The biocompatibility of biomaterials is essentially for its application. The aim of current study was to evaluate the biocompatibility of poly(lactic-co-glycolic acid) (PLGA)/gelatin/nanohydroxyapatite (n-HA) (PGH) nanofibers systemically to provide further rationales for the application of the composite electrospun fibers as a favorable platform for bone tissue engineering. The PGH composite scaffold with diameter ranging from nano- to micrometers was fabricated by using electrospinning technique. Subsequently, we utilized confocal laser scanning microscopy (CLSM) and MTT assay to evaluate its cyto-compatibility in vitro. Besides, real-time quantitative polymerase chain reaction (qPCR) analysis and alizarin red staining (ARS) were performed to assess the osteoinductive activity. To further test in vivo, we implanted either PLGA or PGH composite scaffold in a rat subcutaneous model. The results demonstrated that PGH scaffold could better support osteoblasts adhesion, spreading, and proliferation and show better cyto-compatibility than pure PLGA scaffold. Besides, qPCR analysis and ARS showed that PGH composite scaffold exhibited higher osteoinductive activity owing to higher phenotypic expression of typical osteogenic genes and calcium deposition. The histology evaluation indicated that the incorporation of Gelatin/nanohydroxyapatite (GH) biomimetics could significantly reduce local inflammation. Our data indicated that PGH composite electrospun nanofibers possessed excellent cyto-compatibility, good osteogenic activity, as well as good performance of host tissue response, which could be versatile biocompatible scaffolds for bone tissue engineering.
Recent studies have demonstrated that erythropoietin (EPO) has extensive nonhematopoietic biological functions. However, little is known about how EPO regulates bone formation, although several studies suggested that EPO can affect bone homeostasis. In this study, we investigated the effects of EPO on the communication between osteoclasts and osteoblasts through the ephrinB2/EphB4 signaling pathway. We found that EPO slightly promotes osteoblastic differentiation with the increased expression of EphB4 in ST2 cells. However, EPO increased the expression of Nfatc1 and ephrinB2 but decreased the expression of Mmp9 in RAW264.7 cells, resulting in an increase of ephrinB2-expressing osteoclasts and a decrease in resorption activity. The stimulation of ephrinB2/EphB4 signaling via ephrinB2-Fc significantly promoted EPO-mediated osteoblastic differentiation in ST2 cells. EphB4 knockdown through EphB4 shRNA inhibited EPO-mediated osteoblastic phenotypes. Furthermore, in vivo assays clearly demonstrated that EPO efficiently induces new bone formation in the alveolar bone regeneration model. Taken together, these results suggest that ephrinB2/EphB4 signaling may play an important role in EPO-mediated bone formation.
iRoot BP Plus and MTA had similar favourable results when used as pulp-capping agents.
Simvastatin is considered as a stimulator for bone formation. However, the half-life for simvastatin is generally 2 hours, which means, it is difficult to maintain biologically active simvastatin in vivo. To overcome this limitation, we created a system to slowly release simvastatin in vitro and in vivo. We constructed a poly(lactic-co-glycolic acid)/hydroxyapatite nano-fibrous scaffold to carry simvastatin. Releasing assays showed that simvastatin was released from poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin quickly within - 15 days, and small amounts continued to be released through day 56 (experiments terminated). MTT assays demonstrated that both poly(lactic-co-glycolic acid)/hydroxyapatite and poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin promoted MC3T3-E1 cell proliferation. However, Alkaline phosphatase assays showed that only poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin scaffold significantly promoted the osteogenic differentiation of MC3T3-E1 cells in vitro on day 14. To further test in vivo, we created calvaria bone defect models and implanted either poly(lactic-co-glycolic acid)/hydroxyapatite or poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin. After 4 or 8 weeks post-implantation, the results indicated that poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin scaffold induced bone formation more efficiently than poly(lactic-co-glycolic acid)/hydroxyapatite alone. Our data demonstrates that poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin has the potential to aid in healing bone defects and promoting bone regeneration in the future although we still need to optimize this complex to efficiently promote bone regeneration.
Aim To evaluate the expression of Foxp3‐positive lymphocytes around newly formed tissue after regenerative endodontic treatment (RET) in vivo and investigate the effects of stem cells from the apical papilla (SCAP) on the conversion of CD4+CD25− T cells to CD4+CD25+Foxp3+ regulatory T cells (Tregs) in vitro. Methodology Three 6‐month‐old beagles with nine doubled‐rooted premolars in each dog were randomly assigned to the RET group and the control group. RET was performed after apical periodontitis had been induced in the experimental immature teeth. Three months later, the expression of Foxp3 was detected in the histological sections by immunofluorescent staining. Human SCAP and CD4+CD25− T cells from mice spleens (1 : 1 and 1 : 5) were co‐cultured in cell–cell contact or in Transwells, respectively, for 24 and 72 h in vitro. The percentage of Tregs was evaluated by flow cytometry. The results were analysed using the Fisher's exact test and analysis of variance. P < 0.05 was regarded as statistically significant. Results Inflammatory cells were present with tissue regeneration in the RET group, and Foxp3‐positive T cells were enriched around the newly formed tissues. SCAP promoted Treg conversion after 72 h in vitro. Cell–cell contact played an important role after the 24 h co‐culture, whilst soluble factors were also involved after 72 h (P < 0.05). Conclusions SCAP promoted the conversion of pro‐inflammatory T cells to Tregs in vitro. Tregs were enriched around the regenerating tissues in the root canals after RET, which may create a suitable immune microenvironment for the differentiation of SCAP. This study provides an underlying mechanism for tissue regeneration during RET.
ObjectiveThis study investigated the effects of poly lactic‐co‐glycolic acid (PLGA) loaded with plasmid DNA encoding fibroblast growth factor‐2 (pFGF‐2) on human periodontal ligament cells (hPDLCs) in vitro and evaluated the ability of the PLGA/pFGF‐2 scaffold to promote periodontal ligament (PDL) regeneration in a beagle dog teeth avulsion animal model.BackgroundGrowth factor and scaffold play important roles in PDL regeneration. PLGA is a kind of biodegradable and biocompatible polymer that can be used as a carrier to deliver growth factors or genes. FGF‐2 can induce potent proliferative responses, promote cell migration and regulate the production of extracellular matrix. Therefore, a gene‐activated matrix composed of scaffold and genes is supposed to be a superior approach for promoting tissue regeneration.MethodsIn this study, PLGA and PLGA/pFGF‐2 scaffolds were fabricated using electrospinning. The characterization of scaffolds was shown by scanning electron microscope (SEM) and transmission electron microscope (TEM). dsDNA HS was used to test the plasmid release of PLGA/pFGF‐2 scaffold. The viability and proliferation of hPDLCs on the two kinds of scaffolds were evaluated by the CCK‐8 assay, and the expression of collagen I and scleraxis were analysed by RT‐qPCR. The roots of avulsed teeth were covered by the two types of scaffolds and replanted into the alveolar pockets in beagles. Haematoxylin‐eosin and Masson staining were used to evaluate the effects of PLGA/pFGF‐2 scaffold on promoting PDL regeneration.ResultsThe smooth and uniform fibres can be observed in both scaffolds, and the plasmids were randomly distributed in the PLGA/pFGF‐2 scaffold. dsDNA HS analysis demonstrated that the PLGA/pFGF‐2 scaffold released up to 123 ng pFGF‐2 over 21 days in a sustained manner without any obvious burst release. The PLGA/pFGF‐2 scaffold promoted the proliferation of hPDLCs and increased the expression levels of collagen I and scleraxis compared with PLGA scaffold. Animal experiments showed that more regular PDL‐like tissues and less root surface resorption occurred in the PLGA/pFGF‐2 scaffold group compared with the PLGA scaffold group.ConclusionsThe PLGA/pFGF‐2 scaffold promoted hPDLCs proliferation and facilitated periodontal ligament‐related differentiation. The PLGA/pFGF‐2 scaffold possesses excellent biological characteristics and could be used as a promising biomaterial for improving the treatment prognosis of replanted tooth.
iRoot FM exhibited excellent antibacterial activity against P. endodontalis and could improve the proliferation and differentiation of SCAP. The findings provide evidence that iRoot FM has potential as an intracanal medicament for endodontic procedures in immature permanent teeth.
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