Matrix Metalloproteinase-8 Levels in Peri-Miniscrew Crevicular Fluid During Immediate and Delayed Orthodontic Loading--A Split-Mouth Study.

Purpose: To compare the matrix metalloproteinase-8 (MMP-8) levels in the peri-miniscrew implant crevicular fluid (PMCF) of immediate-loaded and delayed-loaded miniscrew implants at different time intervals. Materials and Methods: Titanium orthodontic miniscrews were placed bilaterally in the attached gingiva of 15 patients between the maxillary second premolar and maxillary first molar for en masse retraction. This split-mouth study was designed to have an immediate-loaded miniscrew on one side and a delayed-loaded miniscrew on the other side that was loaded 8 days after miniscrew placement. PMCF was collected from the mesiobuccal aspects of the immediate-loaded implants at 24 hours, 8 days, and 28 days after loading, and from the delayed-loaded miniscrew implants at 24 hours and 8 days before loading and 24 hours and 28 days after loading. An enzyme-linked immunosorbent assay kit was used to assess MMP-8 levels in the PMCF samples. Unpaired t test, ANOVA F-test, and Tukey post hoc test were used to evaluate data at the P < .05 level. Results: Although there were slight alterations in the MMP-8 levels in the PMCF over time, there was no statistically significant difference in the MMP-8 levels between groups. There was a statistically significant decrease in the levels of MMP-8 between 24 hours after miniscrew placement and 28 days after loading on the delayed-loaded side (P < .05). Conclusion: The MMP-8 levels did not vary much between immediate-loaded and delayed-loaded miniscrew implants as a result of the force application. However, there was no significant difference between immediate loading and delayed loading in terms of biologic response to mechanical stress. The increase in MMP-8 levels after 24 hours post-miniscrew insertion, as well as the subsequent gradual reduction over the course of the study period in both immediate and delayed groups after loading, is probably due to the bone adapting to stimuli.

Synovium-Synovial Fluid Axis in Osteoarthritis Pathology: A Key Regulator of the Cartilage Degradation Process.

Failure of conventional anti-inflammatory therapies in osteoarthritis (OA) underlines the insufficient knowledge about inflammatory mechanisms, patterns and their relationship with cartilage degradation. Considering non-linear nature of cartilage loss in OA, a better understanding of inflammatory milieu and MMP status at different stages of OA is required to design early-stage therapies or personalized disease management. For this, an investigation based on a synovium-synovial fluid (SF) axis was planned to study OA associated changes in synovium and SF along the progressive grades of OA. Gene expressions in synovial-biopsies from different grades OA patients (N = 26) revealed a peak of IL-1ß, IL-15, PGE2 and NGF in early OA (Kellgren-Lawrence (KL) grade-I and II); the highest MMP levels were found in advanced stages (KL grade-III and IV). MMPs (MMP-1, 13, 2 and 9) abundance and FALGPA activity estimated in forty SFs of progressive grades showed the maximum protein levels and activity in KL grade-II and III. In an SF challenge test, SW982 and THP1 cells were treated with progressive grade SFs to study the dynamics of MMPs modulation in inflammatory microenvironment; the test yielded a result pattern, which matched with FALGPA and the protein-levels estimation. Inflammatory mediators in SFs served as steering factor for MMP up-regulation. A correlation-matrix of IL-1ß and MMPs revealed expressional negative correlation.

Evaluation and comparison of Vitamin D receptors in periodontal ligament tissue of Vitamin D-deficient chronic periodontitis patients before and after supplementation of Vitamin D3.

BACKGROUND: Vitamin D, an important hormone required by the body, exerts its biological effects through Vitamin D receptors (VDRs) present on target cells. Vitamin D is ineffective in tissues which lack VDR. Various tissues show the presence of VDRs. However, evidence for the presence of VDRs in human periodontal ligament tissue in fully erupted teeth in adults is lacking. The present study intends to evaluate the presence of VDRs in periodontal ligament (PDL) tissue and assess their response to serum Vitamin D3 levels in chronic periodontic patients. MATERIALS AND METHODS: A total of 19 chronic periodontitis patients were enrolled in the study and tested for serum 25(OH)D3 levels. Deficient patients were supplemented with Vitamin D3. PDL tissue of these patients was isolated after tooth extraction before and after supplementation of Vitamin D3 and analyzed for the presence of VDR in PDL tissue by using enzyme-linked immunosorbent assay. RESULTS: All the chronic periodontitis patients were found to be deficient in Vitamin D3. The mean serum 25(OH)D3 level before supplementation was 13.96 ng/mL which significantly increased to 35.12 ng/mL after supplementation of Vitamin D3 for 6 weeks. VDR analysis determined mean VDR conc. in PDL tissue to be -1.443 ng/mL, which increased to 2.38 ng/mL after supplementation. A concentration dependent correlation was seen between serum 25(OH)D3 levels and VDR conc. in PDL tissue after supplementation. CONCLUSIONS: The study determined Vitamin D Receptors (VDR) in PDL tissue after supplementation of Vitamin D. Thus in addition to the standard treatment modalities, Vitamin D3 supplementation would be an important factor for generation of adequate immune response.

Reduced synovial inflammation and inhibition of matrix metalloproteinases explicates anti-osteoarthritis activity of polyherbal formulations.

OBJECTIVES: Current osteoarthritis (OA) research experiences an incline toward Ayurveda to attain a complete cure without notable adverse effects. Ayurveda uses natural products, which are known to perform the multi-faceted role, a much demanding approach for OA management. However, lack of scientific evidence is a major drawback hindering their wider use. The present work investigated the anti-arthritic potential of Ashwagandharishta, Balarishta, Dashmoolarishta, and Triphala-extract to establish molecular-evidence for their clinical use. MATERIALS AND METHODS: Rabbit synoviocytes were induced using interleukin-1 beta (IL-1 ß) and lipopolysaccharide (LPS) separately and were further treated with study formulations to test anti-inflammatory and anti-oxidant potential, using nitric oxide (NO) and malondialdehyde (MDA) assays. Collagenase inhibition activity was estimated with N-(3-[2-Furyl] acryloyl)-Leu-Gly-Pro-Ala (FALGPA)-substrate and gelatinase spot assays. Data were analyzed with GraphPad Prism using one-way ANOVA followed by Bonferroni's multiple comparison. RESULTS: The study formulations were effective against synovitis, oxidative-stress, and inhibiting collagenase. They caused NO reduction in selected concentrations. DA showed the maximum NO decline of 0.02 ± 0 and 0.97 ± 0.62 µM/ml with IL-1 ß and LPS induction at 5 and 20 µg/ml concentrations, respectively. Estimated by FALGPA assay, increasing collagenase inhibition was observed as the function of concentration. All formulations showed a significant MDA decline, in dose-dependent manner. CONCLUSION: We assessed the anti-OA efficacy of conventionally prescribed Ayurvedic drugs using relevant biochemical assays. The studied formulations revealed potential to restrain synovitis, cartilage degeneration and to reduce oxidative stress, and the signature OA features. With established molecular authenticity, Ayurvedic drugs can offer a safer and affordable therapeutic option for OA.

Probing Regenerative Potential of Moringa oleifera Aqueous Extracts Using In vitro Cellular Assays.

BACKGROUND: Molecules stimulating regeneration and proliferation of cells are of significance in combating ailments caused due to tissue injury, inflammation, and degenerative disorders. Moringa oleifera is one of the most valued food plants having the profile of important nutrients and impressive range of medicinal uses. OBJECTIVE: To evaluate the potential of M. oleifera aqueous leaf and flower extracts to promote the proliferation of cells and explore their effect on cancer cell lines for assessment of safety. MATERIALS AND METHODS: Aqueous leaf and flower extracts of M. oleifera were investigated for effect on rat-derived primary fibroblast, mesenchymal stem cells (MSCs), and cancer cell lines using cell proliferation assay. They were also tested and compared for wound healing, angiogenesis, and hepatoprotective effect using in vitro assays. RESULTS: Statistically significant increase in the proliferation of primary rat fibroblast, MSCs, and angiogenesis was observed after treatment with aqueous flower extract. The aqueous leaf extract determined a comparatively moderate increment in the proliferation of MSCs and angiogenesis. It however showed prominent cytotoxicity to cancer cell lines and a significant hepatoprotective effect. CONCLUSION: A very clear difference in response of the two extracts to different types of cells was detected in this study. The aqueous flower extract exhibited a higher potential to stimulate cell proliferation while not exerting the same effect on cancer cell lines. The leaf extract on the other hand, had a prominent antitumor and hepatoptotective effects. SUMMARY: Moringa oleifera flower extract showed significant ability to promote proliferation of rat fibroblast and mesenchymal stem cells. The extract also had prominent angiogenic and hepatoprotective effects.The extract did not influence proliferation of cancer cell lines indicating its safety for human consumption and use in pharmaceuticals.The Moringa oleifera leaf extract showed relatively less potential to stimulate cells but had prominent cytotoxic effect on cancer cell lines. Abbreviations Used: ALT: Alanine transaminase, AST: Asparatate amino transferase, ATCC: American type culture collection, BMMSC: Bone marrow mesenchymal stem cells (used in this paper), CAM: Chick chorioallantoic membrane, CCl4: Carbon tetra chloride, DMEM: Dulbecco's modified Eagle medium, DMSO: Dimethyl sulfoxide, EDTA: Ethylene diamine tetraacetic acid, HBL 100: Human breast epithelial cell line, Mcf-7: Human breast adenocarcinoma cell line, aMEM: Minimum Essential Medium Eagle alpha modification, MOF: Moringa oleifera aqueous flower extract (used in this paper), MOL: Moringa oleifera aqueos leaf extract (Used in this paper), OD: Optical density, PBS: Phosphate buffered saline.

Characterization of Anticancer Principles of Celosia argentea (Amaranthaceae).

BACKGROUND: An Indian origin, Celosia argentea is a weed growing during rainy season traditionally claimed for treating several ailments. Early researches on C. argentea were focused on the anti-cancer screening of seeds, with few reports on aerial parts. OBJECTIVE: To isolate and characterize bioactive compounds of aerial parts of C. argentea and evaluate their anticancer potential. MATERIALS AND METHODS: The methanolic aerial part extract was fractionated on column chromatography using chloroform: methanol mixture. The fractions; 80:20 and 95:5 were purified on MCI-HP20 HPLC column. Chromatographically pure compounds were pooled, concentrated and characterized spectroscopically. The compounds were further screened for anti-oxidant and cytotoxic potential. RESULTS: Isolated compounds were confirmed as: (1) Luteolin-7-O-glucoside and (2) phenolic, 1-(4-hydroxy-2-methoxybenzofuran-5-yl)-3-phenylpropane-1,3-dione. Both exhibited significant antioxidant potential with IC50 values of 20.80 and 21.30 µg/ml for 2,2-diphenyl-1-picrylhydrazyl assay (***P < 0.001) and significant Trolox equivalent antioxidant capacity (TEAC) values for 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (*P < 0.05) and ferric reducing antioxidant potential assay (****P < 0.0001). In 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide assay, Compound 1 and 2 showed potent cytotoxicity against SiHa, HCT, MCF-7 cancer cell lines at 20 µg/ml (****P < 0.0001) and 18 µg/ml (**P < 0.01), respectively, without affecting the normal Vero cells. Both compounds enabled maximum reduction in cell viability at 50 µg/ml against HT-29 (***P < 0.001) and MCF-7 cell lines (**P < 0.01) in try pan blue viability assay. Apoptosis occurred at concentrations of 47.33 ± 0.8 µg/ml and 56.28 ± 1.2 µg/ml for Compound 1 and 35.15 ± 0.4 µg/ml and 28.05 ± 0.3 µg/ml for Compound 2 for HT-29 and MCF-7 respectively. CONCLUSION: A novel anticancer phenolic compound; (1-(4-hydroxy-2-methoxybenzofuran-5-yl)-3-phenylpropane-1,3-dione), isolated from aerial parts of C. argentea was a valuable finding of the research. SUMMARY: The present study validated the potential of the plant C. argentea as an antioxidant, and anticancer remedy with two valuable isolations. Although one of them is a known compound: Luteolin 7-0 glycoside, the other isolated phenolic compound;-{1-(4-hydroxy-2-methoxybenzofuran-5-yl)-3-phenylpropane-1,3-dione}, is the first to be reported and thus can be considered as a valuable outcome of this research work.

Inflammatory response of cultured rat synoviocytes challenged with synovial fluid from osteoarthritis patients correlates with their radiographic grading: a pilot study.

The inflammatory nature of synovial fluid (SF) of varying grade osteoarthritis (OA) patients was estimated by measuring pro-inflammatory factors and through a unique cell-challenge experiment. SF samples were collected from six OA and one non-OA patient; spanning Kellgren-Lawrence (KL) grades were analyzed for interlukin-1-beta (IL-1ß), nitric oxide (NO) and its derivatives, and glycosaminoglycan (GAG). Levels of IL-1ß, NO, and GAG in SF did not correlate with KL grades of the patients studied. In the cell-challenge experiment, cultured rat synoviocyte fibroblasts (RSFs) were challenged by the patient's SFs with and without pre-treatment of IL-1ß and lipopolysaccharide (LPS). NO released by the cells was taken as an indicator of inflammation. SFs from KL grades 2 and 3 induced maximum inflammation in cultured RSFs (grade 2 64.61 ± 4.8 and 89.51 ± 5.6 µM/ml after 48 and 72 h, grade 3 58.27 ± 2.7 and 64.22 ± 2.8 µM/ml after 48 and 72 h, respectively). Similar trend was observed in RSF pretreated with either recombinant IL-1ß or LPS suggesting that SF from patients KL grades 2 and 3 accumulates more pro-inflammatory factors. IL-1ß-pre-treated RSFs challenged by SF for 72 h showed 234.41 ± 17.6 µM/ml increase (patient 3, grade 3), whereas higher NO after LPS pre-treatment was recorded (118.92 ± 6.2 µM/ml; patient 3, grade 3). Interestingly, SFs from grade 1 and non-OA patient could reduce released NO to 27.10 ± 2.2 µM/ml showing potency to alleviate inflammation. These interesting findings, however, need to be confirmed on a wider number of patients, which may offer significant therapeutic application in treatment of OA.

Novel Aza-resveratrol analogs: synthesis, characterization and anticancer activity against breast cancer cell lines.

Novel Aza-resveratrol analogs were synthesized, structurally characterized and evaluated for cytotoxic activity against MDA-MB-231 and T47D breast cancer cell lines, which exhibited superior inhibitory activity than parent resveratrol compound. The binding mechanism of these compounds with estrogen receptor-α was rationalized by molecular docking studies which indicated additional hydrogen binding interactions and tight binding in the protein cavity. Induction of Beclin-1 protein expression in breast cancer cell lines after treatment with newly synthesized resveratrol analogs indicated inhibition of growth of these cell lines through autophagy. The study highlighted the advantage of introducing the imino-linkage in resveratrol motif in enhancing the anticancer potential of resveratrol suggesting that these analogs can serve as better therapeutic agents against breast cancer and can provide starting point for building more potent analogs in future.