Photobiomodulation inhibits the expression of chondroitin sulfate proteoglycans after spinal cord injury via the Sox9 pathway

Both glial cells and glia scar greatly affect the development of spinal cord injury and have become hot spots in research on spinal cord injury treatment.The cellular deposition of dense extracellular matrix proteins such as chondroitin sulfate proteoglycans inside and around the glial scar is known to affect axonal growth and be a major obstacle to autogenous repair.These proteins are thus candidate targets for spinal cord injury therapy.Our previous studies demonstrated that 810 nm photo biomodulation inhibited the formation of chondroitin sulfate proteoglycans after spinal cord injury and greatly improved motor function in model animals.However,the specific mechanism and potential targets involved remain to be clarified.In this study,to investigate the therapeutic effect of photo biomodulation,we established a mouse model of spinal cord injury by T9 clamping and irradiated the injury site at a power density of 50 mW/cm~2 for 50 minutes once a day for 7 consecutive days.We found that photobiomodulation greatly restored motor function in mice and down regulated chondroitin sulfate proteoglycan expression in the injured spinal cord.Bioinformatics analysis revealed that photobiomodulation inhibited the expression of proteoglycan-related genes induced by spinal cord injury,and versican,a type of proteoglycan,was one of the most markedly changed molecules.Immunofluorescence staining showed that after spinal cord injury,versican was present in astrocytes in spinal cord tissue.The expression of versican in primary astrocytes cultured in vitro increased after inflammation induction,whereas photobiomodulation inhibited the expression of ve rsican.Furthermore,we found that the increased levels of p-Smad3,p-P38 and p-Erk in inflammatory astrocytes were reduced after photobiomodulation treatment and after delivery of inhibitors including FR 180204,(E)-SIS3,and SB 202190.This suggests that Sma d 3/Sox9 and MAP K/Sox9 pathways may be involved in the effects of photobiomodulation.In summary,our findings show that photobiomodulation modulates the expression of chondroitin sulfate proteoglycans,and versican is one of the key target molecules of photo biomodulation.MAPK/Sox9 and Smad3/Sox9 pathways may play a role in the effects of photo biomodulation on chondroitin sulfate proteoglycan accumulation after spinal cord injury.

Proteoglycans in liver cancer

Proteoglycans are a group of molecules that contain at least one glycosaminoglycan chain,such as a heparan,dermatan,chondroitin,or keratan sulfate,covalently attached to the protein core.These molecules arecategorized based on their structure,localization,and function,and can be found in the extracellular matrix,on the cell surface,and in the cytoplasm.Cell-surface heparan sulfate proteoglycans,such as syndecans,are the primary type present in healthy liver tissue.However,deterioration of the liver results in overproduction of other proteoglycan types.The purpose of this article is to provide a current summary of the most relevant data implicating proteoglycans in the development and progression of human and experimental liver cancer.A review of our work and other studies in the literature indicate that deterioration of liver function is accompanied by an increase in the amount of chondroitin sulfate proteoglycans.The alteration of proteoglycan composition interferes with the physiologic function of the liver on several levels.This article details and discusses the roles of syndecan-1,glypicans,agrin,perlecan,collagen XVIII/endostatin,endocan,serglycin,decorin,biglycan,asporin,fibromodulin,lumican,and versican in liver function.Specifically,glypicans,agrin,and versican play significant roles in the development of liver cancer.Conversely,the presence of decorin could potentially provide protective effects.

Inhibition and enhancement of neural regeneration by chondroitin sulfate proteoglycans

The current dogma in neural regeneration research implies that chondroitin sulfate proteoglycans(CSPGs) inhibit plasticity and regeneration in the adult central nervous system(CNS). We argue that the role of the CSPGs can be reversed from inhibition to activation by developmentally expressed CSPG-binding factors. Heparin-binding growth-associated molecule(HB-GAM; also designated as pleiotrophin) has been studied as a candidate molecule that might modulate the role of CSPG matrices in plasticity and regeneration. Studies in vitro show that in the presence of soluble HB-GAM chondroitin sulfate(CS) chains of CSPGs display an enhancing effect on neurite outgrowth. Based on the in vitro studies, we suggest a model according to which the HB-GAM/CS complex binds to the neuron surface receptor glypican-2, which induces neurite growth. Furthermore, HB-GAM masks the CS binding sites of the neurite outgrowth inhibiting receptor protein tyrosine phosphatase sigma(PTPσ), which may contribute to the HB-GAM-induced regenerative effect. In vivo studies using two-photon imaging after local HB-GAM injection into prick-injury of the cerebral cortex reveal regeneration of dendrites that has not been previously demonstrated after injuries of the mammalian nervous system. In the spinal cord, two-photon imaging displays HB-GAM-induced axonal regeneration. Studies on the HB-GAM/CS mechanism in vitro and in vivo are expected to pave the way for drug development for injuries of brain and spinal cord.

In Silico Study on Sulfated and Non-Sulfated Carbohydrate Chains from Proteoglycans in Cnidaria and Their Interaction with Collagen

Proteoglycans and collagen molecules are interacting with each other thereby forming various connective tissues. The sulfation pattern of proteoglycans differs depending on the kind of tissue and/or the degree of maturation. Tissues from Cnidaria are suitable examples for exploration of the effects in relation to the presence and the absence of sulfate groups, when studying characteristic fragments of the long proteoglycan carbohydrate chains in silico. It has been described that a non-sulfated chondroitin appears as a scaffold in early morphogenesis of all nematocyst types in Hydra. On the other hand, sulfated glucosaminoglycans play an important role in various developmental processes of Cnidaria. In order to understand this biological phenomenon on a sub-molecular level we have analysed the structures of sulfated and non-sulfated proteoglycan carbohydrate chains as well as the structure of diverse collagen molecules with computational methods including quantum chemical calculations. The strong interactions between the sulfate groups of the carbohydrates moieties in proteoglycans and positively charged regions of collagen are essential in stabilizing various Cnidaria tissues but could hinder the nematocyst formation and its proper function. The results of our quantum chemical calculations show that the sulfation pattern has a significant effect on the conformation of chondroitin structures under study.

Proteoglycans and their functions in esophageal squamous cell carcinoma

Esophageal squamous cell carcinoma(ESCC)is a highly malignant disease that has a poor prognosis.Its high lethality is mainly due to the lack of symptoms at early stages,which culminates in diagnosis at a late stage when the tumor has already metastasized.Unfortunately,the common cancer biomarkers have low sensitivity and specificity in esophageal cancer.Therefore,a better understanding of the molecular mechanisms underlying ESCC progression is needed to identify novel diagnostic markers and therapeutic targets for intervention.The invasion of cancer cells into the surrounding tissue is a crucial step for metastasis.During metastasis,tumor cells can interact with extracellular components and secrete proteolytic enzymes to remodel the surrounding tumor microenvironment.Proteoglycans are one of the major components of extracellular matrix.They are involved in multiple processes of cancer cell invasion and metastasis by interacting with soluble bioactive molecules,surrounding matrix,cell surface receptors,and enzymes.Apart from having diverse functions in tumor cells and their surrounding microenvironment,proteoglycans also have diagnostic and prognostic significance in cancer patients.However,the functional significance and underlying mechanisms of proteoglycans in ESCC are not well understood.This review summarizes the proteoglycans that have been studied in ESCC in order to provide a comprehensive view of the role of proteoglycans in the progression of this cancer type.A long term goal would be to exploit these molecules to provide new strategies for therapeutic intervention.

EXTRACTION AND ISOLATION OF PROTEOGLYCANS FROM RAT BRAIN

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Interaction of arterial proteoglycans with low density lipoproteins(LDLs):From theory to promising therapeutic approaches

Although atherosclerosis is a multifactorial process,proteoglycans mediated lipoprotein(LDL)retention at the subendothelial space is a necessary and sufficient event in provoking lesion initiation.Proteoglycans(PGs)are usually composed of one core protein backbone with one or more glycosaminoglycan chains(GAGs)covalently linked,mainly include perlecan,biglycan,versican,and decorin.The interaction between LDL and proteoglycans is apparently mediated by the basic amino acids in apoB-100,the moiety of LDL,electrostatic interacting with the negatively charged GAGs(sulfate or carbohydrate groups)of proteoglycans or though some bridge molecules like sphingomyelinase(SMase)or lipoprotein lipase(LpL).In the later section,we collate the promising therapeutic approaches that have been proposed up to now,targeting LDL-PGs interaction.It should be concluded that previous studies on interaction between LDL and PGs mainly focused on perlecan,biglycan,decorin,and versican that all located in the extracellular matrix(ECM),future studies should pay more attention to the endothelial surface glycocalyx and its interaction with LDLs,seeking promising therapeutic targets more specifically.

Abnormal Proteoglycans Secreted by Chondrocytes in Actions of Fulvic Acid and Reactive Oxygen Species

Articular cartilage is an avascular and non-nervous tissue. The transport of metabolites to and from the chondrocytes has to take place through the matrix and hence depends on its composition and structure. A major parameter controlling the properties of cartilage is the proteoglycan (PG) content of the matrix, whose small

Axonal growth inhibitors and their receptors in spinal cord injury:from biology to clinical translation

Axonal growth inhibitors are released during traumatic injuries to the adult mammalian central nervous system, including after spinal cord injury. These molecules accumulate at the injury site and form a highly inhibitory environment for axonal regeneration. Among these inhibitory molecules, myelinassociated inhibitors, including neurite outgrowth inhibitor A, oligodendrocyte myelin glycoprotein, myelin-associated glycoprotein, chondroitin sulfate proteoglycans and repulsive guidance molecule A are of particular importance. Due to their inhibitory nature, they represent exciting molecular targets to study axonal inhibition and regeneration after central injuries. These molecules are mainly produced by neurons, oligodendrocytes, and astrocytes within the scar and in its immediate vicinity. They exert their effects by binding to specific receptors, localized in the membranes of neurons. Receptors for these inhibitory cues include Nogo receptor 1, leucine-rich repeat, and Ig domain containing 1 and p75 neurotrophin receptor/tumor necrosis factor receptor superfamily member 19(that form a receptor complex that binds all myelin-associated inhibitors), and also paired immunoglobulin-like receptor B. Chondroitin sulfate proteoglycans and repulsive guidance molecule A bind to Nogo receptor 1, Nogo receptor 3, receptor protein tyrosine phosphatase σ and leucocyte common antigen related phosphatase, and neogenin, respectively. Once activated, these receptors initiate downstream signaling pathways, the most common amongst them being the Rho A/ROCK signaling pathway. These signaling cascades result in actin depolymerization, neurite outgrowth inhibition, and failure to regenerate after spinal cord injury. Currently, there are no approved pharmacological treatments to overcome spinal cord injuries other than physical rehabilitation and management of the array of symptoms brought on by spinal cord injuries. However, several novel therapies aiming to modulate these inhibitory proteins and/or their receptors are under investigation in ongoing clinical trials. Investigation has also been demonstrating that combinatorial therapies of growth inhibitors with other therapies, such as growth factors or stem-cell therapies, produce stronger results and their potential application in the clinics opens new venues in spinal cord injury treatment.

Galectin-14 promotes hepatocellular carcinoma tumor growth via enhancing heparan sulfate proteoglycan modification

Hepatocellular carcinoma(HCC)is a highly heterogeneous malignancy and lacks effective treatment.Bulk-sequencing of different gene transcripts by comparing HCC tissues and adjacent normal tissues provides some clues for investigating the mechanisms or identifying potential targets for tumor progression.However,genes that are exclusively expressed in a subpopulation of HCC may not be enriched or detected through such a screening.In the current study,we performed a single cell-clone-based screening and identified galectin-14 as an essential molecule in the regulation of tumor growth.The aberrant expression of galectin-14 was significantly associated with a poor overall survival of liver cancer patients with database analysis.Knocking down galectin-14 inhibited the proliferation of tumor growth,whereas overexpressing galectin-14 promoted tumor growth in vivo.Non-targeted metabolomics analysis indicated that knocking down galectin-14 decreased glycometabolism;specifically that glycoside synthesis was significantly changed.Further study found that galectin-14 promoted the expression of cell surface heparan sulfate proteoglycans(HSPGs)that functioned as co-receptors,thereby increasing the responsiveness of HCC cells to growth factors,such as epidermal growth factor and transforming growth factor-alpha.In conclusion,the current study identifies a novel HCC-specific molecule galectin-14,which increases the expression of cell surface HSPGs and the uptake of growth factors to promote HCC cell proliferation.

Epidural electrical stimulation for spinal cord injury

A long-standing goal of spinal cord injury research is to develop effective repair strategies,which can restore motor and sensory functions to near-normal levels.Recent advances in clinical management of spinal cord injury have significantly improved the prognosis,survival rate and quality of life in patients with spinal cord injury.In addition,a significant progress in basic science research has unraveled the underlying cellular and molecular events of spinal cord injury.Such efforts enabled the development of pharmacologic agents,biomaterials and stem-cell based therapy.Despite these efforts,there is still no standard care to regenerate axons or restore function of silent axons in the injured spinal cord.These challenges led to an increased focus on another therapeutic approach,namely neuromodulation.In multiple animal models of spinal cord injury,epidural electrical stimulation of the spinal cord has demonstrated a recovery of motor function.Emerging evidence regarding the efficacy of epidural electrical stimulation has further expanded the potential of epidural electrical stimulation for treating patients with spinal cord injury.However,most clinical studies were conducted on a very small number of patients with a wide range of spinal cord injury.Thus,subsequent studies are essential to evaluate the therapeutic potential of epidural electrical stimulation for spinal cord injury and to optimize stimulation parameters.Here,we discuss cellular and molecular events that continue to damage the injured spinal cord and impede neurological recovery following spinal cord injury.We also discuss and summarize the animal and human studies that evaluated epidural electrical stimulation in spinal cord injury.

Arylsulfatase,β-galactosidase and lysozyme in gastric cancer cells and its relationship to invasion

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Glycosaminoglycan remodeling during diabetes and the role of dietary factors in their modulation

Glycosaminoglycans(GAGs) play a significant role in various aspects of cell physiology.These are complex polymeric molecules characterized by disaccharides comprising of uronic acid and amino sugar.Compounded to the heterogeneity,these are variously sulfated and epimerized depending on the class of GAG.Among the various classes of GAG,namely,chondroitin/dermatan sulfate,heparin/heparan sulfate,keratan sulfate and hyaluronic acid(HA),only HA is non-sulfated.GAGs are known to undergo remodeling in various tissues during various pathophysiological conditions,diabetes mellitus being one among them.These changes will likely affect their structure thereby impinging on their functionality.Till date,diabetes has been shown to affect GAGs in organs such as kidney,liver,aorta,skin,erythrocytes,etc.to name a few,with deleterious consequences.One of the mainstays in the treatment of diabetes is though dietary means.Various dietary factors are known to play a significant role in regulating glucose homeostasis.Furthermore,in recent years,there has been a keen interest to decipher the role of dietary factors on GAG metabolism.This review focuses on the remodeling of GAGs in various organs during diabetes and their modulation by dietary factors.While effect of diabetes on GAG metabolism has been worked out quite a bit,studies on the role of dietary factors in their modulation has been few and far between.We have tried our best to give the latest reports available on this subject.

Promotion of axon regeneration and inhibition of astrocyte activation by alpha A-crystallin on crushed optic nerve

AIM:To explore the effects of αA-crystallin in astrocyte gliosis after optic nerve crush(ONC) and the mechanism of α-crystallin in neuroprotection and axon regeneration.METHODS:ONC was established on the SpragueDawley rat model and αA-crystallin(10^(-4) g/L,4 μL) was intravitreously injected into the rat model.Flash-visual evoked potential(F-VEP) was examined 14 d after ONC,and the glial fibrillary acidic protein(GFAP) levels in the retina and crush site were analyzed 1,3,5,7 and 14 d after ONC by immunohistochemistry(IHC) and Western blot respectively.The levels of beta Tubulin(TUJ1),growth-associated membrane phosphoprotein-43(GAP-43),chondroitin sulfate proteoglycans(CSPGs) and neurocan were also determined by IHC 14 d after ONC.RESULTS:GFAP level in the retina and the optic nerve significantly increased 1d after ONC,and reached the peak level 7d post-ONC.Injection of αA-crystallin significantly decreased GFAP level in both the retina and the crush site 3d after ONC,and induced astrocytes architecture remodeling at the crush site.Quantification of retinal ganglion cell(RGC) axons indicated αAcrystallin markedly promoted axon regeneration in ONC rats and enhanced the regenerated axons penetrated into the glial scar.CSPGs and neurocan expression also decreased 14 d after αA-crystallin injection.The amplitude(N1-P1) and latency(P1) of F-VEP were also restored.CONCLUSION:Our results suggest α-crystallin promotes the axon regeneration of RGCs and suppresses the activation of astrocytes.

蛋白多糖阻抑:涎腺肿瘤治疗的新思路

涎腺肿瘤性肌上皮细胞(neoplastic myoepithelial cells,NMCs)的存在、增殖和分化,导致了涎腺肿瘤组织学形态和临床表现的多样性和复杂性,给临床诊断和治疗带来困难。NMCs是涎腺肿瘤中主要增生的细胞成份,NMCs具有分泌产生蛋白多糖(proteoglycan,PG)的功能。PG是一类大分子糖蛋白,由葡胺多糖与核心蛋白结合构成。研究表明,PG在肿瘤基质中的表达明显高于正常组织或肿瘤周围组织。PG作为细胞外基质的组成部分,对涎腺肿瘤发生、增殖、迁移、侵袭和转移等方面都具有重要作用。通过抑制涎腺NMCs的PG合成与分泌,可以达到抑制涎腺肿瘤增殖、迁移、侵袭和转移的目的,为涎腺肿瘤的治疗提供新思路和新策略。

Glypican-3在胚胎及肿瘤中的生物学研究进展

磷脂酰肌醇聚糖-3蛋白（Glypican-3,GPC3,也称为DGSX,GTR2-2,MXR7,OCI-5,SDYS,SGB,SGBS和SGBS1）是硫酸乙酰肝素糖蛋白（heparan sulfate proteoglycan,HSPG）家族的成员之一,可通过共受体结合相关配基,如生长因子、细胞黏附分子、酶和酶抑制剂、基质成分等,参与调节细胞增殖、分化、黏附和迁移等过程,同时参与调节大部分中胚层组织和器官形成过程。近年来大量研究表明,

Perineuronal nets increase inhibitory GABAergic currents during the critical period in rats

AIM: To investigate inhibitory γ-aminobutyric acid (GABA) ergic postsynaptic currents (IPSCs) and postsynaptic currents (PSCs) in layer IV of the rat visual cortex during the critical period and when plasticity was extended through dissolution of the perineuronal nets (PNNs). METHODS: We employed 24 normal Long-Evans rats to study GABA A-PSC characteristics of neurons within layer IV of the visual cortex during development. The animals were divided into six groups of four rats according to ages at recording: PW3 (P21 -23d), PW4 (P28 -30d), PW5 (P35-37d), PW6 (P42-44d), PW7 (P49-51d), and PW8 (56-58d). An additional 24 chondroitin sulfate proteoglycan (CSPG) degradation rats (also Long-Evans) were generated by making a pattern of injections of chondroitinase ABC (chABC) into the visual cortex 1 week prior to recording at PW3, PW4, PW5, PW6, PW7, and PW8. Immunohistochemistry was used to identify the effect of chABC injection on CSPGs. PSCs were detected with whole-cell patch recordings, and GABA A receptor-mediated IPSCs were pharmacologically isolated. RESULTS: IPSC peak current showed a strong rise in the age-matched control group, peaked at PW5 and were maintained at a roughly constant value thereafter. Although there was a small increase in peak current for the chABC group with age, the peak currents continued to decrease with the delayed highest value at PW6, resulting in significantly different week-by-week com-parison with normal development. IPSC decay time continued to increase until PW7 in the control group, while those in the chABC group were maintained at astable level after an initial increase at PW4. Compared with normal rats, the decay times recorded in the chABC rats were always shorter, which differed significantly at each age. We did not observe any differences in IPSC properties between the age-matched control and penicillinase (P-ase) group. However, the change in IPSCs after chABC treatment was not reflected in the total PSCs or in basic membrane properties in layer IV of the rat visual cortex. CONCLUSION: Our results demonstrate that rather than rapidly increasing during the critical period for neuronal plasticity, IPSCs in layer IV of rat visual cortex are maintained at an immature level when PNNs are removed by chABC. This suggests that GABA receptor maturation involves the conformation of the CSPGs in PNNs.

细胞因子与肾小球细胞外基质

肾小球系膜细胞和细胞外基质(extra-cellular matrix,ECM)是肾小球重要的结构和功能单位。肾小球基底膜是特殊的ECM,它主要由IV型胶原(type IV colla-gen)、层粘蛋白(laminin)和硫酸肝素糖蛋白(heparan sulphate proteoglycan)组成。此外,ECM还包括纤维连接蛋白(fi-bronectin)和硫酸软骨素糖蛋白(chondr-

TLR5 involvement in attenuated IL-8 production in nuclear decorin silenced oral mucosal dysplastic keratinocytes and squamous cell carcinoma

Head and neck cancer is one of the most prevalent cancers in the world. Roughly half of these malignancies originate from oral mucosa and constitute Oral squamous cell carcinomas. Despite many advances in diagnostic and therapeutic regimens, five year survival rate remains at roughly 50 %, indicating the need for in depth understanding of the oral squamous cell carcinoma immunobiology. We have previously shown that in human dysplastic oral keratinocytes (DOK) and malignant squamous cells carcinoma (SCC-25), multifunctional proteoglycan decorin is aberrantly expressed and localized in the nucleus bound to nuclear EGFR. In vitro nuclear decorin knockdown significantly reduced IL-8 and IL8-dependent migration, invasion and angiogenesis in these cells. Since toll-like receptor (TLR) signalling leads to IL-8 production we examined here if these receptors play a role in decorin silencing mediated reduction in IL-8 levels. We have used immunological and molecular techniques to study toll-like receptors involvement in attenuated IL-8 production in nuclear decorin silenced (stable knockdown) oral mucosal dysplastic keratinocytes and squamous carcinoma cells. We show that nuclear decorin silenced DOK and SCC-25 cells show marked diminution of TLR5 mRNA and protein expression compared with respective controls that translated into loss of function in response to appropriate TLR ligand. In these mucosal oral epithelia, decorin stable knockdown significantly down-regulated IL-8 production following activation with TLR5 ligand flagellin. These data suggest that decorin silencing interferes with IL-8 production, in part, by altering TLR5 expression and signaling in dysplastic and malignant oral epithelia. This study highlights the significance of TLR5 expression and signaling in mucosal cancers.

Manifestation of Pathological States of Numerous Diseases in the Largest Organ of the Human Body: (I) Basics and the Diseases of Tendon

We analyze the crucial biochemical and biophysical properties of the basic constituents—connective tissues (CT), and interstitial fluid (IF) constituting the non-cellular part of the fascia. We provide ample evidence that the resident cells and cells in transit in the fascia are continuously interacting with the non-cellular constituents to form an active organ with well-defined functions. We show evidence that pathological states of diseases of internal organs, as well as that of the constituents of the fascia itself, manifest in certain CTIF domains of the fascia. Numerous diseases originate from imbalance of the digestion and synthesis of the native collagen triple helices. Review on the scanning electron microscopy examination of cross-section of tendons indicates that micro-fibrils of collagen I form regular geometrical structures, supporting the hypothesis that the collagen fibrils assemble like liquid crystals. Information on the age of Achilles tendons has been reported, based on dating of the 14C atoms generated from the nuclear bomb tests in 1955-1963. The causes of spontaneous tendon rupture and tendinopathy are analyzed. Plausible clinical measures to treat tendinopathy are briefly discussed, including the application of synthetic mechano-growth factor, glyceryl trinitrate patch (to supply nitric oxide), platelet rich plasma, proteomic profile analysis and microRNA 29a therapy.