Characterization of a bioscaffold containing polysaccharide acemannan and native collagen for pulp tissue regeneration.

Dental pulp regeneration exploits tissue engineering concepts using stem cells/scaffolds/growth-factors. Extracted collagen is commonly used as a biomaterial-scaffold due to its biocompatibility/biodegradability and mimics the natural extracellular matrix. Adding biomolecules into a collagen-scaffold enhanced pulp regeneration. Acemannan, ß-(1-4)-acetylated-polymannose, is a polysaccharide extracted from aloe vera. Acemannan is a regenerative biomaterial. Therefore, acemannan could be a biomolecule in a collagen-scaffold. Here, acemannan and native collagen were obtained and characterized. The AceCol-scaffold's physical properties were investigated using FTIR, SEM, contact angle, swelling, pore size, porosity, compressive modulus, and degradation assays. The AceCol-scaffold's biocompatibility, growth factor secretion, osteogenic protein expression, and calcification were evaluated in vitro. The AceCol-scaffolds demonstrated higher hydrophilicity, swelling, porosity, and larger pore size than the collagen scaffolds (p < 0.05). Better cell-cell and cell-scaffold adhesion, and dentin extracellular matrix protein (BSP/OPN/DSPP) expression were observed in the AceCol-scaffold, however, DSPP expression was not detected in the collagen group. Significantly increased cellular proliferation, VEGF and BMP2 expression, and mineralization were detected in the AceCol-scaffold compared with the collagen-scaffold (p < 0.05). Computer simulation revealed that acemannan's 3D structure changes to bind with collagen. In conclusion, the AceCol-scaffold synergistically provides better physical and biological properties than collagen. The AceCol-scaffold is a promising material for tissue regeneration.

Lipid biomarker profiling of adult Brugia malayi using mass spectrometry detection.

In this article, we first reported the lipid profile of adult Brugia malayi using ultra-performance liquid chromatography electrospray ionization mass spectrometry (UPLC-ESI-MS) to provide a promising drug target for lymphatic filariasis (LF). The MS and MS/MS data analysis indicated that there were target lipids, phosphatidylcholines (PC), at m/z 769.5612 and 831.5767, which were found only in adult males, while at m/z 811.6072, they were found only in females, which is interesting for use as a biomarker in LF disease. In addition, the lipid profile showed that three membrane lipid classes, glycerophospholipids, glycerolipids and sphingolipids, were discovered. Glycerophospholipids were the main components in adult parasites, especially phosphatidylcholine (60%) and phosphatidylethanolamine (27%). Phosphatidylglycerol (5%), phosphatidylserine (4%), phosphatidylinositol (4%) and phosphatidic acid (3%). Consequently, the lipid profile of adult B. malayi is significant and not only provides a promising drug target for LF but also assists in a better understanding of the biological process and mechanism by which parasites interfere with LF disease in the future. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03296-y.

Global Analysis of Protein Expression in A549 Cells After Prolonged Nicotine Exposure by Using Label-free Quantification.

BACKGROUND/AIM: Lung cancer is the leading cause of cancer death worldwide. Cigarette smoke is the most important risk factor for cancer development. Growing evidence indicates that prolonged nicotine exposure is a potential factor associated with tumorigenesis. Here, the effect of prolonged nicotine exposure on A549 cells was investigated, using label-free quantitative proteomics. MATERIALS AND METHODS: Selection of an invasive subpopulation from the A549 cell line was performed to reveal the differential expression of proteins in relation to prolonged nicotine exposure, using Boyden chamber assays in combination with a proteomics approach. RESULTS: One hundred proteins from the NicoA549-L5 subline showed significant change in expression compared to those from the A549-L5 subline and their A549 parental cell line. Heat shock protein, protein disulfide isomerase A3, profilin-1 and legumain were expressed at higher levels in A549 cells after prolonged nicotine exposure. CONCLUSION: These aberrant proteins might serve as novel cancer biomarkers for cigarette smokers.

Acemannan-induced tooth socket healing: A 12-month randomized controlled trial.

BACKGROUND/PURPOSE: Natural compounds have become alternatives for bone regeneration. Acemannan, the main polysaccharide extracted from Aloe vera, has been demonstrated as a promising osteoinductive material in vitro and in vivo. This clinical study investigated the effect of acemannan on tooth socket healing. MATERIALS AND METHODS: Thirty-five otherwise healthy patients, 18-25 years old and diagnosed with horizontal or vertical partial impaction of the lower third molars, were enrolled in this randomized controlled trial. After removing the teeth, the sockets randomly received one of the following treatments: spontaneous blood-clotting (control), 20 mg acemannan sponge, or 50 mg acemannan sponge. Cone-beam computed tomography of the mandible was performed immediately (baseline), and at 3-, 6-, and 12-months postoperatively; the data were analyzed using the OsiriX MD program. Bone healing in the socket was determined measuring the socket volume. One-way ANOVA was used to analyze the differences within each group and between groups. RESULTS: Thirty-five patients with 43 partially impacted lower third molars participated in this study. No patients exhibited alveolar osteitis or secondary infection. Compared with baseline, all groups showed significant reduction in socket volume at all observation time-points (p < 0.05). The 50 mg acemannan group had a significantly greater reduction in socket volume compared with the control at all postoperative time-points (p < 0.05). The 20 mg group had a significantly greater reduction in socket volume compared with the control at 3-months postoperatively (p < 0.05). CONCLUSION: We conclude that acemannan increases bone healing at 3-, 6-, and 12-months after removal of partially impacted mandibular third molars.

Acemannan induces rapid early osseous defect healing after apical surgery: A 12-month follow-up of a randomized controlled trial.

BACKGROUND/PURPOSE: Acemannan is an osteoinductive material. This study's objective was to compare the outcomes of bone defect healing using 3-dimensional images after apical surgery with or without adding acemannan sponges. MATERIALS AND METHODS: Twenty-two anterior teeth from 9 males and 13 females requiring apical surgery were included in this randomized controlled trial. Post-surgery, the bone defects were randomly divided into three groups: blood clot control, 5-, or 10-mg acemannan sponge groups. CBCT scans were taken immediately (baseline), 3-, 6-, and 12-month post-surgery. Sagittal serial sections (1 mm thick slices parallel to the long axis of the tooth) of the defect image were created. The defect boundary was located and the total bone defect volume (BDV) was calculated from the sum of the volume of the serial defect sections. The bone healing was assessed by the percentage of total bone defect volume reduction (%ΔBDV). The paired t-test and one-way ANOVA were used to analyze the differences within each group and between groups, respectively. RESULTS: The baseline mean BDV of the control, 5-, and 10-mg acemannan groups were not significantly different (p > 0.05). After treatment, the mean BDV for each group was reduced in a time-dependent manner. Compared with the control group, the 5- and 10-mg acemannan groups had a significantly greater %ΔBDV (approximately 2- and 1.89-fold) at 3-months post-surgery, respectively (p < 0.05). However, at the 6- and 12- month follow-up, the %ΔBDV was not significantly different between the groups. CONCLUSION: These data suggest acemannan enhanced early bone healing after apical surgery.

Acemannan Used as an Implantable Biomaterial for Vital Pulp Therapy of Immature Permanent Teeth Induced Continued Root Formation.

Direct pulp-capping, a vital pulp therapy, is used to protect and preserve pulp vitality by applying a biomaterial on the pulp exposure site. Acemannan, a polysaccharide extracted from Aloe vera, induces osteodentin-bridge formation to cover the exposure site in vivo. We evaluated the effect of acemannan sponges on partial pulpotomized permanent teeth with caries or accident-induced pulp exposure (n = 50). After removing infected dentin and inflamed pulp tissue, the teeth were randomly divided into acemannan or control (mineral trioxide aggregate (MTA) groups (n = 25). The teeth were examined immediately after treatment (baseline) and at 6- and 12-month follow-ups for clinical and cone beam computed tomography (CBCT) examination. The three-dimensional tooth length and root apex area were simulated to determine treatment success. We found that the overall success rate in the acemannan and MTA groups from baseline to 12-month follow-up was 90.91% and 95.65%, respectively, with no significant difference between the two groups (p > 0.05). In the success teeth in both groups, the root length increased, and the apex area significantly decreased (p < 0.05), indicating continued root formation. Our results suggest that acemannan is a promising low-cost biomaterial for partial pulpotomy treatment for immature permanent teeth requiring vital pulp therapy.

Acemannan Induced Bone Regeneration in Lateral Sinus Augmentation Based on Cone Beam Computed Tomographic and Histopathological Evaluation.

Acemannan, the major polysaccharide extracted from Aloe vera, is biomaterial that has demonstrated osteoinductive effects in vitro and in vivo. However, the effect of acemannan sponges on bone formation in open-type sinus augmentation has not evaluated. Here, we report a case study using radiographic and histological analyses to investigate the effect of acemannan on bone formation after lateral sinus lift surgery. The case was a 57-year-old female patient with an atrophic left posterior maxilla who underwent lateral sinus lift using an acemannan sponge using the two-stage procedure. In the first stage, an acemannan sponge was inserted through the bony window and placed between the antral floor and the elevated sinus membrane. Cone beam computed tomography (CBCT) images were taken immediately as baseline and 6-month postoperation for evaluation. A bone core specimen was also obtained for histological examination at the time of implant placement. The histological results revealed new bone formation, and the CBCT images demonstrated increased alveolar bone height at 6-month postoperation. Our findings suggest that an acemannan sponge could be a biomaterial for inducing bone formation in sinus lift surgery.

Variation in the Protein Composition and Biological Activity of King Cobra (Ophiophagus hannah) Venoms.

The biochemical properties and biological activities of the venom from three individual Ophiophagus hannah (King cobra) specimens was compared. The toxicity against mice, the cytotoxicity against five cell lines, and the antioxidant activity were measured. The KV2 venom showed a higher cytotoxicity than the KV6 and the non-cytotoxic KV9 venoms. Comparative analysis of the O. hannah venom proteins was performed after 2-dimensional (2-D) denaturing gel electrophoresis and reverse phase high performance liquid chromatography (RP-HPLC). 2-D analysis by isoelectric focusing (IEF) Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) resolution of the venoms revealed significant differences between all three venoms, with most spots being unique to that venom. Only 2 out of the 13-16 distinct spots were common to all three venoms, and four spots were common to KV6 and KV9. KV2 had the highest proportion of low molecular mass spots, and KV6 and KV9 appeared more related to each other than to KV9. From peptide mass mapping by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and MASCOT-based amino acid sequence database searching, the two venom proteins that were common to all three specimens are likely to be ophanin and acidic phospholipase A2 (PLA2), whilst the proteins unique to the cytotoxic KV2 venom, included three other PLA2 proteins. The RP-HPLC pattern of KV2 was different from the other two venoms with a higher protein concentration eluting in the 31-41% (v/v) acetonitrile (ACN) fraction than for the other two venoms.

Collagen and mPCL-TCP scaffolds induced differential bone regeneration in ovary-intact and ovariectomized rats.

BACKGROUND: The data on biomaterial-mediated bone regeneration directly comparing commercially available scaffolds in aging and osteoporotic conditions remain sparse. OBJECTIVE: To investigate the effects of an absorbable collagen sponge (ACS) and a medical grade polycaprolactone-tricalcium phosphate (mPCL-TCP) scaffold on calvarial defect healing in ovary-intact and ovariectomized rats. METHODS: Forty-two, 5-month old female Sprague-Dawley rats were divided into sham (OVI) or ovariectomy (OVX) groups (n=21). When rats reached 6 months old, 7 mm diameter calvarial defects were created and treated, further dividing each group into blood clot control, mPCL-TCP, or ACS subgroups (n=7). After four weeks, the calvarial specimens were evaluated using micro-computed tomography for bone volume fraction (BVF), and histopathology. RESULTS: The effects of ovariectomy were confirmed by changes in body, uterine, and vaginal weight, and osteopenia in the femur. A significant increase in BVF was observed in ACS-subgroups compared with their respective control groups (p<0.05). Histopathological analysis revealed no cellular inflammatory infiltrate in any group. Fibrous tissue encapsulated the mPCL-TCP, while the ACS was well-integrated with the bone matrix. The OVX groups presented more osteoid and enlarged marrow cavities compared with the OVI groups. CONCLUSION: ACS scaffold enhanced calvarial bone regeneration in OVI and OVX rats after four weeks.

Acemannan increased bone surface, bone volume, and bone density in a calvarial defect model in skeletally-mature rats.

BACKGROUND/PURPOSE: Acemannan, a ß-(1-4)-acetylated polymannose extracted from Aloe vera gel, has been proposed as biomaterial for bone regeneration. The aim of this study was to investigate the effect of acemannan in calvarial defect healing. MATERIALS AND METHODS: Acemannan was processed to freeze-dried sponge form and disinfected by UV irradiation. Thirty-five female Sprague-Dawley rats were used in the in vivo study. Seven-mm diameter mid-calvarial defects were created and randomly allocated into blood clot control (C), acemannan 1 mg (A1), 2 mg (A2), 4 mg (A4), and 8 mg (A8) groups (n = 7). After four weeks, the calvarial specimens were subjected to microcomputed tomography (microCT) and histopathological analysis. RESULTS: MicroCT revealed a significant increase in bone surface and bone volume in the A1 and A2 groups, and tissue mineral density in the A4 and A8 groups compared with the control group (p < 0.05). Histologically, the acemannan-treated groups had denser bone matrix compared with the control group. CONCLUSION: Acemannan is an effective bioactive agent for bone regeneration, enhancing bone growth as assayed in two- and three-dimensions.

Acemannan increases NF-κB/DNA binding and IL-6/-8 expression by selectively binding Toll-like receptor-5 in human gingival fibroblasts.

Acemannan, an acetylated polymannose from Aloe vera, has immunomodulatory effects. We investigated whether acemannan induces IL-6 and -8 expression and NF-κB/DNA binding in human gingival fibroblasts. IL-6 and -8 expression levels were assessed via RT-PCR and ELISA. The NF-κB p50/p65-DNA binding was determined. The structures of acemannan mono-pentamers and Toll-like receptor 5 (TLR5) were simulated. The binding energies between acemannan and TLR5 were identified. We found that acemannan significantly stimulated IL-6/-8 expression at both the mRNA and protein level and significantly increased p50/DNA binding. Preincubation with an anti-TLR5 neutralizing antibody abolished acemannan-induced IL-6/-8 expression and p50/DNA binding, and co-incubation of acemannan with Bay11-7082, a specific NF- κB inhibitor, abolished IL-6/-8 expression. The computer modeling indicated that monomeric/dimeric single stranded acemannan molecules interacted with the TLR5 flagellin recognition sites with a high binding affinity. We conclude that acemannan induces IL-6/-8 expression, and p50/DNA binding in gingival fibroblasts, at least partly, via a TLR5/NF-κB-dependent signaling pathway. Furthermore, acemannan selectively binds with TLR5 ectodomain flagellin recognition sites.

Effects of protein hydrolysate from chicken feather meal on tyrosinase activity and melanin formation in B16F10 murine melanoma cells.

Tyrosinase is a copper-containing enzyme that controls mammalian melanogenesis. Tyrosinase inhibitors are important for their potential application in cosmetic products. Chicken feather meal is a rich source of amino acids, which have been linked with tyrosinase inhibition activity. This study investigated the tyrosinase inhibitory properties of protein hydrolysates prepared from chicken feather meal. Protein hydrolysates prepared by pepsin-pancreatin with MW <3 kDa exhibited strong tyrosinase inhibition activity for both monophenolase (IC50 5.780 ± 0.188 µg/mL) and diphenolase activities (IC50 0.040 ± 0.024 µg/mL) in a cell-free mushroom tyrosinase system. These samples were uncompetitive inhibitors with Ki values of 18.149 and 27.189 µg/mL in monophenolase and diphenolase activities, respectively. A cell culture model showed that this hydrolysate had the strongest inhibition on the viability of B16F10 cells (IC50 1.124 ± 0.288 µg/mL) and 0.210 µg/mL of the sample exhibited inhibition of tyrosinase activity by 50.493% and melanin synthesis by 14.680% compared to the control.

Serum lipidomics analysis of ovariectomized rats under Curcuma comosa treatment.

ETHNOPHARMACOLOGICAL RELEVANCE: Curcuma comosa Roxb. (C. comosa) or Wan Chak Motluk, Zingiberaceae family, has been used in Thai traditional medicine for the treatment of gynecological problems and inflammation. AIM OF THE STUDY: This study aimed to investigate the therapeutic potential of C. comosa by determining the changes in the lipid profiles in the ovariectomized rats, as a model of estrogen-deficiency-induced hyperlipidemia, after treatment with different components of C. comosa using an untargeted lipidomics approach. MATERIALS AND METHODS: Lipids were extracted from the serum of adult female rats subjected to a sham operation (SHAM; control), ovariectomy (OVX), or OVX with 12-week daily doses of estrogen (17ß-estradiol; E2), (3R)-1,7-diphenyl-(4E,6E)-4,6-heptadien-3-ol (DPHD; a phytoestrogen from C. comosa), powdered C. comosa rhizomes or its crude ethanol extract. They were then analyzed by liquid chromatography-mass spectrometry, characterized, and subjected to the orthogonal projections to latent structures discriminant analysis statistical model to identify tentative biomarkers. RESULTS: Levels of five classes of lipids (ceramide, ceramide-1-phosphate, sphingomyelin, 1-O-alkenyl-lysophosphatidylethanolamine and lysophosphatidylethanolamine) were elevated in the OVX rats compared to those in the SHAM rats, while the monoacylglycerols and triacylglycerols were decreased. The E2 treatment only reversed the levels of ceramides, whereas treatments with DPHD, C. comosa extract or powder returned the levels of all upregulated lipids back to those in the SHAM control rats. CONCLUSIONS: The findings suggest the potential beneficial effects of C. comosa on preventing the increased ceramide levels in OVX rats, a possible cause of metabolic disturbance under estrogen deficiency. Overall, the results demonstrated the power of untargeted lipidomics in discovering disease-relevant biomarkers, as well as evaluating the effectiveness of treatment by C. comosa components (DPHD, extract or powder) as utilized in Thai traditional medicine, and also providing scientific support for its folklore use.

Discovery, isolation, and structural characterization of cyclotides from Viola sumatrana Miq.

Cyclotides are cyclic peptides from plants in the Violaceae, Rubiaceae, Fabaceae, Cucurbitaceae, and Solanaceae families. They are sparsely distributed in most of these families, but appear to be ubiquitous in the Violaceae, having been found in every plant so far screened from this family. However, not all geographic regions have been examined and here we report the discovery of cyclotides from a Viola species from South-East Asia. Two novel cyclotides (Visu 1 and Visu 2) and two known cyclotides (kalata S and kalata B1) were identified in V. sumatrana. NMR studies revealed that kalata S and kalata B1 had similar secondary structures. Their biological activities were determined in cytotoxicity assays; both had similar cytotoxic activity and were more toxic to U87 cells compared with other cell lines. Overall, the study strongly supports the ubiquity of cyclotides in the Violaceae and adds to our understanding of their distribution and cytotoxic activity.

Comparative protein profiles of Butea superba tubers under seasonal changes.

Seasonal changes are major factors affecting environmental conditions which induce multiple stresses in plants, leading to changes in protein relative abundance in the complex cellular plant metabolic pathways. Proteomics was applied to study variations in proteome composition of Butea. superba tubers during winter, summer and rainy season throughout the year using two-dimensional polyacrylamide gel electrophoresis coupled with a nanoflow liquid chromatography coupled to electrospray ionization quadrupole-time-of-flight tandem mass spectrometry. A total of 191 protein spots were identified and also classified into 12 functional groups. The majority of these were mainly involved in carbohydrate and energy metabolism (30.37 %) and defense and stress (18.32 %). The results exhibited the highest numbers of identified proteins in winter-harvested samples. Forty-five differential proteins were found in different seasons, involving important metabolic pathways. Further analysis indicated that changes in the protein levels were due mainly to temperature stress during summer and to water stress during winter, which affected cellular structure, photosynthesis, signal transduction and homeostasis, amino-acid biosynthesis, protein destination and storage, protein biosynthesis and stimulated defense and stress mechanisms involving glycolytic enzymes and relative oxygen species catabolizing enzymes. The proteins with differential relative abundances might induce an altered physiological status within plant tubers for survival. The work provided new insights into the better understanding of the molecular basis of plant proteomes and stress tolerance mechanisms, especially during seasonal changes. The finding suggested proteins that might potentially be used as protein markers in differing seasons in other plants and aid in selecting B. superba tubers with the most suitable medicinal properties in the future.

Proteome analysis of Pueraria mirifica tubers collected in different seasons.

Pueraria mirifica-derived tuberous powder has been long-term consumed in Thailand as female hormone-replacement traditional remedies. The protein profiles of tubers collected in different seasons were evaluated. Phenol extraction, 2D-PAGE, and mass spectrometry were employed for tuberous proteome analysis. Out of the 322 proteins detected, over 59% were functionally classified as being involved in metabolism. The rest proteins were involved in defense, protein synthesis, cell structure, transportation, stress, storage, and also unidentified function. The proteins were found to be differentially expressed with respect to harvest season. Importantly, chalcone isomerase, isoflavone synthase, cytochrome p450, UDP-glycosyltransferase, and isoflavone reductase, which are all involved in the biosynthesis pathway of bioactive isoflavonoids, were most abundantly expressed in the summer-collected tubers. This is the first report on the proteomic patterns in P. mirifica tubers in relevant with seasonal variation. The study enlights the understanding of variance isoflavonoid production in P. mirifica tubers.

Clinical, radiographic, and histologic analysis of the effects of acemannan used in direct pulp capping of human primary teeth: short-term outcomes.

Acemannan has been previously reported as a direct pulp-capping agent in animal study. This natural material demonstrated its biocompatibility and enhanced reparative dentin formation. The objective of this study was to investigate the action of acemannan as a direct pulp-capping material in human primary teeth with deep caries. Forty-two deeply carious mandibular primary molars from 37 children, aged 7-11 years old diagnosed with reversible pulpitis were studied. After completely removing the infected dentine, teeth with a pinpoint pulpal exposure were randomly divided into two treatment groups: acemannan or calcium hydroxide. A glass-ionomer cement base was applied to all teeth prior to restoration with stainless steel crowns. Clinical and radiographic evaluation was performed 6 months post-treatment. The teeth due to exfoliate were extracted and histopathologically evaluated for inflammation, dentine bridge formation, and soft tissue organization. At 6 months, the overall clinical and radiographic success rates of direct pulp capping with acemannan and calcium hydroxide at 6 months were 72.73 and 70.0 %, respectively. The histopathological results indicated that the acemannan-treated group had significantly better histopathological responses compared with the calcium hydroxide-treated group (p < 0.05). These data suggest acemannan offers a valuable alternative biomaterial for vital pulp therapy in primary teeth.

Antioxidant and antiproliferative activities of protein hydrolysate from the rhizomes of Zingiberaceae plants.

Plant proteins have been investigated for their antioxidant activities, but there are still no reports detailing the antioxidant activity levels of plants in the Zingiberaceae family, which are popular food agents and used in folklore medicine. In this study, the crude rhizome protein extract and associated pepsin/pancreatin protein hydrolysate of 15 plants in the Zingiberaceae family were screened using the DPPH method for antioxidant activity. The protein hydrolysate of C. zedoaria possessed the highest antioxidant activity (IC50of 25.7±6.3µg/mL), which was close to that of the reference ascorbic acid (IC50of 22.3±1.8µg/mL). After enrichment by Q Sepharose ion exchange chromatography using a five step elution gradient of increasing NaCl concentration (0, 0.25, 0.5, 0.75 and 1M), the fraction eluting in the 0.5M NaCl (F50) showed the highest antioxidant activity (IC50 of 41.78±2.9µg/mL), and was found to have weak in vitro cytotoxicity against the HEP-G2 and SW620 cell lines (IC50 of 200.8±11.8 and 241.0±9.3µg/mL, respectively), but not the BT474, CHAGO and KATO-3 cell lines. F50 had an estimated molecular weight by MALDI-TOF mass spectrometry of 12,400-12,800 Da.

Deacetylation affects the physical properties and bioactivity of acemannan, an extracted polysaccharide from Aloe vera.

Acemannan, an acetylated polymannose from Aloe vera, induces tissue repair. We investigated the role of acemannan's acetyl-groups on its physical and biological properties. Deacetylated acemannan (DeAcAM) was prepared and characterized. The physical properties and microscopic structure of DeAcAM were evaluated using water solubility, contact angle, X-ray diffraction, and scanning-electron microscopy. The activity of DeAcAM on cell proliferation and gene expression were assessed. Acemannan and DeAcAM structures were simulated and the acemannan tetramer diad and its completely deacetylated structure were also determined. Increased acemannan deacetylation reduced its water solubility and hydrophilicity. Complete deacetylation altered acemannan's conformation to a partial crystal structure. The bioactivity of acemannan was reduced corresponding to its deacetylation. Acemannan induced cell proliferation, and VEGF and Collagen I expression; however, 100% DeAcAM did not. The simulated structures of the acemannan diad and the completely deacetylated diad were different. We conclude acetyl-groups affect acemannan's structure and physical/biological properties.

Proteomics profiling of cholangiocarcinoma exosomes: A potential role of oncogenic protein transferring in cancer progression.

Cholangiocarcinoma (CCA), a common primary malignant tumor of bile duct epithelia, is highly prevalent in Asian countries and unresponsive to chemotherapeutic drugs. Thus, a newly recognized biological entity for early diagnosis and treatment is highly needed. Exosomes are small membrane bound vesicles found in body fluids and released by most cell types including cancer cells. The vesicles contain specific subset of proteins and nucleic acids corresponding to cell types and play essential roles in pathophysiological processes. The present study aimed to assess the protein profiles of CCA-derived exosomes and their potential roles. We have isolated exosomes from CCA cells namely KKU-M213 and KKU-100 derived from Thai patients and their roles were investigated by incubation with normal human cholangiocyte (H69) cells. Exosomes were internalized into H69 cells and had no effects on viability or proliferation of the host cells. Interestingly, the exosomes from KKU-M213 cells only induced migration and invasion of H69 cells. Proteomic analysis of the exosomes from KKU-M213 cells disclosed multiple cancer related proteins that are not present in H69 exosomes. Consistent with the protein profile, treatment with KKU-M213 exosomes induced ß-catenin and reduced E-cadherin expressions in H69 cells. Collectively, our results suggest that a direct cell-to-cell transfer of oncogenic proteins via exosomal pathway may be a novel mechanism for CCA progression and metastasis.