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    08 November 2020, Volume 24 Issue 34 Previous Issue    Next Issue
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    Research hotspots in tissue decellularization method for manufacturing extracellular matrices
    He Jing, Ao Qiang
    2020, 24 (34):  5413-5420.  doi: 10.3969/j.issn.2095-4344.2331
    Abstract ( 512 )   PDF (56873KB) ( 42 )   Save

    BACKGROUND: Biological scaffold materials made of extracellular matrix are currently widely used in clinical treatment and basic studies in vivo and in vitro. The quality of decellularization technique is of great importance since unsuccessful decellularized effects will lead to a series of immunological rejection after implantation, which would finally result in treatment failure.

    OBJECTIVE: To introduce the research advancement of tissue decellularized method of extracellular matrix materials.

    METHODS: The articles published from January 2005 to December 2019 were retrieved from PubMed database. The key words were “extracellular matrix, tissue decellularization, decellularized method, tissue engineering”. The articles published from January 2014 to December 2019 were retrieved from Google Patent database; the key words were “acellular tissue, enzymatic modification”. References closely related to the decellularized method of extracellular matrix materials were included and repetitive studies were excluded.

    RESULTS AND CONCLUSION: Preparation methods of acellular matrix materials vary widely, which mainly include physical, chemical and biological methods. Selecting appropriate decellularized method according to different tissue types is vital for thoroughly removing cell components and retaining the structure and constituent of extracellular matrix to the fullest extent. Severe immunoinflammatory responses can occur under the condition of insufficient decellularized effects. Enzymatic modification of acellular tissue can improve tissue flexibility, elasticity and resistance to friction, and further remove tissue immunogenicity. Additionally, proper selection of sterilization method can help to better preserve the ultrastructural and mechanical properties of extracellular matrix materials. Simultaneously, acellular biological scaffolds of xenogenic sources should strictly comply with the provisions of the food and drug supervision and administration measures, which require that the final method of sterilization for extracellular matrix products is selected according to the guidelines for different bacterial inoculations. 

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    Treatment of meniscus injury or degeneration: the effect and function of stem cells and artificial polymer scaffolds to form tissue engineering system
    Jiang Zongrui, Zhang Zhiqi
    2020, 24 (34):  5421-5427.  doi: 10.3969/j.issn.2095-4344.2334
    Abstract ( 504 )   PDF (38162KB) ( 38 )   Save

    BACKGROUND: As a necessary anatomic structure of knee joint, meniscus injury or meniscectomy could possibly contribute to the initiation of early osteoarthritis. Therefore, it is very important to use tissue engineering technology to find a substitute that can completely imitate normal physiological meniscus.

    OBJECTIVE: To summarize the research progress of cell, scaffold and stimulus of meniscus tissue engineering.

    METHODS: The authors searched PubMed, Web of science, CNKI and Wanfang database to discover eligible articles. The key words were “meniscus; tissue engineering; mesenchymal stem cell; scaffold; synthetic polymer scaffolds; polycaprolactone; hydrogel; ECM component scaffold; tissue; small intestine submucosa; decellularized; growth factor; dynamic compression; tensile; cyclic hydrostatic pressure” in English; and “meniscus, tissue engineering, stem cells, scaffolds, artificial polymer scaffolds, hydrogel, extracellular matrix scaffolds, small intestinal submucosa, acellular technology, growth factors, dynamic compression test, tensile load, and cyclic hydrostatic pressure test” in Chinese. Finally, 61 articles were included.

    RESULTS AND CONCLUSION: Seed cell, scaffold and biological and physical stimulus are three significant important factors in meniscus tissue engineering. Mesenchymal stem cell is so far the most commonly applied seed cell in tissue engineering. Stem cells can be implanted on synthetic polymer scaffold or nature scaffold like extracellular matrix component scaffold and tissue-derived scaffold, or wrapped by hydrogel. With the endogenous or exogenous growth factors and biomechanical stimulation, meniscus regeneration will proceed. Nowadays, researchers tend to systematically integrate seed cell, scaffold and stimulus into a system to study. Therefore, the construction of a system which mostly fits the anatomic structure, physiologic function and biocompatibility of meniscus has become a current hotspot in tissue engineering.

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    Sustained-release effect and clinical application of thermosensitive gels
    Chen Yongjia, Li Yanlin, Liu Dejian, He Yinghong, Yang Xiao
    2020, 24 (34):  5428-5433.  doi: 10.3969/j.issn.2095-4344.2325
    Abstract ( 751 )   PDF (36618KB) ( 39 )   Save

    BACKGROUND: Thermosensitive gel is a newly emerging drug type, which has been a popular area of research due to its advantages including sustained release, controlled release, and targeted drug delivery.

    OBJECTIVE: To summarize the various mechanisms underlying the sustained release of thermosensitive gels published in the literature, and its clinical application outcomes.

    METHODS: A retrieval of Wanfang, CNKI, VIP, PubMed, and Google Scholar was conducted using the


    search terms “thermosensitive gel, sustained release, administration route” in Chinese and English, respectively for the articles concerning sustained release mechanisms of thermosensitive gel and clinical applications.

    RESULTS AND CONCLUSION: (1) The thermosensitive gel possesses the lowest critical solution temperature and can change with the environment temperature changing to some extent. It has more advantages than other gels, and can lead to in vivo sustained release of drugs and reduce the toxicity, prevent drug extravasation, improve the stability of the drug, also has good clinical application prospects. (2) Thermosensitive gels have a variety of administration ways, such as injection, oral, in-ear, nasal cavity, intraocular, and skin, which has wide prospects. However, different kinds of gel materials have different shortcomings. Native biomaterials hold poor plasticity and mechanical strength, and are easy to be contaminated by pathogenic microorganisms, and cannot be prepared in large amounts. Besides, the hydrophilicity of synthetic materials is poor. Moreover, the effect of metabolic pathway of thermosensitive gel on tissues and organs is still under investigation.

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    Prevention and treatment of implant periapical lesions
    Shi Qianhui, Wu Chao, Zhou Qian, Cheng Yuting, Li Fang, Huo Hua, Qi Yuhan, Huang Xiaolin, Wang Yong, Liao Jian
    2020, 24 (34):  5434-5440.  doi: 10.3969/j.issn.2095-4344.2326
    Abstract ( 614 )   PDF (40302KB) ( 49 )   Save

    BACKGROUND: At present, the understanding of periapical lesions of implants is not sufficient.

    OBJECTIVE: Based on the latest research trends related to the clinical symptoms, pathogenic factors and treatment methods of implant periapical lesions, this paper reviews the research progress of the etiology and treatment methods of implant periapical lesions.

    METHODS: The first author searched PubMed, VIP, Wanfang and CNKI databases using computer for relevant articles published from January 1985 to November 2019. The search terms were “apical peri-implantitis, retrograde peri-implantitis, dental implant, implants periapical lesion” in English and Chinese, respectively. The etiology, clinical manifestation, and treatment plan of implant periapical lesions were summarized.

    RESULTS AND CONCLUSION: The implant periapical lesions may be caused by infection, bone burning, poor bone quality, systemic and local factors. The clinical manifestations were redness and swelling of the mucosa in the periapical area of the implant, tenderness, or fistula formation. Imaging results showed a low-density radiation shadow in the periapical area of the implant. The treatment methods include root canal therapy of adjacent teeth, surgical debridement, implant root tip resection, guided bone regeneration, and systemic application of antibiotics. Therefore, to be familiar with the etiology and treatment methods of implant periapical lesions can provide positive guidance for the prevention and treatment of implant periapical lesions in clinical practice.

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    Application of nanocomposite hydrogels in bone tissue engineering
    Yu Xingge, Lin Kaili
    2020, 24 (34):  5441-5446.  doi: 10.3969/j.issn.2095-4344.2312
    Abstract ( 479 )   PDF (38222KB) ( 248 )   Save

    BACKGROUND: Nanocomposite hydrogels are biomimetic materials with good biocompatibility and bone conductive properties and are widely used in bone tissue engineering.

    OBJECTIVE: To summarize the classification of nanoparticles and the application of nanocomposite hydrogels in bone tissue engineering.

    METHODS: The literatures published in the CNKI, PubMed, and Web of science from 2000 to 2019 were searched by computer. The key words were “Nanocomposites; Bone Tissue Engineering; Nanoparticle; Hydrogels”. According to the inclusion and exclusion criteria, 69 articles were included for final analysis.

    RESULTS AND CONCLUSION: (1) Nanoparticles currently used as nanofillers mainly include carbon-based nanoparticles, metal and metal oxide nanoparticles, polymer nanoparticles, and calcium phosphate-based


    nanoparticles. (2) The combination of nanoparticles and hydrogels can improve the mechanical properties of hydrogels. (3) Nanocomposite hydrogels can promote the adhesion and osteogenic differentiation of seed cells and thus be applied to bone tissue engineering. (4) At present, the application of nanocomposite hydrogels in bone tissue engineering still has some problems such as weak dispersion of nanoparticles, low vascularization capacity in bone tissue engineering, and the difficulty in the control of degradation rate. 

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    Effects of calcined bone/chitosan composite materials on proliferation and adhesion in osteoblasts
    Liao Jian, Huang Xiaolin, Huo Hua, Zhou Qian, Cheng Yuting, Qi Yuhan, Wu Chao, Yang Tongjing, Liao Yunmao, Liang Xing
    2020, 24 (34):  5447-5453.  doi: 10.3969/j.issn.2095-4344.2357
    Abstract ( 334 )   PDF (28566KB) ( 200 )   Save

    BACKGROUND: The different ratios of calcined bone/chitosan composites were prepared in previous studies, and in vitro cell experiments showed that the composites were safe and non-toxic.

    OBJECTIVE: To observe the effect of calcined bone/chitosan composite on the proliferation and adhesion of osteoblasts.  

    METHODS: The composite materials with calcined bone and chitosan mass ratios of 1/2, 1/1, and 2/1 were prepared by solution blending method. Neonatal rat osteoblasts were inoculated on three kinds of composite materials, and the cells cultured alone were used as control. CCK-8 kit was used to detect cell proliferation. DAPI fluorescence staining was used to observe the growth of cells on the material. Scanning electron microscopy was used to observe the adhesion of cells on the surface of the composite material.

    RESULTS AND CONCLUSION: (1) The CCK-8 test showed that the osteoblasts of the four components proliferated well, proliferated slowly from the first to third days of culture, accelerated after the third day, entered the logarithmic growth phase from the third to seventh days, and entered the plateau phase after the seventh day. There was no difference in cell proliferation between the composite materials group and the control group. (2) After 3 days of culture, DAPI fluorescent staining showed that osteoblasts grew well on the surface of the three kinds of composites, and there was no significant difference between the groups. (3) Scanning electron microscopy showed that after 3 days of culture, the osteoblasts adhered tightly to the surface of the three kinds of composites, stretched completely; the cell body was full and protruded the pseudopod that was embedded in the material. After 7 days of culture, the cells covered the surface of the material and grew densely, among which the cells on the surface of calcined bone/chitosan composite with a mass ratio of 2/1 were most dense. (4) The results showed that osteoblasts could proliferate and adhere on the surface of the calcined bone/chitosan composite material.

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    In vivo biosafety and histocompatibility of absorbable poly-D,L-lactic acid screws implanted with ultrasound-assisted technology
    Lei Senlin, Liu Hongyuan, Yang Hongsheng, Xiong Yan, Duan Hong
    2020, 24 (34):  5454-5460.  doi: 10.3969/j.issn.2095-4344.2324
    Abstract ( 366 )   PDF (26245KB) ( 42 )   Save

    BACKGROUND: According to the characteristics of low glass transition temperature of poly-D,L-lactic acid, we developed a new ultrasonic welding device to treat limb fractures with restorable poly-D,L-lactic acid screws. But whether the physical properties of poly-D,L-lactic acid changed by ultrasound has impact on material safety is unknown. In addition, under high intensity ultrasound action, the effects of absorbable screws on tissues around the absorbable screws remain unclear.

    OBJECTIVE: To evaluate the biosafety and histocompatibility of ultrasound-assisted absorbable poly-D,L-lactic acid screws in rats.

    METHODS: Twenty-four adult SD rat models of 1.6 mm-sized bone defects were made in the anterior side of the right tibial shaft, and then randomly divided into two groups. Absorbable poly-D,L-lactic acid screws were implanted in the experimental group and nothing was implanted in the control group. At postoperative 1, 3, 7 and 14 days and 2, 4, 6, and 8 weeks, routine blood tests, blood biochemical and serum inflammatory indicators were detected. At postoperative 2 weeks and 1, 3 and 6 months, gross observation, X-ray imaging, and histological observation of the bone defects were performed. This study was approved by the Animal Ethics Committee, West China Hospital of Sichuan University, China.  

    RESULTS AND CONCLUSION: (1) In both experimental and control groups, after surgery, wounds healed well without signs of infection and rejection, and there were no abnormal fluctuations in routine blood tests, biochemical and inflammatory indicators. (2) Gross observation revealed that by 1 month after surgery, absorbable poly-D,L-lactic acid screws began to degrade with a small amount of cartilage callus crawling towards the center; by 6 months after surgery, there were a large amount of new bone around the screws, and no abnormal reactive hyperplasia was observed in the surrounding tissues. (3) X-ray showed that the bone defect area in the experimental group was gradually reduced, and by 6 months after surgery, there were a large amount of new bone, and no abnormal reaction of the surrounding bone matrix was observed. (4) Histological observation revealed that at 2 weeks after surgery, the interface between the screws and the bone was clear, adjacent bone trabeculae did not grow in the screws, and fibrous tissue proliferated and covered the surface of the materials. The screws gradually degraded, new bone tissue gradually increased and matured. (5) These results suggest that ultrasound-assisted absorbable poly-D,L-lactic acid screws exhibit good biosafety and histocompatibility and can degrade gradually in vivo. 

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    3D-printed icariin/decalcified bone matrix material promotes the repair of femoral condyle defects in rabbits
    Zhang Huxiong, Li Wei, Yang Wupeng, New Suyaratu
    2020, 24 (34):  5461-5466.  doi: 10.3969/j.issn.2095-4344.2313
    Abstract ( 398 )   PDF (27852KB) ( 33 )   Save

    BACKGROUND: Icariin can promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, and has good osteogenic properties. Decalcified bone matrix material exhibits good bone conduction and bone induction. It can be used in bone repair alone or in combination with other materials.

    OBJECTIVE: To investigate the sustained-release performance, cytocompatibility and osteogenesis of 3D-printed icariin/decalcified bone matrix material.  

    METHODS: Icariin/decalcified bone matrix materials and decalcified bone matrix materials were prepared by 3D printing technology. The in vitro sustained-release performance of the icariin/decalcified bone matrix material was detected. Bone marrow mesenchymal stem cells were inoculated on the surfaces of the two materials respectively. The cells cultured separately were used as the control. At the designated time points, the live/dead staining and MTT assay were performed, and alkaline phosphatase activity and osteocalcin content were determined. Thirty New Zealand rabbit models of femoral condyle defect were prepared and then divided into three groups. In the control group, no implant was used. In the simple decalcified bone matrix group, decalcified bone matrix and allogeneic rabbit bone marrow mesenchymal stem cells composite were implanted. In the icariin/decalcified bone matrix group, icariin/decalcified bone matrix and allogeneic rabbit bone marrow mesenchymal stem cells were implanted. At 4 and 12 weeks after surgery, micro-CT and histological and mechanical properties evaluation were performed.

    RESULTS AND CONCLUSION: (1) 3D-printed icariin/decalcified bone matrix material exhibited sustained-release performance. At 28 days, the release of icariin was (54.9±7.9)% of total release amount. (2) The live/dead staining revealed that the number of cells on the material surface in simple decalcified bone matrix and icariin/decalcified bone matrix groups were decreased after 1 day of inoculation and significantly increased with the prolongation of culture time. At 7 days, cells grew well, and the cells on the surface of icariin/decalcified bone matrix material were more evenly distributed and increased in number. (3) MTT test showed that cells in the icariin/decalcified bone matrix group proliferated faster after 7, 10, and 14 days of culture than those in the other two groups (P < 0.05). (4) The alkaline phosphatase activity and osteocalcin content in the icariin/decalcified bone matrix group were higher than those in the other two groups after 7, 10, and 14 days of culture (P < 0.05). (5) At 12 weeks after surgery, micro-CT revealed that a large amount of bone trabeculae were observed in both simple decalcified bone matrix and icariin/decalcified bone matrix groups. The number of bone trabeculae was higher, bone trabeculae were thicker, and defect cavity was smaller in the icariin/decalcified bone matrix group than those in the simple decalcified bone matrix group. (6) At 12 weeks after surgery, histological results revealed that a large number of bone tissue was formed in both simple decalcified bone matrix and icariin/decalcified bone matrix groups, but the amount of newly formed bone was higher in the latter group. (7) The compressive strength of bone defects in the icariin/decalcified bone matrix group was higher than that in the other two groups (P < 0.05). (8) These results suggest that 3D-printed icariin/decalcified bone matrix material exhibits good sustained-release performance, cytocompatibility, and osteogenesis. 

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    Icariin deproteinized inorganic bovine bone composite and deproteinized inorganic bovine bone material in repairing mandibular defects
    Dong Wenjie, Zhang Shiyang, Zhao Lei, Wang Yukun
    2020, 24 (34):  5467-5472.  doi: 10.3969/j.issn.2095-4344.2878
    Abstract ( 358 )   PDF (20957KB) ( 33 )   Save

    BACKGROUND: Deproteinized inorganic bovine bone is similar to human bone in structure. It has been widely used in oral bone regeneration surgery, but it lacks osteogenic induction ability. In recent years, it has been found that icariin has the effect on preventing and treating osteoporosis.

    OBJECTIVE: To observe the effect of icariin deproteinized inorganic bovine bone composite in the repair of mandibular defects.

    METHODS: MC3T3-E1 mouse osteoblasts were inoculated on icariin deproteinized inorganic bovine bone composite (observation group) and deproteinized inorganic bovine bone material (control group) respectively. After 7 days of culture, the survival of the cells on the surface of the material was observed by live-dead cell staining. The adhesion of the cells on the surface of the material was observed by scanning electron microscopy. After 5 and 10 days of culture, the alkaline phosphatase activity of the cells was detected. A 13 mm×6 mm×4 mm full-thickness defect of mandible was made in 30 New Zealand white rabbits. The experimental side (right) was implanted with icariin deproteinized inorganic bovine bone composite, and the control side (left) was implanted with deproteinized inorganic bovine bone material. The bilateral mandible tissues were obtained at 4, 8, and 12 weeks after the operation. The cone beam CT detection, histological observation and modified Gomori staining were performed respectively. The experiment was approved by the Ethics Committee of Experimental Animal Center of Mudanjiang Medical College.

    RESULTS AND CONCLUSION: (1) Live-dead cell staining demonstrated that the osteoblasts on the surface of the two groups grew well. (2) Scanning electron microscopy showed that osteoblasts adhered to the surfaces of the two materials. The cells on the surface of observation group had more adhesion and more uniform distribution than those in the control group. (3) The alkaline phosphatase activity of observation group cultured for 10 days was higher than that of control group (P < 0.05). (4) Cone-beam CT showed that the bone defect on the experimental side healed at 12 weeks after operation, while the bone defect on the control side could be seen at 12 weeks after operation. (5) Histological observation showed that at 12 weeks after operation, a large number of mature bone tissue could be seen in the defect area on the experimental side, and only a small amount of residual material and adipose tissue could be seen. On the control side, although the material was partially degraded and more new bone tissue could be seen, but the bone maturity was lower than that on the experimental side. (6) Modified Gomori staining showed that at 12 weeks after operation, a large number of new bone tissue with higher maturity could be seen on the experimental side, and more mature bone tissue could be seen on the control side, but the bone mass and maturity were not as good as those on the experimental side. (7) Results confirmed that compared with deproteinized inorganic bovine bone material, icariin deproteinized inorganic bovine bone composite can promote the repair of mandibular defects.

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    Injectable bone graft for the repair of nonunion of midshaft clavicular fracture after internal fixation
    Chen Shaobo, Shuang Feng, Hu Wei, Li Hao, Shao Yinchu
    2020, 24 (34):  5473-5477.  doi: 10.3969/j.issn.2095-4344.2356
    Abstract ( 550 )   PDF (16271KB) ( 415 )   Save

    BACKGROUND: Treatment of nonunion of clavicular fractures usually requires the insertion of bone material to facilitate bone healing. Injectable bone grafts have been successfully used in the treatment of limb fractures and lumbar fusion. However, there are few studies on its use in the treatment of nonunion.

    OBJECTIVE: To investigate the clinical effect of injection bone graft in the treatment of nonunion of clavicle fracture after internal fixation.

    METHODS: Totally 13 patients (9 males and 4 females aged 19-54 years old) with nonunion after internal fixation of clavicular fractures in the middle segment of the clavicle, who were treated in the No. 908 Hospital of Joint Logistic Support Force of Chinese PLA from January 2015 to December 2016, were selected. All of them were treated with ALLOMATRIX® injection bone graft. Postoperative follow-up was conducted for 12 months. The fracture healing quality was evaluated by Lane-Sandhu X-ray score, and the shoulder joint function was evaluated by Constant score. This experiment was approved by the Ethics Committee of No. 908 Hospital of Joint Logistic Support Force of Chinese PLA.

    RESULTS AND CONCLUSION: (1) All the 13 patients received osseous healing, which lasted 8-16 weeks, and no complications such as infection, dislocation, or fixator loosening occurred. (2) The total Lane-Sandhu X-ray score of 13 patients at 3 months and the last follow-up was higher than that before the operation (P < 0.05). (3) The total Constant score of shoulder joint function in 13 patients was higher than that before surgery at 3 months after surgery and at the last follow-up (P < 0.05). (4) The results showed that ALLOMATRIX® injection bone graft could be used to treat clavicular nonunion.

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    Clinical efficacy of calcium sulphate/demineralized bone matrix versus bone allograft in anterior cervical discectomy and fusion
    Li Yongtao, Song Wenhui, Liu Changwen, Liang Kaiheng, Zhang Kaixi, Liu Changfeng
    2020, 24 (34):  5478-5485.  doi: 10.3969/j.issn.2095-4344.2323
    Abstract ( 503 )   PDF (29923KB) ( 74 )   Save

    BACKGROUND: Different bone graft substitutes are widely used in clinical bone repair and tissue reconstruction. However, it remains unclear what bone graft substitute can acquire good bone fusion.

    OBJECTIVE: To investigate the clinical efficacy of different bone grafts filled in Zero-P cage for single-segment anterior cervical discectomy and fusion to treat cervical radiculopathy.

    METHODS: Seventy-seven patients with cervical radiculopathy who received treatment in the Second Hospital of Shanxi Medical University between January 2017 and January 2019 were included in this study. All included patients underwent single-segment anterior cervical discectomy and fusion and were divided into three groups according to bone graft materials: calcium sulphate/demineralized bone matrix group (n=27), allogeneic bone group (n=24), and autologous bone group (n=26). After surgery, 1-year follow up was performed. The Japanese Orthopedic Association (JOA) score, Visual Analog Scale (VAS) score, and Neck Disability Index (NDI) were evaluated. Changes in bone fusion, cervical curvature, and the height of intervertebral space were observed by imaging examination. This study was approved by the Hospital Ethics Committee, Second Hospital of Shanxi Medical University, China.   

    RESULTS AND CONCLUSION: (1) Compared with before surgery, JOA score was significantly increased, VAS score, and NDI were significantly decreased in each group at 1 year after surgery (P < 0.05). There were no significant differences in JOA score, VAS score, and NDI among three groups at 1 year after surgery (P > 0.05). (2) Compared with before surgery, cervical curvature and the height of intervertebral space were significantly increased in each group at 1 year after surgery (P < 0.05). There were no significant differences in cervical curvature and the height of intervertebral space between groups (P > 0.05). (3) At 6 months after surgery, bone fusion was observed in each group, and the degree of bone fusion in the calcium sulphate/demineralized bone matrix group was similar to that in the autologous bone group (P > 0.05), and the degree of bone fusion in the allogeneic bone group was lower than that in the autologous bone group (P < 0.05). (4) These results suggest that calcium sulphate/demineralized bone matrix for anterior cervical discectomy and fusion can acquire good bone fusion, and the clinical efficacy of calcium sulphate/demineralized bone matrix is similar to that of autologous bone and superior to that of allogenic bone. 

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    Long-term histological changes of diced cartilage grafts wrapped in advanced platelet-rich fibrin membrane
    Yang Yinhui, Yang Fan, Fan Xu, Xu Fengrui, Zheng Liang, Liu Wei, He Mingwu
    2020, 24 (34):  5486-5492.  doi: 10.3969/j.issn.2095-4344.2330
    Abstract ( 353 )   PDF (31143KB) ( 54 )   Save

    BACKGROUND: Advanced platelet-rich fibrin has been studied and prepared by scholars. Advanced platelet-rich fibrin has been proven to promote the regeneration and repair of autologous soft and hard tissues through animal experiments.

    OBJECTIVE: To prepare advanced platelet-rich fibrin membrane and observe the histological changes of the diced cartilage grafts wrapped in advanced platelet-rich fibrin membrane and autogenous fascia.

    METHODS: The following same procedures were carried out in 12 New Zealand rabbits: preparation of advanced platelet-rich fibrin membrane, collection of cartilage tissue of the left ear and the fascia tissue of the right hind limb of the rabbit. There were three types of cartilage grafts: diced cartilage; diced cartilage wrapped with autogenous fascia; diced cartilage grafts wrapped with advanced platelet-rich fibrin membrane. The grafts then were placed into subcutaneous pockets prepared on the backs of the same rabbits. The grafts were removed, and subjected to general observation and histopathologic examination 4 months after surgery.

    RESULTS AND CONCLUSION: (1) The thickness of diced cartilage grafts wrapped with autogenous fascia and advanced platelet-rich fibrin membrane was thicker than diced cartilage grafts; the shape of the grafts was more rounded; and the surface was flatter. (2) The chondrocyte survival rate and regeneration potential of diced cartilage grafts wrapped with advanced platelet-rich fibrin membrane were significantly better than the other two grafts (P < 0.05). There was no statistical difference in the other histological parameters among the three types of cartilage grafts. (3) The results show that advanced platelet-rich fibrin membrane can improve the survival rate of diced cartilage and graft shaping and reduce the resorption of diced cartilage after transplantation.

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    Potential of corn husk fibers reinforced chitosan-based hydrogels in cartilage tissue engineering scaffold
    Fang Yulu, Yi Bingcheng, Shen Yanbing, Tang Han, Zhang Yanzhong
    2020, 24 (34):  5493-5501.  doi: 10.3969/j.issn.2095-4344.2328
    Abstract ( 607 )   PDF (37378KB) ( 75 )   Save

    BACKGROUND: How to enhance the mechanical properties of the chitosan-based hydrogels has become a focus in the field of cartilage tissue engineering. In our previous research, chitosan was modified by citric acid, which led to significant improvement in the tensile and compression strength of the modified hydrogen in wet state.

    OBJECTIVE: To further enhance the mechanical performance of the citric acid modified chitosan hydrogel (i.e., CC) using the cellulose fibers derived from the natural corn husk fibers (CHFs), and to explore the potential of CHFs/CC composite hydrogel for cartilage tissue engineering scaffold.

    METHODS: A freeze-thawing method was employed to prepare CC and CHFs/CC (mass ratios of cellulose fiber to citric acid modified chitosan: 1:1 and 2:1). The structure and mechanical properties of the prepared hydrogels were characterized. Rabbit chondrocytes were seeded onto the hydrogel surface, and the cytocompatibility of the prepared hydrogels was analyzed by live/dead cell staining, CCK-8 assay, and scanning electron microscopy. The ability of CHFs/CC composite hydrogels in promoting functional expression of chondrocytes was determined by collagen immunofluorescence staining and quantitative analysis of glycosaminoglycans.

    RESULTS AND CONCLUSION: (1) With the increase in CHFs loading fractions, the porosity, equilibrium swelling rate, and degradation rate of CHFs/CC composite hydrogels were gradually reduced, but the compressive strength and Young’s modulus in dry and wet states were significantly enhanced. (2) Live/dead cell staining revealed that at 7 days of culture, chondrocytes proliferated well on the surface of three types of hydrogels. (3) The CCK-8 assay showed that with the increase in CHF loading fractions, the proliferative ability of chondrocytes on CC was gradually decreased. (4) Scanning electron microscope revealed that the chondrocytes on CC remained elliptic and were sparsely distributed, whereas most of the chondrocytes on CHFs/CC showed completely spread morphology. (5) After 28 days of culture, with the increase of CHFs loading fractions within CC, the chondrocytes showed gradually decreased expression of type I collagen, gradually increased expression of type II collagen, and there was no significant change in the content of glycosaminoglycans secreted. (6) These results demonstrate a great potential of the CHFs/CC composite hydrogels with good mechanical strength for cartilage tissue engineering scaffold.

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    Defatting effect of gradient alcohol treatment on cancellous bone allograft and the biomechanical properties of defatted cancellous bone allograft
    Hua Kunchi, Hu Yongcheng
    2020, 24 (34):  5502-5507.  doi: 10.3969/j.issn.2095-4344.2322
    Abstract ( 447 )   PDF (21952KB) ( 225 )   Save

    BACKGROUND: Defatting is considered to be the primary procedure for obtaining bone allograft. However, the current defatting method cannot avoid the problem of residual organic solvents.

    OBJECTIVE: To investigate the defatting effect of gradient alcohol treatment and whether this defatting method can reduce the mechanical properties of bone.  

    METHODS: Sixty pieces of fresh cancellous bone mass were obtained from human femoral condyles. They were divided into three groups according to different defatting treatments: gradient alcohol group, acetone group, and fresh group (control group). The general shape of the bone mass and the morphology and color of the cancellous bone allograft were observed with the naked eye. Soxhlet extraction was used to determine the residual lipid content of the bone mass after defatting. Infrared spectroscopy was used to observe the changes of various components in the bone mass after different defatting treatments. The maximum stress and elastic modulus were used to evaluate the effect of defatting treatment on the biomechanical properties of bone mass.

    RESULTS AND CONCLUSION: (1) Gross observation revealed that fresh bone mass was yellow with a large amount of fat in the pores, and the defated bone mass was white with a clear porous network structure. (2) There was no statistical difference in residual lipid content between the gradient alcohol and the acetone groups (P=0.385). (3) Infrared spectroscopy showed that the defatting effects of the two defatting treatments were similar and had no effect on other components in the bone, such as PO43-, carbonate, and hydroxyapatite. (4) Biomechanical tests showed that there were no significant differences in the maximum stress and elastic modulus between groups. (5) These results suggest that the defatting effect of the gradient alcohol treatment is similar to that of the traditional acetone solvent extraction, and gradient alcohol treatment cannot reduce the biomechanical properties of bone

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    Effect of different retentive grooves and adhesives on the retentive force between crown and post and core
    Li Zheng, Wang Yuan, Tian Mengting, He Huiyu
    2020, 24 (34):  5508-5513.  doi: 10.3969/j.issn.2095-4344.2319
    Abstract ( 495 )   PDF (25261KB) ( 256 )   Save

    BACKGROUND: Post and core crowns are used in the clinic under some conditions, including root fractures, broken posts, and post-core detachment caused by large-scale dental caries and defects. Dentists mainly increase the retentive force by selecting a bonding material with stronger adhesion or by making a retentive groove parallel to the long axis on the surface of the core, but the effect is not obvious.

    OBJECTIVE: To investigate the effect of different types of retentive grooves and two different adhesives on the retentive force after the post-core crown restoration.

    METHODS: Sixty cobalt-chromium alloy metal crowns and post-core modules simulating the morphology of mandibular premolars were prepared. The axial cohesion angle of the cores was 8°. These crowns and post-core modules were randomly divided into six groups with 10 crowns/pieces per group. In the smooth group, there was no auxiliary retentive groove on the post-core surface. In the horizontal and vertical groups, there were retentive grooves with equal spacing parallel to (horizontal group) or perpendicular to (vertical group) the long axis of the post and core. In the oblique group, the retentive grooves were placed with an oblique angle of 45° with the long axis of the post-core. In the mesh group, the retentive grooves were at an oblique angle of 45° with the long axis of the post and core and crossed. In the point group, the retentive grooves were dot-shaped. In all six groups, glass ionomer cement and self-curing resin adhesive were used to bond post-and-core with crown. The retentive force in each group was tested by the tensile test, and the displacement change of three-dimensional finite element model during the same time was analyzed by the finite element analysis.

    RESULTS AND CONCLUSION: (1) Regardless of whether it was bonded with glass ionomer cement or self-curing resin adhesive, the retentive force was highest in the mesh group and lowest in the point group. There was no interaction between adhesives and retentive grooves used to increase retentive force (F=0.26, P=0.91). The effect of different adhesives on retentive force was not significant (F=1.04, P=0.31). The effect of different auxiliary retentive grooves on retentive force was significant (F=14.74, P < 0.05). (2) When the same force was applied at the same time, the displacement of three-dimensional finite element model was smallest in the mesh group. (3) These results suggest that preparation of retentive grooves on the post-core surface can enhance the retentive force. Mesh-shaped retentive grooves provide the greatest retentive force. Different types of adhesives do not provide obviously different effect on the retentive force between the crown and the post and core.

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    Short-term effect comparison of a modified socket shield technique and conventional flapless immediate implant and immediate restoration in maxillary aesthetic area
    Zhang Bin, Sun Lihua, Zhang Junhua, Liu Tongbin, Liu Yusan, Cui Caiyun, Li Jun
    2020, 24 (34):  5514-5519.  doi: 10.3969/j.issn.2095-4344.2344
    Abstract ( 473 )   PDF (23028KB) ( 263 )   Save

    BACKGROUND: The conventional shield technique can reduce the absorption of labial fasciculate bone after immediate implantation of anterior teeth, but it is highly demanded by the operator. When implants are implanted, it is easy to loosen the remaining teeth on the labial side, resulting in failure of the operation.

    OBJECTIVE: To evaluate the clinical effects of a modified socket shield technique for immediate implant and immediate restoration and flapless immediate implant and immediate restoration in maxillary aesthetic area.

    METHODS: Sixty patients were randomly chosen with failing tooth from September 2016 to August 2017 in Department of Stomatology in Affiliated Hospital of Binzhou Medical College. The samples included 33 males and 27 females, aged from 20 to 81 years. There were 38 cases of central incisor and 22 cases of lateral incisor. The patients were randomly divided into two groups. The 30 patients in the experimental group received a modified socket shield technique for immediate implant and immediate restoration. The 30 patients in the control group received conventional flapless immediate implant and immediate restoration. During return visit after 1 year, the retention rate of the implant and pink esthetic score were recorded; the buccal bone thicknesses of the implant neck and roots 2, 4 mm were observed by cone beam computed tomography; and the patient satisfaction was analyzed. The study was approved by the Medical Ethics Committee of the Affiliated Hospital of Binzhou Medical College (Hospital Ethics Approval Number: 2019-LW-025).

    RESULTS AND CONCLUSION: (1) The survival rate of both groups was 100%. (2) One year after operation, the pink esthetic score of the experimental group was higher than the preoperative score and that in the control group (P < 0.05). The pink esthetic score of the control group was lower 1 year after surgery than preoperative score (P < 0.05). (3) One year after operation, the buccal bone thickness loss of the implant neck and roots 2, 4 mm in the experimental group was lower than that in the control group (P < 0.05). (4) The satisfaction rate of the experimental group was higher than that in the control group (P < 0.05). (5) The results showed that the modified socket shield technique used for immediate implant and immediate restoration can maintain the buccal bone thickness and improve the aesthetic effect.

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    Granule cartilage plus n-butyl α-cyanoacrylate glue combined with 3D printing technology for preparation of individualized ear scaffolds
    Zhang Fan, Li Gaofeng, Hu Yigao, Cai Lihong
    2020, 24 (34):  5520-5525.  doi: 10.3969/j.issn.2095-4344.2321
    Abstract ( 301 )   PDF (24382KB) ( 50 )   Save

    BACKGROUND: The manufacture of ear brackets in traditional ear reconstruction is likely to cause ear distortion, and 3D printing technology makes it possible to prepare individualized and highly simulated ears.

    OBJECTIVE: To explore the feasibility of making individualized ear scaffolds using granule cartilage plus n-butyl α-cyanoacrylate glue with 3D printing technology.

    METHODS: Male and female modes of 3D ear scaffolds were prepared for the affected ears according to the ears on the non-injured side. Granule cartilage and n-butyl α-cyanoacrylate glue were placed in the female modes to prepare ear cartilage scaffolds through slight adjustment based on male modes. Then the ear cartilage scaffolds were implanted into the back of New Zealand rabbits. The morphology of constructed ear scaffolds was dynamically observed. At postoperative 4, 8, and 12 weeks, the ear scaffolds were taken out in batches. The connection between granule cartilage and cartilage survival was histologically observed. Immunohistochemistry was performed to detect the expression of type II collagen. The animal experiments were approved by the Animal Ethics Committee of Hunan Normal University, China (approval No. 201517).  

    RESULTS and CONCLUSION: (1) Gross observation: compared with before surgery, the length, width and height of the reconstructed ears were not significantly changed, but the weight of the reconstructed ears was significantly increased at 4, 8, and 12 weeks after surgery (P < 0.05). (2) Histological observation: Inflammatory cell infiltration was observed around granular cartilage. With the prolongation of ear scaffold implantation time, inflammatory cell count gradually decreased, and there were more newly formed vessels in the fascia attached to the scaffold. With the prolongation of implantation time, the number of new chondrocytes in the cartilage membrane gradually increased, cartilage volume gradually increased, and mature chondrocytes gradually increased. Obvious decomposition of n-butyl α-cyanoacrylate between cartilage pieces was not observed. Most of the cartilage pieces were still separated by n-butyl α-cyanoacrylate, and a few of cartilage pieces had fiber connections. With the prolongation of implantation time, the connections between cartilage pieces increased. (3) Immunohistochemistry observation: With the prolongation of ear scaffold implantation time, the expression of type II collagen gradually increased. (4) These results suggest that granule cartilage plus n-butyl α-cyanoacrylate glue combined with 3D printing technology make preparation of individualized ear scaffolds feasible. 

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    Canine dental pulp stem cells-polyglycolic acid scaffold complex for canine periodontal tissue defect
    Zhou Pengfei, Lin Jing, Chen Yuying, Lin Minkui
    2020, 24 (34):  5526-5531.  doi: 10.3969/j.issn.2095-4344.2350
    Abstract ( 417 )   PDF (23224KB) ( 40 )   Save

    BACKGROUND: Our previous studies have constructed polyglycolic acid-human dental pulp stem cells complex, and it is shown that mechanical stimulation of the complex can be effective in the construction of tissue-engineered tendons or ligaments.

    OBJECTIVE: We assume that tissue-engineered tendons or ligaments constructed by polyglycolic acid and canine dental pulp stem cells may promote the healing of periodontal tissue defect, so we verify the promoting effect of the complex for repairing canine one-wall periodontal defects.

    METHODS: One-wall acute periodontal defect models of six Beagle dogs were prepared. The models were randomly assigned to experimental group (canine dental pulp stem cells-polyglycolic acid complex) and control group (without any implanting). Periodontal regeneration was observed by hematoxylin-eosin staining and AZAN staining at 12 weeks after implantation. The animal experiment was approved by the experimental animal welfare and ethics committee of Fuzhou General Hospital of Nanjing Military Area Command.

    RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining: There were numerous new bones at the site of the periodontal defect; there were still differences between the new bone structure in the distal part of the root surface defect and the normal bone structure; the fibrous connective tissue above was thick in the experimental group. There was only a small amount of new bone at the site of the defect, and the gingiva and the root surface did not combine to form long junctional epithelium structure in the control group. (2) AZAN staining: Compact new bone formation, periodontal membrane regeneration and thin cementum formation could be seen at the site of the periodontal defect; new periodontal membrane was close to the normal periodontal membrane tissue in the experimental group. There was only a small amount of new bone formation at the site of the defect, almost no new periodontal membrane, and the upper part was gingival connective tissue, and the gingiva and the root surface did not combine to form long junctional epithelium structure in the control group. (3) Histological quantitative analysis results showed that the length of junctional epithelium, new cementum, new periodontal membrane, new bone height, and new bone area were more in the experimental group than in the control group (P < 0.05). (4) These results confirm that canine dental pulp stem cells combined with polyglycolic acid contribute to periodontal regeneration.

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    Porous beta-tricalcium phosphate-polypyrrole-biotin-icariin composite scaffold promotes recruitment of bone marrow mesenchymal stem cells
    Liu Xin, Du Bin, Sun Guangquan, Cao Jinxing, Jiang Xiaohong
    2020, 24 (34):  5532-5537.  doi: 10.3969/j.issn.2095-4344.2347
    Abstract ( 358 )   PDF (25521KB) ( 413 )   Save

    BACKGROUND: Icaritin, as an osteogenic inducible active substance, has been widely loaded in orthopedic scaffold materials. However, in most of the existing studies, it is mainly placed in the pores of the scaffold directly in the form of polymerized microspheres, which makes it difficult to release icaritin in step with scaffold degradation, and the local effective utilization rate is low.

    OBJECTIVE: To prepare porous β-tricalcium phosphate-polypyrrole-biotin-icariin microspheres composite scaffolds, and preliminarily explore the biological characteristics of coculture of the scaffolds and bone marrow mesenchymal stem cells.

    METHODS: FeCl3 was used as oxidant, and polypyrrole was synthesized by oxidative chemical synthesis. Polypyrrole, biotin and β-tricalcium phosphate were mixed and electrochemical synthesis was carried out. After that, the porous β-tricalcium phosphate-polypyrrole-biotin composite scaffold was prepared by three-dimensional printing technology. Icariin-biotin-polylactic acid microspheres were prepared by HDDD reaction and loaded on the scaffold above. The ordinary porous β-tricalcium phosphate scaffold prepared by three-dimensional printing was set as the control group and loaded with icaritin-polylactic acid microspheres. The compressive strength, porosity, drug loading, drug binding force and drug sustained release performance of the two groups of scaffolds were detected and compared, and the curve of icaritin release was drawn. The biological effects of the scaffolds on bone marrow mesenchymal stem cells were observed by scanning electron microscopy.

    RESULTS AND CONCLUSION: The drug loading capacity, drug binding force and drug sustained release performance of the porous β-tricalcium phosphate-polypyrrole-biotin composite scaffold were significantly better than those in the control group (P < 0.05). The number of bone marrow mesenchymal stem cells growing on the surface of the scaffold under scanning electron microscopy was significantly higher than that of the control group (P < 0.05). There was no significant difference in the compressive strength and porosity of the scaffolds between the two groups (P > 0.05). The results confirmed that compared with the traditional sustained-release scaffolds, the porous β-tricalcium phosphate-polypyrrole-biotin composite scaffold further improves the drug loading capacity and sustained-release performance, and has good mechanical strength. Simultaneously, it may have a better role in recruiting bone marrow mesenchymal stem cells to participate in the bone repair around the scaffolds.

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    Collagen/heparin sulfate scaffolds loaded with brain-derived neurotrophic factor promote neurological and locomotor function recovery in rats after traumatic brain injury
    Zhang Jian, Chen Miao, Li Weixin, Ye Yichao, Xu Huiyou, Ma Ke, Chen Xuyi, Sun Hongtao, Zhang Sai
    2020, 24 (34):  5538-5544.  doi: 10.3969/j.issn.2095-4344.2316
    Abstract ( 354 )   PDF (31479KB) ( 36 )   Save

    BACKGROUND: The porous scaffold fabricated with collagen and heparin sulfate can effectively immobilize neurotrophic factor and greatly improve neurological and locomotor function recovery.  

    OBJECTIVE: To investigate the effect of collagen/heparan sulfate scaffold loaded with brain-derived neurotrophic factor in the treatment of traumatic brain injury (TBI).

    METHODS: Collagen scaffolds, collagen/heparan sulfate scaffolds and collagen/heparan sulfate scaffolds loaded with brain-derived neurotrophic factor were prepared. The porosity, water absorption, compression modulus and compression stress of collagen scaffolds and collagen/heparan sulfate scaffolds were measured. The in vitro sustained-release performance of collagen/heparan sulfate scaffolds loaded with brain-derived neurotrophic factor was measured. The cytotoxicity and cytocompatibility of collagen/heparin sulfate scaffolds and collagen/heparin sulfate scaffolds loaded with brain-derived neurotrophic factor were detected. Forty-eight Sprague-Dawley rats were randomly assigned to undergo corresponding interventions: opening bone window followed by suture (control group), induction of craniocerebral trauma (model group), induction of craniocerebral trauma + implantation of collagen/heparin sulfate scaffolds (scaffold group), and induction of craniocerebral trauma + implantation of collagen/heparin sulfate scaffolds loaded with brain-derived neurotrophic factor (brain-derived neurotrophic factor group). After surgery, modified neurological severity scoring and Morris water maze test were performed to evaluate the recovery of neurological and locomotor function. Rat cerebral morphology was performed. This study was approved by the Animal Ethics Committee of Characteristic Medical Center of Chinese People's Armed Police Force, China.

    RESULTS AND CONCLUSION: (1) The porosity, compression modulus and compression stress of the collagen/heparin sulfate scaffold were significantly higher than those of the collagen scaffold (P < 0.05), and the water absorption rate of the collagen/heparin sulfate scaffold was lower than that of the collagen scaffold (P < 0.05). (2) Collagen/heparin sulfate scaffolds had no cytotoxicity, and collagen/heparin sulfate scaffolds loaded with brain-derived neurotrophic factor were more conducive to the proliferation of cerebral microvascular endothelial cells; cerebral microvascular endothelial cells grew well and distributed evenly in the micropores of the collagen/heparin sulfate scaffolds loaded with brain-derived neurotrophic factor. (3) There were a large number of newly formed nerve cells and nerve fibers in the lesions in the scaffold and brain-derived neurotrophic factor groups. The number of nerve cells and the density of nerve fibers in the scaffold group were higher compared with the brain-derived neurotrophic factor group. (4) In the brain-derived neurotrophic factor group, escape latency was shorter (P < 0.01, P < 0.05), quadrant stay time and the number of crossing the platform were higher (P < 0.01, P < 0.05) compared with the scaffold and model groups. (5) The modified neurological severity score in the brain-derived neurotrophic factor group was significantly lower than that in the scaffold and model groups, respectively at postoperative 3-7 weeks (P < 0.05, P < 0.01). (6) These results suggest that collagen/heparin sulfate scaffolds loaded with brain-derived neurotrophic factor can promote neurological and locomotor function recovery in rats after traumatic brain injury. 

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    Comparison of 3Dmax light-weight hernia mesh and Prolene weight mesh for the repair of inguinal hernia
    Fan Zhongbao, Shen Jianfen, Wang Jian, Yu Hao, Liu Yanfeng
    2020, 24 (34):  5545-5551.  doi: 10.3969/j.issn.2095-4344.2851
    Abstract ( 592 )   PDF (26831KB) ( 242 )   Save

    BACKGROUND: Laparoscopic total extraperitoneal anterior peritoneal repair of inguinal hernia has a good clinical effect. However, it should be further discussed how to choose the right mesh in surgery can accelerate the recovery time and improve the postoperative recovery quality of patients.

    OBJECTIVE: To compare the effect of 3Dmax light-weight hernia mesh and Prolene weight mesh on the repair of inguinal hernia.

    METHODS: A total of 110 patients with inguinal hernia treated in the People’s Hospital of Liaoning Province from January 2016 to December 2018 were selected, including 97 males and 13 females, at the age of 18-70 years old. All the patients underwent laparoscopic total extraperitoneal anterior peritoneal hernia repair. 3Dmax light-weight hernia meshes were used in 67 patients and Prolene heavy mesh were used in 43 patients. Postoperative complications were recorded. The patients were postoperatively followed up for 12 months. Visual analogue scale score and SF-36 quality of life score were compared between the two groups. The experiment was approved by the Ethics Committee of the People’s Hospital of Liaoning Province.

    RESULTS AND CONCLUSION: (1) The number of cases of non-infectious fever in the 3Dmax mesh group was less than that in the Prolene mesh group (P < 0.05). There was no statistically difference between the two groups in incision infection, urinary retention, and hematoma (P > 0.05). (2) The visual analogue scale score of the 3Dmax mesh group was lower than that of the Prolene mesh group at 3, 6 and 12 months after surgery (P < 0.05). (3) The SF-36 quality of life score 7 days after surgery showed that the health status, living ability and social function scores and total scores of the 3Dmax mesh group were higher than those of the Prolene mesh group (P < 0.05). (4) The SF-36 quality of life score at 3, 6 and 12 months after surgery showed that the 3Dmax mesh group had less restrictions on daily activities such as walking, lifting heavy objects and physical activities than those in the Prolene mesh group (P < 0.05). (5) The results show that compared to the Prolene weight mesh, the use of 3Dmax light-weight hernia mesh for repair of laparoscopic total extraperitoneal anterior peritoneal hernia can reduce adverse reactions, reduce the degree of pain, accelerate the recovery time of patients, and improve the quality of postoperative recovery. 

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    Preparation of injectable tantalum oxide-loaded brushite bone cement and investigation on its radiocapacity performance
    Xing Yuying, He Yuying, Jiang Ningning, Zhu Ruijie, He Ran, Qi Yangkun, Wang Kejing, Li Guangda
    2020, 24 (34):  5552-5558.  doi: 10.3969/j.issn.2095-4344.2318
    Abstract ( 376 )   PDF (25304KB) ( 42 )   Save

    BACKGROUND: Brushite bone cement has similar chemical composition to human bone tissue, and thus it is difficult to differentiate the cement and the bone on X-ray images due to their similar radiocapicity, limiting the clinical application of the material. However, as far as we know, few researches focus on the improvement of the radiocapacity of brushite bone cement.

    OBJECTIVE: To prepare injectable tantalum oxide-loaded brushite bone cement and investigate the effects of tantalum on injectability, microstructure, radiopacity, compressive strength, and cyto-biocompatibility of tantalum-loaded brushite bone cement.

    METHODS: The powder of brushite bone cement was prepared by mixing β-calcium phosphate and dicalcium phosphate at the molar ratio of 1:1. Then 0 wt%, 1 wt%, 3 wt%, and 5 wt% tantalum oxide was added to the powder of brushite bone cement. Solid phase bone cement was mixed with chitosan-containing citric acid solidified solution to prepare injectable brushite bone cement loaded with different amounts of tantalum oxide. The solidification time, injectability, micromorphology, crystal phase composition, chemical group, radiopacity, compressive strength, biodegradability and cytocompatibility of the tantalum-loaded brushite bone cement were tested.

    RESULTS AND CONCLUSION: (1) With the increase of the proportion of tantalum oxide, the solidification time of bone cement gradually prolonged and the bone cement maintained good anti-washout ability and injectability. (2) The addition of tantalum oxide did not greatly change the crystal structure and group composition of brushite bone cement, but obviously changed the surface appearance of the material. (3) The addition of tantalum oxide increased the compressive strength of brushite bone cement, but decreased its compressive strength. (4) With the increase of the proportion of tantalum oxide added, the radiopacity of brushite bone cement was gradually enhanced. (5) The injectable brushite bone cement loaded with different amounts of tantalum oxide had no cytotoxicity, and MG63 cells normally adhered to and proliferated on its surface. (6) These findings suggest that injectable tantalum oxide-loaded brushite bone cement has good injectability, radiopacity, compressive strength, biodegradability, and cytocompatibility.   

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    Combination of hyaluronic acid hydrogel and bone marrow mesenchymal stem cells promotes cardiac function after myocardial infarction in rats (II)
    Shang Qingqing, Zhou Jianye
    2020, 24 (34):  5559-5563.  doi: 10.3969/j.issn.2095-4344.2106
    Abstract ( 499 )   PDF (24815KB) ( 280 )   Save

    BACKGROUND: Our previous experimental results have shown that hyaluronic acid (HA) hydrogel can act as a vehicle for bone marrow mesenchymal stem cell (BMSC) delivery to improve the cardiac function of rats with myocardial infarction.

    OBJECTIVE: To explore the effects of BMSCs encapsulated in HA hydrogel on histological changes of the rat myocardium after myocardial infarction.

    METHODS: BMSCs from male Sprague-Dawley rats were isolated and cultured, and then HA-encapsulated BMSCs were cultured in vitro in the three-dimensional manner. A model of myocardial infarction was made by ligating the left anterior descending artery of female Sprague-Dawley rats. After 1 week, the model rats were screened by ultrasonic testing and then eligible ones were randomly divided into four groups: PBS group (n=8), HA group (n=8), BMSCs group (n=29), and HA-encapsulated BMSCs group (n=29). At 1 week after ligation, the model rats underwent the secondary thoracotomy followed by corresponding injections into the infarcted region and its marginal zone. At 4 week after injection, cardiovascular regeneration, myocardial protection, and ventricular remodeling were evaluated using hematoxylin-eosin staining, Masson’s staining and immunohistochemical staining, respectively.

    RESULTS AND CONCLUSION: At 4 weeks after transplantation, compared with the PBS group, the infarcted ventricular wall was thickened in the HA group, cardiomyocyte hypertrophy was relieved and neovascularization was increased in the BMSCs group, and the largest benefits were indicated in the HA-encapsulated BMSCs group by reversing cardiac remodeling, increasing neovascularization and viable myocardium tissues in the infarct zone. Therefore, the HA-encapsulated BMSCs can achieve the best effect in the treatment of myocardial infarction.

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    Immunogenicity of heterogeneous acellular biosurgical patch
    Cheng Xiang, Guo Huan, Liu Yanping, Wei Xiaohui, Lei Dan, Wu Tao, Huang Ting, Huang Ming, Rong Zuyuan, Ling You
    2020, 24 (34):  5564-5569.  doi: 10.3969/j.issn.2095-4344.2343
    Abstract ( 429 )   PDF (23752KB) ( 35 )   Save

    BACKGROUND: Since the removal of immunogenicity of heterogeneous acellular patch is directly related to the success of the implantation into human body, it is vital to evaluate the immunogenicity of the material.

    OBJECTIVE: To evaluate the immunogenicity of heterogeneous acellular surgical path.

    METHODS: Twenty Balb/c mice were randomly divided into four groups (n=5 per group). Mice in the experimental group and control group were subcutaneously implanted with heterogeneous acellular biosurgical patch and pericardium on the back. Negative controls received sham operation. Mice in the positive control group received subcutaneous injection of the mixture of Freund’s adjuvant and bovine serum albumin in equal volumes. At 4 weeks after implantation, the body weight of mice was recorded; the coefficients of spleen and thymus were calculated; the level of total serum IgG and IgM, the proliferation activity of lymphocytes in vitro and the distribution of lymphocyte subtypes in the spleen were measured; and the histopathology of the skin, spleen and thymus was observed. The experiment was approved by the Laboratory Animal Management and Use Committee of Safety Evaluation Center, Sichuan Institute for Food and Drug Control (approval No. IACUC-2018-KYYL-008).  

    RESULTS and CONCLUSION: (1) There were no significant changes in the mouse body weight between the experimental group and negative control group (P > 0.05). Body weight was higher in the control group than in the negative control group (P < 0.05). (2) There was no significant difference in the brain ratios of spleen and thymus between the experimental group and the negative control group (P > 0.05). The brain ratio of spleen was higher in the control group than in the negative control group (P < 0.05). (3) There was no significant difference in lymphocyte proliferation between the experimental group and the negative control group (P > 0.05). Lymphocyte proliferation was higher in the control group than in the negative control group (P < 0.05). (4) Compared with the negative control group, the percentage of CD3+CD8+ cells was significantly decreased in the experimental group (P < 0.05). Compared with the negative control group, the percentages of CD3+ cells, CD3+CD4+ cells, CD3+CD8+ cells, and CD45+SSClow cells were decreased (P < 0.05) and the percentage of CD3-CD19+ cells was increased (P < 0.05) in the control group. (5) There was no significant difference in serum IgM and IgG antibody levels between the experimental and control groups and the negative control group (P < 0.05). (6) The histological examination results showed that there were no obvious pathological changes in the spleen and thymus in the experimental group and the control group, and no obvious inflammatory reaction in the experimental group. In the control group, there were severe granulomatous inflammation, fibrous tissue hyperplasia, wrapping, necrosis and disintegration of implants. (7) The results demonstrated that compared with the raw material, the immunogenicity of acellular treated biosurgical patch could be reduced effectively.

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    Human acellular amniotic membrane and acellular subamniotic membrane in repairing skin defects
    Wang Dan, Yin Jing, Wang Yuying, Du Yong, Li Jingling, Yu Shun
    2020, 24 (34):  5570-5576.  doi: 10.3969/j.issn.2095-4344.2355
    Abstract ( 423 )   PDF (27671KB) ( 218 )   Save

    BACKGROUND: Amniotic membrane and subamniotic membrane are homologous and belong to medical waste. They have low immunogenicity and certain toughness. Amniotic membrane as tissue-engineered scaffold material has been reported more, but the research of subamniotic membrane as tissue-engineered scaffold is less.

    OBJCETIVE: To compare the difference between acellular amniotic membrane and acellular subamniotic membrane in combination with bone marrow mesenchymal stem cells for skin repair.

    METHODS: The acellular subamniotic membrane materials were prepared by sodium dodecyl sulfate + nuclease method, and the acellular amniotic membrane materials were prepared by Triton X-100 + pancreatin method. Bone marrow mesenchymal stem cells were cultured with two kinds of acellular materials. The cytotoxicity of the two kinds of acellular materials was detected by CCK-8 method. Bone marrow mesenchymal stem cells were seeded on the surfaces of two kinds of acellular materials. The growth and adhesion of cells were observed under light microscope and scanning electron microscope. Skin defects deep into the dermis were made on the back of one spine of each Sprague-Dawley rat, and 36 rats were randomly divided into three groups (n=12). No materials were implanted in the blank group. Acellular amniotic membrane and bone marrow mesenchymal stem cell complex were implanted in the control group. Acellular subamniotic membrane and bone marrow mesenchymal stem cell complex were implanted in the observation group. Wound healing rate, wound pathology, wound gene and protein were detected at 7, 14 and 21 days after operation, respectively. This study was approved by the Animal Ethics Committee of Jiangnan University.  

    RESULTS AND CONCLUSION: (1) In the 3-7 days of culture, two kinds of acellular materials could promote the proliferation of bone marrow mesenchymal stem cells. (2) The results of light microscope and scanning electron microscope showed that bone marrow mesenchymal stem cells grew well on the surfaces of two kinds of acellular materials, and the morphology of bone marrow mesenchymal stem cells was the same as that of cultured in culture bottle. (3) The wound healing rate of the observation and control groups was higher than that of the blank group at various time points after surgery (P < 0.05); and that in the observation group was higher than that in the control group (P < 0.05). (4) At 14 days after surgery, pathological observation showed that the skin repair effect of the observation group was better than that of the blank group, and the skin repair of the observation group was better than that in the control group. (5) The expression of type III collagen, CK18, type I collagen and vascular endothelial growth factor gene in the observation and control groups was higher than that in the blank group at various time points after surgery (P < 0.05); and the expression was higher in the observation group than that in the control group (P < 0.05). (6) The expression of CK18 and vascular endothelial growth factor protein in the observation and control groups was higher than that in the blank group at various time points (P < 0.05); and the expression was higher in the observation group than that in the control group (P < 0.05). (7) Compared with acellular amniotic membrane, the lower layer of acellular subamniotic membrane can promote skin repair.

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