[Study of cytokines in peripheral blood and lung of rats exposed to hard metal dust].

Objective: To study the dynamic changes of cytokines in bronchoalveolar lavage fluid (BALF) and serum of hard metal lung disease (HMLDR) rats. Methods: In March 2019, the rats were randomly divided into 6 groups, each group included 8 rats: control (C) group include 3 groups, hard metal (HM) group include 3 groups. 10 mg HM were administered in HM group by using the pulmonary endotracheal tube. After 4, 8 and 12 week, the BALF and serum were collected for the enzyme-linked immunosorbent assay (ELISA) of matrix metalloproteinase-1 (MMP-1) , tissue inhibitor of metalloproteinase-1 (TIMP-1) and tumor necrosis factor-alpha (TNF-α) . Results: There was no abnormality in behavior, diet and fur of rats in C and HM group at each exposure time. There was no significant difference in body weight between the two groups of rats (P>0.05) . Compared with the C group, the expression of MMP-1 in BALF of rats in HM group were significantly higher in all stages (4, 8 and 12 weeks after exposure) (P<0.05) , the expression of TIMP-1 in BALF of rats in HM group were significantly higher in 8 and 12 weeks after exposure (P<0.05) . However, there was no significant difference in serum MMP-1 and TIMP-1 levels between the two groups in each stage (P>0.05) . There was no significant difference in TNF-α. level in BALF and serum between C and HM group in all stages (P>0.05) . Conclusion: Expression of MMP-1 and TIMP-1 in BALF have reference value in the HMLD auxiliary diagnosis.

Protective effect of an egg yolk-derived immunoglobulin (IgY) against Prevotella intermedia-mediated gingivitis.

AIMS: To investigate the effects of an egg yolk-derived immunoglobulin (IgY) specific to Prevotella intermedia in vitro and in vivo. METHODS AND RESULTS: An IgY specific to P. intermedia was produced by immunizing hens with formaldehyde-inactivated P. intermedia and showed high titres when subjected to an ELISA. The obtained IgY inhibited the growth of P. intermedia in a dose-dependent manner at concentrations from 1 to 20 mg ml(-1) in Center for Disease Control and Prevention liquid medium. Forty rats were challenged with P. intermedia on gingivae and then randomly divided into four groups, which were syringed respectively with phosphate-buffered saline, 1 mg ml(-1) of tinidazole, 20 mg ml(-1) of nonspecific IgY and 20 mg ml(-1) of the IgY specific to P. intermedia at a dosage of 300 µl per day. Gingival index (GI), plaque index (PI), bleeding on probing (BOP), counts of white blood cell (WBC) and histopathological slide of the gums were measured after treatment for 15 days. The gingivitis rats treated with the IgY specific to P. intermedia showed significantly decreased GI, PI, BOP and WBC (P < 0·05). Gum histopathology of the treated rats demonstrated a superior protective effect of the specific IgY on P. intermedia-mediated gingivitis. CONCLUSIONS: A new immunoglobulin specific to P. intermedia was developed from egg yolk. This specific IgY can dose-dependently inhibit the growth of P. intermedia and protect rats from gingivitis induced by P. intermedia. SIGNIFICANCE AND IMPACT OF THE STUDY: The new IgY has potential for the treatment of P. intermedia-mediated gingivitis.

Preparation and characterisation of andrographolide niosomes and its anti-hepatocellular carcinoma activity.

OBJECTIVE: In this study, a novel andrographolide (AG) preparation formulation, niosomes, was prepared to improve the bioavailability and tissue distribution of AG. METHODS: The niosomal formulation of AG was prepared by film hydration/sonication method and tissue distribution was measured by liquid chromatography-mass spectrometry (LC-MS) method in mice, and anti-hepatocellular carcinoma (anti-HCC) activity was examined by MTT method in HepG2. RESULTS: Entrapment efficiency, drug-loading ratio and average particle size of AG niosomes were 72.36%, 5.90% and 206 nm, respectively. The tissue distribution in mice demonstrated that the AG niosomes were absorbed in liver much more than the free AG. Furthermore, the anti-HCC activity in HepG2 cells showed that there was no significant difference between free AG and AG niosomes. CONCLUSION: The present results suggest that AG niosomes may have a significant potential of liver targeting, which is valuable in chemotherapy of HCC.

[Application of retroauricular sulcus incision in the operation of benign tumors in the deep lobe of parotid gland].

Objective: To investigate the application of retroauricular groove incision in the resection of benign tumors in the deep lobe of parotid. Methods: From January 2017 to January 2022, 19 patients (11 males and 8 females, age ranged from 17 to 69 years, with a median age of 48) with benign tumor in the deep lobe of parotid gland underwent parotidectomy through retroauricular sulcus incision in Linyi People's Hospital. Among them, 17 cases with tumor diameter≤4.0 cm underwent simple retroauricular groove incision, and 2 cases were dumbbell type with tumor diameter>4.0 cm on the medial side of mandible protruding into the parapharyngeal space, in which the deep lobe and tumor of parotid gland were resected through retroauricular sulcus incision combined with intraoral incision. Results: Tumors were completely removed through retroauricular sulcus incision in 17 cases, and dumbbell type tumors were removed through retroauricular sulcus incision combined with intraoral incision in 2 cases. Postoperative pathological examinations showed pleomorphic adenoma in 13 cases, basal cell adenoma in 4 cases and Warthin's tumor in 2 cases. Temporary mandibular marginal branch paralysis occurred in 2 patients and returned to normal 3 weeks after operation. All incisions healed in Phase I. By following-up of 1-5 years with a median follow-up time of 3.1 years, none of the patients had Frey syndrome, salivary fistula, other complications and tumor recurrence. The patients and their families were satisfied with the postoperative facial appearances. Conclusion: The retroauricular groove approach can not only preserve the function of parotid superficial lobe and facial nerve, but also has less trauma, less tissue defect and hidden scar. As the advantages of less complication, low recurrence rate and good cosmetic effect, the incision is worthy of clinical application.

An indirect ELISA for serodiagnosis of cattle footrot caused by Fusobacterium necrophorum.

A serodiagnostic ELISA (rL-ELISA) using recombinant truncated leukotoxin protein PL2 (aa 311-644) of Fusobacterium necrophorum as antigen was developed for detection of antibodies against F. necrophorum from cattle footrot. In rL-ELISA, the recombinant diagnostic antigen showed no cross-reaction with antisera against bovine foot and mouth disease virus, bovine rhinotracheitis virus, bovine viral diarrhea virus, bovine rotavirus type A, bovine Escherichia coli, and bovine Salmonella. The rL-ELISA could confirm the existence of antibodies against F. necrophorum at day 7 after infection. Detection of the field samples indicated relative sensitivity of rL-ELISA to nL-ELISA using the purified native leukotoxin A as antigen was 96.43%, and relative specificity of rL-ELISA to nL-ELISA was 94.26%. These data demonstrated the rL-ELISA would have a potential use for early diagnosis of cattle footrot caused by F. necrophorum.

[Application of dental floss traction-assisted endoscopic submucosa dissection to rectal neuroendocrine neoplasm].

Objective: To evaluate the safety and efficacy of dental floss traction-assisted endoscopic submucosal dissection (DFS-ESD) for rectal neuroendocrine neoplasm (NEN). Methods: A retrospective cohort study was performed. Clinical data of rectal NEN patients undergoing ESD at Endoscopy Center of Zhongshan Hospital, Fudan University from January 2016 to December 2017 were retrospectively analyzed. Inclusion criteria: 1) age of 18 to 80 years old; 2) maximal diameter of lesions <1.5 cm; 3) tumor locating in the submucosa without invasion into the muscularis propria; 4) no enlarged lymph nodes around bowel and in abdominal cavity; 5) ESD requested actively by patients. A total of 37 patients were enrolled, including 23 male and 14 female cases with mean age of (56.0±11.3) years. All the lesions were single tumor of stage T1, and the mean size was 0.8±0.2(0.5-1.2) cm. Postoperative pathology revealed all samples as neuroendocrine tumors (NET). Seventeen patients received DFS-ESD treatment (DFS-ESD group) and 20 patient received conventional ESD treatment (conventional ESD group). In DFS-ESD group, after the mucosa was partly incised along the marker dots, the endoscopy was extracted, and the dental floss was tied to one arm of the metallic clip. When the endoscope was reinserted, the hemoclip was attached onto the incised mucosa; another hemoclip was attached onto normal mucosa opposite to the lesion in the same way. The submucosa was clearly exposed with the traction of dental floss and the resection could proceed. The conventional ESD group received the traditional ESD operation procedure. The operation time, modified operation time (remaining time after excluding the assembly time of dental floss traction in DFS-ESD group), en bloc resection rate, R0 resection rate, morbidity of operative complication, recurrence and metastasis were compared between two groups. Results: The average tumor size was (0.8±0.2) cm in DFS-ESD group and (0.7±0.2) cm in conventional ESD group (t=0.425, P=0.673). According to postoperative pathological grading of rectal neuroendocrine neoplasm, 13 were G1 and 4 were G2 in DFS-ESD group, while 17 cases were G1 and 3 cases were G2 in conventional ESD group without significant difference (P=0.680). There were no significant differences in baseline data between in the two groups (all P>0.05). All the basal resection margins were negative, the en bloc resection rate was 100% and the R0 resection rate was 100%. Pathological results showed tumor tissue close to the burning margin in 5 cases of conventional ESD group and in 2 cases of DFS-ESD group (P=0.416). The operation time was (17.9±6.6) minutes in conventional ESD group and (14.7±3.3) minutes in DFS-ESD group (t=1.776, P=0.084). The modified operation time of DFS-ESD group was (11.9±2.8) minutes, which was significantly shorter than (17.9±6.6) minutes in conventional ESD group (t=3.425, P=0.002). The hospital stay was (2.3±0.6) days and (2.0±0.5) days in conventional ESD group and DFS-ESD group, respectively, without significant difference (t=1.436, P=0.160). No patient was transferred to surgery, and no delayed bleeding or perforation occurred in either group. There was no recurrence or primary tumor-related death, and all the patients recovered well during a follow-up period of 14(1-24) months. Conclusion: Dental floss traction-assisted ESD for rectal neuroendocrine neoplasm can simplify operation and ensure negative basal margin.

Emergence and whole-genome sequence of Senecavirus A in Colombia.

In 2015 and 2016, Senecavirus A (SVA) emerged as an infectious disease in Brazil, China and the United States (US). In a Colombian commercial swine farm, vesicles on the snout and coronary bands were reported and tested negative for foot-and-mouth disease virus (FMDv), but positive for SVA. The whole-genome phylogenetic analysis indicates the Colombian strain clusters with the strains from the United States, not with the recent SVA strains from Brazil.

Effect of 200 days' sludge retention time on performance of a pilot scale submerged membrane bioreactor for high strength industrial wastewater treatment.

A high strength industrial wastewater was treated using a pilot scale submerged membrane bioreactor (MBR) at a sludge retention time (SRT) of 200 d. The MBR was operated at a high sludge concentration of 20 g/L and a low F/M ratio of 0.11 during 300 d of operation. It was found that the MBR could achieve COD and TOC overall removal efficiencies at more than 99 and 98% TN removal. The turbidity of the permeate was consistently in the range of 0.123 to 0.136 NTU and colour254 absorbance readings varied from 0.0912 to 0.0962 a.u. cm(-1). The sludge concentration was inversely proportional to the hydraulic retention time (HRT), yielded excellent organic removal and extremely low sludge production (0.0016 kgVSS/day).

[Surveillance on drinking-water-born endemic fluorosis in China, 2013].

OBJECTIVE: To investigate the prevalence of fluorosis and related control measures on drinking water type of endemic fluorosis in China. METHODS: According to the national program- "Surveillance Scheme of Drinking-Water-Borne Endemic Fluorosis" , 136 counties were selected in 29 provinces, autonomous regions and municipalities. Three epidemic villages were randomly selected as fixed monitoring sites in each county. Dental fluorosis of all the children aged 8-12 living in the villages under the monitoring program, was identified under the ariteria from "Diagnosis of dental fluorosis" (WS/T 208-2011). Operating conditions and contents of fluoride in all the'water-improved projects' were investigated. Contents of fluoride in drinking water were tested in villages without the 'water-improved projects'. "Standard Test Method for Drinking Water" (GB/T 5750.5-2006) was used to detect the water fluoride. RESULTS: The overall prevalence of dental fluorosis among children aged 8-12 in all the villages under monitor program, was 28.58% (7 950/27 817), with the dental fluorosis index (DFI) as 0.58. Among them, the prevalence was 22.28% (3 917/17 583) and DFI was 0.44 in the'water-improved projects' villages that under normal operation and with qualified fluoride contents. The prevalence appeared as 38.74% (1 926/4 971) with DFI as 0.84 in those villages with 'water-improved projects' but mal-operated or with excessive fluoride. The prevalence was 40.03% (2 107/5 263), and DFI was 0.81 in those villages without 'water-improved projects'. The prevalence rates of dental fluorosis in children from the three types of endemic areas were significantly different. For 'water-improved projects', the normal opration rate was 93.77% (286/305) and the qualification rate of fluoride content was 76.77% (228/297). CONCLUSIONS: Dental fluorosis in children living in the drinking-water-born endemic fluorosis areas was on the edge of epidemics in China. Effective improvement on the quality of drinking water can significantly reduce the severity of dental fluorosis in children. The rate of proper operation on 'water-improved projects' was near to 95% in the endemic area. However, rate that met the criteria on qualified fluoride contents of these projects was still below 80%.

Crystal structures of an N-terminal fragment from Moloney murine leukemia virus reverse transcriptase complexed with nucleic acid: functional implications for template-primer binding to the fingers domain.

Reverse transcriptase (RT) serves as the replicative polymerase for retroviruses by using RNA and DNA-directed DNA polymerase activities coupled with a ribonuclease H activity to synthesize a double-stranded DNA copy of the single-stranded RNA genome. In an effort to obtain detailed structural information about nucleic acid interactions with reverse transcriptase, we have determined crystal structures at 2.3 A resolution of an N-terminal fragment from Moloney murine leukemia virus reverse transcriptase complexed to blunt-ended DNA in three distinct lattices. This fragment includes the fingers and palm domains from Moloney murine leukemia virus reverse transcriptase. We have also determined the crystal structure at 3.0 A resolution of the fragment complexed to DNA with a single-stranded template overhang resembling a template-primer substrate. Protein-DNA interactions, which are nearly identical in each of the three lattices, involve four conserved residues in the fingers domain, Asp114, Arg116, Asn119 and Gly191. DNA atoms involved in the interactions include the 3'-OH group from the primer strand and minor groove base atoms and sugar atoms from the n-2 and n-3 positions of the template strand, where n is the template base that would pair with an incoming nucleotide. The single-stranded template overhang adopts two different conformations in the asymmetric unit interacting with residues in the beta4-beta5 loop (beta3-beta4 in HIV-1 RT). Our fragment-DNA complexes are distinct from previously reported complexes of DNA bound to HIV-1 RT but related in the types of interactions formed between protein and DNA. In addition, the DNA in all of these complexes is bound in the same cleft of the enzyme. Through site-directed mutagenesis, we have substituted residues that are involved in binding DNA in our crystal structures and have characterized the resulting enzymes. We now propose that nucleic acid binding to the fingers domain may play a role in translocation of nucleic acid during processive DNA synthesis and suggest that our complex may represent an intermediate in this process.

Roles of alpha 114 and beta 87 amino acid residues in the polymerization of hemoglobin S: implications for gene therapy.

Three novel recombinant mutants of sickle hemoglobin (Hb S, beta 6Glu-->Val) have been constructed to assess the role of proline at alpha 114 and threonine at beta 87 in the polymerization of deoxygenated Hb S. Using the hemoglobin expression system (pHE2) designed in our laboratory, four plasmids were expressed separately in Escherichia coli to produce the four recombinant hemoglobins: r Hb S (beta 6Glu-->Val); r Hb S-Chiapas (beta 6Glu-->Val, alpha 114Pro-->Arg); r Hb S-D-Ibadan (beta 6Glu-->Val, beta 87Thr-->Lys); and r Hb S-Chiapas-D-Ibadan (beta 6Glu-->Val, alpha 114Pro-->Arg, beta 87Thr-->Lys). The structural features of these four recombinant hemoglobins were analyzed by proton nuclear magnetic resonance spectroscopy, and were found to be similar to those of human normal adult hemoglobin (Hb A) under identical conditions. The recombinant hemoglobins were further investigated by measuring the oxygen-binding properties, which were found to be comparable to those of Hb A. Delay-time gelation studies of the three mutants of r Hb S were carried out in 1.8 M potassium phosphate (pH 7.34) by a temperature jump from 4 degrees C to 30 degrees C and an increase in delay time over that of r Hb S was observed, as well as an overall decrease in the polymerization of these three mutants of Hb S. A more detailed and quantitative investigation has also been carried out to determine the equilibrium solubility (Csat) in 0.1 M potassium phosphate (pH 7.35) at 25 degrees C of the three Hb S mutants as well as of mixtures of these mutants with Hb S versus mixtures of fetal hemoglobin (Hb F) and Hb A with Hb S. The inhibition of polymerization demonstrated in these experiments suggests that the interactions involving the two amino acid residues alpha 114Pro and beta 87Thr are very important to the formation of Hb S polymer, and modification of these amino acids results in an anti-sickling potential. Of particular interest is the inhibitory effect of alpha 114Pro-->Arg, which offers a novel opportunity to use an alpha-chain construct, in addition to a beta-chain construct in the same vector, in gene therapy for sickle cell anemia, with the objective of modifying a larger number of hemoglobin tetramers at a given level of expression.

Removal of humic acid foulant from ultrafiltration membrane surface using photocatalytic oxidation process.

The experimental results indicated that without the TiO2 particles and PCO treatment, the permeate flux of ultrafiltration (UF) membrane declined to 40% of the initial permeate flux after 8 hours filtration. Feeding the humic acid solution with TiO2 particles dosage of 1 g/L with calcium ions into UF membrane, after the same filtration time and PCO reaction at 120 minutes, the permeate flux was increased to about 90% of the initial permeate flux. At longer PCO reaction times, a better water quality of UF permeate was observed. It has been found that with the coexistence of calcium ions in humic acid solution, the smaller molecular fragments of humic acid (HA) generated by PCO reaction may be transferred to the surface of TiO2 by means of adsorption. The humic acid adsorption by TiO2 in the presence of Ca2+ is also pH dependent. The adsorption rates were 21.0, 14.9 and 10.8 ppmTOC/gTiO2 for pH value of 4, 7 and 10 respectively. The combination of effects of PCO mineralization of humic acid into CO2 and adsorption of humic acid by TiO2 through the forming of HA-Ca(2+)-TiO2 aggregate particles were responsible for the removal of humic acid foulant from UF membrane surface.

Simultaneous loss of expression of syndecan-1 and E-cadherin in the embryonic palate during epithelial-mesenchymal transformation.

Epithelial-mesenchymal transformation (EMT) is the key mechanism for fusion and confluence of the rodent palate. During this process, medial edge epithelia (MEE) form a midline seam that subsequently transforms to mesenchymal cells. We studied syndecan-1 and E-cadherin, two molecules which have been shown to promote the epithelial phenotype, to determine their fate during palatal EMT. We found that both syndecan-1 and E-cadherin are expressed on basolateral surfaces of the MEE at day 14. Twelve hours later, when a midline seam has formed, syndecan-1 and E-cadherin are still present on its basal and lateral epithelial surfaces and they persist after the seam breaks up into epithelial islands. Then, expression of both molecules is lost simultaneously and abruptly when EMT occurs. On the contrary, previous in vitro studies of cell lines transfected with antisense cDNAs suggested that loss of syndecan-1 would lead to loss of E-cadherin or vice versa. We conclude that in vivo, synthesis of both E-cadherin and syndecan-1 is downregulated synchronously by the initiation of EMT, leading to an effective and correctly timed conversion of the epithelial cells to mesenchyme.

Mesenchymal dental pulp cells attenuate dentin resorption in homeostasis.

Dentin in permanent teeth rarely undergoes resorption in development, homeostasis, or aging, in contrast to bone that undergoes periodic resorption/remodeling. The authors hypothesized that cells in the mesenchymal compartment of dental pulp attenuate osteoclastogenesis. Mononucleated and adherent cells from donor-matched rat dental pulp (dental pulp cells [DPCs]) and alveolar bone (alveolar bone cells [ABCs]) were isolated and separately cocultured with primary rat splenocytes. Primary splenocytes readily aggregated and formed osteoclast-like cells in chemically defined osteoclastogenesis medium with 20 ng/mL of macrophage colony-stimulating factor (M-CSF) and 50 ng/mL of receptor activator of nuclear factor κB ligand (RANKL). Strikingly, DPCs attenuated osteoclastogenesis when cocultured with primary splenocytes, whereas ABCs slightly but significantly promoted osteoclastogenesis. DPCs yielded ~20-fold lower RANKL expression but >2-fold higher osteoprotegerin (OPG) expression than donor-matched ABCs, yielding a RANKL/OPG ratio of 41:1 (ABCs:DPCs). Vitamin D3 significantly promoted RANKL expression in ABCs and OPG in DPCs. In vivo, rat maxillary incisors were atraumatically extracted (without any tooth fractures), followed by retrograde pulpectomy to remove DPCs and immediate replantation into the extraction sockets to allow repopulation of the surgically treated root canal with periodontal and alveolar bone-derived cells. After 8 wk, multiple dentin/root resorption lacunae were present in root dentin with robust RANKL and OPG expression. There were areas of dentin resoprtion alternating with areas of osteodentin formation in root dentin surface in the observed 8 wk. These findings suggest that DPCs of the mesenchymal compartment have an innate ability to attenuate osteoclastogenesis and that this innate ability may be responsible for the absence of dentin resorption in homeostasis. Mesenchymal attenuation of dentin resorption may have implications in internal resorption in the root canal, pulp/dentin regeneration, and root resorption in orthodontic tooth movement.

Salivary antibodies to human immunodeficiency virus type 1 in a phase I AIDS vaccine trial.

HIV-1-specific secretory antibodies may be a desirable outcome in individuals receiving AIDS vaccines. We investigated parotid and whole saliva samples for HIV-specific antibodies collected from five volunteers who received a recombinant HIV-1 envelope glycoprotein (rgp160) vaccine. Ten healthy, adult volunteers received intramuscularly either three doses of rgp160 (40 or 80 micrograms), a hepatitis B vaccine, or a placebo on days 0, 30, and 180. Saliva samples were collected on days 0, 28, 60, 120, 194, and 270 from the volunteers. All volunteers were negative for serum HIV antibodies by ELISA (Abbott). By Western blotting, serum antibodies to envelope antigens were demonstrated in one of three individuals who received the low dose vaccine and two of two who received the high dose. Antibodies to gp160 were detected in whole saliva on day 194 from one of these individuals by Western blotting. Parotid saliva collected on all dates did not contain detectable HIV-specific antibodies. The finding of HIV-1-specific antibodies in whole saliva following vaccination may indicate that development of mucosal immunity is possible.

Variation of secretory antibodies in parotid saliva to human immunodeficiency virus type 1 with HIV-1 disease stage.

The secretory immune response to pathogens of the gut-associated lymphoid tissue is often independent of the systemic response. We investigated and compared the presence of antibodies to human immunodeficiency virus type 1 (HIV-1) antigens in parotid saliva and serum by Western blotting in 22 HIV-1-infected individuals. Antibodies to the HIV-1 envelope antigen gp160 were detected in saliva samples from 21 of 22 individuals and in serum from all individuals who were classified as CDC Group II, III, or IV. Antibody titers to gp160 were approximately 3000 times higher in serum than in saliva. Antibodies to viral core antigen p24 were detected in 6 of 7 Group II individuals in saliva and in 7 of 7 in serum. Antibodies to p24 were not found in the parotid saliva, but were detected in the sera of 3 of 3 Group III and 11 of 12 Group IV patients. The absence of secretory antibodies to HIV-1 core antigen p24 was correlated with CD4+ cell counts of less than 200/mm3. The results suggest that loss of secretory anti-p24 antibodies may be an early sign of progression to higher CDC clinical stages in HIV-1-infected individuals.

Inhibition of HIV replication by liposomal encapsulated amphotericin B.

This report shows the potential of using a liposomal encapsulated preparation of amphotericin B (a polyene macrolide antibiotic) for the in vitro inhibition of HIV. There was no significant difference between the effective doses of the free form of drug when compared to the liposomal encapsulated preparation in inhibiting the growth of HIV. Virus expression was suppressed at a concentration of 5-10 micrograms/ml of the drugs. The liposomal preparation showed greatly reduced cytotoxicity in experiments using cultures of murine leukocytes. These results show the potential usefulness of liposomal encapsulated drugs in the treatment of patients with AIDS or AIDS related complex.

In vitro reaction to orthopaedic biomaterials by macrophages and lymphocytes isolated from patients undergoing revision surgery.

Periprosthetic tissues observed at sites of loose total joint implants exhibit abundant macrophages, lymphocytes, fibroblasts and particulate debris. Macrophages phagocytose orthopaedic debris and release proinflammatory cytokines, chemokines, matrix metalloproteinases and other substances. In addition, other cell types present in tissues harvested from the bone-implant interface are thought to influence periprosthetic bone resorption. The present study examined the effects of polymethylmethacrylate (PMMA), cobalt chrome molybdenum alloy (CoCr), and titanium-alloy particle challenge on macrophages co-cultured with lymphocytes in vitro. Potential synergistic effects of lymphocytes on macrophage activation were determined by measuring interleukin-6 and tumor necrosis factor-alpha release following exposure to orthopaedic biomaterial particles. Exposure of macrophages or macrophages co-cultured with lymphocytes to all three types of particles resulted in increased release of interleukin-6 and tumor necrosis factor-alpha at 48 h, when compared to macrophages or macrophages co-cultured with lymphocytes, respectively, cultured in the absence of particles. Lymphocytes isolated from periprosthetic tissues secreted increased basal levels of cytokines relative to peripheral blood lymphocytes. Higher doses of PMMA and titanium-alloy particles stimulated increased levels of cytokine release in the macrophage and macrophage/lymphocyte groups. In contrast, a higher dose of CoCr particles (0.075% v/v) was not as effective as the 0.015% v/v dose, indicating probable CoCr toxicity. The macrophage/lymphocyte co-culture did not show synergism between the two types of cells with respect to cytokine release. T-cells at the bone-implant interface may alter the biological response to particulate debris.

Interleukin-10 inhibits polymethylmethacrylate particle induced interleukin-6 and tumor necrosis factor-alpha release by human monocyte/macrophages in vitro.

Periprosthetic membranes commonly observed at sites of total joint implant loosening exhibit abundant macrophages and particulate debris. Macrophages phagocytose orthopedic debris and release the pro-inflammatory mediators interleukin-1, interleukin-6, tumor necrosis factor-alpha, and prostaglandin E2. Populations of activated lymphocytes are often seen in periprosthetic membranes. These lymphocytes may modulate the monocyte/macrophage response to particulate debris and influence aseptic loosening. In addition, other immunologic agents, such as interleukin-10, are present in tissues harvested from the bone-implant interface of failed total joint arthroplasties. The present study examined the effects of interleukin-10 on polymethylmethacrylate (PMMA) particle challenged human monocyte/macrophages in vitro. Human monocyte/macrophages isolated from buffy coats of five healthy individuals were exposed to 1-10 microm PMMA particles. Interleukin-10 was added to the monocyte/macrophages with and without the addition of PMMA particles. Interleukin-10-induced alterations in monocyte/macrophage metabolism were determined measuring interleukin-6 and tumor necrosis factor-alpha release by the cells following exposure to PMMA particles. Exposure of the monocyte/macrophages to PMMA particles resulted in a dose-dependent release of interleukin-6 and tumor necrosis factor-alpha at 48 h. Interleukin-10 reduced the levels of interleukin-6 and tumor necrosis factor-alpha release by macrophages in response to PMMA particles in a dose-dependent manner. At 48 h, particle-induced interleukin-6 release was inhibited by 60 and 90% with 1.0 and 10.0 ng/ml treatments of interleukin-10, respectively. At 48 h, particle-induced tumor necrosis factor-alpha release was inhibited by 58 and 88% with 1.0 and 10.0 ng/ml treatments of interleukin-10, respectively. Interleukin-10 challenge alone did not significantly alter basal interleukin-6 or tumor necrosis factor-alpha release relative to control cultures. The data presented in this study demonstrate that the anti-inflammatory cytokine, interleukin-10, inhibits monocyte/macrophage release of the pro-inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha in response to PMMA particle challenge in vitro.

Interleukin-4 inhibits granulocyte-macrophage colony-stimulating factor, interleukin-6, and tumor necrosis factor-alpha expression by human monocytes in response to polymethylmethacrylate particle challenge in vitro.

The outcome of total joint arthroplasty is determined by biological events at the bone-implant interface. Macrophages phagocytose implant or wear debris at the interface and release proinflammatory mediators such as interleukins 1 and 6, tumor necrosis factor-alpha, and prostaglandin E2. These mediators are thought to contribute to the resorption of periprosthetic bone. Previous studies of tissues harvested from the bone-implant interface of failed orthopaedic implants demonstrated a possible role for two other cytokines, granulocyte-macrophage colony-stimulating factor and interleukin-4. The present study examined the effects of in vitro challenge with polymethylmethacrylate particles on the expression of granulocyte-macrophage colony-stimulating factor by primary human monocytes/macrophages and the role of interleukin-4 in regulating this expression. The polymethylmethacrylate particles caused a dose-dependent release of granulocyte-macrophage colony-stimulating factor at 48 hours. This release was accompanied by increased expression of interleukins 6 and 1beta and tumor necrosis factor-alpha. Release of the lysosomal enzyme hexosaminidase also increased in response to the particles. Interleukin-4 inhibited the expression of granulocyte-macrophage colony-stimulating factor, interleukin-6, and tumor necrosis factor-alpha at 48 hours in a dose-dependent manner. The data presented in this study confirm the hypothesis that interleukin-4 downregulates particle-induced activation of macrophages, as demonstrated by the decreased release of proinflammatory mediators.