation combined with diminished cell cycle checkpoint arrest (p21) and anti-apoptotic (bcl-2) signaling. Radiosensitization by PBT was however not observed in wt (mean SF4Gy proton/SF4Gy photon = 1.14), corroborated by gene and protein expression of DDR-related pathways. We observed dysregulation of multiple pathways relating to EMT and DNA repair in our panel of RR-cancer cell lines, compared to the wt-counterparts. Additionally, we show the potential for PBT to overcome radioresistance in these models by targeting the DNA repair machinery. Due to the high cost of PBT, precise patient stratification is needed to ensure that PBT is applied only to patients who benefit significantly from this procedure over photon-RT. Our preliminary data represents a proof-of-concept for feasibility of patient stratification to PBT. Citation Format: Pek Lim Chu, Eugenia L.L. Yeo, Dennis J.J. Poon, Ce-belle Chen, Minqin Ren, Saumitra Vajandar, Dewi Susanti, Hong Qi Tan, Sung Yong Park, Thomas Osipowicz, Kwok Wai Lo, Stanley K. Liu, Khee Chee Soo, Melvin Lk Chua. Proton irradiation sensitizes cancer cell lines with acquired radioresistance by exploiting differentially dysregulated DNA damage response (DDR) pathways [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 3062.

Synthesis of a novel nanocomposite containing chitosan as a three‐dimensional printed wound dressing technique: Emphasis on gene expression

In this study, a highly porous three-dimensional (3D)-printed wound healing core/shell scaffold fabricated using poly-lactic acid (PLA). The core of scaffold was composed of hyaluronic acid (HA), copper carbon dots (Cu-CDs), rosmarinic acid, and chitosan hydrogel. Cu-CDs were synthesized using ammonium hydrogen citrate under hydrothermal conditions. Formulation containing 1mgml-1 concentration of Cu-CDs showed an excellent antibacterial activity against gram bacteria. At 0.25 mgml-1 of Cu-CDs concentration, scaffold had a good biocompatibility as confirmed by cytotoxicity assay on L929 fibroblast stem cells. in vivo wound healing experiments on groups of rats revealed that after 15 days of treatment, the optimal formulation of composite scaffold significantly improves the wound healing process compared to the PLA scaffold. This finding was confirmed by histological analysis and the relative expression of PDGF, TGF-β, and MMP-1 genes. The biocompatible antibacterial CU-CDS/PLA/HA/chitosan/rosmarinic acid nanocomposite is a promising wound healing scaffold which highly accelerates the process of skin regeneration.

Synthesis and characterization of new 1,2,4-triazole anticancer scaffold derivatives: In Vitro study

Four Schiff bases were obtained via one direct click reaction between 3-amino-1H-1,2,4-triazole and different substituted benzaldehyde. The synthesized compounds were characterized on the basis of their spectral data including IR spectra and 1HNMR. The synthesized compounds were tested against three human cancer cell lines to evaluate their in vitro anticancer activity, and also tested on Vero cells extracted from African green monkey kidney to investigate their side effect. The screening showed that the Schiff bases TB-NO2 and TB-OCH3 produced an effective anticancer activity against HEPG2, HCT-116, and MCF-7 cell lines. Further investigations were performed to determine the apoptotic and antifibrotic ability of TB-NO2 and TB-OCH3 on MCF-7 cells by determining the expression of Bax, Bcl-2, CTGF, and PDGF. Results of Bax/Bcl-2 ratio for TB-NO2 and TB-OCH3 confirmed the apoptotic effect of these compounds on MCF-7 cells. The reduction in CTGF and PDGF genes expression confirmed the regulatory effect of these compounds on MCF-7 cells growth.

Application of Spirulina-Chitosan Nano Hydrogel for Enhanced Wound Healing through Alteration of Expression Pattern of TGF-andszlig; and PDGF Genes

Background: The wound healing process comprises of highly complex and dynamic events which allow the reestablishment of skin's structural integrity. To overcome the drawbacks associated with this process, researchers have been focused on the development of new therapeutics. To meet this demand, an interesting option is Chitosan based biopolymers. This study develops and evaluates a combination of Chitosan and bioactive Spirulina microalgae, extract hydrogel as skin serum to enhance the wound healing process. Methods: Crude extract of dry S. platensis was prepared using three solvents (methanol, ethanol and water). Spirulina Extract-Chitosan Hydrogels (SECH) was synthesized using an ionic dilution method. SECH was characterized using Dynamic Light Scattering (DLS) and TEM microscopy. In vitro cytotoxicity of nano hydrogels on HFF-1 fibroblast Skin Normal cells was determined using MTT assay. Scratch assay was performed to evaluate the wound healing properties of SECH. QReal Time PCR was applied to evaluate the effect of SECH on gene expression of TNF-s and PDGF. Results: Aqueous SECH (SECH-W) showed higher proliferation activity compared with the control group by 1.78- fold and 2.55-fold increase of cell viability after 24 and 48 hr respectively with 3 mg/ml concentration. Aqueous SECH enhanced wound healing process on HFF-1 cell line with 100% wound closure after 48 hrs. TNF-s and PDGF gene expressions were up-regulated by 3-fold and 4.2-fold after 24 h incubation, respectively. Conclusions: Our findings showed that the SECH-W increased the proliferation of HFF-1 cells significantly and promotes the wound healing process in vitro. Therefore, the Spirulina-chitosan hydrogel could be of potential value in cosmeceutical and biomedical applications.

Effect of PDGF-B Gene-Activated Acellular Matrix and Mesenchymal Stem Cell Transplantation on Full Thickness Skin Burn Wound in Rat Model.

Full thickness burn wounds are lack of angiogenesis, cell migration, epithelialisation and finally scar tissue formation. Tissue engineered composite graft can provide sustained release of growth factor and promote the wound healing by cell migration, early angiogenesis and proliferation of extracellular matrix and wound remodeling. The objective of this study was to evaluate the gene embedded (pDNA-platelet-derived growth factor, PDGF-B) porcine acellular urinary bladder matrix with transfected mesenchymal stem cells (rBMSC) on healing of full thickness burn wound in rat model. Full thickness burn wound of 2 × 2cm size was created in dorsum of rat model under general anesthesia. Burn wounds were treated with silver sulfadiazine; porcine acellular urinary bladder matrix (PAUBM); PAUBM transfected with pDNA-PDGF-B; PAUBM seeded with rBMSC; PAUBM seeded with rBMSC transfected with pDNA-PDGF-B in groups A, B, C, D and E respectively. The wound healing was assessed based on clinical, macroscopically, immunologically, histopathological and RT-qPCR parameters. Wound was significantly healed in group E and group D with early extracellular matrix deposition, enhanced granulation tissue formation and early angiogenesis compared to all other groups. The immunologic response against porcine acellular matrix showed that PDGF-B gene activated matrix along with stem cell group showed less antibody titer against acellular matrix than other groups in all intervals. PDGF gene activated matrix releasing the PDGF-B and promote the healing of full thickness burn wound with neovascularization and neo tissue formation. PDGF gene also enhances secretion of other growth factors results in PDGF mediated regenerative activities. This was confirmed in RT-qPCR at various time intervals. Gene activated matrix encoded for PDGF-B protein transfected stem cells have been clinically proven for early acceleration of angiogenesis and tissue regeneration in burn wounds in rat models. Evaluation of PDGF-B gene-activated acellular matrix and mesenchymal stem cell in full thickness skin burn wound in rat.

Mechanisms underlying the wound healing and tissue regeneration properties of Chenopodium album.

In the current study, aerial parts (leaves, stem and shoots) of C. album were extracted with methanol and subjected to phytochemical and HPLC analysis. Agar well diffusion method was used for anti-bacterial activity against Gram-negative strains Escherichia coli, Salmonella typhi, Klebsiella, Pseudomonas aeruginosa and Gram-positive Bacillus cereus, Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus. Burn was induced through flame heated metal rod on mice. C. album ointment (2% w/w), Vaseline (vehicle) and silver sulfadiazine (standard) were topically applied thrice daily for 15days. Wound area was measured on day 0, 5, 10 and 15. On last day, the wound tissues were excised and subjected to histopathological, quantitative PCR and immunohistochemical analysis. Phenols, alkaloids, phytosterols, tannins, saponins, flavonoids, carbohydrates and glycosides were detected in phytochemical analysis. HPLC chromatogram displayed peaks for rutin, quercetin, ascorbic acid, gallic acid and various other phyto-constituents. The extract exhibited zone of inhibition in millimeter (mm) against E. coli (12.3 ± 0.57), S. typhyi (14.6 ± 1.52), Klebsiella (11.8 ± 0.76), P. aeruginosa (12.3 ± 0.57), B. cereus (12.5 ± 1.29), S. aureus (18.3 ± 2.08), and MRSA (11.8 ± 0.76). The wound area in C. album group was significantly (60%) reduced as compared to vehicle group (11%). Histological analysis showed complete re-epithelialization and fine tissue in extract treated group. qPCR data revealed up-regulation of EGF, PDGF and TGF-β1 genes in extract treated group. Similarly, immunohistochemistry results confirmed heightened EGFR expression in extract treated group. Our findings suggest that C. album can promote wound healing and tissue regeneration through control of burns related infection and modulation of growth factors and its receptors.