Isolation and Characterization of Multipotent Postnatal Stem Cells from the Alveolar Bone and Gingival Cervical Margin

Periodontitis is a destructive inflammatory disorder of the periodontium branded by the destruction of periodontal tissues namely the PDL, cementum, alveolar bone, and gingiva. Once these tissues are lost, the foremost goal of periodontal therapy is to regenerate the diseased tissues if possible to their original form, architecture, and function. This a demanding task, that requires the harmonization of many actions at both cellular and molecular levels. Successful periodontal regeneration encompasses the formation of new gingival connective tissue, the restoration of bone, and most decisively, the insertion of new connective tissue fibres into newly formed cementum on the formerly diseased root surfaces. A variety of procedures to regenerate the lost tissue have been suggested, including the utilization of the principles of guided tissue regeneration and the application of an assortment of growth factors as well as bone and enamel matrix proteins on the root surfaces. However, these strategies showed only limited regeneration of periodontal tissue and were associated with incoherent and unpredictable clinical outcomes. Stem cells are progenitor cells characterized by their ability to self-renew and differentiate to produce specialized cells. There are two main categories of stem cells, the pluripotent embryonic stem cells and the multipotent adult ones. Recently, dental tissues such as the PDL, the dental pulp and the tooth follicle have been recognized as readily available sources of adult stem cells. Employing the criteria defined by Bartold et al. (2006) as well as the minimal standards accepted by the International Society for Cellular Therapy (Dominici et al. 2006) to define human MSCs and bearing in mind that the PDL shares the same developmental path with the cervical gingival margin and alveolar bone, the aim of this study was to present two protocols for the isolation and characterization of populations of mesenchymal multipotent stem cells-periodontal stem cells (PDSCs) derived from the alveolar bone and the cervical gingival margin of the periodontium. In this study the sample consisted of eleven partially impacted wisdom teeth, which were surgically extracted together with a piece of alveolar bone and cervical gingival margin attached to them. Cells isolated from the alveolar bone and the cervical gingival margin of the periodontium showed adherence to tissue culture plastics under standard in vitro culture conditions. They both exhibited the ability to form colony-forming units and were highly positive for the surface markers CD73, CD90, CD105 (> 95%), moderately positive for the markers CD146 and STRO-1, and negative for CD14, CD34, and CD45. On the mRNA level unstimulated cells from both sources expressed the osteogenic markers alkaline phosphatase, type I and III collagen, osteonectin, osteopontin, and osteocalcin. In addition, they both expressed type V collagen. Finally, cells from both sources demonstrated in vitro multilineage differentiation potential as evidenced by histological staining and PCR assays. It is concluded from this study that adult postnatal multipotent PDSCs can be readily isolated from the alveolar bone as well as from the cervical gingival margin

Toll-Like Receptors: The Key of Immunotherapy in MSCs

Human mesenchymal stem cells (MSCs) are potential candidates for various applications in the fields of immunotherapy. Their multilineage differentiation capability and immune modulatory features allow their prospective application for the management of different immunological circumstances. However, the local microenvironment, in addition to the source of the MSCs can control diverse biological features of the cells. Indeed, throughout their therapeutic application, MSCs may interact with their microenvironment through their expressed toll-like-receptors (TLRs), producing immune modulating reactions. Stimulation of MSCs before or within the potential treatment procedures with distinct TLR ligands may assist as an effective step controlling the biological function of the MSCs as needed in different therapeutic stages of the disease

The Effect of Instructional Guidelines on Pregnant Women’s Perception Regarding Urinary Tract Infection

Background: Urinary tract diseases (UTIs) are the foremost predominant diseases in pregnancy, counting for about 20% of all pregnancies. Moreover, they are responsible for 10% of all hospitalizations during pregnancy. Aim: The study aimed to evaluate effect of instructional guidelines on pregnant women’s perception regarding urinary tract infection. Design: A quasi-experimental research design was used at this study. Setting: The study was conducted in Obstetric and Gynecological clinic in El Hussein Hospital Al-azar University. Sample: A purposive sample of ninety-six pregnant women with urinary tract infection. Tools: The study utilized two assessment tools: a Structured Interviewing Questionnaire and Likert scale of women attitudes for pregnant women regarding urinary tract infection (UTI). Results: The findings of the study indicated that prior to the application of instructional guidelines, more than two-thirds of the pregnant women had an unsatisfactory level of knowledge about UTI. However, after the application of the guidelines, more than three-thirds of them demonstrated satisfactory knowledge. Similarly, nearly three-quarters of the participants held a negative attitude towards UTI before the guidelines, whereas after the guideline's application, more than three-thirds of them displayed a positive attitude. Conclusion: Based on these results, it can be concluded that the instructional guidelines effectively improved both the knowledge and attitude of the pregnant women towards UTI, thereby supporting the research hypothesis. Recommendations: The study recommends the implementation of routine screening for UTIs in all pregnant women during each trimester of pregnancy, including bacteriuria examination and urine culture, to ensure early diagnosis and appropriate treatment

Can vitamins improve periodontal wound healing/regeneration?

Periodontitis is a complex inflammatory disorder of the tooth supporting structures, associated with microbial dysbiosis, and linked to a number if systemic conditions. Untreated it can result in an irreversible damage to the periodontal structures and eventually teeth loss. Regeneration of the lost periodontium requires an orchestration of a number of biological events on cellular and molecular level. In this context, a set of vitamins have been advocated, relying their beneficial physiological effects, to endorse the biological regenerative events of the periodontium on cellular and molecular levels. The aim of the present article is to elaborate on the question whether or not vitamins improve wound healing/regeneration, summarizing the current evidence from in vitro, animal and clinical studies, thereby shedding light on the knowledge gap in this field and highlighting future research needs. Although the present review demonstrates the current heterogeneity in the available evidence and knowledge gaps, findings suggest that vitamins, especially A, B, E, and CoQ10 , as well as vitamin combinations, could exert positive attributes on the periodontal outcomes in adjunct to surgical or nonsurgical periodontal therapy

Root damage induced by intraosseous anesthesia'An in vitro investigation

Objectives: The principle of the intraosseous anesthesia (IOA) relies on the perforation of the cortical plate of the bone for direct application of the local anesthetic solution into the underlying cancellous structures. During this procedure, IOA needles might accidentally come in contact with the tooth roots. The aim of the current in vitro study was to examine the consequences of this 'worst case scenario' comparing five commercially available IOA systems. Material and Methods: Extracted human roots were randomly perforated using five different IOA systems with a drilling time ?5s. To simulate normal in vivo conditions, the roots were kept humid during the drilling procedure. Data was statistically evaluated using F-test (SPSS16, SPSS Inc., Chicago, USA) and the significance level was set at p?0.05. Results: All examined systems resulted in root perforation. Drill fractures occurred in either none 0% (Quicksleeper®, Anesto®, Intraflow®, Stabident®) or 100% (X-Tip®) of the applications. Excessive heat generation, as evident by combustion odor as well as metal and tooth discoloration, appeared in 30% (Quicksleeper®), 40% (Anesto®), 60% (Intraflow®), 90% (Stabident®) and 100% (X-Tip®) of all perforations. Conclusion: Within the limits of in-vitro studies, the results show a potential for irreversible root damage that might be inflicted by an improper use of IOA systems

TlR expression profile of human gingival margin-derived stem progenitor cells

Background: Gingival margin-derived stem/progenitor cells (G-MSCs) show remarkable periodontal regenerative potential in vivo. During regeneration, G-MSCs may interact with their inflammatory environment via toll-likereceptors (TLRs). The present study aimed to depict the G-MSCs TLRs expression profile. Material and Methods: Cells were isolated from free gingival margins, STRO-1-immunomagnetically sorted and seeded to obtain single colony forming units (CFUs). G-MSCs were characterized for CD14, CD34, CD45, CD73, CD90, CD105, CD146 and STRO-1 expression, and for multilineage differentiation potential. Following G-MSCs’ incubation in basic or inflammatory medium (IL-1β, IFN-γ, IFN-α, TNF-α) a TLR expression profile was generated. Results: G-MSCs showed all stem/progenitor cells’ characteristics. In basic medium G-MSCs expressed TLRs 1, 2, 3, 4, 5, 6, 7, and 10. The inflammatory medium significantly up-regulated TLRs 1, 2, 4, 5, 7 and 10 and diminished TLR 6 (p≤0.05, Wilcoxon-Signed-Ranks-Test). Conclusions: The current study describes for the first time the distinctive TLRs expression profile of G-MSCs under uninflamed and inflamed conditions

Synthesis and reactions of (Z)-2-imino-5-(3,4,5-trimethoxy benzylidene)thiazolidin-4(H)one

5-Arylmethylene-2-imino-4-oxo-2-thiazolidine 3 was obtained as the sole product from the reaction of α-cyano-3,4,5-trimethoxy cinnamonitrile and/or ethyl-α-cyano-3,4,5-trimethoxy cinnamate (1a,b) with 2-imino-4-oxo-2-thiazolidine 2. The reaction of 3 with benzyl amine gave the imidazolidin-4(H)one derivative 4 while with hydrazine hydrate afforded the dimeric product 5. Also, reaction of thiazolidinone derivative 3 with piperidine gave thiazol-4(5H)one derivative 6 which on treatment with Grignard reagent and active methylene compounds afforded thiazolidin-4-one derivatives 7-9, respectively. Compound 6 was converted to the potassium salt 10 which treated with acetic acid, ethyl chloroacetate and furoyl chloride to give the compounds 11-13, respectively. The structures of all new compounds were evidenced by microanalytical data and spectral data

Effect of total sonicated Aggregatibacter actinomycetemcomitans fragments on gingival stem/progenitor cells

Aggregatibacter-actinomycetemcomitans (A.actinomycetemcomitans) are strongly associated with localized-aggressive-periodontitis (LAgP). The study?s aim was to test for the first time the effect of total sonicated A.actinomycetemcomitans-bacterial-fragments on gingival mesenchymal stem/progenitor cells? (G-MSCs) proliferation and regenerative gene expression in-vitro. G-MSCs were isolated, characterized, expanded and stimulated by total sonicated A.actinomycetemcomitans-bacterial-fragments (0 (negative-control), 15, 60, 120 and 240µg/ml; serovar-b; n=6/group). Cellular proliferation and NF-?? (NFKB1), Alkaline Phosphatase (ALPL), Collagen-I (COL1A1), Collagen-III (COL3A1), Osteonectin (SPARC) and Osteopontin (SPP1) m-RNA expression were assessed via reverse-transcription-polymerase-chain-reaction (RT-PCR) at 24, 48 and 72 hours and CFUs-ability evaluated at twelve days. G-MSCs demonstrated stem/progenitor cells? characteristics. A.actinomycetemcomitans-bacterial-fragments (up to 72 hours) resulted in marked G-MSCs? proliferation over-time (p0.05; Friedman-test). Longer-term stimulation for twelve days reduced G-MSCs? CFUs. Sonicated A.actinomycetemcomitans-bacterial-fragments? exert beneficial short-term effects on G-MSCs? proliferative and non-mineralized tissue forming aptitude. Results shed new light on the importance of periodontal treatment for LAgP patients, using power driven sonic/ultrasonic devices, which, in addition to reducing the subgingival microbial load, produces cell-stimulatory A.actinomycetemcomitans-bacterial-fragments, with positive attributes on tissue reparative/regenerative responses of tissue resident stem/progenitor cells in their niche

The Changes in the Lipid Composition of Mung Bean Seeds as Affected by Processing Methods

Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG geförderten) Allianz- bzw. Nationallizenz frei zugänglich.This publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively.This study was conducted to assess in detail the possible effects of some technological processes such as soaking, germination, cooking, soaking + cooking, and germination + cooking on the lipid composition of mung bean seeds of Giza 1 variety. TLC analysis of mung bean lipids showed that the phospholipids and triglycerides recorded the highest percentage among lipid fractions (32.26 and 30.10%), while the 1,3 diglycerides constituted the least percentage (2.80%) in mung bean seeds. The soaking, germination and cooking processes caused a decrease in the phospholipids, triglycerides and hydrocarbons accompanied with an increase in monoglycerides, 1,2-(2,3)-diglycerides, sterols and free fatty acids. Eleven fractions were separated from phospholipids class of the studied samples; seven of these fractions were identified. The major component of phospholipids was phosphatidyl choline, amounting to 21.30, 17.84, 16.21, 13.87, 13.20 and 11.47% of the total phospholipids in raw, soaked, germinated, raw-cooked, soaked-cooked and germinated-cooked mung bean seeds, respectively. Gas liquid chromatography of the total lipids of mung bean seeds showed that the unsaturated fatty acids represented 69.58, 64.35, 63.3, 63.16, 61.84 and 61.12%, while the levels of saturated fatty acids were low being 30.37, 34.05, 35.66, 34.64, 37.93 and 38.75% of the total fatty acids in raw, soaked, germinated, raw-cooked, soaked-cooked and germinated-cooked, respectively. The total essential fatty acids (linoleic and linolenic) represented the highest proportion of fatty acids (50.10% of the total fatty acids)

Synthesis of novel pyrimidine and fused pyrimidine derivatives

4-Amino-2-(benzylthio)-6-(4-methoxyphenyl)pyrimidine-5-carbonitrile (1) was prepared by treatment of s-benzylthiuronium chloride with 2-(4-methoxybenzylidene)malononitrile in ethanolic sodium hydroxide with hydrazine hydrate to afford the hydrazino derivative 2, which was allowed to react with different electrophilic reagents to give the pyrimidine derivatives 3-12. The proclivity of (E)-2-cyano-3-(4-nitrophenyl)acrylamide (13) towards carbon and nitrogen nucleophiles was also investigated. IR, 1H NMR and mass spectra for all the synthesized compounds were discussed. All derived compounds were investigated for anti avian influenza (H5N1) virus activity and compared with zanamivir as control drug. All the synthesized compounds didn’t possess any antiviral activity