Total Flavonoids of Rhizoma Drynariae Promotes Differentiation of Osteoblasts and Growth of Bone Graft in Induced Membrane Partly by Activating Wnt/β-Catenin Signaling Pathway

Total flavonoids of Rhizoma drynariae (TFRD), a Chinese medicine, is widely used in the treatment of fracture, bone defect, osteoporosis and other orthopedic diseases, and has achieved good effects. Purpose of this trial was to explore efficacy of TFRD on bone graft’s mineralization and osteoblasts’ differentiation in Masquelet induced membrane technique in rats. Forty male Sprague-Dawley rats were randomly divided into high dose group (H-TFRD), middle dose group (M-TFRD), low dose group (L-TFRD) and control group (control). The critical size bone defect model of rats was established with 10 rats in each group. Polymethyl methacrylate (PMMA) spacer was implanted into the defect of right femur in rats. After the formation of the induced membrane, autogenous bone was implanted into the induced membrane. After 12 weeks of bone graft, bone tissues in the area of bone graft were examined by X-ray, Micro-CT, hematoxylin-eosin (HE) and Masson trichrome staining to evaluate the growth of the bone graft. The β-catenin, c-myc, COL1A1, BMP-2 and OPN in bone graft were quantitatively analyzed by Western blot and Immunohistostaining. Osteoblasts were cultured in the medium containing TFRD. Cell Counting Kit-8 (CCK-8) method, Alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining, Western blot, RT-PCR and other methods were used to detect the effects of TFRD on the proliferation of osteoblasts and the regulation of Wnt/β-catenin signaling pathway. In vivo experiments showed that the growth and mineralization of bone graft in TFRD group was better. Moreover, the expression of Wnt/β-catenin and osteogenesis-related proteins in bone tissue of TFRD group was more than that in other groups. In vitro experiments indicated that osteoblasts proliferated faster, activity of ALP was higher, number of mineralized nodules and proteins related to osteogenesis were more in TFRD group. But blocking Wnt/β-catenin signaling pathway could limit these effects. Therefore, TFRD could promote mineralization of bone graft and differentiation of osteoblasts in a dose-dependent manner during growing period of the bone graft of induced membrane technique, which is partly related to the activation of Wnt/β-catenin signaling pathway.

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Corrigendum: Total Flavonoids of Rhizoma Drynariae Promotes Differentiation of Osteoblasts and Growth of Bone Graft in Induced Membrane Partly by Activating Wnt/β-Catenin Signaling Pathway

Frontiers in Pharmacology ◽

10.3389/fphar.2021.726831 ◽

2021 ◽

Vol 12 ◽

Author(s):

Shuyuan Li ◽

Hongliang Zhou ◽

Cheng Hu ◽

Jiabao Yang ◽

Jinfei Ye ◽

...

Keyword(s):

Bone Graft ◽

Signaling Pathway ◽

Total Flavonoids ◽

Induced Membrane ◽

Rhizoma Drynariae

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Modified Masquelet Technique Using Allogeneic Graft for a Gustilo-Anderson Type III-A Open Fracture of the Femur with an 8 cm Bone Defect

Case Reports in Orthopedics ◽

10.1155/2021/8829158 ◽

2021 ◽

Vol 2021 ◽

pp. 1-7

Author(s):

Hichem Issaoui ◽

Mohammed Reda Fekhaoui ◽

Moheddin Jamous ◽

Alain-Charles Masquelet

Keyword(s):

Bone Graft ◽

Bone Defect ◽

Bony Union ◽

Induced Membrane ◽

Masquelet Technique ◽

Membrane Technique ◽

Defect Area ◽

Induced Membrane Technique ◽

Large Bone

The induced membrane technique was initially described by Masquelet et al. in 1986 as a treatment for tibia nonunion; then, it became one of the established methods in the management of bone defects. Several changes have been made to this technique and have been used in different contexts and different methodologies. We present the case of a 16-year-old girl admitted to our department for a polytrauma after a motorcycle accident. She presented a Gustilo III-A open fracture of the right femoral shaft with a large bone defect of 8 centimeters that we treated with a modified Masquelet technique. In the first stage, an Open Reduction and Internal Fixation of the fracture was made using a 4,5 mm Dynamic Compression Plate and a PMMA cement was inserted at the bone defect area. The second stage was done after 11 weeks, and the defect area was filled exclusively with bone allograft from a bone bank. Complete bony union was seen at 60 weeks of follow-up. After the removal of the implants by another surgeon, the patient presented an atraumatic fracture of the neoformed bone that we treated with intramedullary femoral nailing associated with a local autograft using reaming debris. A complete bony union was achieved after 12 weeks with a complete range of motion of the hip and knee. The stability given to the fracture is essential because it influences the quality of the induced membrane and Masquelet has recommended high initial fixation rigidity to promote incorporation of the graft. It is recommended to delay the second stage of this technique after 8 weeks, especially in femoral reconstruction, to optimize the quality of the induced membrane. Several studies used a modified induced membrane technique to recreate a traumatic large bone defect, and all of them used an autologous bone graft alone or an enriched bone graft. In this case, the use of allograft exclusively seems to be as successful as an autologous or enriched bone graft. Now, with the advent of bone banks, it is possible to get an unlimited amount of allograft, so additional research and large studies are necessary before giving recommendations.

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Bone-Graft Resorption Reduced by the Induced Membrane Technique and Factors Affecting Volumetric Changes

The Journal of Bone and Joint Surgery (American) ◽

10.2106/jbjs.19.00804 ◽

2020 ◽

Vol 102 (14) ◽

pp. 1269-1278

Author(s):

Jae-Woo Cho ◽

William T. Kent ◽

Chang-Wug Oh ◽

Beom-Soo Kim ◽

Won-Tae Cho ◽

...

Keyword(s):

Bone Graft ◽

Factors Affecting ◽

Induced Membrane ◽

Membrane Technique ◽

Induced Membrane Technique ◽

Graft Resorption

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Induced membrane technique in the treatment of infectious bone defect: A clinical analysis

Orthopaedics & Traumatology Surgery & Research ◽

10.1016/j.otsr.2019.01.007 ◽

2019 ◽

Vol 105 (3) ◽

pp. 535-539 ◽

Cited By ~ 5

Author(s):

Jianbing Wang ◽

Qudong Yin ◽

Sanjun Gu ◽

Yongwei Wu ◽

Yongjun Rui

Keyword(s):

Bone Defect ◽

Clinical Analysis ◽

Induced Membrane ◽

Membrane Technique ◽

Induced Membrane Technique

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Induced membrane technique combined with antibiotic-loaded calcium sulfate–calcium phosphate composite as bone graft expander for the treatment of large infected bone defects: preliminary results of 12 cases

Annals of Translational Medicine ◽

10.21037/atm-20-1932 ◽

2020 ◽

Vol 8 (17) ◽

pp. 1081-1081

Author(s):

Zihou Zhao ◽

Guoliang Wang ◽

Yong Zhang ◽

Wen Luo ◽

Shiyu Liu ◽

...

Keyword(s):

Calcium Phosphate ◽

Bone Graft ◽

Calcium Sulfate ◽

Bone Defects ◽

Induced Membrane ◽

Preliminary Results ◽

Membrane Technique ◽

Induced Membrane Technique

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Post-traumatic forearm bone defect reconstruction using the induced membrane technique

Orthopaedics & Traumatology Surgery & Research ◽

10.1016/j.otsr.2021.103036 ◽

2021 ◽

pp. 103036

Author(s):

Paul Commeil ◽

Arthur Seguineau ◽

Alison Delesque ◽

Alexandra Erbland ◽

Edouard Harly ◽

...

Keyword(s):

Bone Defect ◽

Induced Membrane ◽

Defect Reconstruction ◽

Membrane Technique ◽

Induced Membrane Technique ◽

Post Traumatic ◽

Forearm Bone ◽

Bone Defect Reconstruction

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Assemble Flavone of Rhizoma Drynariae Promotes Differentiation of Osteoblasts and Growth of Bone Graft in Induced Membrane Partly by Activating Wnt/β-Catenin Signal Pathway

10.21203/rs.3.rs-117503/v1 ◽

2020 ◽

Author(s):

Shuyuan Li ◽

Hongliang zhou ◽

Chenghu Hu ◽

Jiabao Yang ◽

Yue Li ◽

...

Keyword(s):

Bone Graft ◽

Signal Pathway ◽

Autogenous Bone ◽

Control Group ◽

Sprague Dawley ◽

Induced Membrane ◽

Membrane Technique ◽

Rhizoma Drynariae

Abstract Background: Assemble Flavone of Rhizoma Drynariae (AFRD) is not only the extract of Rhizoma Drynariae, but also the effective component of Qianggu capsule, which is widely used in the treatment of fracture, bone defect, osteoporosis and other orthopedic diseases, and has achieved good results. Purpose of this trial was to explore effects of AFRD on bone graft mineralization and osteoblast differentiation in Masquelet induced membrane in rats.Methods: Twenty male Sprague-Dawley rats aged 10-12 weeks were randomLy divided into AFRD group (n=10) and control group (n=10). Critical-sized defects model of rats was established with 10 rats in each group. Polymethyl methacrylate (PMMA) was implanted into the defect of femur in rats. After the formation of the induced membrane, autogenous bone was implanted into the induced membrane. After 10 weeks of bone grafting, bone tissue in the bone graft area was examined by X-ray, Micro-CT and hematoxylin-eosin (HE) staining to evaluate the growth of the bone graft. Serums of the two groups of rats were extracted respectively, and these serums were used to culture osteoblasts in vitro. CKK8 method, Alkaline phosphatase (ALP) staining, Western blot and RT-PCR and other methods were used to evaluate the effects of AFRD on the proliferation of osteoblasts and the regulation of Wnt/β-catenin signal pathway.Results: In vivo experiment showed that the growth and mineralization effect of bone graft in AFRD group was better. In vitro experiment showed that osteoblasts proliferated faster, ALP activity was higher, number of mineralized nodules was more, and expression of proteins related to Wnt/β-catenin signal pathway and osteogenesis were more in AFRD group.Conclusions: AFRD can promote mineralization of bone graft and differentiation of osteoblasts during the bone graft growth period of induced membrane technique, which is related to the activation of Wnt/ β-catenin signal pathway.

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