Reduced Wound Healing Research Articles

Effectiveness of maggot extractions and secretion (E/S) of Lucilia sericata in reducing wound surface in experimental scalding burn injury

Abstract. Amiri ZS, Akbarzadeh K, Douraghi M, Abdi KM, Afshar AA, Ghaffari J, Kordshouli RS, Akbari M, Rafinejad J. 2021. Effectiveness of maggot extractions and secretion (E/S) of Lucilia sericata in reducing wound surface in an experimental scalding burn injury. Nusantara Bioscience 13: 11-15. Lucilia sericata larvae have been successfully used as medicinal larvae for wound healing. This study was performed to evaluate the therapeutic effects of the primary ointment made from excretory-secretory substances of L.sericata larvae on experimental burn wounds on rabbits under laboratory conditions. Extraction of ES from the third stage of L.sericata larvae and antibiogram testing was done. Four rabbits were randomly divided into four groups; three standard third-grade burn wounds were created on the back of each rabbit for intervention groups (B, D) of wounds were used respectively from the original ointment made from ES and ES without accompanying material. Nitrofurazone ointment was used for the positive (C) and placebo ointment was used for the negative (A). On the 21st day, all scars of wounds in groups B and D were separated and the wound was free of infectious tissue, whereas in groups A and C that the scars were clinging to the base. The level of the ulcer was significantly lower in the E/S intervention group and the infectious tissue was not observed during the treatment of group B and D wounds. Methicillin-resistant Staphylococcus aureus (MRSA) did not show resistance to excretory-secretory substances of L. sericata larvae. The debridement and recovery process was faster in the ES-treated group and the initial ointment than in the control group. The results showed that maggot extractions and secretion (E/S) of L. sericata in reducing wound healing is appropriate for treatment.

Effect of Selenium nanoparticles in wound healing

Background: Wound healing is a complex process necessary to repairing damaged tissues and preventing infection. Selenium nanoparticles were known by their antioxidant and antimicrobial effect that is important in wound healing. Aim: the aim of this study was to investigate the effect of selenium nanoparticles in reducing and accelerating the wound healing time in mice. Methods: Se NPs were synthesized by a simple wet chemical method. Then the experimental animals were divided into the following groups: Negative control group, Positive control group, selenium nanoparticles control group, wounded mice treated with selenium nanoparticles group. At the end of experiment, mice were scarified& skin tissue samples were collected for biological analysis [Vascular Endothelial cell Growth Factor, Collagenase I and Nitric oxide] and histopathological study. Results: The results showed that the percentage of wound area has been significantly reduced in Se NPs treated group compared to the positive control. The level of Vascular Endothelial Growth Factor and Collagenase I in Se NPs groups were significantly increased compared to positive control while nitric oxide was significantly decreased in Se NPs treated group. Skin tissue showed effect of selenium nanoparticles by regenerating and recavering skin layer. Conclusion: Selenium nanoparticles have an important role in accelerating and reducing wound healing time.

Novel Decompressive Hemicraniectomy Technique for Traumatic Brain Injury: Technical Note

Traumatic brain injury (TBI) is a significant cause of morbidity and mortality across all age groups. Decompressive hemicraniectomy is the treatment for TBI-related refractory intracranial hypertension. The traditional technique for this procedure can result in wound complications due to injury of the scalp flap's vascular supply, namely the superficial temporal and postauricular arteries. In this technical note we describe our experience using a novel technique that preserves both vascular territories by placing the inferior aspect of the incision posterior to the ear as opposed to anterior to it. This modification has the potential to reduce wound healing complications, especially in those at higher risk, while also reducing operative time by avoiding temporalis muscle incision and closure during procedure. After performing hospital chart review, a total of 7 patients were found who underwent this hemicraniectomy technique for severe TBI. Of these, 5 patients had this performed on the left side, and 2 patients had this performed on the right side. Six of the patients had an accompanying subdural hematoma, whereas 1 patient had no intracranial hemorrhage present. In each case, both the superficial temporal and postauricular arteries were preserved, and rapid healing of the scalp flap occurred. In addition to providing a large bone window to allow the brain to swell, this technique has the potential to reduce complications of wound healing by preserving the vascular supply of the scalp flap and reduce operative times by minimizing temporalis muscle dissection.

Anti-Oxidant, Anti-Hemolytic Effects of Crataegus aronia Leaves and Its Anti- Proliferative Effect Enhance Cisplatin Cytotoxicity in A549 Human Lung Cancer Cell Line.

Objective:For Arabian traditional medicine, Crataegus aronia syn. Azarolus (L) Bosc. ex DC (Rosaceae) is widely used to treat diabetes, sexual weakness, cardiovascular diseases and cancer. The anti-cancerous and anti-hemolysis effects of the hydroalcoholic extract of this plant have never been investigated before. The present study aims to evaluate the biological activities of the hydroalcoholic extract of Crataegus aronia leaves in combination with cisplatin, one of the most widely employed chemotherapeutics, on A549 human lung cancer cell line. Methods:The anti-oxidant and anti-proliferative activities of leaves, fruits, seeds of C. aronia were investigated by DPPH method and MTT assay; respectively. Cell migration activity was investigated by wound healing and by cell aggregation assays. The effect of C. aronia in inducing cell cycle arrest along with activating cell apoptosis was evaluated by flow cytometry and Western blot assays, respectively. Results:Our results showed that C. aronia leaves (C. aronia L.) had the highest anti-oxidant and anti-proliferative activities. The leaves extract was potent against hemolysis of the human erythrocytes and showed elevated decrease in migration by reducing wound healing migration and by increasing cell aggregation. Finally, C. aronia L. treatment exhibited apoptotic activity on A549 cells by the down-regulation of PARP-1, caspase-3 and Bcl-2 proteins and by increasing the percentage of A549 cells in sub G0 cell cycle. Moreover, the co-treatment of C. aronia L. and cisplatin remarkably sensitised A549 cells to cisplatin. Conclusion:The results suggested that C. aronia L. could be used as a potential treatment against human lung cancer exhibiting minimal side effects on human health.

Effectiveness of recombinant human granulocyte macrophage colony-stimulating factor for treating deep second-degree burns: a systematic review and meta-analysis

IntroductionIt is uncertain whether treatment by recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) can promote healing of deep second-degree burns. This meta-analysis aimed to systematically review and assess randomised controlled...

Abstract 5970: Function and regulatory mechanism of GPM6A in hepatocellular carcinoma development and progression

Abstract BACKGROUND: GPM6A is a glycoprotein found in endothelial cells and is considered to be a malignant lymphoma oncogene. However, the role of GPM6A in the development of hepatocellular carcinoma (HCC) is unknown. Through Affymetrix gene expression microarray and bioinformatics analysis, very low GPM6A expression was found in HCC tissue. The present study aims to explore the function and regulatory mechanism of GPM6A in HCC development and progression. METHODS: Levels of GPM6A expression was determined in various hepatoma cell lines and HCC specimens from different base disorders, and its role on cell proliferation was investigated in GPM6A stably over-expressing cell lines. Modulating effects of a specific miRNA were evaluated with miRNA mimetic transfection. RESULTS: Much lower GPM6A levels were found in HCC occuring in HBV-infection or nonalcoholic steatohepatitis. TCGA data confirmed that low GPM6A expression in HCC correlates with a poor overall survival and progression-free survival rate (p=0.018 and 0.004). Over-expression of GPM6A inhibited proliferation in SMMC-7721 and MHCC-97H cells by 31% and 30%, respectively (p<0.01), and led to decreased colony formation by 40%. Overexpression of GPM6A was found to significantly reduce wound healing and migration rates in these two cell types. miR-96-5p was expressed at 6 to 61-fold higher in HCC tissues than adjacent tissues (p<0.01); whereas GPM6A gene expression was at 3 to 55-fold lower (p<0.01) in a negative correlation between miR-96-5p levels and GPM6A levels in HCC specimens (r=-0.64, p<0.05). Further experiments demonstrated that miR-96-5p acted directly on the 3'UTR of the GPM6A gene, and transfection of miR-96-5p significantly decreased GPM6A mRNA expression by 50% in two hepatoma cell lines. Moreover, MiR-96-5p transfection promoted proliferation (66%, 59%, p<0.01), migration (10%, 9%, p<0.05) and invasion (557±66 vs. 226±19; 235±36 vs. 153±17, p<0.05-0.01) of SMMC-7721 and MHCC-97H hepatoma cells; whereas the function of oncogenic microRNA-96 was significantly inhibited in GPM6A-overexpressed hepatoma cells. CONCLUSIONS: GPM6A expression in HCC is commonly suppressed, and correlated to a poor prognosis. Its low expression in HCC tissues is most likely attributed to upregulated miR-96-5p, which may function as an oncolytic regulator through suppressed GMP6A that may block proliferation, cloning formation, migration and invasion of hepatoma cells. Citation Format: Rui Zong Li, Gang Xu, Min Ji Zhu, Jian Wu, Jian Wu. Function and regulatory mechanism of GPM6A in hepatocellular carcinoma development and progression [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5970.

The effect of the macrophage migration inhibitory factor (MIF) on excisional wound healing in vivo

Background: The macrophage migration inhibitory factor (MIF) has been determined as a cytokine exerting a multitude of effects in inflammation and angiogenesis. Earlier studies have indicated that MIF may also be involved in wound healing and flap surgery. Methods: We investigated the effect of MIF in an excisional wound model in wildtype, Mif-/- and recombinant MIF treated mice. Wound closure rates as well as the macrophage marker Mac-3, the pro-inflammatory cytokine tumor necrosis factor α (TNFα) and the pro-angiogenic factor von Willebrand factor (vWF) were measured. Finally, we used a flap model in Mif-/- and WT mice with an established perfusion gradient to identify MIF’s contribution in flap perfusion. Results: In the excision wound model, we found reduced wound healing after MIF injection, whereas Mif deletion improved wound healing. Furthermore, a reduced expression of Mac-3, TNFα and vWF in Mif-/- mice was seen when compared to WT mice. In the flap model, Mif-/- knockout mice showed mitigated flap perfusion with lower hemoglobin content and oxygen saturation as measured by O2C measurements when compared to WT mice. Conclusions: Our data suggest an inhibiting effect of MIF in wound healing with increased inflammation and perfusion. In flaps, by contrast, MIF may contribute to flap vascularization.

Review Effect of Aloe vera on Wound Healing

Skin acts as a protective barrier of body. It keeps the internal body organs safe from external environmental conditions which can prove harmful for them. So, maintenance of skin integrity is very important for normal functioning of body. In our daily life body of both animals and humans encounter to different types of violence, some of them are of such a severity that they break the skin continuity. This breakage of skin continuity is termed as wound. Whenever, the skin continuity is broken, internal organs of body are exposed. This is a dangerous situation and can be fatal for life. Immediate response of skin after wound is the start of wound healing process which starts automatically. This process can be assisted in a number of ways by keeping the wound free from bacterial contamination, moisture and dirt. For this purpose a variety of antiseptic dressings are available which when applied topically support wound healing process. They prevent wound to be contaminated by any micro-organism and reduce wound healing time. Aloe vera is one of them, it possess significant wound healing properties. Herbs have been integral to both traditional and non-traditional forms of medicine dating back at least 5000 years. Herbal medicines, especially in wound management involve disinfection, debridement and provision of a suitable environment for aiding the natural course of healing. The super focus of this review is to summarize the findings of earlier scientists with authentic concluding verse for utilization of herbal medicines (Aloe vera) being safe, readily available and cheaper.

Effects of Vitamin D Receptor Knockout and Vitamin D Deficiency on Corneal Epithelial Wound Healing and Nerve Density in Diabetic Mice.

Diabetic keratopathy occurs in ∼70% of all people with diabetes. This study was designed to examine the effects of vitamin D receptor knockout (VDR-/-) and vitamin D deficiency (VDD) on corneal epithelial wound healing and nerve density in diabetic mice. Diabetes was induced using the low-dose streptozotocin method. Corneal epithelial wounds were created using an Algerbrush, and wound healing was monitored over time. Corneal nerve density was measured in unwounded mice. VDR-/- and VDD diabetic mice (diabetic for 8 and 20 weeks, respectively) had slower healing ratios than wild-type diabetic mice. VDR-/- and VDD diabetic mice also showed significantly decreased nerve density. Reduced wound healing ratios and nerve densities were not fully rescued by a supplemental diet rich in calcium, lactose, and phosphate. We conclude that VDR-/- and VDD significantly reduce both corneal epithelial wound healing and nerve density in diabetic mice. Because the supplemental diet did not rescue wound healing or nerve density, these effects are likely not specifically related to hypocalcemia. This work supports the hypothesis that low vitamin D levels can exacerbate preexisting ophthalmic conditions, such as diabetes.

Telocytes inhibited inflammatory factor expression and enhanced cell migration in LPS-induced skin wound healing models in vitro and in vivo

BackgroundCell proliferation and death are key components of wound healing and tissue repair. Telocytes (TCs) represent a newly discovered cell type that can protect tissue from acute injury via cell–cell communication with adjacent cells. The aim of this study was to use a mouse model of skin wound healing and lipopolysaccharide (LPS)-induced cell injury to evaluate the effects of TCs on skin wound healing in vivo and in vitro.Material/methodsImmunohistochemical staining was performed to evaluate the alteration of TCs in tissues from normal and chronic wound patients. Then, a male C57BL/6 mouse wound model of the back was established. The mice were divided randomly into three groups, and wound healing was estimated according to the wound healing rate and histology. An LPS-induced co-culture model of a mouse lung telocyte cell line (TCs) with human keratinocyte (HaCaT), human dermal microvascular endothelial cell (HDMEC) or murine fibroblast (L929) cell lines was established to analyse the effects of TCs on constitutive cell types of the skin. Cell proliferation, migration and apoptosis were examined, and reactive oxygen species (ROS) and inflammatory factors in HaCaT cells, HDMECs, and L929 cells were detected to study the mechanisms involved in TC protection in skin wounds.ResultsTCs were significantly increased in tissues from chronic wound patients compared with healthy controls. Wound healing was significantly improved in wound mouse models treated with exogenous TCs compared with LPS-induced models. TCs reversed the LPS-induced inhibition of HaCaT cells and HDMECs and reduced the LPS-induced apoptosis of HaCaT cells and the death ratios of HDMECs and L929 cells. TCs reversed LPS-induced ROS in HDMECs and L929 cells and decreased inflammatory factor mRNA levels in HaCaT cells, HDMECs and L929 cells.ConclusionsTCs reduce wound healing delay, and inflammatory responses caused by LPS might be mediated by inflammatory inhibition, thus restricting apoptosis and promoting migration of the main component cell types in the skin.

Nicotine suppresses proliferation and mineralized tissue-associated gene expressions of cementoblasts.

Previous studies reported that nicotine, which is the prominent constituent of tobacco, has negative effects on periodontium cells. However, the precise role of nicotine in cementoblast functions remains unclear. In the present study, we investigated the effects of nicotine on the functions of cementoblasts (OCCM-30) in terms of proliferation, migration, and mineralized tissue-associated gene expression. Immortalized murine cementoblasts were exposed to various concentrations (0, 10-6 , 10-5 , 10-4 , 10-3 , 10-2 , 10-1 , 1, 2.5, 5, and 10mM) of nicotine, and cementoblast proliferation was then evaluated using a real-time cell analyzer for 142 hours. Using an in vitro wound healing assay, cell migration was evaluated 2, 4, 6, and 24 hours after exposure to different concentrations of nicotine (1, 2.5, 5, and 10mM). The mRNA expressions of bone sialoprotein (BSP), collagen type I (COL-I), osteocalcin (OCN), runt-related transcription factor 2 (Runx2), and alkaline phosphatase (ALP) were assessed in the nicotine-treated (0, 10-3 , 10-2 , 10-1 , 1, 2.5, 5, and 10mM) OCCM-30 cells by reverse transcription quantitative polymerase chain reaction at 8 and 24 hours exposure. At concentrations of 1 to 10mM, nicotine significantly reduced cementoblast proliferation (P<0.01). Exposure to nicotine at other concentrations (1, 2.5, and 5mM) significantly reduced wound healing rates, whereas nicotine at a concentration of 10mM immediately decreased the viability of OCCM-30 cells. Similar results were observed in inverted microscopy images at the highest nicotine concentrations. All concentrations of nicotine decreased the transcripts of BSP and COL-I in a dose- and time-dependent manner (P<0.001). Nicotine concentrations higher than 1mM reduced the expression of OCN, RunX2, and ALP in a time-dependent manner (P<0.001). This study indicated that nicotine inhibited the proliferation, migration, and mineralized tissue-associated gene expression of OCCM-30 cells. These findings suggest that nicotine negatively affects cementoblast function and the formation of new cementum, which is critical for new attachment.

Effect of Carica papaya Extract toward Incised Wound Healing Process in Mice (Mus musculus) Clinically and Histologically.

Wound healing entails a sequence of complex biological processes, which is a protective function of the body that focuses on a quick recovery. Reducing wound healing time is crucial in a wound as it lowers the chance of infection and decreases complications and costs. Papaya extract was obtained by a maceration method. It contains papain enzyme, flavonoid, saponin, and alkaloid, which act as an anti-inflammatory, astringent substance, vasodilator, antioxidant, analgesic, with antifungal, and antibacterial properties, and increase the collagen synthesis. This study aimed to assess the effect of Carica papaya extract application toward incised wound healing process in mice (Mus musculus) clinically and histologically. This experimental laboratory study was performed using 24 mice weighing between 30 and 40 grams and 12 and 14 weeks of age as experimental animals. Each group was incised along 5 mm at the labial gingiva under both of mandible anterior teeth with a depth reaching alveolar bone. Papaya extract was applied topically in the morning and evening for 14 days. The clinical result was obtained by assessing the length of wound closure measured every day for 14 days. Based on the statistic test result, it showed that the papaya extract has a significant effect (p ≤ 0.05) toward the healing process of an incised oral wound in mice. Histologically at day 14, 75% concentration papaya extract showed perfect epithelial layer and fibrillation.

Closure of radial forearm free flap donor site: A comparative study between keystone flap and skin graft.

The aim was to investigate the feasibility of radial forearm free flap (RFFF) donor site closure by keystone flap (KF) and compare its outcomes to those of skin graft (SG) closure. One hundred and one patients who underwent RFFF for head and neck reconstruction were included (35 KF closure and 65 SG closure). Duration of wound healing and donor site complications was collected. After a minimal follow-up of 1 year, patients were questioned about functional and esthetic impairment. Coverage of donor site by KF was successful in all cases. The duration of wound healing was longer after SG than after KF (32 days vs 18 days, P < .001). Healing complications, esthetic and functional results were not statistically different. Forearm donor site closure by KF is a feasible alternative to the traditional SG. Its main advantages are the reduced wound healing time and the avoidance of a second donor site.

Ocean acidification reduces net calcification and wound healing in the tropical crustose coralline alga, Porolithon onkodes (Corallinales, Rhodophyta)

Reef dwelling algae employ a variety of physical and chemical defenses against herbivory, and the response to wounding is extremely important in algal communities. Wound healing mechanisms in crustose coralline algae (CCA) are related to skeletal growth and net calcification rate. Ocean acidification (OA) is known to affect rates of net calcification in a number of calcifying organisms, including CCA. Reduced rates of net calcification in CCA are likely to alter wound healing, and thus affect the consequences of herbivore-CCA interactions on coral reefs. The response of the tropical CCA Porolithon onkodes to OA and artificial wounding was quantified in a 51-day laboratory experiment. Eight artificially wounded (cut to a mean depth of 182 μm) and eight non-wounded samples of P. onkodes were randomly placed into each of four treatments (n = 64 samples total). Each treatment was maintained at a different pCO2 level representative of either ambient conditions or end-of-the-century, predicted conditions (IPCC, 2014); 429.31 ± 20.84 (ambient), 636.54 ± 27.29 (RCP4.5), 827.33 ± 38.51 (RCP6.0), and 1179.39 ± 88.85 μatm (RCP8.5; mean ± standard error). Elevated pCO2 significantly reduced rates of net calcification in both wounded and non-wounded samples of P. onkodes (slopes = −6.4 × 10−4 and −5.5 × 10−4 mg cm−2 d−1 per μatm pCO2, respectively over 51 days). There also was a significant reduction in the rate of vertical regeneration of thallus tissue within the wounds as pCO2 increased (slope = −1.5 × 10−3 μm d−1 per μatm pCO2 over 51 days). This study provides evidence that elevated pCO2 could reduce the ability of this important alga to recover from wounding. Because wounding by herbivores plays an important role in determining CCA community structure, we propose reduced wound healing as a mechanism by which OA might affect the structure and functional roles of CCA communities on coral reefs.

Breath-Hold Paradigm to Assess Variations in Oxygen Flow in Diabetic Foot Ulcers Using a Noncontact Near-Infrared Optical Scanner.

Objective: Diabetic foot ulcers (DFUs) occur in almost 25% of all patients with diabetes in their lifetime, with oxygen being the key limiting factor in healing. Identifying regions of compromised oxygenated flow can help clinicians cater the wound treatment process, possibly reducing wound healing time. Herein, a handheld, noncontact near-infrared optical scanner (NIROS) was developed and used to measure temporal changes in hemoglobin concentrations in response to a breath-hold (BH) paradigm. Approach: Noncontact imaging studies were carried out on DFU subjects and control subjects in response to a 20-s BH paradigm. Continuous-wave-based multiwavelength diffused reflective signals were acquired to generate effective oxy-hemoglobin, deoxy-hemoglobin, total hemoglobin, and oxygen saturation concentration maps using modified Beer-Lambert's law. Pearson's correlation analysis was carried out to determine variations in oxygen flow from hemoglobin concentration maps and the extent of variation observed in controls versus DFU subjects. Results: Temporal changes in hemoglobin concentration maps were observed in controls and DFU subjects. However, the oxygen flow in response to BH varied within 10% in all controls but significantly varied between wound and background regions in subjects with DFUs. Innovation: A method to assess variations in oxygen supply in and around DFUs was demonstrated using NIROS. This approach has potential to better cater DFU treatment process. Conclusion: Changes in all hemoglobin parameters due to 20 s of BH was observed. Pearson's analysis indicates that oxy-hemoglobin, deoxy-hemoglobin, and oxygen saturation fluctuations are synchronous in controls. In DFUs, changes are asynchronous with blood flow between the wound region and background region being significantly different.

Abstract 193: Antiangiogenic effect of conditioned media derived from triple negative breast cancer cells in brain microvascular endothelial cells in vitro

Abstract Breast cancer is the most common malignancy among females and is a leading cause of cancer mortality. Brain metastasis is associated with poor prognosis and reduced survival among breast cancer patients. For brain metastasis to occur, cancer cells are required to remain in direct contact with endothelial cells that line brain blood vessels. Data regarding the interaction between brain endothelial cells and breast cancer is limited. The aim of this work was to understand the interaction between breast cancer cells and brain endothelial cells. Human brain microvascular endothelial cells (hBMECs) were treated with conditioned media from MDA-MB-231 cell line and their angiogenic characteristics were evaluated. The involvement of the glycogen synthase kinase-3β (GSK-3β), vascular endothelial growth factor (VEGF), brain derived neurotrophic factor (BDNF), and thrombospondin-1 (TSP-1) was evaluated using release levels and inhibitor-based approaches. Levels of secreted proteins were assayed using ELISA. Results of the study showed that conditioned media from MDA-MB-231 breast cancer cells reduced angiogenic potential of hBMECs. Conditioned media significantly suppressed migration and tube formation of hBMECs compared to control group. In these experiments, control group represents hBMECs which were incubated with regular serum-free RPMI-1640 culture media. Inhibiting GSK-3β in conditioned media-treated hBMECs by its selective inhibitor (SB 216763; 10nM), restored the migratory potential of endothelial cells. In addition, MDA-MB-231 cells released VEGF at a rate that exceeded VEGF release from endothelial cells (6342 vs. 67.6 pg/ml). The release of VEGF from conditioned media-treated hBMECs was not altered by the high levels of VEGF presented in MDA-MB-231 cells conditioned media. GSK-3β inhibition in hBMECs significantly increased VEGF release from endothelial cells by about 1000 pg/ml compared to conditioned media-treated endothelial cells. MDA-MB-231 cells also released high levels of BDNF into the media and treating hBMECs with conditioned media from MDA-MB-231 cells did not alter the endogenous release of total BDNF from endothelial cells. GSK-3β inhibition did not alter BDNF release from endothelial cells. BDNF inhibition using TrkB-Fc (4µg/ml) did not restore the migratory potential of endothelial cells and the anti-angiogenic effect of conditioned media was maintained despite the inhibition of BDNF. Breast cancer cells released high levels of TSP-1 into the media. hBMECs treated with TSP-1 (5µg/ml) reduced wound healing by 40%, an effect that was restored by GSK-3β inhibitor. In conclusion, conditioned media from breast cancer cells induced anti-angiogenic effects in hBMECs which was found to be GSK-3β-dependent. The anti-angiogenic effect was further illustrated in terms of increased TPS-1 levels released by breast cancer cells. Citation Format: Nehad M. Ayoub, Ahmed Alhusban, Laila Alhusban. Antiangiogenic effect of conditioned media derived from triple negative breast cancer cells in brain microvascular endothelial cells in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 193.

Bioassay-Guided Fractionation of Wound Healing Active Compounds From Piper nigrum L. Berries Extract in Malaysia

The study aimed to further evaluate the wound healing property of P. nigrum L. using bioassay-guided fractionation method. The ethanolic extract of Piper nigrum L. was fractionated in two stages using column chromatography and preparative reversed phase C18 HPLC, respectively. Significant wound healing properties screened using the scratch wound assay was observed through the cell migration assay in fraction number three (PNE3) out of the 14 fractions, exhibiting 36.7% and 43.8% closure of wound gap within 20 hours at concentration of 0.3 and 1.0 µg/mL, respectively. Sub-fractions which were further fractionated from PNE3 showed comparatively reduced wound healing activity using the same bioassay. The sub-fractions were also compared to Piperine, a major component of P. nigrum L. and the results were comparable. This experimental study revealed that Piperine works together with other compounds in P. nigrum L. to improve wound healing as claimed by those home remedy.

Nociceptin/Orphanin FQ (NOP) receptor is differentially expressed on glial cells

Glial cells constitute the majority of the cellular components of the CNS. They are involved in a wide range of physiological and pathological conditions including immunomodulation, neuropathic pain, and opioid tolerance1. Opioid receptors are classified into classical (MOP, DOP and KOP) and non-classical (NOP). NOP receptor has been found to play a central role in pain processing and opioid tolerance2. In this study, we screened for the expression and function of the NOP receptor on glial cell lines. The pattern of NOP receptor expression at the level mRNA (qPCR)3 and protein ([leucyl-3H] N/OFQ saturation binding)4 was studied in a range of glial cell lines including 1321N1 human astrocytes, C6 rat astrocytes, MO3.13 human oligodendrocytes, HOG human oligodendrocytes, and EOC-20 mouse microglia. NOP receptor activity was assessed using a scratch wound healing/cell migration assay at a fixed time of 30 hours5. 1231N1, MO3.13, and HOG but not EOC-20 cells expressed NOP mRNA6. In addition, C6 cell lines were found to express NOP receptor mRNA. Furthermore, 1231N1, MO3.13 and HOG expressed NOP protein in a radioligand binding assay. Highest expression of mRNA was in MO3.13 and in radioligand binding was in HOG and 1321N16. In lines that expressed NOP receptor protein, we assessed the activity of N/OFQ, a structurally inactive derivative (desPhe1-N/OFQ) and the metabolically stable derivative PWT-N/OFQ. In both astrocytes (1321N1) and oligodendrocytes (MO3.13 and HOG) N/OFQ and PWT-N/OFQ but not desPhe1-N/OFQ (all at 1μM) reduced wound healing (migration), Table 1. NOP receptor is differentially expressed in a range of glial cell lines where it was functionally active. Interestingly there is no expression on microglia. Further studies in primary glial cell cultures are underway.Table 1The effect of NOP ligands on wound healing /cell migration of glial cells, data (% wound healing) from n=5, * = P≤0.05 after 30 hrs compared to control (ANOVA)Cell typeControlN/OFQPWT-N/OFQdesPhe1-N/OFQ1321N1 astrocytes55.6±2.042.5±5.8*35.8±1.8*50.0±1.8MO3.13 oligodendrocytes31.1±2.722.0±0.7*21.0±1.9*29.0±2.9HOG oligodendrocytes33.5±3.226.1±2.5*24.8±3.4*30.6±3.7 Open table in a new tab This work is sponsored by the Higher Committee for Education Development in Iraq HCED/Office of the Prime Minister/Iraq. Peptides were provided by Drs R Guerrini and G Calo/University of Ferrara /Italy. 1.Giaume et al. Cell Death Differ. 2007; 14: 1324–35.2.Lambert. Nat Rev Drug Discov. 2008; 7: 694–710.3.McDonald et al. Br J Anaesth. 2010; 104: 698–704.4.McDonald et al. Br J Pharmacol. 2003; 140: 61–70.5.Available from https://www2.le.ac.uk/colleges/medbiopsych/facilities-and-services/cbs/AIF/tips-and-tricks-1/wound-healing-assay.6.Available from https://www.bna.org.uk/static/uploads/resources/BNA2017_ABSTRACT_BOOK_SAGE_FINAL.pdf.

Preparation and evaluation of poly glycerol sebacate/poly hydroxy butyrate core‐shell electrospun nanofibers with sequentially release of ciprofloxacin and simvastatin in wound dressings

Biodegradable wound dressing of poly glycerol sebacate/poly hydroxy butyrate was fabricated via the coaxial electrospinning process. Simvastatin and ciprofloxacin were loaded in the core and shell of the fibers, respectively. Scanning electron microscopy and transmission electron microscopy images showed a uniform core/shell structure. Introducing drugs into the polymers would cause the dressing samples to become more hydrophilic and degradation to occur faster. Drugs release would face no interventions, in which, approximately 60% of ciprofloxacin was released during the first 24 hours. Simvastatin exhibited a slower and controlled release behavior, with its release peak recorded after 2 days. The drug‐containing samples showed a proper bactericidal activity against both Gram‐positive and Gram‐negative bacteria. It may be concluded that the drug‐laden wound dressing fabricated in this study is capable of releasing the 2 drugs sequentially and that it is the ideal conditions for controlling infections and reducing wound healing duration.

Small molecule inhibition of the CBFβ/RUNX interaction decreases ovarian cancer growth and migration through alterations in genes related to epithelial-to-mesenchymal transition

ObjectiveOvarian cancer survival and treatment have improved minimally in the past 20years. Novel treatment strategies are needed to combat this disease. This study investigates the effects of chemical inhibition of the CBFβ/RUNX protein-protein interaction on ovarian cancer cell lines. MethodsOvarian cancer cell lines were treated with CBFβ/RUNX inhibitors, and the effects on proliferation, DNA replication, wound healing, and anchorage-independent growth were measured. RNA-Seq was performed on compound-treated cells to identify differentially expressed genes. Genes altered by compound treatment were targeted with siRNAs, and effects on DNA replication and wound healing were measured. ResultsChemical inhibition of the CBFβ/RUNX interaction decreases ovarian cancer cell proliferation. Inhibitor treatment leads to an S-phase cell cycle delay, as indicated by an increased percentage of cells in S-phase, and a decreased DNA replication rate. Inhibitor treatment also reduces wound healing and anchorage-independent growth. RNA-Seq on compound-treated cells revealed changes in a small number of genes related to proliferation and epithelial-to-mesenchymal transition. siRNA-mediated knockdown of INHBA and MMP1 – two genes whose expression decreases with compound treatment – slowed DNA replication and impaired wound healing. ConclusionsChemical inhibition of the CBFβ/RUNX interaction is a viable strategy for the treatment of ovarian cancer.