Molecular mechanism of hypoxia and alpha-ketoglutaric acid on collagen expression in scleral fibroblasts

AIM:To investigate the molecular mechanisms underlying the influence of hypoxia and alpha-ketoglutaric acid(α-KG)on scleral collagen expression.METHODS:Meta-analysis and clinical statistics were used to prove the changes in choroidal thickness(ChT)during myopia.The establishment of a hypoxic myopia model(HYP)for rabbit scleral fibroblasts through hypoxic culture and the effects of hypoxia andα-KG on collagen expression were demonstrated by Sirius red staining.Transcriptome analysis was used to verify the genes and pathways that hypoxia andα-KG affect collagen expression.Finally,real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used for reverse verification.RESULTS:Meta-analysis results aligned with clinical statistics,revealing a thinning of ChT,leading to scleral hypoxia.Sirius red staining indicated lower collagen expression in the HYP group and higher collagen expression in the HYP+α-KG group,showed that hypoxia reduced collagen expression in scleral fibroblasts,whileα-KG can elevated collagen expression under HYP conditions.Transcriptome analysis unveiled the related genes and signaling pathways of hypoxia andα-KG affect scleral collagen expression and the results were verified by RT-qPCR.CONCLUSION:The potential molecular mechanisms through which hypoxia andα-KG influencing myopia is unraveled and three novel genes TLCD4,TBC1D4,and EPHX3 are identified.These findings provide a new perspective on the prevention and treatment of myopia via regulating collagen expression.

Osteogenic lithium-doped brushite cements for bone regeneration

This study investigated the osteogenic performance of new brushite cements obtained from Li+-dopedβ-tricalcium phosphate as a promising strategy for bone regeneration.Lithium(Li+)is a promising trace element to encourage the migration and proliferation of adipose-derived stem cells(hASCs)and the osteogenic differentiation-related gene expression,essential for osteogenesis.In-situ X-ray diffraction(XRD)and in-situ 1H nuclear magnetic resonance(1H NMR)measurements proved the precipitation of brushite,as main phase,and monetite,indicating that Li+favored the formation of monetite under certain conditions.Li+was detected in the remaining pore solution in significant amounts after the completion of hydration.Isothermal calorimetry results showed an accelerating effect of Li+,especially for low concentration of the setting retarder(phytic acid).A decrease of initial and final setting times with increasing amount of Li+was detected and setting times could be well adjusted by varying the setting retarder concentration.The cements presented compressive mechanical strength within the ranges reported for cancellous bone.In vitro assays using hASCs showed normal metabolic and proliferative levels.The immunodetection and gene expression profile of osteogenic-related markers highlight the incorporation of Li+for increasing the in vivo bone density.The osteogenic potential of Li-doped brushite cements may be recommended for further research on bone defect repair strategies.