A minocycline-releasing PMMA system as a space maintainer for staged bone reconstructions— in vitro antibacterial, cytocompatibility and anti-inflammatory characterization
Tiago SilvaLiliana GrenhoJoana BarrosJosé Carlos SilvaRosana V. PintoAna C. MatosBruno ColaçoMaria Helena FernandesAna BettencourtPedro S. Gomes
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In the present work, we study the development and biological characterization of a polymethyl methacrylate (PMMA)-based minocycline delivery system, to be used as a space maintainer within craniofacial staged regenerative interventions. The developed delivery systems were characterized regarding solid state characteristics and assayed in vitro for antibacterial and anti-inflammatory activity, and cytocompatibility with human bone cells. A drug release profile allowed for an initial burst release and a more sustained and controlled release over time, with minimum inhibitory concentrations for the assayed and relevant pathogenic bacteria (i.e., Staphylococcus aureus, slime-producer Staphylococcus epidermidis and Escherichia coli) being easily attained in the early time points, and sustained up to 72 h. Furthermore, an improved osteoblastic cell response-with enhancement of cell adhesion and cell proliferation-and increased anti-inflammatory activity were verified in developed systems, compared to a control (non minocycline-loaded PMMA cement). The obtained results converge to support the possible efficacy of the developed PMMA-based minocycline delivery systems for the clinical management of complex craniofacial trauma. Here, biomaterials with space maintenance properties are necessary for the management of staged reconstructive approaches, thus minimizing the risk of peri-operative infections and enhancing the local tissue healing and early stages of regeneration.Keywords:
Minocycline
Bone cement
Debridement (dental)
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Aqueous solutions of tri-minocycline (equivalent to 20–50 mg/ml minocycline HCl) and preconstituted ready-for-use injections were prepared at the pH of blood; these exert high antibiotic activities equal to those of the parent substance. The chloroform/water distribution coefficients of tri-minocycline in the different types of injections were somewhat lower, but in the same order of magnitude as that of the parent substance. The MIC values of tri-minocycline proved to be very low and the LD50 values very high, their separation covering more than 3 orders of magnitude.
Minocycline
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Compared with maintainer lines,17 kinds of free amino acids in anthers of CMS peppers(Capsicum annuum L.) with different nuclear background were analyzed.The results showed that the contents of free proline and free lysine in sterile lines ZTA,17A and 8A were obviously lower than those in their maintainer lines ZTB,17B and 8B.Free threonine and arginine contents in sterile lines were obviously higher than those in maintainer lines.Total content of free amino acids in the anthers of sterile lines was significantly lower than that of maintainer lines.The result also showed that different nuclear backgrounds among the three sweet pepper CMS lines led to differences of most free amino acids in their anthers.
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Capsicum annuum
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Objective To evaluate the in vitro antibacterial activity of minocycline and doxycycline against Staphylococci epidermidis.Methods Deterimination of antibacterial activity of minocycline and doxycycline against 124 of Staphylococcus epidermidis strains was performed by using agar dilution method,and the antibacterial activity of minocycline was compared with doxycycline.Results The antibacterial efficiency of minocycline and doxycycline against Staphylococcus epidermidis was both good,and the MIC90 of 81 strains of methicillin-sensitive Staphylococcus epidermidis and 43 strains of methicillin-resistent Staphylococcus epidermidis were 1 mg/L and 2 mg/L respectively.Conclusion Minocycline and doxycycline have strong antibacterial activity against Staphylococcus epidermidis,and the sensitive rate of minocycline and doxycycline are both 100%.
Minocycline
Agar Dilution Method
Agar dilution
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8 ISSR primers and 7 chloroplast SSR(cpSSR)primer pairs were employed to study genetic diversity among 9 pairs of kenaf UG93 cytoplasmic male sterile/maintainer lines and 5 restorer lines.Nucleus genome genetic similarity coefficients(SC) based on ISSR ranged from 0.333 to 1.000,with the average of 0.583,0.689 and 0.812 among maintainer lines,maintainer and restorer lines,and restorer lines respectively.Chloroplast genome SC based on cpSSR ranged from 0 to 0.286.The 23 tested lines were clustered into 4 haplotypes,with all 9 sterile lines were classified into a single type.5 of 9 couple of sterile/maintainer lines showed some degree diversity which indicated that replacement of nucleus genome between sterile line and maintainer line was not complete.The results will help generating new kenaf sterile resource and excellent cross combinations.
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Kenaf
Cytoplasmic male sterility
Hibiscus
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The assessment of genetic diversity among nineteen maintainer lines was studied based on thirteen qualitative traits. The single linkage clustering, morphological dendogram were performed to assess the traits. Most of the traits showed variation in different maintainer lines except auricle colour. On the basis of flag leaf attitude, a maximum four groups were formed with erect, semi-erect, horizontal and descending type leaf angle. The maintainer line BRRI20B has awn tip, which is unique from the rest of the studied maintainer lines. Nineteen maintainer lines were grouped into four different clusters and a considerable level of variability was displayed for most of the traits examined. The clustering pattern revealed, cluster I was the largest and consisted seven maintainer lines. Among them maintainer lines BRRI52B and BRRI60B were the most closely associated. Cluster II represent diverse sources materials and its revealed non-correspondence of geographic diversity with genetic divergence. Thus the cluster analysis has revealed the genetic variation and the traits contributing for the variation. Hence, this maintainer lines can be utilized for trait improvement in breeding programmes for the traits contributing for major variation.
Bangladesh Rice j. 2018, 22(2): 79-87
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Dendrogram
Trait
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The mitochondrial DNA of 2 CMS tobacco lines and their homolog maintainer lines were analysed by the method of random amplified polymorphic DNA using 124 random 10-base primers.The result showed that 112 primers, 92.56% of total primers,did not amplified ploymorphic band between CMS and maintainer lines.This indicated the homolog of mitochondria between CMS and maintainer line.Meanwhile,some obvious differences in mitochondrial genome were detected between CMS and maintainer line.The relationship of these different bands in CMS to male sterility needs farther study.
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Cytoplasmic male sterility
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Objectives:The special protein which was concern with cytoplasmic male sterility was to be founded through A-PAGE analysis of Gliadins in seeds of K, V,T Type Cytoplasmic male sterile wheat,maintainer lines and hybrid wheat.Methods:A-PAGE analysis,consistence of gel is 6.72%.Conclusion: Differences were founded between the pattern of Tai911289 male sterility and its maintainer lines.There is a special band in β-section of the pattern of maintainer lines.But this difference was not founded in the pattern of Ji5418 male sterility and its maintainer lines.The result showed that the differences between male sterility and their maintainer lines were dependent on the cultivar.We can also found that there are great differences between the pattern of hybrid wheat and that of their male parent.The effect of female parent was more than that of male parent.Some differences was founded among different type hybrid wheat.
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Cytoplasmic male sterility
Common wheat
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Primary teeth play a pivotal role in preventing malocclusion of permanent dentition. When there is premature tooth loss, the space should be maintained for preserving the arch form and preventing space loss. Space maintainer is the treatment of choice in such conditions. When the conventional method is not applicable, we plan for modifications. In this clinical scenario, we have modified the crown and loop space maintainer with a single loop known as the Mayne's space maintainer.
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Five CMS lines were crossed with 49 genotypes as ‘testers‘ to get 245 hybrids. The 245 hybrids were subjected to pollen and spikelet fertility analysis. Among the 245 hybrids 21 hybrids were expressed as restorers, 24 as maintainers, and 200 intermediate types. Six tester‘s viz., BR6592-4-6-4, BR6839-41-5-1, IR74052-184-3-3, IR72593-B-3-2-2-2, BR7011-37-1-2, and IR72049-B-R-22-3-1-1 were identified as restorers for highest 2 lines out of the five CMS lines, differently. None of them were found to be restorer for all the five CMS lines. BAU581 was found to be maintainer for four lines except for D.ShanA. Purbachi was found to be maintainer for three lines except for D.ShanA and IR73328A out of the five CMS lines. Out of 245 crosses only 45 crosses contributed directly to the identification of maintainer and restorer. Other crosses were more or, less of intermediate types which indicated neither maintainer nor restorer. It is well known that, pollen fertility is controlled by one dominant gene (RfRf). If pollen fertility is governed by only one gene the product would be either restorer or, maintainer where will be no existence of intermediate types. So, there might be modifier genes in different genotypes which interacted with male sterile nuclear genes that resulted in intermediate male sterility in the crosses.
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Cytoplasmic male sterility
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Proline content of leaves from three cytoplasmic male sterile lines and their maintainer lines in sugarbeet were studied during their vegetative growth stage. The results showed that the proline content of leaves from the maintainer lines were higher than those from the sterile lines. This result also indicated that there were significant differences between maintainer lines and the sterile lines on physiological and biochemical properties during their vegetative growth stage.
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