The importance of osteoclast-mediated bone resorption in the process of osseointegration has not been widely considered. In this study, cell culture was used to investigate the hypothesis that the function of implant-adherent bone marrow stromal cells (BMSCs) in osteoclastogenesis is influenced by surface topography. BMSCs isolated from femur and tibia of Sprague-Dawley rats were seeded onto 3 types of titanium surfaces (smooth, micro, and nano) and a control surface (tissue culture plastic) with or without osteogenic supplements. After 3 to 14 d, conditioned medium (CM) was collected. Subsequently, rat bone marrow-derived macrophages (BMMs) were cultured in media supplemented with soluble receptor activator of NF-κB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) as well as BMSC CM from each of the 4 surfaces. Gene expression levels of soluble RANKL, osteoprotegerin, tumor necrosis factor α, and M-CSF in cultured BMSCs at different time points were measured by real-time polymerase chain reaction. The number of differentiated osteoclastic cells was determined after tartrate-resistant acid phosphatase staining. Analysis of variance and t test were used for statistical analysis. The expression of prominent osteoclast-promoting factors tumor necrosis factor α and M-CSF was increased by BMSCs cultured on both micro- and nanoscale titanium topographies (P < 0.01). BMSC CM contained a heat-labile factor that increased BMMs osteoclastogenesis. CM from both micro- and nanoscale surface-adherent BMSCs increased the osteoclast number (P < 0.01). Difference in surface topography altered BMSC phenotype and influenced BMM osteoclastogenesis. Local signaling by implant-adherent cells at the implant-bone interface may indirectly control osteoclastogenesis and bone accrual around endosseous implants.
Surface topography broadly influences cellular responses. Adherent cell activities are regulated, in part, by RhoA, a member of the Rho-family of GTPases. In this study, we evaluated the influence of surface topography on RhoA activity and associated cellular functions. The murine mesenchymal stem cell line C3H10T1/2 cells (osteoprogenitor cells) were cultured on titanium substrates with smooth topography (S), microtopography (M), and nanotopography (N) to evaluate the effect of surface topography on RhoA-mediated functions (cell spreading, adhesion, migration, and osteogenic differentiation). The influence of RhoA activity in the context of surface topography was also elucidated using RhoA pharmacologic inhibitor. Following adhesion, M and N adherent cells developed multiple projections, while S adherent cells had flattened and widespread morphology. RhoA inhibitor induced remarkable longer and thinner cytoplasmic projections on all surfaces. Cell adhesion and osteogenic differentiation was topography dependent with S < M and N surfaces. RhoA inhibition increased adhesion on S and M surfaces, but not N surfaces. Cell migration in a wound healing assay was greater on S versus M versus N surfaces and RhoA inhibitor increased S adherent cell migration, but not N adherent cell migration. RhoA inhibitor enhanced osteogenic differentiation in S adherent cells, but not M or N adherent cells. RhoA activity was surface topography roughness dependent (S < M, N). RhoA activity and -mediated functions are influenced by surface topography. Smooth surface adherent cells appear highly sensitive to RhoA function, while nano-scale topography adherent cell may utilize alternative cellular signaling pathway(s) to influence adherent cellular functions regardless of RhoA activity.
An efficient propagation system of pitaya with diverse genetic background was established by using tender stem as explants. Various types of plant growth regulators (PGRs) were used to determine the most effective hormone combination for shoot proliferation. Zeatin (ZT), thidiazuron (TDZ), 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 6-benzylaminopurine (6-BA) alone could induce multiple shoots. In that case, the most shoots per explant i.e. 4.6 (13.68 lM ZT), 4.8 (0.11 lM TDZ), 4.3 (17.76 lM 6-BA) and 3.2 (0.23 lM 2, 4-D) were obtained. The best PGRs combination was MS medium with 3.0 lM ZT and 0.5 lM IBA producing the most shoots per explant and the most vigorous shoots. Of the two varieties and six superior selections cultured on the best PGRs combination, more than 6.0 shoots per explant were obtained. No polymorphism was detected among in vitro-derived plantlets selected at random after 11 sub-cultures. Those results showed that MS media with 13.68 lM ZT and 2.46 lM IBA was suitable for shoot propagation of diverse pitaya varieties and selections. The protocol can be used for largescale propagation of pitaya to meet the demand of increasing commercial cultivation.
The seroprevalence of toxoplasmosis in pigs was examined in China's southern Guangdong Province from March 2008 through May 2009. A total of 1,022 serum samples was collected from 12 administrative cities and assayed for Toxoplasma gondii antibodies by enzyme-linked immunosorbent assay test. The seroprevalence ranged from 0 to 58.1% and the average T. gondii antibody-positive prevalence was 27.0% (276/1,022), with breeding boars having the highest level at 39.6% (38/96) followed by breeding sows at 36.9% (52/141). The results of the present survey indicate that T. gondii infection is a significant health problem in pigs and represents a public health concern in southern China.
Enzootic pneumonia caused by Mycoplasma hyopneumoniae is a severe disease of pigs, causing significant economic losses to the pig industry worldwide, including the tropical and subtropical regions. In order to obtain the baseline prevalence of M. hyopneumoniae in pigs from intensive farms in southern China, double-sandwich enzyme-linked immunosorbent assay (ELISA) was used to detect M. hyoneumoniae antibodies in 460 pig serum samples collected from 12 administrative cities in China's southern Guangdong province. According to the proportions of the infected animals, among the 12 intensive farms, only two of them showed no infection of M. hyoneumoniae and the seroprevalence ranged from 0% to 90%, with an averaged prevalence of 45.7%. The highest prevalence was found in breeding boars (68.8%), followed by sows (54.5%). These data showed that the infection of pigs with M. hyopneumoniae is severe, and boars might be more important carriers and transfers of M. hyoneumoniae than sows. Integrated strategies and measures should be taken to control the infection of pigs with M. hyopneumoniae in southern China.
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