Loss of physiological function, uncomfortable symptoms and various disease processes are thought to be directly related to the formation of scar tissue following tissue damage. Between ten and thirty percent of patients requiring spinal surgery suffer from failed back surgery syndrome. The pain and instability resulting from failed back syndrome often requires medical treatment and may even require additional surgeries to alleviate its associated symptoms. Following surgery, scar tissue forms that often becomes adherent to the dura and entangled in the ganglia and nerve fibers of the spinal nerves. This scar tissue is considered to play a major role in the development of failed back syndrome. Following tissue injury, excessive oxidative stress and inflammation are considered to be the primary stimulators behind increased fibroblast proliferation and activation, resulting in abundant extracellular matrix deposition. The excessive laying down of extracellular matrix ultimately leads to abundant scar tissue formation. I hypothesized that reducing oxidative stress and inflammation will mitigate scar tissue formation and produce a better outcome after spinal surgeries. Quercetin is a dietary flavonoid with anti-oxidant and anti-inflammatory properties that has been shown to improve the outcome following injury to the spinal cord and reduce the proliferation of fibroblasts. L-2-Oxothiazolidine-4-carboxylate (OTC) also minimizes inflammation and protects against oxidative stress by promoting the synthesis of the potent antioxidant and anti-inflammatory agent glutathione. OTC reduces airway inflammation in asthma models and is potentially capable of modulating extracellular matrix production. <p>Treatment with these two agents was hypothesized to decrease oxidative stress and inflammation, thereby causing an amelioration of scar tissue formation following spinal surgery and improve the outcome. Morphological changes observed initially indicated that improvements in wound healing were occurring in the experimentally treated tissues. In addition, the scar tissue area and the lateral widths of the peridural scar forming between the muscular tissue areas suggested a reduction in the scar size. Although inflammatory cell numbers increased slightly in the experimental treatment groups, particularly during the initial three day post laminectomy time point, this increase was not statistically significant. <p>While quercetin and OTC did not appear to inhibit the influx of inflammatory cells following laminectomy, they did appear to induce a more beneficial wound healing environment. It is possible that these agents are affecting parameters of wound healing not considered by these studies. For instance the myriad of processes mediated by growth factors and cytokines involved in wound healing process may play a much greater role than the inflammatory cells themselves. In conclusion, reducing oxidative stress and inflammation by these agents to ameliorate scar tissue formation following spinal laminectomy was supported by the observed morphology, but not supported by the quantification of inflammatory cells. Additional studies investigating the efficacy of quercetin and OTC on the wound healing process are needed to further understand the role they play in repair and scar tissue formation.

This project was carried out in the Cell Signaling Research Group headed by Dr. Carme Caelles at IRB Barcelona. As a part of the research-line that deals with physiological and pharmacological (anti-inflammatory and/or anti-diabetic) actions conducted by some nuclear receptor (NR) ligands through negative interference with the c-Jun N-terminal kinase (JNK) signaling pathway, this project was focused on studying the mechanism of cross-talk between those pathways. The results of the study show the ligand-dependent LXR inhibition of the LPS-activated SAPK (JNK and p38MAPK) pathways. Moreover, PP5, a serine/threonine phosphatase previously shown to regulate MAPK pathways, is suggested as a novel target of LXR that negatively regulates LPS-induced activation of SAPK pathways. Furthermore, it is proposed that through the inhibition of SAPK activity, and thereby cJun/AP-1 activity, PP5 is mediating negative regulation of LPS-induced Mmp13 gene expression by LXR in murine primary macrophages. / Este proyecto se llevó a cabo en el Grupo de Investigación en Señalización Celular del IRB Barcelona y fue dirigido por la Dra. Carme Caelles. El trabajo se centra en el estudio del mecanismo de interferencia entre las vías de los receptores nucleares (NR) y la señalización de la quinasa c-Jun N-terminal Kinase (JNK). Esta inhibición forma parte de la línea investigación sobre las acciones fisiológicas y farmacológicas (anti-inflamatorias y / o anti-diabéticas) realizadas por los ligandos de algunos NR. El estudio demuestra la inhibición de las vías SAPK (JNK y p38MAPK) en respuesta a LPS a través de la activación dependiente de ligando de LXR. Además, PP5, una fosfatasa serina/treonina que previamente se demostró que regula las vías de las MAPKs, se sugiere como el mediador de esta inhibición. Esta interacción estaría inhibiendo la expresión en respuesta a LPS del gen Mmp13 en macrófagos de ratón.

Loss of physiological function, uncomfortable symptoms and various disease processes are thought to be directly related to the formation of scar tissue following tissue damage. Between ten and thirty percent of patients requiring spinal surgery suffer from failed back surgery syndrome. The pain and instability resulting from failed back syndrome often requires medical treatment and may even require additional surgeries to alleviate its associated symptoms. Following surgery, scar tissue forms that often becomes adherent to the dura and entangled in the ganglia and nerve fibers of the spinal nerves. This scar tissue is considered to play a major role in the development of failed back syndrome. Following tissue injury, excessive oxidative stress and inflammation are considered to be the primary stimulators behind increased fibroblast proliferation and activation, resulting in abundant extracellular matrix deposition. The excessive laying down of extracellular matrix ultimately leads to abundant scar tissue formation. I hypothesized that reducing oxidative stress and inflammation will mitigate scar tissue formation and produce a better outcome after spinal surgeries. Quercetin is a dietary flavonoid with anti-oxidant and anti-inflammatory properties that has been shown to improve the outcome following injury to the spinal cord and reduce the proliferation of fibroblasts. L-2-Oxothiazolidine-4-carboxylate (OTC) also minimizes inflammation and protects against oxidative stress by promoting the synthesis of the potent antioxidant and anti-inflammatory agent glutathione. OTC reduces airway inflammation in asthma models and is potentially capable of modulating extracellular matrix production. <p>Treatment with these two agents was hypothesized to decrease oxidative stress and inflammation, thereby causing an amelioration of scar tissue formation following spinal surgery and improve the outcome. Morphological changes observed initially indicated that improvements in wound healing were occurring in the experimentally treated tissues. In addition, the scar tissue area and the lateral widths of the peridural scar forming between the muscular tissue areas suggested a reduction in the scar size. Although inflammatory cell numbers increased slightly in the experimental treatment groups, particularly during the initial three day post laminectomy time point, this increase was not statistically significant. <p>While quercetin and OTC did not appear to inhibit the influx of inflammatory cells following laminectomy, they did appear to induce a more beneficial wound healing environment. It is possible that these agents are affecting parameters of wound healing not considered by these studies. For instance the myriad of processes mediated by growth factors and cytokines involved in wound healing process may play a much greater role than the inflammatory cells themselves. In conclusion, reducing oxidative stress and inflammation by these agents to ameliorate scar tissue formation following spinal laminectomy was supported by the observed morphology, but not supported by the quantification of inflammatory cells. Additional studies investigating the efficacy of quercetin and OTC on the wound healing process are needed to further understand the role they play in repair and scar tissue formation.

In recent years, studies have increasingly recognized that many natural products with biological activity have been isolated from marine organisms, while the chemical structures are very different from those of land-based organisms. Therefore, the ocean is a natural drug source. Regarding drug screening, anti-inflammatory activity has become a key point, and many studies confirm that inflammation plays an important role in many human diseases. Many different compounds are now in the clinical evaluation stage. However, the inflammation-related diseases being closely linked, there is an urgent need to study the anti-inflammatory effects as well as screen the therapeutic drugs for research and development. In this study, we isolated and purified compounds from Formosan gorgonian (Briareum excavatum) and Formosan soft coral (Lobophytum sarcophytoides) and investigated biological activities. We confirmed that the natural compound Brei from B. excavatum and the compounds Sac-1 and Sac-2 from L. sarcophytoides produced significant inhibition of the proinflammatory proteins inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in the lipopolysaccharide (LPS)-induced murine macrophages (RAW 264.7) cell model. We examined in vivo whether the B. excavatum Brei has anti-inflammatory and antinociceptive effects by using the carrageenan-induced inflammation model. Using the paw-edema assay, we performed several important investigations such as the plantar analgesia test, mechanical hyperalgesia test (allodynia), and weight-bearing analysis of animal behavior to evaluate the degree of pain and inflammation. Our results demonstrate that the natural product Brei can reduce paw-pad swelling, thermal hyperalgesia, threshold latency, and improve the affected limb in the carrageenan-induced inflammatory model. In the histopathology analysis, we showed that Brei significantly inhibited the aggregation and infiltration of inflammation-related blood cells and improved the inflammatory status of the tissues. Therefore, the marine natural compound Brei has anti-inflammatory activity and it can be used as a therapeutic compound for acute inflammation in the near future.

Soft corals of the genus Briareum (Briareidae) have been well known as a rich source for marine natural products with novel structural features. Briarane-related natural products attracted the attentions of researchers because of the structural complexity and interesting biological activity associated with numerous compounds of this type. Previous studies on the secondary metabolites of wild-type and cultured Formosan octocoral Briareum excavatum were collected around the sea area of Kenting. In the thesis of our studies on secondary metabolites from marine organisms, the acetone-soluble of the Formosan octocoral B. excavatum collected at Orchid Island has led to the isolation of eleven briarane-type diterpenoids (1−11), compounds 3, 4, and 6−10 are new compounds. The structures of these compounds were determined on the basis of their spectroscopic analysis (1H NMR, 13C NMR, 1H−1H COSY, HSQC, HMBC, NOESY, IR and mass spectra) and physical data by comparison of the physical and spectral data with those of the related literatures. The antiviral activity against HCMV (human cytomegalovirus) cells of these secondary metabolites was evaluated. Metabolite 8 exhibited significant activity against HCMV cells and compound 11 showed anti-inflammatory activity.−

Placental vessels and the umbilical circulatory network function to carry oxygen and nutrients to the fetus. It is at this level that placental lesions such as villitis, obliterative vasculopathy, and thrombotic vasculopathy have been observed in association with fetal inflammatory response syndrome (FIRS) and cerebral palsy. We used human umbilical vein endothelial cells (HUVECs) as a model to study the regulation of inflammation and thrombosis in fetal vessels by microbial products. In this thesis we measured interleukin-8 (IL-8) and tissue factor (TF) expression by HUVECs treated with lipopolysaccharide (LPS), poly (I:C) (PIC), and peptidoglycan (PG). Our results show a profound induction of IL-8 by PIC, a TLR-3 ligand. We also show a moderate induction of tissue factor expression in PIC-treated HUVECs. These results show that HUVECs are exquisitely sensitive to PIC and suggests an important role for viral infection in umbilical vessel inflammation. We additionally treated HUVECs with dexamethasone (DEX), an anti-inflammatory steroid, and melatonin (MT), a pineal gland product with immunomodulatory and anti-oxidant properties. DEX reduced the level of both IL-8 and TF expression in PIC-treated cells. MT, however, further enhanced IL-8 expression in PIC-treated cells. Our results indicate a potential role for glucocorticoid therapy in reducing placental vessel inflammation and thrombosis. Thus, intervention with GC in pregnancies with FIRS may reduce the severity of placental lesions associated with cerebral palsy.

Diss. (sammanfattning) Göteborg : Göteborgs universitet, 2009. / Härtill 7 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2010.

Thesis (Ph. D.)-- State University of New York at Binghamton, Department of Biological Sciences, 2009.

Dissertation (Ph.D.)--University of South Florida, 2008. / Includes vita. Includes bibliographical references. Also available online.