Publications by authors named "Haohao Ren"

12 Publications

  • Page 1 of 1

Effects of bovine cancellous bone powder/poly amino acid composites on cellular behaviors and osteogenic performances.

Biomed Mater 2021 Jul 6;16(5). Epub 2021 Jul 6.

School of Automation and Information Engineering, Sichuan University of Science and Engineering, Zigong, People's Republic of China.

Xenogeneic bone has good biological activity, but eliminating immunogenicity, while retaining osteogenic abilities, is a challenge. By combining xenogeneic bone with poly amino acid (PAA) that has an amide bond structure, a new type of composite conforming to bionics and low immunogenicity may be obtained. In this study, according to the principles of component bionics, three composites of delipidized cancellous bone powder (DCBP) and PAA were designed and obtained by anpolycondensation method, an extrusion molding (EM) method, and a solution-blend method. The three composites were all macroscopically uniform, non-cytotoxic, and demonstrated low immunogenicity by effective removal of residual antigens during preparation. Compared with PAA, mouse bone marrow mesenchymal stem cells (BMSCs) on the surfaces of three composites showed different cellular morphologies. The effects of different preparation methods and cellular morphology on cellular differentiation were confirmed by alkaline phosphatase activity, calcium nodule formation and the expression levels of osteogenic differentiation-related genes (bone morphogenetic protein 2, runt-related transcription factor 2, osteopontin and osteocalcin). Among these composites, DCBP/PAA EM showed best cell proliferation and osteogenic differentiation, and possessed greater bone formation than PAA in a rabbit femoral condyle study. This study may provide a new method for preparing bioactive bone repair materials with low immunogenicity and superior ability to stimulate differentiation of BMSCsand osteogenesis. DCBP/PAA EM might be a promising bone repair material for bone defect treatment.
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http://dx.doi.org/10.1088/1748-605X/ac0d94DOI Listing
July 2021

Effects of pullulan on the biomechanical and anti-collapse properties of dicalcium phosphate dihydrate bone cement.

J Biomater Appl 2021 Jun 1:8853282211020158. Epub 2021 Jun 1.

School of Chemical Engineering, Sichuan University, Chengdu, China.

In this work, a modified dicalcium phosphate dihydrate (DCPD) bone cement with unique biodegradable ability in a calcium phosphate cement system was prepared by the hydration reaction of monocalcium phosphate monohydrate and calcium oxide and integration with pullulan (Pul), a non-toxic, biocompatible, viscous, and water-soluble polysaccharide that has been successfully used to improve defects in DCPD bone cement, especially its rapid solidification, fragile mechanical properties, and easy collapse. The effect of different contents of Pul on the structure and properties of DCPD were also studied in detail. The modified cement was characterised by X-ray diffraction, Fourier-transform infrared spectroscopy, scanning electron microscopy, ultraviolet-visible absorption, X-ray photoelectron spectroscopy analysis, and rheological property measurements. The results indicated that Pul promoted the hydration formation of DCPD, and interface bonding occurred between Pul and DCPD. With increasing content of Pul, the setting time of the DCPD bone cement increased from 2.6 min to 42.3 min, the compressive strength increased from 0 MPa to 20.4 MPa, and the anti-collapse ability also improved owing to the strong interface bonding, implying that the DCPD bone cement improved by Pul has better potential for application in the field of non-loading bone regenerative medicine compared to unmodified DCPD bone cement.
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http://dx.doi.org/10.1177/08853282211020158DOI Listing
June 2021

Developing a novel magnesium calcium phosphate/sodium alginate composite cement with high strength and proper self-setting time for bone repair.

J Biomater Appl 2021 Aug 30;36(2):346-357. Epub 2021 May 30.

College of Physics, Sichuan University, Chengdu, China.

In this work, novel magnesium calcium phosphate/sodium alginate composite cements were successfully fabricated with a proper setting time (5-24 min) and high compressive strength (91.1 MPa). The physicochemical and biological properties of the cement in vitro were fully characterized. The composite cements could gradually degrade in PBS as the soaking time increase, and the weight loss reached 20.74% by the end of 56th day. The cements could induce the deposition of Ca-P layer in SBF. Cell experiments proved that the extracts of the composite cements can effectively promote the proliferation and differentiation of the mouse bone marrow mesenchymal stem cells (MSCs). These preliminary results indicate that the magnesium calcium phosphate/sodium alginate composite cements could be promising as potential bone repair candidate materials.
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http://dx.doi.org/10.1177/08853282211021535DOI Listing
August 2021

First report of Heterodera filipjevi on winter wheat from Hebei Province in North China.

Plant Dis 2021 Jan 28. Epub 2021 Jan 28.

Henan Agricultural University, 70573, College of Plant Protection, Zhengzhou, China;

Three of the cereal cyst nematodes, Heterodera avenae, H. filipjevi and H. latipons are considered to be the most economically important cyst nematodes that affect cultivated cereals around the world. H. filipjevi was first detected in China from Xuchang, Henan Province in 2010 (Peng et al. 2010) and now has been recorded in the Central China of Henan, Shandong and Anhui provinces and the Northwest China of Xinjiang Uygur Autonomous Region (Cui et al. 2020). In June 2019, 42 samples consisting of roots and soil were collected from winter wheat fields in Hebei Province of North China. Cysts were detected in 37 soil samples with a mean of 6.4 ± 1.67 cysts per 100 ml of soil. Cysts and second-stage juveniles (J2s) were extracted from root and soil following Cobb's sieving gravity method. Morphological and molecular studies of J2s and cysts confirmed its identity with H. filipjevi in 5 samples from Handan (N36°10'052" and E114°35'056"; N36°37'054" and E114°22'052"), Xingtai (N36°53'060" and E114°30'011") and Shijiazhuang (N 37°26'048" and E 116°05'039") in Hebei Province, China. Morphologically, the cysts are lemon-shaped, light or dark brown in color. The vulval cone is bifenestrate with horseshoe-shaped semifenestrae, strongly globular bullae, and well-developed underbridge. Measurements (mean +_ sd (range)) of cysts (n=10), body length not including neck is 743.0 ± 36.1 μm (665 - 780 μm), body width is 559.0 ± 50.0 μm (455 - 639 μm), length / width ratio is 1.33 ± 0.07 (1.20 - 1.46); neck length is 99.3 ± 8.8 μm (85 - 122 μm); fenestrae length is 56.8 ± 5.0 μm (49 - 65 μm) and width is 25.5 ± 1.8 μm (21.1 - 27.8 μm); underbridge length is 84.0 ± 8.1 μm (62 - 93 μm); and vulval slit length is 8.6 ± 0.5 μm (7.2 - 9.1 μm). Measurements of J2s (n = 12), body length is 541 ± 11.4 μm (490 - 578 μm); stylet length is 22.3 ± 0.5 μm (22.0 - 25.0 μm) with anchor-shaped basal knobs; tail length is 57.7 ± 3.7 μm (52.7 - 65.2 μm), and hyaline tail terminal length is 36.5 ± 2.8 μm (32 - 39.8 μm). The tail had a sharp terminus. Morphology of the cysts and J2s were consistent with the record of H. filipjevi (Peng et al. 2010; Subbotin et al. 2010). The amplifications of rDNA-internal transcribed spacer (ITS) fragments were generated with a PCR fragment of 1054 bp from single cysts of each population, using primers TW81 and AB28 (Joyce et al. 1994). The PCR tests for each sample were repeated five times. The PCR product was purified and sequenced. All nucleotide sequences of ITS-rDNA were submitted to GenBank under accession numbers MW282843-6. Sequences from the ITS region were more than 99.5% identical to those of H. filipjevi from Egypt (KF225725), Turkey (KR704308, KR704293 and MN848333) and China (KT314234, MT254744 and KY448473). These results from ITS supported its identity as H. filipjevi. The results were also confirmed by species specific sequence characterized amplified region primers of H. filipjevi (Peng et al. 2013). Pathogenicity of the H. filipjevi was confirmed by infection of winter wheat (Triticum aestivum L cv. 'Aikang58') and examination of the nematode development and reproduction. Wheat seeds were germinated in petri dishes and then transplanted into five polyvinyl chloride tubs (3 cm in diameter, 25 cm in length) that contained 150 cm3 of a sterile soil mixture (loamy soil: sand = 1:1), each with 5 cysts (mean of 252.0 eggs/cyst). Plants were grown in an artificial climate box for one week at 14/18°C, two weeks at 16/20°C, five weeks at 18/25°C and two weeks at 22/30°C, under 8 h of darkness/16 h light and normal culturing practices (Cui et al. 2015). The parasitic J2s, third and fourth-stage juveniles, and adult females were observed in roots stained with acid fuchsin at 10, 20, 30, and 50 days after inoculation (DAI), and an average of 32.0 cysts per tubes were extracted 70 DAI. The new cyst' morphological and molecular characteristics were identical to the H. filipjevi cysts from the original soil samples. Three other tubes without cysts were set as control and there were no newly formed cysts. Heterodera avenae and H. filipjevi had been detected in a total of 16 wheat-producing provinces in China, which resulted in losses of 1.9 billion CNY year-1 (Cui et al. 2015). To our knowledge, this is the first report of H. filipjevi in Hebei Province of North China. Cereal cyst nematodes are easily transferred to non-infested areas by many avenues, resulting in increased species and pathotype complexity (Cui et al. 2020). Once H. filipjevi continues to spread in main wheat producing area of China, it could become be a new threat to cereals production. It is time to take effective control methods to prevent H. filipjevi further dispersal, especially through the farming machinery transmission. Hebei Province is one of the most important major grain-producing areas, our findings will be very beneficial for H. filipjevi management and further research on winter wheat in Hebei Province, North China.
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http://dx.doi.org/10.1094/PDIS-11-20-2519-PDNDOI Listing
January 2021

Biological evaluation of the modified nano-amorphous phosphate calcium doped with citrate/poly-amino acid composite as a potential candidate for bone repair and reconstruction.

J Mater Sci Mater Med 2021 Jan 25;32(1):16. Epub 2021 Jan 25.

College of Physics, Sichuan University, Chengdu, 610064, China.

Large numbers of research works related to fabricating organic-inorganic composite materials have been carried out to mimic the natural structure of bone. In this study, a new modified n-ACP doped with citrate (n-ACP-cit)/poly (amino acids) (PAA) composite (n-ACP-cit/PAA) was synthesized by employing high bioactive n-ACP-cit and the biodegradable and biocompatible PAA copolymer. Its basic structure was characterized by X-ray diffraction spectroscopy, Fourier transformed infrared spectroscopy, and X-ray photoelectron spectroscopy. Moreover, the degradability, bioactivity, biocompatibility, and osteoconductivity of n-ACP-cit/PAA composite were evaluated in vitro and in vivo, using simulated body fluid (SBF) solution soaking test, mouse bone marrow mesenchymal stem cells proliferation and differentiation, morphological observation test, expression of genes associated with osteogenesis, and bone defect model repair test, respectively. The modified n-ACP-cit/PAA composite exhibited a much higher weight loss rate (36.01 wt.%) than that of PAA (23.99 wt.%) after immersing in SBF solution for 16 weeks and the pH values of local environment restored to neutral condition. Moreover, cells co-culturing with composites exhibited higher alkaline phosphatase activity, more calcium nodule-formation, and higher expression levels of osteogenic differentiation-related genes (Bmp-2, Colla I, OCN, OPN, and Runx-2) than that of PAA. Furthermore, the bone defect model repair test revealed that the composite could be intimately incorporated with the surrounding bone without causing any deleterious reaction and capable of guiding new bone formation. Together, these results indicated that the new modified bone repair n-ACP-cit/PAA composite material with specific characteristics may be designed for meeting diverse requirements from biomedical applications.
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http://dx.doi.org/10.1007/s10856-020-06482-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7829244PMC
January 2021

Pathotype, Resistance Classification, and Seed-Coating Control of and in the North China Plain.

Plant Dis 2020 Dec 20;104(12):3230-3238. Epub 2020 Oct 20.

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

and are cereal cyst nematodes (CCNs) that infect cereals in 16 provinces of China. CCN populations from Xuchang, Tangyin, Qihe, and Juye were tested using 23 barley, oat, and wheat entries of the International Test Assortment for Defining Cereal Cyst Nematode Pathotypes. populations from Tangyin, Qihe, and Juye were classified as pathotype Ha91, and from Xuchang was classified as a new pathotype similar to pathotype West. Among 42 other winter wheat cultivars, 29 and 30 were differentially susceptible, 13 and 12 were differentially resistant to and , respectively. Three entries were resistant to both species, and three other entries were resistant to and moderately resistant to . Coating wheat seed with abamectin + isopycnic imidacloprid or methylene (bis) thiocyanate + thiamethoxam reduced the number of and cysts by 46 to 56%, increased wheat yield by 9 to 27%, and improved net income by 660 to 2,640 Chinese Yuan ha, respectively. Resistant wheat cultivars are scarce in China, and seed coating is considered the most suitable method for controlling CCNs in the North China Plain, where crop rotation cannot be practiced.
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http://dx.doi.org/10.1094/PDIS-02-20-0258-REDOI Listing
December 2020

First record of Heterodera elachista infecting rice in Henan Province of central China.

Plant Dis 2020 Sep 10. Epub 2020 Sep 10.

Henan Agricultural University, 70573, College of Plant Protection, Zhengzhou, China;

From June 2018 to November 2019, a survey for cyst-forming nematodes was conducted in rice fields in Henan Province of central China. Cysts were recovered from two rice fields (N32° 14' 048″8 and E115° 4' 008″) at Huangchuan County, leading to more intensive sampling. A further 25 soil samples were then collected with a valve bag from each of these two locations. Cysts and second-stage juveniles (J2) were recovered from roots and soil following Cobb's gravity sieving method. Live cysts were detected in all soil samples with a mean of 6.7±1.5 cysts per 100 ml of soil. Morphologically, the cysts were spherical to lemon-shaped, light to dark brown in color with subcrystalline layer. The vulval cone was well developed, cone terminus with a few large, peripheral, dark brown bullae lacking finger-like projections, and the ambifenestrae were almost rounded with two semifenestrae; width and length of the semifenestrae were similar. The vulval bridge was narrow, with a medium sized underbridge. Cyst measurements (n = 8) determined a mean body length of 431.1 ± 47.23 (351.0 - 516.0) µm, body width 304.3 ± 47.40 (240.0 - 381.0) µm; body length to width ratio 1.42 ± 0.10 (1.2 to 1.6); fenestrae length 39.4 ± 7.06 (26.0 - 47.0) µm; fenestrae width 36.5 ± 5.96 (25.0 - 43.0) µm; vulva slit length 37.1 ± 3.62 (30.0 - 42.0) µm; and the mean underbridge length 75.0 ± 3.39 (70.0 - 81.0) µm. Morphometric J2 measurements (n = 10) included a body length of 432.3 ± 53.26 (379.0 - 512.0) µm; stylet length 20.8 ± 1.87 (18.0 - 24) µm with rounded knobs; tail length 63.1 ± 7.92 (52.0 - 75.0) µm with a hyaline terminal tail length of 35.8 ± 6.14 (28.0 - 45.0) µm. The key morphometrics of this isolate were intermediate to those of the Japanese isolates (Nobbs et al. 1992.) and Chinese isolates (Ding et al. 2012), and other morphological character values were within the range of those reported for Heterodera elachista (Nobbs et al. 1992; Tanha Maafi et al. 2003). Amplification of DNA from single cysts (n = 7) was conducted using the protocol described by Ding et al. (2012). rDNA - ITS sequences were amplified with the universal primers TW81 and AB28 (Joyce et al. 1994). The PCR product was purified and sequenced. The ITS sequences were submitted to GenBank under accession numbers MT579616. Comparisons showed a sequence identity of more than 99.9% for H. elachista sequence MN720080 from Korea and 99.5% for H. elachista sequences JN864884 and JN202916 from China. Species identification was also confirmed using sequence characterized amplified region (SCAR) methods with H. elachista-specific primers He-F/He-R (Qi, 2012). An expected PCR fragment of approximately 434 bp was obtained, which was consistent with those previously reported for H. elachista. Pathogenicity was confirmed by infection and reproduction on rice (Oryza sativa cv. 'Nipponbare'). Seeds were sown into three tubes containing 150 ml of a sterile soil mixture (loamy soil: sand = 1:1), each with 5 cysts (mean of 185 eggs/cyst) and cultivated in an artificial climate box at 25/30°C, under a 12-h dark/12-h light cycle. Three other tubes without cysts were set as control. Two weeks after sowing, stunting and reduction of leaf length were observed and third- and fourth-stage juveniles were observed in roots stained with acid fuchsin. On average, 142 cysts per 150 ml soil were recovered at 5 weeks after sowing. The newly formed cysts corresponded morphologically and molecularly to the cysts from the original soil samples. The globally recognized and economically important rice-damaging cyst nematodes include H. oryzae, H. oryzicola, H. elachista, H. sacchari and H. graminophila (Zhuo et al. 2014). Ohshima (1974) first reported H. elachista, which was originally recorded as H. oryzae in Japan by Luc and Brizuela (1961). H. elachista was then detected from a rice field at Mazandaran Province in Iran (Tanha Maafi et al., 2003), and in upland rice fields in Hunan (Ding et al., 2012) and Guangxi, China (Zhuo et al. 2014). To the best of our knowledge, this is the first report of H. elachista as a pathogen on rice in Henan Province, in central China. According to our field observations, H. elachista was much more serious in direct-seeded rice field than in the transplanted rice fields. H. elachista was also reported attacking corn (Xiao et al., 2019). Henan is the most important corn-producing province in China, thus H. elachista is a potential threat to corn production in Henan. Our findings will be very beneficial for H. elachista management and further research on direct-seeded rice and corn in Henan Province, central China.
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http://dx.doi.org/10.1094/PDIS-06-20-1293-PDNDOI Listing
September 2020

Enhancement of osteoblast cells osteogenic differentiation and bone regeneration by hydroxyapatite/phosphoester modified poly(amino acid).

Mater Sci Eng C Mater Biol Appl 2020 Jun 22;111:110769. Epub 2020 Feb 22.

College of Physics, Sichuan University, Chengdu, Sichuan 610065, China. Electronic address:

Hydroxyapatite/poly(amino acid) (HA/PAA) has been used to treat a variety of long bone and vertebral bony defects, and a further biocompatibility improvement is a key for better application. Phosphoester (PE) contained materials are highly biocompatible but could hardly treat massive bone defects due to its fast-degradation-derived mechanical instability. To address the problems of the two materials, we have incorporated PE molecule into the main chain of PAA by chemical bonding. As a result, the compressive strength of HA/PAA with 1 wt% and 2.5 wt% PE maintained in the range of 80-150 MPa after soaking in PBS for 12 weeks, which could be attributed to the amplified hydrogen-bonding inside composites. Besides, the PE-containing HA/PAAs with increased hydrophilic function groups (O=P-O bonds and O=P-N), created a more favourable surface for cell adhesion. Meanwhile, compared with HA/PAA, the PE-containing HA/PAAs had a fast minerlization speed and promoted cell osteogenic differentiation. Furthermore, the in vivo study indicated that PE-containing HA/PAAs could facilitate bone formation (4 weeks), and form a complete bone bridging (12 weeks) in a rabbit cranial bone defect. In summary, the HA/PE-m-PAAs possessed good mechanical stability, improved cytocompatibility and osteoconductivity, so the composites have a great potential for massive bone defect treatment.
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http://dx.doi.org/10.1016/j.msec.2020.110769DOI Listing
June 2020

Preparation, Characterization and In Vitro Biological Evaluation of a Novel Pearl Powder/Poly-Amino Acid Composite as a Potential Substitute for Bone Repair and Reconstruction.

Polymers (Basel) 2019 May 8;11(5). Epub 2019 May 8.

College of Physical Science and Technology, Sichuan University, Chengdu 610064, China.

Many studies about fabricating organic-inorganic composite materials have been carried out in order to mimic the natural structure of bone. Pearl, which has a special block-and-mortar hierarchical structure, is a superior bone repair material with high osteogenic activity, but it shows few applications in the clinical bone repair and reconstruction because of its brittle and uneasily shaped properties. In this work, pearl powder (P)/poly (amino acid) (PAA) composites were successfully prepared by a method of in situ melting polycondensation to combine the high osteogenic activity of the pearl and the pliability of the PAA. The mechanical properties, in vitro bioactivity and biocompatibility as well as osteogenic activity of the composites were investigated. The results showed that P/PAA composites have both good mechanical properties and bioactivity. The compressive strength, bending strength and tensile strength of the composites reached a maximum of 161 MPa, 50 MPa and 42 MPa, respectively; in addition, apatite particles successfully deposited on the composites surface after immersion in simulated body fluid (SBF) for 7 days indicated that P/PAA composites showed an enhanced mineralization capacity and bioactivity due to incorporation of pearl powder and PAA. The cell culture results revealed that higher cell proliferation and better adhesion morphology of mouse bone marrow mesenchymal stem cells (MSCs) appeared on the composite surface. Moreover, cells growing on the surface of the composites exhibited higher alkaline phosphatase (ALP) activity, more calcium nodule-formation, and higher expression levels of osteogenic differentiation-related genes (COL 1, RunX2, OCN, and OPN) than cells grown on PAA surface. The P/PAA composites exhibited both superior mechanical properties to the pearl powder, higher bioactivity and osteogenic capability compared with those of PAA.
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http://dx.doi.org/10.3390/polym11050831DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6572604PMC
May 2019

Mechanics, degradability, bioactivity, in vitro, and in vivo biocompatibility evaluation of poly(amino acid)/hydroxyapatite/calcium sulfate composite for potential load-bearing bone repair.

J Biomater Appl 2016 Mar 2;30(8):1261-72. Epub 2015 Dec 2.

College of Physical Science and Technology, Sichuan University, Chengdu, China

A ternary composite of poly(amino acid), hydroxyapatite, and calcium sulfate (PAA/HA/CS) was prepared using in situ melting polycondensation method and evaluated in terms of mechanical strengths, in vitro degradability, bioactivity, as well as in vitro and in vivo biocompatibility. The results showed that the ternary composite exhibited a compressive strength of 147 MPa, a bending strength of 121 MPa, a tensile strength of 122 MPa, and a tensile modulus of 4.6 GPa. After immersion in simulated body fluid, the compressive strength of the composite decreased from 147 to 98 MPa for six weeks and the bending strength decreased from 121 to 75 MPa for eight weeks, and both of them kept stable in the following soaking period. The composite could be slowly degraded with 7.27 wt% loss of initial weight after soaking in phosphate buffered solution for three weeks when started to keep stable weight in the following days. The composite was soaked in simulated body fluid solution and the hydroxyapatite layer, as flower-like granules, formed on the surface of the composite samples, showing good bioactivity. Moreover, it was found that the composite could promote proliferation of MG-63 cells, and the cells with normal phenotype extended and spread well on the composite surface. The implantation of the composite into the ulna of sheep confirmed that the composite was biocompatible and osteoconductive in vivo, and offered the PAA/HA/CS composite promising material for load-bearing bone substitutes for clinical application.
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http://dx.doi.org/10.1177/0885328215620711DOI Listing
March 2016

[Comparative studies on the material performances of natural bone-like apatite from different bone sources].

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi 2014 Apr;31(2):352-6

The compressive strength of the original bone tissue was tested, based on the raw human thigh bone, bovine bone, pig bone and goat bone. The four different bone-like apatites were prepared by calcining the raw bones at 800 degrees C for 8 hours to remove organic components. The comparison of composition and structure of bone-like apatite from different bone sources was carried out with a composition and structure test. The results indicated that the compressive strength of goat bone was similar to that of human thigh bone, reached (135.00 +/- 7.84) MPa; Infrared spectrum (IR), X-ray diffraction (XRD) analysis results showed that the bone-like apatite from goat bone was much closer to the structure and phase composition of bone-like apatite of human bones. Inductively Coupled Plasma (ICP) test results showed that the content of trace elements of bone-like apatite from goat bone was closer to that of apatite of human bone. Energy Dispersive Spectrometer (EDS) results showed that the Ca/P value of bone-like apatite from goat bone was also close to that of human bone, ranged to 1.73 +/- 0.033. Scanning electron microscopy (SEM) patterns indicated that the macrographs of the apatite from human bone and that of goat bone were much similar to each other. Considering all the results above, it could be concluded that the goat bone-like apatite is much similar to that of human bone. It can be used as a potential natural bioceramic material in terms of material properties.
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April 2014
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