Project description:Purpose: Hyperlipidemia has been shown to contribute to reduced implant osseointegration, but the underlying mechanism remains poorly understood. Recently, non-coding RNAs (ncRNAs) have been established as key regulators of various pathophysiological processes. Therefore, the current study aims to examine the expression profiles of mRNAs and ncRNAs in femoral tissues around surgical implants in a murine hyperlipidemia model via RNA sequencing.Materials and method: Twenty adult male Wistar rats were equally divided between an experimental group and a control group, which were fed a high-fat diet and a normal diet, respectively. Titanium root-form implants was inserted into both femoral shafts of each rat in eight weeks. Bone tissues were collected at day 10 after the insertion of the implants and subsequently used for RNA extraction and sequencing. The sequenced reads were refined, assembled and annotated to obtain mRNA, microRNA (miRNA), long non-coding RNA (lncRNA) and circular RNA (circRNA) transcripts. The levels of all types of RNA molecules in both animal groups were quantified and differentially expressed candidates were identified. The function of the identified RNAs were predicted by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. LncRNA-miRNA-mRNA and circRNA-miRNA-mRNA co-expression network of differentially expressed RNAs were constructed. Real-time polymerase chain reactions (RT-PCR) were performed to validate the expression levels of selected candidates.Results: The serum lipid levels of the experimental group were significant higher than those of the control group (p < 0.05). In total, 389 mRNAs, 73 miRNAs, 770 lncRNAs and 7 circRNAs were shown to be differentially expressed (p < 0.05, fold change > 2). GO analysis found that regulation of bone mineralization, negative regulation of chondrocyte differentiation and bone development changed significantly. The network that maybe related to osteogenesis included 6 mRNAs, 8 miRNAs, 6 circRNAs, 119 lncRNAs and 362 associations. Expression level of most RNAs detected by RT-PCR was consistent with that by RNA sequencing. Conclusion: Functional analysis predicted a panel of differentially expressed mRNAs and ncRNAs associated with osteogenesis. The results of the current study will greatly contribute to the elucidation of the mechanism for implant osseointegration under hyperlipidemia.

Project description:Bone development and regeneration is associated with the Wnt signaling pathway that, according to literature, can be modulated by lithium ions (Li+). The aim of this study was to evaluate the gene expression profile during peri-implant healing of poly(lactic-co-glycolic acid) (PLGA) implants with incorporated Li+, while PLGA without Li+ was used as control, and a special attention was then paid to the Wnt signaling pathway. The implants were inserted in rat tibia for 7 or 28 days and the gene expression profile was investigated using a genome-wide microarray analysis. The results were verified by qPCR and immunohistochemistry. Histomorphometry was used to evaluate the possible effect of Li+ on bone regeneration. The microarray analysis revealed a large number of significantly differentially regulated genes over time within the two implant groups. The Wnt signaling pathway was significantly affected by Li+, with approximately 34% of all Wnt-related markers regulated over time, compared to 22% for non-Li+ containing (control; Ctrl) implants. Functional cluster analysis indicated skeletal system morphogenesis, cartilage development and condensation as related to Li+. The downstream Wnt target gene, FOSL1, and the extracellular protein-encoding gene, ASPN, were significantly upregulated by Li+ compared with Ctrl. The presence of β-catenin, FOSL1 and ASPN positive cells was confirmed around implants of both groups. Interestingly, a significantly reduced bone area was observed over time around both implant groups. The presence of periostin and calcitonin receptor-positive cells was observed at both time points. This study is to the best of the authors’ knowledge the first report evaluating the effect of a local release of Li+ from PLGA at the fracture site. The present study shows that during the current time frame and with the present dose of Li+ in PLGA implants, Li+ is not an enhancer of early bone growth, although it affects the Wnt signaling pathway. PLGA implants with +/- incorporated Li were inserted in rat tibia for 7 or 28 days, peri-implant bone was harvested and RNA extracted using Qiazol lysis reagent and TissueLyser followed by Qiagen's Rneasy Micro Kit. The microarray experiment was conducted at SCIBLU Genomics (www.lu.se/sciblu) according to Affymetrix guidelines (n=6).

Project description:Bone development and regeneration is associated with the Wnt signaling pathway that, according to literature, can be modulated by lithium ions (Li+). The aim of this study was to evaluate the gene expression profile during peri-implant healing of poly(lactic-co-glycolic acid) (PLGA) implants with incorporated Li+, while PLGA without Li+ was used as control, and a special attention was then paid to the Wnt signaling pathway. The implants were inserted in rat tibia for 7 or 28 days and the gene expression profile was investigated using a genome-wide microarray analysis. The results were verified by qPCR and immunohistochemistry. Histomorphometry was used to evaluate the possible effect of Li+ on bone regeneration. The microarray analysis revealed a large number of significantly differentially regulated genes over time within the two implant groups. The Wnt signaling pathway was significantly affected by Li+, with approximately 34% of all Wnt-related markers regulated over time, compared to 22% for non-Li+ containing (control; Ctrl) implants. Functional cluster analysis indicated skeletal system morphogenesis, cartilage development and condensation as related to Li+. The downstream Wnt target gene, FOSL1, and the extracellular protein-encoding gene, ASPN, were significantly upregulated by Li+ compared with Ctrl. The presence of β-catenin, FOSL1 and ASPN positive cells was confirmed around implants of both groups. Interestingly, a significantly reduced bone area was observed over time around both implant groups. The presence of periostin and calcitonin receptor-positive cells was observed at both time points. This study is to the best of the authors’ knowledge the first report evaluating the effect of a local release of Li+ from PLGA at the fracture site. The present study shows that during the current time frame and with the present dose of Li+ in PLGA implants, Li+ is not an enhancer of early bone growth, although it affects the Wnt signaling pathway.

			Action	DRS
	SRR7056880.fastq.gz	Fastqsanger.gz
	SRR7056881.fastq.gz	Fastqsanger.gz
	SRR7056882.fastq.gz	Fastqsanger.gz
	SRR7056883.fastq.gz	Fastqsanger.gz
	SRR7056884.fastq.gz	Fastqsanger.gz

Dataset Information

Co-expression Networks of mRNAs and non-coding RNAs of bones around implants in hyperlipidemic rats (miRNA-seq data sets)

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