Reduced Matrix Metalloproteinase Research Articles

All-Trans Retinoic Acid Reduces Matrix Metalloproteinase-2 and Increases E-Cadherin Levels in BeWo Choriocarcinoma Cells

Background: Choriocarcinoma is a malignant trophoblastic tumor that can degrade the uterine basement membrane to facilitate local and distant metastasis. Matrix metalloproteinase-2 (MMP-2) is a vital proteinase produced by trophoblasts and can cleave type IV collagen in the basement membrane of the uterine epithelium. In addition, E-cadherin controls cell adhesion, which is associated with tumorprogression, and loss of E-cadherin function is associated with metastasis. Vitamin A analogues, including all-trans retinoic acid (ATRA), exhibit anticancer properties, including metastasis inhibition.This study aimed to investigate the ability of ATRA to prevent trophoblast invasion and metastasis in choriocarcinoma by assessing MMP-2 and E-cadherin activity. Methods: This study administered various doses of ATRA to the BeWo (American Type Culture Collection CCL-98) choriocarcinoma cell line and assessed MMP-2 and E-cadherin activity through flow cytometry. The research was performed at the Biomedical Laboratory of the Faculty of Medicine of Universitas Brawijaya Malang, East Java, Indonesia. Results: BeWo cells were exposed to 0, 50, 100, 200, 400, or 800 μg/mL ATRA. MMP-2 was downregulated after ATRA treatment. The greatest reduction in MMP-2 level was observed after 200 μg/mL ATRA treatment, but this result was not statistically significant (P = 0.550). However, ATRA treatment significantly reduced the number of BeWo cells with low E-cadherin levels (P = 0.012). Conclusion: ATRA downregulates MMP-2 and upregulates E-cadherin. Further in vitro and animal studies are required to evaluate the ability of ATRA to inhibit metastasis. Bangladesh Journal of Medical Science Vol. 22 No. 02 April’23 Page : 336-340

Liquiritigenin Attenuated Collagen-Induced Arthritis and Cardiac Complication via Inflammation and Fibrosis Inhibition in Mice.

Rheumatoid arthritis (RA) is a common autoimmune disease with increased cardiovascular disease risk. Liquiritigenin (LG) is a triterpene with anti-inflammatory properties. Our study aimed to explore the effect of LG on RA and the cardiac complication. Collagen-induced arthritis (CIA) mice with LG treatment exhibited obvious alleviation in histopathological changes, accompanied by the decreased expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-17A in synovium and serum. LG attenuated cartilage destruction by reducing matrix metalloproteinase (MMP)-3 and MMP-13 expression in the synovium of CIA mice. The echocardiography results proved the alleviation of cardiac dysfunction in CIA mice. The electrocardiogram, biochemical, and histochemical analysis proved the cardioprotection effect of LG against RA. The decreased expression of inflammatory factors (TNF-α, IL-1β, and IL-6) and fibrotic markers (fibronectin, Collagen I, and Collagen III) in cardiac tissues of CIA mice further corroborated the attenuation of myocardial inflammation and fibrosis by LG. Mechanistic studies showed that LG could inhibit transforming growth factor β-1 (TGF-β1) and phos-Smad2/3 expression in cardiac tissues of CIA mice. Our study suggested that LG could relieve RA and its cardiac complication probably by inhibiting the TGF-β1/Smad2/3 pathway. All these suggested that LG might be a potential candidate for RA and its cardiac complication therapy.

Trimetazidine alleviates paclitaxel-induced peripheral neuropathy through modulation of TLR4/p38/NF-κB and klotho protein expression

Chemotherapy-induced peripheral neuropathy is a common adverse effect associated with a number of chemotherapeutic agents including paclitaxel (PTX) which is used in a wide range of solid tumors. Development of PTX-induced peripheral neuropathy (PIPN) during cancer treatment requires dose reduction which limits its clinical benefits. This study is conducted to investigate the role of toll like receptor-4 (TLR4) /p38 signaling and Klotho protein expression in PIPN and the role of trimetazidine (TMZ) in this pathway. Sixty-four male Swiss albino mice were divided into 4 groups (n = 16); Group (1) injected intraperitoneally (IP) with ethanol/tween 80/saline for 8 successive days. Group (2) received TMZ (5 mg/kg, IP, day) for 8 successive days. Group (3) treated with 4 doses of PTX (4.5 mg/kg, IP) every other day over a period of 7 days. Group (4) received a combination of TMZ as group 2 and PTX as group 3. The Effect of TMZ on the antitumor activity of PTX was studied in another set of solid Ehrlich carcinoma (SEC)-bearing mice that was similarly divided as the above-mentioned set. TMZ mitigated tactile allodynia, thermal hypoalgesia, numbness and fine motor discoordination associated with PTX in Swiss mice. The results of the current study show that the neuroprotective effect of TMZ can be attributed to inhibition of TLR4/p38 signaling which also includes a reduction in matrix metalloproteinase-9 (MMP9) protein levels as well as the proinflammatory interleukin-1β (IL-1β) and preserving the levels of the anti-inflammatory IL-10. Moreover, the current study is the first to demonstrate that PTX reduces the neuronal levels of klotho protein and showed its modulation via cotreatment with TMZ. In addition, this study showed that TMZ neither alter the growth of SEC nor the antitumor activity of PTX. In conclusion, we suggest that (1) Inhibition of Klotho protein and upregulation of TLR4/p38 signals in nerve tissues may contribute to PIPN. (2) TMZ attenuates PIPN by modulating TLR4/p38 and Klotho protein expression without interfering with its antitumor activity.

Momordica cochinchinensis Suppresses the Human Breast Cancer Cells Growth and Migration by Inhibiting Mevalonate Pathway

Background: Momordica cochinchinensis (MC) is shown with high antioxidation and is used as herbal medicine in South-East countries. Objectives: This study aims to determine the four parts (aril, fruit, leaf, and seed) of MC extract against human breast cancer cells with underlying mechanisms of actions. Materials and Methods: The effects of MC extract were examined by sulforhodamine B, colony formation, wound healing, real time polymerase chain reaction (RT-PCR), and Western blotting method. Results: Four parts of MC extracts significantly increased MCF-7 cells death, interestingly, seed extract had the greatest activity with IC50 values of 412.3 ± 49.8 µg/mL for 24 h and 113.3 ± 6.4 µg/mL for 48 h. MC extracts potentiated the anticancer drug effect, doxorubicin, with lower IC50 values when compared with MC treatment alone. Moreover, MC extracts decreased colony formation and accompanied by inhibited cell migration. Furthermore, Seed extract inhibited migration by reducing matrix metalloproteinases (MMP) 2, MMP 9, and vascular endothelial growth factor A (VEGFA) inhibition. The molecular analyses revealed that seed extract inhibited cell viability by decreasing cyclin D1 consistent with activating p21, cytochrome c, and caspase-3 protein levels along with reduction of mevalonate (MVA) pathway, Rac1, and RhoA levels, it appeared to inhibit growth and migration. Conclusion: Taken together, MC extract induces MCF-7 cell death and decreases cell migration relatively via attenuation of MVA pathway.

Synergistic prevention and reparative effects of sesquiterpene farnesol in a rabbit model of surgical resection-induced osteoarthritis.

Articular cartilage may regenerate poorly after injury or during aging. In vitro, farnesol can modulate extracellular matrix synthesis and restore chondrocyte phenotypes by increasing type II collagen (COL II) and glycosaminoglycan (GAG) production. Here, we evaluated farnesol's preventive and reparative effects against osteoarthritis (OA) in vivo. We induced OA in rabbits through resection of the lateral collateral ligament and meniscus. After 2 weeks, the affected limb was treated with 0.5 ml of 0.4 mM farnesol, hyaluronan (HA) nanoparticle-encapsulated 0.8 mM farnesol (Farn/HA), or HA nanoparticles intra-articularly. After 2 and 6 treatment weeks, synovial inflammatory cytokine levels were analyzed. We also removed the entire joint cartilage from lateral femoral condyles for histological investigation. The half-maximum inhibitory concentration of farnesol was 0.5 mM. Farn/HA had relatively low cytotoxicity showing cells remained viable after being treated with 1 mM a concentration Farn/HA. Untreated lateral condyle exhibited extensive wear. By contrast, 0.4 mM farnesol or 0.8 mM Farn/HA led to a relatively transparent and bright appearance. After 2 and 6 treatment weeks, farnesol, particularly 0.8 mM Farn/HA, reduced matrix metalloproteinase 1 and 13 levels considerably. Therefore, 0.8 mM Farn/HA, which enabled slow drug release, demonstrated the highest anti-inflammatory and OA preventive effects. After 6 treatment weeks, farnesol also promoted COL II and GAG synthesis and, thus, aided healing.

The Effect of Chayote (Sechium Edule) Extract Cream Can Reduce Matrix Metalloproteinase-2 (MMP-2) Levels and Increase Collagen Density in The Wound Healing Process of Male Wistar Rats (Rattus Novergicus) With Diabetes Mellitus

Introduction: The main complication in Diabetes Mellitus patients is delayed wound healing resulting in chronic wounds. Chayote is known as a vegetable, which has anti-inflammatory properties. The tannin and saponin compounds in chayote produce antibacterial agents. This study aims to prove the role of chayote extract cream on the wound healing process of diabetic rats through MMP-2 markers and collagen density. Methods: This study was an experimental post-test only control group design using male rats. Samples were induced hyperglycaemia with 46 mg of nicotinamide and 10 mg of streptozotocin one week before the experiment. The sample was divided into 2 groups: the placebo group (P0) and the treatment group (P1) with a total of 11 samples per group. The treatment group received cream of chayote extract 60% for 1 week. On the 8th day, both groups were taken by punch biopsy to assess collagen density, then examined for MMP-2 levels in the blood. Results: The average MMP-2 level in the control group was 4.97 ± 0.36 ng/ml, while the average MMP-2 in the treatment group was 4.59 ± 0.42 ng/ml. The average collagen level in the control group was 37.08 ± 12.92 %/ body surface, while the mean collagen in the treatment group was 54.21 ± 15.76 %/ body surface. Conclusion: Administration of ethanol extract of chayote (Sechium edule) significantly reduced matrix metalloproteinase-2 (MMP-2) levels and increased collagen density in the wound healing process of rats.

Talar Arthroscopic Reduction and Internal Fixation (TARIF): A Novel All-Inside Soft-Tissue-Preserving Technique.

Talar Arthroscopic Reduction and Internal Fixation (TARIF): A Novel All-Inside Soft-Tissue-Preserving Technique.

LQB-118 Suppresses Migration and Invasion of Prostate Cancer Cells by Modulating the Akt/GSK3β Pathway and MMP-9/Reck Gene Expression.

Prostate cancer (PCa) is one of the most common malignancies in adult men. LQB-118 is a pterocarpanquinone with antitumor activity toward prostate cancer cells. It inhibits cell proliferation by down-regulating cyclins D1 and B1 and up-regulating p21. However, the effects of LQB-118 on PCa cell migration are still unclear. Herein, the LQB-118 effects on PCa metastatic cell migration/invasion and its mechanism of action were evaluated. PC3 cells were treated with LQB-118 or Paclitaxel (PTX), and cell migration (wound healing and Boyden chamber assays) and invasion (matrigel assay) were determined. The LQB-118 mechanisms were evaluated by αVβIII protein expression (flow cytometry), protein phosphorylation (Western blot), and mRNA expression (qPCR). LQB-118 impaired PCa cell migration and invasion, down-regulated Akt phosphorylation, and also reduced GSK3β phosphorylation, through a FAK-independent pathway. Also, it was observed that LQB-118 controlled the invasiveness behavior by reducing matrix metalloproteinase-9 (MMP-9) and up-regulating reversion-inducing cysteine rich protein with Kazal motifs (Reck) mRNA levels. Interestingly, LQB-118 increased integrin αvβIII expression, but this effect was not related to its activation, since the cell adhesion ability was reduced after LQB-118 treatment. These data highlight novel LQB-118 mechanisms in prostate cancer cells. LQB-118 acts as a negative regulator of the Akt/GSK3 signaling pathway and can modulate PCa cell proliferation, death, and migration/invasion. The results also support the use of LQB-118 for the treatment of metastatic PCa, alone or combined with another chemotherapeutic agent, due to its demonstrated pleiotropic activities.

Effects of L-carnitine on healing of Achilles tendon in rats.

In this experimental study, we aimed to analyze the effects of levocarnitine (L-carnitine) on tendon healing after surgical repair of Achilles tendon rupture in a rat model. The study included 40 Wistar Albino rats divided into four groups: Group 1, neither surgical intervention nor substance applications were performed for the Achilles tendons. In the other groups, the right Achilles tendons were cut using a scalpel and repaired with a modified Kessler-type technique with 3/0 monofilament polydioxanone suture. In Group 2, the rats did not receive any additional treatment, except for surgical repair. In Group 3, the same volume similar to Group 4 of saline solution was administered intraperitoneally for seven days. In Group 4, each rat received 300 mg/kg of L-carnitine intraperitoneally for seven days. At Week 6, all rats were sacrificed. All right Achilles tendons were used for biomechanical tests and histopathological evaluations. Biochemical analysis of the matrix metalloproteinase was also performed using the blood specimens. There were no significant differences among the groups in terms of the histopathological parameters. Although the mean matrix metalloproteinase level was low in the L-carnitine group, it did not reach statistical significance. A significant increase in maximum force, tensile strength, and strength to 2-mm gap was observed in the L-carnitine group. The significant effects of L-carnitine on biomechanical parameters may indicate favorable effects on Achilles tendon healing in rats by reducing matrix metalloproteinase 2 and 9. To improve Achilles tendon healing, further investigation for these markers is needed. Since the effects of L-carnitine on the Achilles tendon cannot be clearly distinguished histopathologically, further studies involving L-carnitine-induced effects are warranted.

Abstract 15700: ALK1-Mediated Smad1 Activation in a Myofibroblast Subset Protects the Infarcted Heart From Adverse Remodeling and Excessive Fibrosis

Cardiac fibroblasts acquire inflammatory, proangiogenic, or matrix-synthetic phenotypes following MI and play a pivotal role in repair and remodeling of the infarcted heart. Members of the Transforming Growth Factor (TGF)β superfamily (including TGFβs and BMPs) are activated following MI and regulate fibroblast phenotype and cardiac fibrosis. Canonical TGFβ signaling is mediated through the receptor-activated Smads: TGF-βs activate Smad2/3, whereas BMPs predominantly stimulate Smad1/5 cascade. Smad3 signaling plays an important role in activation of infarct myofibroblasts; however, the role of the Smad1 pathway remains unknown. We hypothesized that fibroblast-specific actions of Smad1 regulate repair and remodeling in the infarcted myocardium. In a mouse model of non-reperfused infarction, Smad1 was activated in a subset of cardiac myofibroblasts not only by BMPs, but also by all three TGF-β isoforms. TGF-β activation of Smad1 was dependent on both ALK1 and ALK5 signaling. Analysis of single cell RNA-seq data suggested that ALK1 is a marker of a myofibroblast subset exhibiting anti-fibrotic properties. In order to examine the role of fibroblast Smad1 in vivo, we generated mice with myofibroblast-specific Smad1 loss (MFS1KO). Echocardiography showed that MFS1KO mice have accentuated systolic dysfunction and LV dilation 7-28 days after MI (p<0.01, N=13-49) . Worse adverse remodeling in infarcted MFS1KO hearts was associated with scar expansion, higher myofibroblast density and accentuated fibrosis. Smad1 knockdown in cardiac fibroblasts increased mRNA levels of structural and matricellular genes while reducing matrix metalloproteinase synthesis. RNA-seq showed that Smad1 loss activates pathways involved in matrix regulation and collagen biosynthesis, and suggested that Smad1 may exert anti-fibrotic actions through inhibition of TGF-β/Smad3 axis, p53, and IL4/STAT6 pro-fibrotic cascades. In conclusion, Smad1 activation in a subset of anti-fibrotic infarct myofibroblasts restrains cardiac fibrosis and protects from adverse remodeling by inhibiting key pro-fibrotic pathways. These findings suggest that Smad1 activation in fibroblasts may be a promising strategy to restrain cardiac fibrosis in infarcted and failing hearts.

Mesenchymal stem cell transplantation improvesbiomechanical properties of vaginal tissuefollowing full-thickness incision in aged rats.

Pelvic organ prolapse (POP) is common among post-menopausal women and is associated with bladder, bowel, and sexual dysfunction. Surgical repair with the patients' native tissues is sub-optimal with high reoperation rates, potentially due to diminished age-related healing. We demonstrate that systemic transplantation of mesenchymal stem cells (MSCs) improves healing of full-thickness vaginal incision in the vaginal wall of old rats, as suggested by both histological and functional analysis. Transplanted MSCs homed and survived at the surgical vaginal site. Attenuation of the injury-induced inflammatory response, increased angiogenesis, and reduced matrix metalloproteinase 9 expression were observed at the surgical site of transplanted rats. Most importantly, the functional biomechanical properties of the healed vagina, at day 30 post-injury, were improved in MSC-transplanted, compared with sham-operated non-transplanted, old rats. These results may pave the way to further translational studies toward clinical transplantation of MSCs adjuvant to POP repair for the improvement of surgical outcome.

Trilinolein, a Natural Triacylglycerol, Protects Cerebral Ischemia through Inhibition of Neuronal Apoptosis and Ameliorates Intimal Hyperplasia via Attenuation of Migration and Modulation of Matrix Metalloproteinase-2 and RAS/MEK/ERK Signaling Pathway in VSMCs.

Cerebrovascular disease is one of the leading causes of disability and death worldwide, and seeking a potential treatment is essential. Trilinolein (TriL) is a natural triacylglycerol presented in several plants. The effects of TriL on cerebrovascular diseases such as cerebral ischemia and carotid stenosis have never been studied. Accordingly, we investigated the protection of TriL on cerebral ischemia/reperfusion (I/R) and vascular smooth muscle cell (VSMC) migration in vivo and in vitro. The cerebral infarction area, the intima to media area (I/M ratio), and proliferating cell nuclear antigen (PCNA)-staining of the carotid artery were measured. Platelet-derived growth factor (PDGF)-BB-stimulated A7r5 cell migration and potential mechanisms of TriL were investigated by wound healing, transwell, and Western blotting. TriL (50, 100, and 200 mg/kg, p.o.) reduced: the cerebral infarction area; neurological deficit; TUNEL-positive apoptosis; intimal hyperplasia; and PCNA-positive cells in rodents. TriL (5, 10, and 20 µM) significantly inhibited PDGF-BB-stimulated A7r5 cell migration and reduced matrix metalloproteinase-2 (MMP-2), Ras, MEK, and p-ERK protein levels in PDGF-BB-stimulated A7r5 cells. TriL is protective in models of I/R-induced brain injury, carotid artery ligation-induced intimal hyperplasia, and VSMC migration both in vivo and in vitro. TriL could be potentially efficacious in preventing cerebral ischemia and cerebrovascular diseases.

Cardiac sympathetic afferent denervation reduces matrix metalloproteinase expression and improves cardiac remodeling in rats post myocardial infarction

Abstract In a previous study from our laboratory, we demonstrated that chronic and selective cardiac sympathetic afferent (spinal) denervation at the time of myocardial infarction (MI) using epicardial application of resiniferatoxin (RTX), a neuronal toxin capable of inducing rapid degeneration of transient receptor potential vanilloid 1 (TRPV1)-expressing afferent neurons and fibers, markedly reduced the cardiac remodeling process 9–11 weeks post-MI in rats. This included reduced cardiac hypertrophy, fibrosis and apoptosis. Here, we further investigated the effect of epicardial application of RTX at the time of MI on cardiac extracellular matrix (ECM) and remodeling post-MI. Echocardiographic and morphologic data demonstrated that, compared to MI+vehicle, MI+RTX exhibited a significantly slower LV chamber dilation (6-week echocardiographic data: left ventricular end-diastolic diameter, 10.7±0.2 vs. 9.6±0.3 mm; left ventricular end-systolic diameter: 8.8±0.2 vs. 7.8±0.3 mm; MI+vehicle vs. MI+RTX, n=18, p<0.05; mean±SE). Scanning electron microscopy showed that RTX reduced collagen deposition in the peri-infarct area in post-MI rats. Western blot and zymography were used to further evaluate the effect of RTX application on matrix metalloproteinase (MMP) expression and activity, which is responsible for degrading ECM and contributing to cardiac dilation post MI. Our data suggest time-dependent increases in MMP expression in infarcted hearts post MI. RTX application largely prevented the increase in MMP9 but not MMP2, 4 weeks post-MI. RTX prevented both increased MMP2 and MMP9 activities in the peri-infarct myocardium at 8–10 weeks post-MI. We further investigated the effect of RTX application on MI-induced cardiac inflammation, which has been reported as an upstream mechanism triggering MMP activation post-MI. The data show that RTX largely abolished MI-induced plasma extravasation and reduced macrophage infiltration and cardiac cytokine content in the peri-infarct or remote myocardium of post-MI rats. These data suggest that cardiac sympathetic afferent denervation at the time of MI exerts a local anti-inflammatory effect and reduces ECM remodeling by preventing excessive MMP activation in post-MI rats. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Institutes of Health Grant HL126796

Modulation of inflammatory M1-macrophages phenotype by valvular interstitial cells

BackgroundAortic valve stenosis involves inflammation, excess deposition of a collagen-rich extracellular matrix, and calcification. Recent studies have shown that M1 or inflammatory macrophages derived from infiltrating monocytes promote calcification of valvular interstitial cells, the most prevalent cell type of the aortic valve. We hypothesized that valvular interstitial cells could modulate inflammatory macrophages phenotype. MethodsWe first assessed macrophage phenotype in human aortic valve stenosis and control aortic valves from donors. Then, we examined profibrotic and inflammatory-related gene expression in valves and valvular interstitial cells. Finally, we investigated whether valvular interstitial cells can modify the phenotype of inflammatory macrophages. ResultsCirculating monocytes and plasma transforming growth factor beta-1 levels of patients with aortic valve stenosis were significantly higher compared with patients without aortic valve stenosis. Histologic analysis of thickened spongiosa of the aortic valve from patients with aortic valve stenosis showed a high macrophage infiltration but a low matrix metalloproteinase-9 expression compared with control aortic valves. On the other hand, valvular interstitial cell culture of aortic valve stenosis exhibited a profibrotic phenotype with a high expression of transforming growth factor beta-1 and transforming growth factor beta-1/transforming growth factor beta-3 ratio but a decreased expression of the peroxisome proliferator-activated receptor gamma nuclear receptor. Valvular interstitial cell–conditioned media of aortic valve stenosis led to a decrease in enzymatic activity of matrix metalloproteinase-9 and an increase in production of collagen in inflammatory macrophages compared with valvular interstitial cell–conditioned media from control aortic valve donors. ConclusionsThese findings indicate that profibrotic valvular interstitial cells promote the imbalance of extracellular matrix remodeling by reducing matrix metalloproteinase-9 production on inflammatory macrophages that lead to excessive collagen deposition observed in aortic valve stenosis. Further investigation is needed to clarify the role of transforming growth factor beta-1/proliferator-activated receptor gamma nuclear receptor/matrix metalloproteinase-9 in aortic valve stenosis.

Retinal Pigment Epithelium-Secreted VEGF-A Induces Alpha-2-Macroglobulin Expression in Endothelial Cells.

Alpha-2-macroglobulin (A2M) is a protease inhibitor that regulates extracellular matrix (ECM) stability and turnover. Here, we show that A2M is expressed by endothelial cells (ECs) from human eye choroid. We demonstrate that retinal pigment epithelium (RPE)-conditioned medium induces A2M expression specifically in ECs. Experiments using chemical inhibitors, blocking antibodies, and recombinant proteins revealed a key role of VEGF-A in RPE-mediated A2M induction in ECs. Furthermore, incubation of ECs with RPE-conditioned medium reduces matrix metalloproteinase-2 gelatinase activity of culture supernatants, which is partially restored after A2M knockdown in ECs. We propose that dysfunctional RPE or choroidal blood vessels, as observed in retinal diseases such as age-related macular degeneration, may disrupt the crosstalk mechanism we describe here leading to alterations in the homeostasis of choroidal ECM, Bruch’s membrane and visual function.

Laser-assisted nanoparticle delivery to promote skin absorption and penetration depth of retinoic acid with the aim for treating photoaging

Retinoic acid (RA) is an approved treatment for skin photoaging induced by ultraviolet (UVA). Topically applied RA is mainly located in the stratum corneum (SC) with limited diffusion into the deeper strata. A delivery system capable of facilitating dermal delivery and cellular internalization for RA is critical for a successful photoaging therapy. Two delivery approaches, namely nanoparticles and laser ablation, were combined to improve RA’s absorption efficacy and safety. The nanoparticle absorption enhancement by the lasers was compared between full-ablative (Er:YAG) and fractional (CO2) modalities. We fabricated poly-L-lactic acid (PLA) and PLA/poly(lactic-co-glycolic acid) (PLGA) nanoparticles by an emulsion-solvent evaporation technique. The mean size of PLA and PLA/PLGA nanocarriers was 237 and 222 nm, respectively. The RA encapsulation percentage in both nanosystems was > 96 %. PLA and PLA/PLGA nanocarriers promoted RA skin deposition by 5- and 3-fold compared to free control. The ablative lasers further enhanced the skin deposition of RA-loaded nanoparticles, with the full-ablative laser showing greater permeation enhancement than the fractional mode. The skin biodistribution assay evaluated by confocal and fluorescence microscopies demonstrated that the laser-assisted nanoparticle delivery achieved a significant dermis and follicular accumulation. The cell-based study indicated a facile uptake of the nanoparticles into the human dermal fibroblasts. The nanoparticulate RA increased type I collagen and elastin production in the UVA-treated fibroblasts. A reduction of matrix metalloproteinase (MMP)-1 was also highlighted in the photoaging cells. The calculation of therapeutic index (TI) by multiplying collagen/elastin elevation percentage and skin deposition predicted better anti-photoaging performance in Er:YAG laser-assisted nanoparticle delivery than CO2 laser. Nanoencapsulation of RA decreased the cytotoxicity against skin fibroblasts. In vivo skin tolerance test on a nude mouse showed less skin damage after topical application of the nanoparticles than free RA. Our results hypothesized that the laser-mediated nanoparticle delivery provided an efficient and safe use for treating photoaging.

ADAM-10 Regulates MMP-12 during Lipopolysaccharide-Induced Inflammatory Response in Macrophages.

A disintegrin and metalloprotease 10 (ADAM-10), a member of the ADAM protease family, has biological activities related to TNF-α activation, cell adhesion, and migration, among other functions. Macrophages are important immune cells that are involved in the inflammatory response of the body. ADAM-10 is involved in inflammatory responses, but the specific regulatory mechanisms are not fully understood. In this study, we investigated the regulatory mechanism of ADAM-10 in the lipopolysaccharide-promoted proliferation (LPS) of the macrophage inflammatory response. Differentially expressed or regulated proteins were identified in interfered ADAM-10 (sh ADAM-10) macrophages using tandem mass tag (TMT) proteomics. The changes and regulatory role of ADAM-10 during LPS-induced inflammatory response in normal, interfering, and overexpressing ADAM-10 (EX ADAM-10) cells were determined. Results indicated that ADAM-10 interference affected inflammation-related pathways and reduced matrix metalloproteinase 12 (MMP-12) protein levels, as identified by TMT proteomics. In normal cells, LPS decreased ADAM-10 gene expression, but promoted ADAM-10 secretion, MMP-12 and TNF-α gene expression, and MMP-12, iNOS, IL-10, and cyclinD1 protein expression. Additionally, ADAM-10 knockdown decreased macrophage viability in sh-ADAM-10 cells. Moreover, an MMP-12 inhibitor had no impact on the viability effect of LPS on cells or the expression of ADAM-10. iNOS expression decreased, whereas IL-10 expression increased after ADAM-10 depletion. ADAM-10 knockdown decreased MMP-12, iNOS, TNF-α, IL-1β, and FKN, while overexpression had an opposite effect. ADAM-10 overexpression further increased MMP-12, iNOS, and TNF-α gene expression in response to LPS. Cell viability was increased in EX ADAM-10 cells, and ADAM-10 secretion was further increased in the EX and LPS groups. Flow cytometry and immunofluorescence staining revealed that EX-ADAM 10 cells had increased iNOS expression, which acted as an IL-6 expression driver. In summary, we found that ADAM-10 is activated by LPS and positively participates in LPS-stimulated macrophage inflammatory responses by positively regulating MMP-12 during the inflammatory process.

Bacillus amyloliquefaciens Lysate Ameliorates Photoaging of Human Skin Fibroblasts through NRF2/KEAP1 and TGF-β/SMAD Signaling Pathways

More and more research in dermatology and cosmetic science is devoted to the development and application of postbiotic raw materials. In order to explore the anti-photoaging efficacy and application prospect of Bacillus amyloliquefaciens lysate (BAL1) on the skin, we used 16 J/cm2 UVA stimulation of human embryonic fibroblasts (CCC-ESF-1) to establish a UVA photodamage model to investigate the anti-photoaging efficacy of BAL1 and its mechanism of action. In this study, we found that BAL1 activated the transcription of downstream antioxidant enzyme genes mainly by promoting the nuclear displacement of NF-E2-related factor 2 (Nrf2) within CCC-ESF-1, thus increasing the antioxidant capacity of antioxidant enzymes to scavenge excessive reactive oxygen species in cells. Meanwhile, BAL1 promoted intracellular TGF-β/Smad signaling pathway and reduced matrix metalloproteinase expression to alleviate the degradation of extracellular matrix. In conclusion, the results of this study demonstrate the potential benefit of BAL1 in protecting the skin against UVA-induced photoaging and highlight the potential of BAL1 in skin photoprotection.

5-Fluorouracil reduces the fibrotic scar via inhibiting matrix metalloproteinase 9 and stabilizing microtubules after spinal cord injury.

Fibrotic scars composed of a dense extracellular matrix are the major obstacles for axonal regeneration. Previous studies have reported that antitumor drugs promote neurofunctional recovery. We investigated the effects of 5-fluorouracil (5-FU), a classical antitumor drug with a high therapeutic index, on fibrotic scar formation, axonal regeneration, and functional recovery after spinal cord injury (SCI). 5-FU administration after hemisection SCI improved hind limb sensorimotor function of the ipsilateral hind paws. 5-FU application also significantly reduced the fibrotic scar formation labeled with aggrecan and fibronectin-positive components, Iba1+ /CD11b+ macrophages/microglia, vimentin, chondroitin sulfate proteoglycan 4 (NG2/CSPG4), and platelet-derived growth factor receptor beta (PDGFRβ)+ pericytes. Moreover, 5-FU treatment promoted stromal cells apoptosis and inhibited fibroblast proliferation and migration by abrogating the polarity of these cells and reducing matrix metalloproteinase 9 expression and promoted axonal growth of spinal neurons via the neuron-specific protein doublecortin-like kinase 1 (DCLK1). Therefore, 5-FU administration impedes the formation of fibrotic scars and promotes axonal regeneration to further restore sensorimotor function after SCI.

Therapeutic effects of statins on osteoarthritis: A review.

Osteoarthritis (OA) is a progressive joint disease. The etiology of OA is considered to be multifactorial. Currently, there is no definitive treatment for OA, and the existing treatments are not very effective. Hypercholesterolemia is considered a novel risk factor for the development of OA. Statins act as a competitive inhibitor of the β-hydroxy β-methylglutaryl-CoA (HMG-CoA) reductase and are widely used to manage hypercholesterolemia. Inhibition of HMG-CoA reductase results in reduced synthesis of a metabolite named mevalonate, thereby reducing cholesterol biosynthesis in subsequent steps. By this mechanism, statins such as atorvastatin and simvastatin could potentially have a preventive impact on joint cartilage experiencing osteoarthritic deterioration by reducing serum cholesterol levels. Atorvastatin can protect cartilage degradation following interleukin-1β-stimulation. Atorvastatin stimulates the STAT1-caspase-3 signaling pathway that was shown to be responsible for its anti-inflammatory effects on the knee joint. Simvastatin had chondroprotective effects on OA in vitro by reducing matrix metalloproteinases expression patterns. In this study, we tried to review the therapeutic effects of statins on OA.