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P17. Effect of bone marrow mesenchymal stem cell combined with concentrate growth factor (CGF) on postmenopausal bone defects
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Proceedings of the 34th Annual Meeting of the North American Spine Society / The Spine Journal 19 (2019) S158−S194
P17. Effect of bone marrow mesenchymal stem cell combined with concentrate growth factor (CGF) on postmenopausal bone defects Xiang Yu, PhD1, Hui Ren1, Qi Shang, MD2, De Liang, MD2, Jiang Xiaobing, PhD3; 1 Guangzhou University of Chinese Medicine, Guangzhou, Baiyun/Guangdong, China; 2 Guangzhou, China; 3 1st Affiliated Hospital of Guangzhou University of Chinese Medicine, Guang Zhou City, China BACKGROUND CONTEXT: Concentrate growth factor (CGF) is a third-generation platelet concentrate that releases a variety of bioactive substances and plays a good role in promoting tissue healing. Bone marrow mesenchymal stem cell (BMSCs) transplantation has been used to treat osteoporosis. However, effect of bone marrow mesenchymal stem cell combined with concentrate growth factor (CGF) on postmenopausal bone defects remains elusive. PURPOSE: To observe the effect of BMSCs combined with concentrate growth factor (CGF) on postmenopausal bone defects and to explore related mechanisms. METHODS: A total of 50 Sprague-Dawley rats were randomly divided into A (SHAM), B (model), C (CGF), D (BMSCs) and E (CGF + BMSCs) group. Group A did not remove ovaries and L6 vertebral body was created a 3mm £ 3mm £ 3mm large bone defect. The B, C, D, E groups experimented ovariectomy and then L6 vertebral body created a 3mm £ 3mm £ 3mm large bone defect model. C and E groups were filled with a suitable concentrate growth factor (CGF). D and E groups were injected Brdu-labeled BMSCs through tail vein. After 4 and 8 weeks of treatment, the vertebral body of the bone defect was taken for micro-CT and the parameters such as TV/BV, Tb.N, Tb.Th, Tb.Sp and vBMD. HE staining of vertebral bodies were to observe the morphology of bone tissue. Brdu immunohistochemistry and Brdu immunofluorescence were used to detect stem cell homing. Immunofluorescence was used to detect the expression of VEGF, BMP2 and NF-KB. Runx2, CTSK and VEGF NFKB were detected by PCR and Western-bolt. RESULTS: The Brdu labeling rate of BMSCs was up to 95.43%. After 4 and 8 weeks of intervention, bone formation in bone defects of groups C, D and E were significantly higher than that of group B. TV/BV, Tb. N, Tb.h and vBMD of C, D group were significantly higher than the model group, and the Tb.Sp of group B was higher than that of group C, D and E. The morphology of bone tissue in groups C, D and E was better than that in model group. The CGF combined BMSCs group showed a good synergistic effect, and their bone micro-structure and bone histomorphology were better than that of CGF or BMSCs alone. In the D and E groups, Brdu immunofluorescence and immunohistochemistry were detected in the bone defect. Compared with the model group, the expression of VEGF, BMP2 and NF-KB was more obvious in the bone defects of group C, D and E. Compared with the SHAM group, the expression of CTSK mRNA and protein in the model group was significantly up-regulated (P<0.01), and Runx2, VEGF and NF-KB were down-regulated. Compared with B group, the expressions of CTSK mRNA and protein in C, D and E groups were significantly down-regulated (P<0.01), and the expressions of Runx2, VEGF and NF-KB mRNA and protein were up-regulated (P<0.01). CONCLUSIONS: BMSCs combined with CGF can effectively treat postmenopausal bone defects, which may promote bone defect repair by regulating inflammation, bone formation and angiogenesis. FDA DEVICE/DRUG STATUS: This abstract does not discuss or include any applicable devices or drugs. https://doi.org/10.1016/j.spinee.2019.05.441
P18. Treatment of osteoporotic vertebral fractures/defect in rats by naringin/hydroxyapatite/silk fibroin composite scaffolds in regulation of osteogenic differentiation, angiogenesis and inflammation Xiang Yu, PhD1, Hui Ren1, Gengyang Shen, MD2, De Liang, MD2, Zhida Zhang, PhD1, Qi Shang, MD3, Jiang Xiaobing, PhD3; 1 Guangzhou University of Chinese Medicine, Guangzhou, Baiyun/Guangdong, China;
2 Guangzhou, China; 3 1st Affiliated Hospital of Guangzhou University of Chinese Medicine, Guang Zhou City, China
BACKGROUND CONTEXT: Biomaterial scaffolds, as one of the three major elements of regenerative medicine, play an important role in bone remodeling and are therefore commonly used in the treatment of osteoporotic vertebral fractures (OVF). Among many biological materials, hydroxyapatite has attracted much attention in the research of bone defect regeneration due to its better bone conduction characteristics and mineralization processes and structures similar to bone tissue. Silk fibroin is a kind of natural biopolymer with high mechanical strength, controlled biodegradability, good biocompatibility and easy handling. The use of modern science and technology to make silk fibroin and hydroxyapatite into a composite scaffold can not only integrate the advantages of the two, but also avoid the disadvantages of low load bearing pressure, rejection reaction and non-degradability of traditional materials, which is an ideal, new tissue engineering material for OVF treatment. In recent years, traditional Chinese medicine has shown its unique advantages in the prevention and treatment of OVF. Traditional Chinese medicine has the effect of strengthening kidney and strengthening bone, promoting blood circulation and removing blood stasis. It is often used to promote fracture healing in clinic. Naringin is the main active ingredient of Rhizoma Drynariae, which can promote bone formation, angiogenesis, and inhibit inflammation. The combination of naringin, hydroxyapatite and silk fibroin as a new composite scaffold may have a synergistic effect and provide new insight on osteoporotic vertebral fractures/bone defect. PURPOSE: To investigate the therapeutic effect and molecular mechanism of a new scaffold with naringin /hydroxyapatite/silk fibroin (NG/ GMs/HA/SF) on vertebral fractures in osteoporotic rats. METHODS: Loaded granules of naringin microspheres (NG/GMs), silk fibroin scaffolds (SF), hydroxyapatite/silk fibroin composite scaffolds (HA/SF), and granule naringin /hydroxyapatite/silk fibroin composite scaffold (NG//HA/SF) and loaded granule naringenin microsphere/hydroxyapatite/silk fibroin composite scaffold (NG/GMs/HA/SF) were conducted. The microscope was used to observe, calculate the volume of GMs and swelling ratio. X-ray diffraction (XRD) was used to analyse HA structure characterization; Fourier transform infrared spectroscopy was used to detect each scaffold structure. Scanning electron microscopy was used to analyse the surface structure, pore size and biocompatibility of each scaffold. Scanning calorimetry (DSC) was used to analyze the thermal properties and stability of each scaffold. UV spectrophotometer was used to detect the drug release rate in vitro. Trypan blue staining was used to detect the activity of MC3T3-E1 osteoblast cell line in the scaffold. We co-cultured bone marrow derived-mesenchymal stem cells (BMSCs) and scaffolds. And then ALP staining was used to detect the osteogenic differentiation potential of BMSCs, alizarin red staining used to detect mineralization ability, qRT-PCR used to detect the expression levels of osteogenic markers OCN, BMP-2 and Runx2 mRNA; AKP reagent used to detect osteoblast activity, Western-blot used to detect OCN, BMP-2, Runx2 protein expression. The rats experimented ovariectomize surgery to construct postmenopausal osteoporosis model, and next osteoporosis model was successfully used to construct lumbar vertebrae bone defect model, and then the scaffold was filled in bone Defects. X-ray was for the detection of bone defect formation, HE staining, Safranin solid staining and Masson staining for detection of bone defect morphological changes. Micro-CT examined three-dimensional structure and bone structure parameters of bone defect. Serum estrogen, CTSK, OCN/BGP, AKP levels were detected by ELISA. qRT-PCR, immunohistochemistry and Western-blot was used to detect osteoclast markers (OCN, Runx2, BMP2, CTSK), Collagen II), angiogenesis markers (VEGFA) and inflammatory markers (IK6, TNF-a, NF-kB) mRNA and protein expression. RESULTS: NG/GMs, SF, HA/SF, NG//HA/SF, NG/GMs/HA/SF scaffolds were successfully constructed, and the scaffolds had good biocompatibility and sustained drug release rate. The loaded granule naringenin microsphere/hydroxyapatite/silk fibroin composite scaffold can promote osteogenic differentiation and mineralization of BMSCs, and the scaffold can up-regulate the mRNA and protein expression of OCN, BMP-2 and
Refer to onsite annual meeting presentations and postmeeting proceedings for possible referenced figures and tables. Authors are responsible for accurately reporting disclosure and FDA device/drug status at time of abstract submission.
Proceedings of the 34th Annual Meeting of the North American Spine Society / The Spine Journal 19 (2019) S158−S194
P17. Effect of bone marrow mesenchymal stem cell combined with concentrate growth factor (CGF) on postmenopausal bone defects Xiang Yu, PhD1, Hui Ren1, Qi Shang, MD2, De Liang, MD2, Jiang Xiaobing, PhD3; 1 Guangzhou University of Chinese Medicine, Guangzhou, Baiyun/Guangdong, China; 2 Guangzhou, China; 3 1st Affiliated Hospital of Guangzhou University of Chinese Medicine, Guang Zhou City, China BACKGROUND CONTEXT: Concentrate growth factor (CGF) is a third-generation platelet concentrate that releases a variety of bioactive substances and plays a good role in promoting tissue healing. Bone marrow mesenchymal stem cell (BMSCs) transplantation has been used to treat osteoporosis. However, effect of bone marrow mesenchymal stem cell combined with concentrate growth factor (CGF) on postmenopausal bone defects remains elusive. PURPOSE: To observe the effect of BMSCs combined with concentrate growth factor (CGF) on postmenopausal bone defects and to explore related mechanisms. METHODS: A total of 50 Sprague-Dawley rats were randomly divided into A (SHAM), B (model), C (CGF), D (BMSCs) and E (CGF + BMSCs) group. Group A did not remove ovaries and L6 vertebral body was created a 3mm £ 3mm £ 3mm large bone defect. The B, C, D, E groups experimented ovariectomy and then L6 vertebral body created a 3mm £ 3mm £ 3mm large bone defect model. C and E groups were filled with a suitable concentrate growth factor (CGF). D and E groups were injected Brdu-labeled BMSCs through tail vein. After 4 and 8 weeks of treatment, the vertebral body of the bone defect was taken for micro-CT and the parameters such as TV/BV, Tb.N, Tb.Th, Tb.Sp and vBMD. HE staining of vertebral bodies were to observe the morphology of bone tissue. Brdu immunohistochemistry and Brdu immunofluorescence were used to detect stem cell homing. Immunofluorescence was used to detect the expression of VEGF, BMP2 and NF-KB. Runx2, CTSK and VEGF NFKB were detected by PCR and Western-bolt. RESULTS: The Brdu labeling rate of BMSCs was up to 95.43%. After 4 and 8 weeks of intervention, bone formation in bone defects of groups C, D and E were significantly higher than that of group B. TV/BV, Tb. N, Tb.h and vBMD of C, D group were significantly higher than the model group, and the Tb.Sp of group B was higher than that of group C, D and E. The morphology of bone tissue in groups C, D and E was better than that in model group. The CGF combined BMSCs group showed a good synergistic effect, and their bone micro-structure and bone histomorphology were better than that of CGF or BMSCs alone. In the D and E groups, Brdu immunofluorescence and immunohistochemistry were detected in the bone defect. Compared with the model group, the expression of VEGF, BMP2 and NF-KB was more obvious in the bone defects of group C, D and E. Compared with the SHAM group, the expression of CTSK mRNA and protein in the model group was significantly up-regulated (P<0.01), and Runx2, VEGF and NF-KB were down-regulated. Compared with B group, the expressions of CTSK mRNA and protein in C, D and E groups were significantly down-regulated (P<0.01), and the expressions of Runx2, VEGF and NF-KB mRNA and protein were up-regulated (P<0.01). CONCLUSIONS: BMSCs combined with CGF can effectively treat postmenopausal bone defects, which may promote bone defect repair by regulating inflammation, bone formation and angiogenesis. FDA DEVICE/DRUG STATUS: This abstract does not discuss or include any applicable devices or drugs. https://doi.org/10.1016/j.spinee.2019.05.441
P18. Treatment of osteoporotic vertebral fractures/defect in rats by naringin/hydroxyapatite/silk fibroin composite scaffolds in regulation of osteogenic differentiation, angiogenesis and inflammation Xiang Yu, PhD1, Hui Ren1, Gengyang Shen, MD2, De Liang, MD2, Zhida Zhang, PhD1, Qi Shang, MD3, Jiang Xiaobing, PhD3; 1 Guangzhou University of Chinese Medicine, Guangzhou, Baiyun/Guangdong, China;
2 Guangzhou, China; 3 1st Affiliated Hospital of Guangzhou University of Chinese Medicine, Guang Zhou City, China
BACKGROUND CONTEXT: Biomaterial scaffolds, as one of the three major elements of regenerative medicine, play an important role in bone remodeling and are therefore commonly used in the treatment of osteoporotic vertebral fractures (OVF). Among many biological materials, hydroxyapatite has attracted much attention in the research of bone defect regeneration due to its better bone conduction characteristics and mineralization processes and structures similar to bone tissue. Silk fibroin is a kind of natural biopolymer with high mechanical strength, controlled biodegradability, good biocompatibility and easy handling. The use of modern science and technology to make silk fibroin and hydroxyapatite into a composite scaffold can not only integrate the advantages of the two, but also avoid the disadvantages of low load bearing pressure, rejection reaction and non-degradability of traditional materials, which is an ideal, new tissue engineering material for OVF treatment. In recent years, traditional Chinese medicine has shown its unique advantages in the prevention and treatment of OVF. Traditional Chinese medicine has the effect of strengthening kidney and strengthening bone, promoting blood circulation and removing blood stasis. It is often used to promote fracture healing in clinic. Naringin is the main active ingredient of Rhizoma Drynariae, which can promote bone formation, angiogenesis, and inhibit inflammation. The combination of naringin, hydroxyapatite and silk fibroin as a new composite scaffold may have a synergistic effect and provide new insight on osteoporotic vertebral fractures/bone defect. PURPOSE: To investigate the therapeutic effect and molecular mechanism of a new scaffold with naringin /hydroxyapatite/silk fibroin (NG/ GMs/HA/SF) on vertebral fractures in osteoporotic rats. METHODS: Loaded granules of naringin microspheres (NG/GMs), silk fibroin scaffolds (SF), hydroxyapatite/silk fibroin composite scaffolds (HA/SF), and granule naringin /hydroxyapatite/silk fibroin composite scaffold (NG//HA/SF) and loaded granule naringenin microsphere/hydroxyapatite/silk fibroin composite scaffold (NG/GMs/HA/SF) were conducted. The microscope was used to observe, calculate the volume of GMs and swelling ratio. X-ray diffraction (XRD) was used to analyse HA structure characterization; Fourier transform infrared spectroscopy was used to detect each scaffold structure. Scanning electron microscopy was used to analyse the surface structure, pore size and biocompatibility of each scaffold. Scanning calorimetry (DSC) was used to analyze the thermal properties and stability of each scaffold. UV spectrophotometer was used to detect the drug release rate in vitro. Trypan blue staining was used to detect the activity of MC3T3-E1 osteoblast cell line in the scaffold. We co-cultured bone marrow derived-mesenchymal stem cells (BMSCs) and scaffolds. And then ALP staining was used to detect the osteogenic differentiation potential of BMSCs, alizarin red staining used to detect mineralization ability, qRT-PCR used to detect the expression levels of osteogenic markers OCN, BMP-2 and Runx2 mRNA; AKP reagent used to detect osteoblast activity, Western-blot used to detect OCN, BMP-2, Runx2 protein expression. The rats experimented ovariectomize surgery to construct postmenopausal osteoporosis model, and next osteoporosis model was successfully used to construct lumbar vertebrae bone defect model, and then the scaffold was filled in bone Defects. X-ray was for the detection of bone defect formation, HE staining, Safranin solid staining and Masson staining for detection of bone defect morphological changes. Micro-CT examined three-dimensional structure and bone structure parameters of bone defect. Serum estrogen, CTSK, OCN/BGP, AKP levels were detected by ELISA. qRT-PCR, immunohistochemistry and Western-blot was used to detect osteoclast markers (OCN, Runx2, BMP2, CTSK), Collagen II), angiogenesis markers (VEGFA) and inflammatory markers (IK6, TNF-a, NF-kB) mRNA and protein expression. RESULTS: NG/GMs, SF, HA/SF, NG//HA/SF, NG/GMs/HA/SF scaffolds were successfully constructed, and the scaffolds had good biocompatibility and sustained drug release rate. The loaded granule naringenin microsphere/hydroxyapatite/silk fibroin composite scaffold can promote osteogenic differentiation and mineralization of BMSCs, and the scaffold can up-regulate the mRNA and protein expression of OCN, BMP-2 and
Refer to onsite annual meeting presentations and postmeeting proceedings for possible referenced figures and tables. Authors are responsible for accurately reporting disclosure and FDA device/drug status at time of abstract submission.