Study on the In Vitro and In Vivo Antibacterial Activity and Biocompatibility of Novel TiN/Ag Multilayers Immobilized onto Biomedical Titanium.

To determine the short- and long-term antibacterial properties of a novel biomedical titanium alloy to ensure excellent biocompatibility of the TiN/Ag multilayers loaded with different doses of Ag+. First, nanosized TiN/Ag multilayers were accumulated onto titanium alloy (Ti-6Al-4V) substrates via multi-arc ion plating. Then, the multilayers were implanted with different doses of silver ions (1×1017 ions/cm², 1×1018 ions/cm², 5×1016 ions/cm², and 5×1017 ions/cm²). Both short- and long-term antibacterial properties against Streptococcus mutans and Staphylococcus aureus were assessed via unique methods. Additionally, the response and behaviors of MC3T3-E1 and L929 cells on the different surfaces were evaluated by a variety of methods through comparison to a normal matched substrate (Ti-6Al-4V). In Vitro and In Vivo analyses revealed that the multilayers containing different doses of Ag ions effectively prevented bacterial adhesion and eliminated the majority of adhered bacteria in the initial period. In addition, the antibacterial activity of each TiN/Ag group improved with time, with the antibacterial rate (Ra) ultimately reaching 99% (antibacterial activity: 1 × 1018 ions/cm² > 5 × 1017 ions/cm² > 1 × 1017 ions/cm² > 5 × 1016 ions/cm²). All of the samples loaded with Ag+ exhibited good compatibility, as well as higher cell proliferation and lower apoptosis than the pure Ti-6Al-4V substrates. Considering both bacteriostasis and biocompatibility, 1 × 1017 ions/cm² and 5 × 1017 ions/cm² are the recommended doses for orthopedic and dental implants. The results indicate that all of the samples loaded with Ag+ possess excellent biocompatibility and antibacterial activity against common bacteria that cause implantation infection. The samples loaded with Ag+ can be implanted into soft and hard growing tissues to greatly improve the survival rate of orthopedic and dental implants.

Molecular cloning, expression profiles, and characterization of a novel polyphenol oxidase (PPO) gene in Hevea brasiliensis.

The polyphenol oxidase (PPO) is involved in undesirable browning in many plant foods. Although the PPOs have been studied by several researchers, the isolation and expression profiles of PPO gene were not reported in rubber tree. In this study, a new PPO gene, HbPPO, was isolated from Hevea brasiliensis. The sequence alignment showed that HbPPO indicated high identities to plant PPOs and belonged to dicot branch. The cis-acting regulatory elements related to stress/hormone responses were predicted in the promoter region of HbPPO. Real-time RT-PCR analyses showed that HbPPO expression varied widely depending on different tissues and developmental stages of leaves. Besides being associated with tapping panel dryness, the HbPPO transcripts were regulated by ethrel, wounding, H2O2, and methyl jasmonate treatments. Moreover, the correlation between latex coagulation rate and PPO activity was further confirmed in this study. Our results lay the foundation for further analyzing the function of HbPPO in rubber tree.

Mechanism underlying effects of cellulose-degrading microbial inoculation on amino acid degradation and biosynthesis during composting.

Amino acids are essential organic compounds in composting products. However, the mechanism underlying the amino acid metabolism during composting remains unclear. This study aims at exploring the impacts of inoculating cellulose-degrading microbes on amino acid metabolism during composting with mulberry branches and silkworm excrements. Cellulose-degrading microbial inoculation enhanced amino acid degradation by 18%-43% by increasing protease and sucrase activities and stimulating eight amino acid degradation pathways from the initial to thermophilic phases, with Enterococcus, Saccharomonospora, Corynebacterium being the dominant bacterial genera, but stimulated amino acid production by 54% by increasing sucrase and urease activities, decreasing ß-glucosidase activities, and stimulating twenty-two amino acid synthesis pathways at the mature phase, with Thermobifida, Devosia, and Cellulosimicrobium being the dominant bacterial genera. The results suggest that cellulose-degrading microbial inoculation enhances amino acid degradation from the initial to thermophilic phases and biosynthesis at the mature phase, thereby improving the quality of organic fertilizer.

Addition of cellulose and hemicellulose degrading microorganisms intensified nitrous oxide emission during composting.

This study aims to clarify the mechanisms underlying effects of inoculating cellulose and hemicellulose-degrading microorganisms on nitrous oxide (N2O) emissions during composting with silkworm excrement and mulberry branches. Inoculation with cellulose and hemicellulose-degrading microorganisms resulted in significant increases of total N2O emission by 10.4 ± 2.0 % (349.1 ± 6.2 mg N kg-1 dw) and 26.7 ± 2.1 % (400.6 ± 6.8 mg N kg-1 dw), respectively, compared to the control (316.3 ± 3.6 mg N kg-1 dw). The stimulation of N2O emission was attributed to the enhanced contribution of ammonia-oxidizing bacteria (AOB) and denitrifying bacteria to N2O production, as evidenced by the increased AOB amoA and denitrifying nirK gene abundances. Moreover, microbial inoculation stimulated N2O reduction to N2 owing to increased abundances of nosZⅠ and nosZⅠⅠ genes. These findings highlight the necessity to develop cost-effective and environmentally friendly strategies to reduce N2O emissions when cellulose and hemicellulose-degrading microorganisms are inoculated during composting.

N-containing functional groups induced superior cytocompatible and hemocompatible graphene by NH2 ion implantation.

Graphene is functionalized with amine by NH2 ion implantation at room temperature in vacuum. The reaction is featured by nucleophilic substitution of C-O groups by the ammonia radicals. The presence of N-containing functional groups in graphene is identified by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. N element was successfully introduced to graphene, the atomic ratio of N to C rose to 3.12 %. NH2 ion implanted graphene (G-NH2) is a better hydrophilic material than pristine grahene according to the contact angle experiment. Mouse fibroblast cells and human endothelial cells cultured on G-NH2 displayed superior cell-viability, proliferation and stretching over that on pristine graphene. Platelet adhesion, hemolysis and Kinetic-clotting time were measured on G-NH2, showing excellent anticoagulation, with as good hemolysis as pristine graphene.

High plant species diversity enhances lignin accumulation in a subtropical forest of southwest China.

Plant species diversity (PSD) benefits soil organic carbon (SOC) accumulation, but mechanisms underlying the stimulative effects of PSD on SOC pools have not been well explored, especially in terms of how PSD impacts plant-derived C accumulation. Here, 45 plots covering a natural gradient of PSD ranging from 0.15 to 3.57 (Shannon's diversity index) were selected in a subtropical forest with calcareous soil to determine the pattern of and controls on the variation of plant-derived C as indexed by lignin phenols along with PSD. The absolute contents of lignin phenols ranged from 1.18 to 6.62 mg g-1 soil with an average of 2.48 ± 1.13 mg g-1 soil across the 45 plots. PSD significantly enhanced soil lignin accumulation via three mechanisms. First, PSD benefited lignin accumulation by stimulating plant detritus inputs. Second, PSD directly and indirectly increased reactive minerals, so that enhanced mineral protection of lignin. Third, decrease in microbial C limitation due to increased soil C availability resulted in lowered peroxidase activity and subsequently lignin degradation, which in turn benefited lignin accumulation. Our study provides mechanisms underlying SOC accumulation in response to increased PSD, which may be integrated into Earth system models in order to better predict SOC dynamics under PSD alteration.

Adjusting the Dose of Ag-Ion Implantation on TiN-Ag-Modified SLA-Ti Creates Different Micronanostructures: Implications on Bacteriostasis, Biocompatibility, and Osteogenesis in Dental Implants.

The prevention of aseptic loosening and peri-implantitis is crucial for the success of dental implant surgery. In this study, different doses of Ag-implanted TiN/Ag nanomultilayers were prepared on the sandblasting with large grit and acid etching (SLA)-Ti surface using a multiarc ion-plating system and an ion-implantation system, respectively. The physical and chemical properties of the samples were assessed using various techniques, including scanning electron microscopy, energy-dispersive spectroscopy, X-ray diffraction, X-ray photoelectron spectroscopy, atomic force microscopy, inductively coupled plasma atomic emission spectrometry, and water contact angle measurements. In addition, the applicability and biosafety of the SLA/1 × 1017-Ag and SLA/1 × 1018-Ag surfaces were determined via biocompatibility testing in vivo and in vitro. The results demonstrated that the physical and chemical properties of SLA/1 × 1017-Ag and SLA/1 × 1018-Ag surfaces were different to some extent. However, compared with SLA-Ti, silver-loaded TiN/Ag-modified SLA-Ti surfaces (SLA/1 × 1018-Ag) with enhanced bacteriostatis, osteogenesis, and biocompatibility have great potential for dental applications.

Deciphering the dominant components and functions of bacterial communities for lignocellulose degradation at the composting thermophilic phase.

The decomposition and transformation of organic matters during composting process are performed by various microorganisms. However, the bacterial communities and their functions usually vary with composting materials. Here the dominant bacterial genera and their functions were identified at the thermophilic phase during composting of mulberry branches with silkworm excrement (MSE), pig manure (MPM) and cow manure (MCD). The activities of ß-glucosidase and endoglucanase were highest for MCD (1.31 and 17.15 µg g-1 min-1) and lowest for MPM (0.92 and 14.22 µg g-1 min-1). Random Forest model and correlation analysis revealed that Stenotrophomonas, Bacillus, and Sinibacillus were the dominant bacterial genera involved in lignocellulose degradation regardless of composting materials. Carbohydrate metabolism, amino acid metabolism, and DNA replication and repair were primary functions of the bacterial communities for the three types of composting. The quantification of lignocellulose degradation genes further verified the dominant functions of the bacterial communities.

RNA-Seq analysis and de novo transcriptome assembly of Hevea brasiliensis.

Hevea brasiliensis, being the only source of commercial natural rubber, is an extremely economically important crop. In an effort to facilitate biological, biochemical and molecular research in rubber biosynthesis, here we report the use of next-generation massively parallel sequencing technologies and de novo transcriptome assembly to gain a comprehensive overview of the H. brasiliensis transcriptome. The sequencing output generated more than 12 million reads with an average length of 90 nt. In total 48,768 unigenes (mean size = 436 bp, median size = 328 bp) were assembled through de novo transcriptome assembly. Out of 13,807 H. brasiliensis cDNA sequences deposited in Genbank of the National Center for Biotechnology Information (NCBI) (as of Feb 2011), 11,746 sequences (84.5%) could be matched with the assembled unigenes through nucleotide BLAST. The assembled sequences were annotated with gene descriptions, Gene Ontology (GO) and Clusters of Orthologous Group (COG) terms. In all, 37,432 unigenes were successfully annotated, of which 24,545 (65.5%) aligned to Ricinus communis proteins. Furthermore, the annotated uingenes were functionally classified according to the GO, COG and Kyoto Encyclopedia of Genes and Genomes databases. Our data provides the most comprehensive sequence resource available for the study of rubber trees as well as demonstrates effective use of Illumina sequencing and de novo transcriptome assembly in a species lacking genomic information.

Identification and characterization of genes associated with tapping panel dryness from Hevea brasiliensis latex using suppression subtractive hybridization.

BACKGROUND: Tapping panel dryness (TPD) is one of the most serious threats to natural rubber production. Although a great deal of effort has been made to study TPD in rubber tree, the molecular mechanisms underlying TPD remain poorly understood. Identification and systematical analyses of the genes associated with TPD are the prerequisites for elucidating the molecular mechanisms involved in TPD. The present study is undertaken to generate information about the genes related to TPD in rubber tree. RESULTS: To identify the genes related to TPD in rubber tree, forward and reverse cDNA libraries from the latex of healthy and TPD trees were constructed using suppression subtractive hybridization (SSH) method. Among the 1106 clones obtained from the two cDNA libraries, 822 clones showed differential expression in two libraries by reverse Northern blot analyses. Sequence analyses indicated that the 822 clones represented 237 unique genes; and most of them have not been reported to be associated with TPD in rubber tree. The expression patterns of 20 differentially expressed genes were further investigated to validate the SSH data by reverse transcription PCR (RT-PCR) and real-time PCR analysis. According to the Gene Ontology convention, 237 unique genes were classified into 10 functional groups, such as stress/defense response, protein metabolism, transcription and post-transcription, rubber biosynthesis, etc. Among the genes with known function, the genes preferentially expressed were associated with stress/defense response in the reverse library, whereas metabolism and energy in the forward one. CONCLUSIONS: The genes associated with TPD were identified by SSH method in this research. Systematic analyses of the genes related to TPD suggest that the production and scavenging of reactive oxygen species (ROS), ubiquitin proteasome pathway, programmed cell death and rubber biosynthesis might play important roles in TPD. Therefore, our results not only enrich information about the genes related to TPD, but also provide new insights into understanding the TPD process in rubber tree.

Study on the osteogenesis of rat mesenchymal stem cells and the long-term antibacterial activity of Staphylococcus epidermidis on the surface of silver-rich TiN/Ag modified titanium alloy.

The main causes of failure of orthopedic implants are infection and poor bone ingrowth. Surface modification of the implants to allow for long-term antibacterial and osteogenic functions is an effective solution to prevent failure of the implants. We developed silver-rich TiN/Ag nano-multilayers on the surface of titanium alloy with different doses of Ag+ . The antibacterial stability and osteogenesis of the silver-rich surface were determined by evaluating the adhesion and proliferation of Staphylococcus epidermidis, and the adhesion, proliferation, alkaline phosphatase activity, extracellular matrix mineralization, and the expression level of genes involved in osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs). The results demonstrated that the antibacterial rates (Ra) of 5 × 1016 -Ag, 1 × 1017 -Ag, 5 × 1017 -Ag, and 1 × 1018 -Ag were respectively 46.21%, 85.66%, 94.99%, 98.48%, and 99.99%. After subcutaneous implantation in rats or immersion in phosphate buffered saline for up to 12 weeks, the silver-rich surface of the titanium alloy showed long-term stable inhibition of Staphylococcus epidermidis. Furthermore, in vitro and in vivo studies indicated that the Ag-implanted titanium did not show apparent cytotoxicity and that lower Ag+ implanted groups (5 × 1016 -Ag, 1 × 1017 -Ag) had better viability and biological safety when compared with higher Ag+ implanted groups. In addition, when compared with the Ti6Al4V-group, all Ag-implanted groups exhibited enhanced osteogenic indicators in rat BMSCs. Regarding osteogenic indicators, the surfaces of the 5 × 1017 -Ag group had better osteogenic effects than those of other groups. Therefore, the proper dose of Ag+ implanted TiN/Ag nano-multilayers may be one of the options for the hard tissue replacement materials with antibacterial activity and osteogenic functions.

Gene-informed decomposition model predicts lower soil carbon loss due to persistent microbial adaptation to warming.

Soil microbial respiration is an important source of uncertainty in projecting future climate and carbon (C) cycle feedbacks. However, its feedbacks to climate warming and underlying microbial mechanisms are still poorly understood. Here we show that the temperature sensitivity of soil microbial respiration (Q10) in a temperate grassland ecosystem persistently decreases by 12.0 ± 3.7% across 7 years of warming. Also, the shifts of microbial communities play critical roles in regulating thermal adaptation of soil respiration. Incorporating microbial functional gene abundance data into a microbially-enabled ecosystem model significantly improves the modeling performance of soil microbial respiration by 5-19%, and reduces model parametric uncertainty by 55-71%. In addition, modeling analyses show that the microbial thermal adaptation can lead to considerably less heterotrophic respiration (11.6 ± 7.5%), and hence less soil C loss. If such microbially mediated dampening effects occur generally across different spatial and temporal scales, the potential positive feedback of soil microbial respiration in response to climate warming may be less than previously predicted.

Biological actions of Cu/Zn coimplanted TiN on Ti-6Al-4V alloy.

Ti-6Al-4V alloy, as a widely used orthopedic and dental implant, has excellent biocompatibility and machinability. However, its poor corrosion resistance and antibacterial property may lead to tissue inflammation and postoperative infection, which hinders its further development. In this paper, to solve the above problems, copper (Cu) and zinc (Zn) ions were coimplanted into a titanium nitride (TiN) coated Ti-6Al-4V alloy via a plasma immersion ion implantation system (PIII). Then, the structure and composition of Cu/Zn coimplanted TiN (Cu/Zn-TiN-PIII) were characterized by scanning electron microscopy and x-ray photoelectron spectroscopy. Also, the results of the corrosion test, the water contact angles test, and the protein electrophoresis experiment showed that the corrosion resistance, hydrophilicity, and the protein adsorption capacity of Cu/Zn-TiN-PIII were improved simultaneously. In addition, compared with TiN-PIII, Cu/Zn-TiN-PIII promoted both cytocompatibility and the antibacterial property according to L929 cells and Escherichia coli assays in vitro. Therefore, Cu/Zn-TiN-PIII may be a good candidate for orthopedic implants.

Involvement of HbMC1-mediated cell death in tapping panel dryness of rubber tree (Hevea brasiliensis).

Tapping panel dryness (TPD) causes a significant reduction in the latex yield of rubber tree (Hevea brasiliensis Muell. Arg.). It is reported that TPD is a typical programmed cell death (PCD) process. Although PCD plays a vital role in TPD occurrence, there is a lack of detailed and systematic study. Metacaspases are key regulators of diverse PCD in plants. Based on our previous result that HbMC1 was associated with TPD, we further elucidate the roles of HbMC1 on rubber tree TPD in this study. HbMC1 was up-regulated by TPD-inducing factors including wounding, ethephon and H2O2. Moreover, the expression level of HbMC1 was increased along with TPD severity in rubber tree, suggesting a positive correlation between HbMC1 expression and TPD severity. To investigate its biological function, HbMC1 was overexpressed in yeast (Saccharomyces cerevisiae) and tobacco (Nicotiana benthamiana). Transgenic yeast and tobacco overexpressing HbMC1 showed growth retardation compared with controls under H2O2-induced oxidative stress. In addition, overexpression of HbMC1 in yeast and tobacco reduced cell survival after high-concentration H2O2 treatment and resulted in enhanced H2O2-induced leaf cell death, respectively. A total of 11 proteins, rbcL, TM9SF2-like, COX3, ATP9, DRP, HbREF/Hevb1, MSSP2-like, SRC2, GATL8, CIPK14-like and STK, were identified and confirmed to interact with HbMC1 by yeast two-hybrid screening and co-transformation in yeast. The 11 proteins mentioned above are associated with many biological processes, including rubber biosynthesis, stress response, autophagy, carbohydrate metabolism, signal transduction, etc. Taken together, our results suggest that HbMC1-mediated PCD plays an important role in rubber tree TPD, and the identified HbMC1-interacting proteins provide valuable information for further understanding the molecular mechanism of HbMC1-mediated TPD in rubber tree.

Enhanced cell growth on 3D graphene scaffolds implanted with nitrogen ions.

One of the key challenges in engineering tissues for cell-based therapies is developing biocompatible scaffold materials to direct cell behavior. In this paper, the cytocompatibilities of a flexible three-dimensional graphene scaffold (3D-G) and the same scaffold implanted with nitrogen ions (N+/3D-G) are compared using an in vitro assay based on 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. The N+/3D-G samples were prepared from low-temperature hydrothermally synthesized flexible 3D-G by ion implantation and were found to display improved adhesion and proliferation of rat osteoblast and mouse fibroblast cells. In particular, the N+/3D-G sample with a nitrogen content of ∼10% showed the highest levels of cell viability and proliferation. The flexible N+/3D-G has potential applications as a biocompatible scaffold material that provides improved surface area and hydrophilic groups for cell growth and proliferation.

N+ implantation induce cytocompatibility of shape-controlled three-dimensional self-assembly graphene.

AIM: The aim of the present research was to synthesize N+ implanted 3D self-assembly graphene (N+/3D-SGHs) to overcome the weaknesses of graphene (small sizes and poor hydrophilicity) in tissue engineering scaffolds. MATERIALS & METHODS: N+/3D-SGHs was achieved by ion implantation on one-step hydrothermal synthesized 3D self-assembly graphene (3D-SGHs), and N+/3D-SGHs with different doses of nitrogen ions (1 × 1016 ions/cm2, 1 × 1018 ions/cm2 and 1 × 1020 ions/cm2), which adjusted by nitrogen ion beam intensity. RESULTS: N+/3D-SGHs, as scaffolds, provide stereo space and hydrophilic groups for mouse-fibroblast cells (L929) growth and proliferation. Notably, N+/3D-SGHs with the N+ injected quantity of 1 × 1020 ions/cm2 displayed the highest protein-adhesion strength, cell viability and proliferation, which supported its good cytocompatibility. CONCLUSION: This study demonstrated N+/3D-SGHs as a promising and effective tissue scaffold that might have applications in biomedicine.

Ag+ implantation induces mechanical properties, cell adhesion and antibacterial effects of TiN/Ag multilayers in vitro.

AIM: This study aims to investigate the effect of Ag+ implantation dose on the structure, hardness, adhesion strength, friction resistance, cell adhesion and antibacterial effects of TiN/Ag multilayers. METHODS: Nanoscale TiN/Ag multilayers were deposited on Ti-6Al-4V substrates using multiarc ion plating. The multilayers were then implanted by Ag ions. RESULTS: A distinct multilayer structure and large titanium nitride grains with better (111) crystallinity were proved. The hardness and elastic modulus of the multilayer reached 32.2 and 318.9 GPa, respectively. The largest critical load was 32.5 mN, and the minimum friction coefficient was 0.092. The mechanical properties, the cell proliferation and antibacterial properties of the multilayers with Ag+ implantation were better than those without Ag+ implantation. CONCLUSION: Our results indicate that a dose of 1 × 1017 ions/cm2 induced an improvement in crystallinity, mechanical properties, as well as preferable cell adhesion and antibacterial effects.

Transcriptome analyses reveal molecular mechanism underlying tapping panel dryness of rubber tree (Hevea brasiliensis).

Tapping panel dryness (TPD) is a serious threat to natural rubber yields from rubber trees, but the molecular mechanisms underlying TPD remain poorly understood. To identify TPD-related genes and reveal these molecular mechanisms, we sequenced and compared the transcriptomes of bark between healthy and TPD trees. In total, 57,760 assembled genes were obtained and analyzed in details. In contrast to healthy rubber trees, 5652 and 2485 genes were up- or downregulated, respectively, in TPD trees. The TPD-related genes were significantly enriched in eight GO terms and five KEGG pathways and were closely associated with ROS metabolism, programmed cell death and rubber biosynthesis. Our results suggest that rubber tree TPD is a complex process involving many genes. The observed lower rubber yield from TPD trees might result from lower isopentenyl diphosphate (IPP) available for rubber biosynthesis and from downregulation of the genes in post-IPP steps of rubber biosynthesis pathway. Our results not only extend our understanding of the complex molecular events involved in TPD but also will be useful for developing effective measures to control TPD of rubber trees.

Genome-wide identification and expression analysis of the metacaspase gene family in Hevea brasiliensis.

Metacaspases, a family of cysteine proteases, have been suggested to play important roles in programmed cell death (PCD) during plant development and stress responses. To date, no systematic characterization of this gene family has been reported in rubber tree (Hevea brasiliensis). In the present study, nine metacaspase genes, designated as HbMC1 to HbMC9, were identified from whole-genome sequence of rubber tree. Multiple sequence alignment and phylogenetic analyses suggested that these genes were divided into two types: type I (HbMC1-HBMC7) and type II (HbMC8 and HbMC9). Gene structure analysis demonstrated that type I and type II HbMCs separately contained four and two introns, indicating the conserved exon-intron organization of HbMCs. Quantitative real-time PCR analysis revealed that HbMCs showed distinct expression patterns in different tissues, suggesting the functional diversity of HbMCs in various tissues during development. Most of the HbMCs were regulated by drought, cold, and salt stress, implying their possible functions in regulating abiotic stress-induced cell death. Of the nine HbMCs, HbMC1, HbMC2, HbMC5, and HbMC8 displayed a significantly higher relative transcript accumulation in barks of tapping panel dryness (TPD) trees compared with healthy trees. In addition, the four genes were up-regulated by ethephon (ET) and methyl jasmonate (MeJA), indicating their potential involvement in TPD resulting from ET- or JA-induced PCD. In summary, this work provides valuable information for further functional characterization of HbMC genes in rubber tree.

Enhancement of interaction of L-929 cells with functionalized graphene via COOH+ ion implantation vs. chemical method.

Low hydrophilicity of graphene is one of the major obstacles for biomaterials application. To create some hydrophilic groups on graphene is addressed this issue. Herein, COOH+ ion implantation modified graphene (COOH+/graphene) and COOH functionalized graphene were designed by physical ion implantation and chemical methods, respectively. The structure and surface properties of COOH+/graphene and COOH functionalized graphene were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), and contact angle measurement. Compared with graphene, COOH+/graphene and COOH functionalized graphene revealed improvement of cytocompatibility, including in vitro cell viability and morphology. More importantly, COOH+/graphene exhibited better improvement effects than functionalized graphene. For instance, COOH+/graphene with 1 × 1018 ions/cm2 showed the best cell-viability, proliferation and stretching. This study demonstrated that ion implantation can better improve the cytocompatibility of the graphene.