Journal: Journal of Orthopaedic Surgery and Research

Article Title: Periostin/Bone Morphogenetic Protein 1 axis axis regulates proliferation and osteogenic differentiation of sutured mesenchymal stem cells and affects coronal suture closure in the TWIST1 +/− mouse model of craniosynostosis

doi: 10.1186/s13018-024-04604-3

Figure Lengend Snippet: Periostin improves coronal craniosynostosis in TWIST1 +/− mice. A – B RT-qPCR and Western blot measurements of Periostin in mice. C HE staining of cranial suture. D IHC staining of Periostin and BMP1 in cranial suture of mice. Data were expressed as mean ± SD (n = 4). * P < 0.05

Article Snippet: siRNA and pcDNA 3.1 overexpression vectors targeting BMP1 and Periostin (RiboBio, Guangzhou, China) were transfected into SMSCs using Lipofectamine 3000 (Invitrogen, USA), and SMSCs were collected 48 h later for evaluating the transfection efficiency by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blot.

Techniques: Quantitative RT-PCR, Western Blot, Staining, Immunohistochemistry

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Periostin/Bone Morphogenetic Protein 1 axis axis regulates proliferation and osteogenic differentiation of sutured mesenchymal stem cells and affects coronal suture closure in the TWIST1 +/− mouse model of craniosynostosis

doi: 10.1186/s13018-024-04604-3

Figure Lengend Snippet: Periostin effectively inhibits SMSC proliferation and osteogenic differentiation. A SMSCs were isolated from cranial suture bone tissues of both TWIST1 +/− and TWIST1 +/+ mice and identified by flow cytometry as CD44 and CD105 positive. TWIST1 +/− SMSCs were transfected with pcDNA 3.1-Periostin. B Western blot assays were performed to assess Periostin protein expression in each group of SMSCs. C Colony formation assay was conducted to evaluate the proliferative capacity of SMSCs. D EdU assay was used to further quantify SMSC proliferation. E Wound healing assays were carried out to measure the wound closure rate of SMSCs. F ALP staining was used to determine ALP activity in SMSCs. G ARS staining was employed to observe calcium deposition in SMSCs. H Western blot analysis was conducted to measure the expression levels of Ki-67, RUNX2, and OPN in SMSCs. Data were presented as mean ± SD (N = 3). * P < 0.05

Article Snippet: siRNA and pcDNA 3.1 overexpression vectors targeting BMP1 and Periostin (RiboBio, Guangzhou, China) were transfected into SMSCs using Lipofectamine 3000 (Invitrogen, USA), and SMSCs were collected 48 h later for evaluating the transfection efficiency by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blot.

Techniques: Isolation, Flow Cytometry, Transfection, Western Blot, Expressing, Colony Assay, EdU Assay, Staining, Activity Assay

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Periostin/Bone Morphogenetic Protein 1 axis axis regulates proliferation and osteogenic differentiation of sutured mesenchymal stem cells and affects coronal suture closure in the TWIST1 +/− mouse model of craniosynostosis

doi: 10.1186/s13018-024-04604-3

Figure Lengend Snippet: Periostin triggers the proliferation and osteogenic differentiation of SMSCs. si-Periostin was transfected into the SMSCs of the TWIST1 +/− group. A Western blot assays were used to assess Periostin protein expression in the SMSCs across the different groups. B Colony formation assays were conducted to evaluate the proliferative capacity of SMSCs. C EdU assays were utilized to further quantify the proliferation of SMSCs. D Wound healing assays were carried out to measure the wound closure rate of SMSCs. E ALP staining was employed to determine the ALP activity in SMSCs. F ARS staining was used to observe calcium deposition in SMSCs. G Western blot analysis was conducted to assess the expression levels of Ki-67, RUNX2, and OPN in SMSCs. Data were represented as mean ± SD (N = 3). * P < 0.05

Article Snippet: siRNA and pcDNA 3.1 overexpression vectors targeting BMP1 and Periostin (RiboBio, Guangzhou, China) were transfected into SMSCs using Lipofectamine 3000 (Invitrogen, USA), and SMSCs were collected 48 h later for evaluating the transfection efficiency by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blot.

Techniques: Transfection, Western Blot, Expressing, Staining, Activity Assay

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Periostin/Bone Morphogenetic Protein 1 axis axis regulates proliferation and osteogenic differentiation of sutured mesenchymal stem cells and affects coronal suture closure in the TWIST1 +/− mouse model of craniosynostosis

doi: 10.1186/s13018-024-04604-3

Figure Lengend Snippet: Periostin regulates BMP1 expression in SMSCs. A Western blot assays were performed to evaluate the protein expression of BMP1 in SMSCs across the different groups. B The interactive network between Periostin and BMP1 was investigated using the bioinformatics website https://cn.string-db.org . C CO-IP experiments were conducted to detect the interaction between Periostin and BMP1. Data were represented as mean ± SD (N = 3). * P < 0.05

Article Snippet: siRNA and pcDNA 3.1 overexpression vectors targeting BMP1 and Periostin (RiboBio, Guangzhou, China) were transfected into SMSCs using Lipofectamine 3000 (Invitrogen, USA), and SMSCs were collected 48 h later for evaluating the transfection efficiency by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blot.

Techniques: Expressing, Western Blot, Co-Immunoprecipitation Assay

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Periostin/Bone Morphogenetic Protein 1 axis axis regulates proliferation and osteogenic differentiation of sutured mesenchymal stem cells and affects coronal suture closure in the TWIST1 +/− mouse model of craniosynostosis

doi: 10.1186/s13018-024-04604-3

Figure Lengend Snippet: Periostin/BMP1 axis regulates SMSC proliferation and osteogenesis. pcDNA 3.1-Periostin and pcDNA 3.1-BMP1 were co-transfected into the SMSCs of the TWIST1 +/− group. A Western blot assays were conducted to measure BMP1 protein expression in the SMSCs across the groups. B Colony formation assays were performed to evaluate the proliferative capacity of SMSCs. C EdU assays were utilized to further quantify the proliferation of SMSCs. D Wound healing assays were carried out to assess the wound closure rate of SMSCs. E ALP staining was employed to determine the ALP activity in SMSCs. F ARS staining was used to observe calcium deposition in SMSCs. G Western blot analysis was conducted to measure the expression levels of Ki-67, RUNX2, and OPN in SMSCs. Data were represented as mean ± SD (N = 3). * P < 0.05

Article Snippet: siRNA and pcDNA 3.1 overexpression vectors targeting BMP1 and Periostin (RiboBio, Guangzhou, China) were transfected into SMSCs using Lipofectamine 3000 (Invitrogen, USA), and SMSCs were collected 48 h later for evaluating the transfection efficiency by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blot.

Techniques: Transfection, Western Blot, Expressing, Staining, Activity Assay