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This study aimed to compare the effectiveness, in terms of viability and quality post‐warming, when vitrifying in vitro‐produced (IVP) pig blastocysts with either a modified Cryotop (n = 161; 20 blastocysts/device) or the conventional Superfine Open Pulled Straw (SOPS; n=160; 5‐6 blastocysts/device systems. Blastocyst viability, morphology, and apoptosis parameters were evaluated after 24 h of in vitro culture. The Cryotop system yields better results in terms of higher embryo viability and total cell numbers (p <.05) and lower apoptosis (p <.05) parameters than the SOPS procedure, defining a high effectiveness to simultaneously vitrify 20 pig IVP blastocysts at one time, thus increasing the yield of IVP blastocysts readily available for embryo transfer. [ABSTRACT FROM AUTHOR]

Contents This study evaluated the effect of three reversible meiotic inhibitors ( MINs) and their interaction with gonadotrophins (Gns) on the meiotic maturation and developmental competence of porcine oocytes. In experiment 1, the oocytes were matured for 22 hr in the presence or absence of dbc AMP (1 mM), cycloheximide (7 μM) or cilostamide (20 μM) with or without Gns, and for an additional 22 hr in the absence of MINs and Gns. At 22 hr of maturation, regardless of the presence of Gns, a higher proportion ( p < .001) of oocytes cultured in the presence of MINs were effectively arrested at the germinal vesicle stage compared with the oocytes cultured without MINs. At 44 hr of maturation, the proportion of oocytes that reached MII was higher ( p < .05) in groups with Gns compared with groups without Gns. In experiment 2, oocytes that were matured as in experiment 1 were inseminated and cultured for 7 days to evaluate fertilization parameters and blastocyst formation. Only oocytes from the dbc AMP + Gns group had higher ( p < .05) efficiency of fertilization compared with the other treatment groups. The presence of dbc AMP during maturation also increased ( p < .05) blastocyst formation and efficiency of blastocyst formation in both the presence and absence of Gns. These results indicate that the interaction of Gns with the tested MINs improved meiotic progression. In addition, regardless of supplementation with Gns, the presence of dbc AMP during the first maturation period increased and even doubled the capacity of oocytes to develop to the blastocyst stage. [ABSTRACT FROM AUTHOR]

Contents More than eighteen years have passed since the first derivation of human embryonic stem cells ( ESCs), but their clinical use is still met with several challenges, such as ethical concerns regarding the need of human embryos, tissue rejection after transplantation and tumour formation. The generation of human induced pluripotent stem cells ( iPSCs) enables the access to patient-derived pluripotent stem cells ( PSCs) and opens the door for personalized medicine as tissues/organs can potentially be generated from the same genetic background as the patient recipients, thus avoiding immune rejections or complication of immunosuppression strategies. In this regard, successful replacement, or augmentation, of the function of damaged tissue by patient-derived differentiated stem cells provides a promising cell replacement therapy for many devastating human diseases. Although human iPSCs can proliferate unlimitedly in culture and harbour the potential to generate all cell types in the adult body, currently, the functionality of differentiated cells is limited. An alternative strategy to realize the full potential of human iPSC for regenerative medicine is the in vivo tissue generation in large animal species via interspecies blastocyst complementation. As this technology is still in its infancy and there remains more questions than answers, thus in this review, we mainly focus the discussion on the conceptual framework, the emerging technologies and recent advances involved with interspecies blastocyst complementation, and will refer the readers to other more in-depth reviews on dynamic pluripotent stem cell states, genome editing and interspecies chimeras. Likewise, other emerging alternatives to combat the growing shortage of human organs, such as xenotransplantation or tissue engineering, topics that has been extensively reviewed, will not be covered here. [ABSTRACT FROM AUTHOR]

Contents With the development of the non-surgical deep uterine (Ns DU) embryo transfer ( ET) technology, the commercial applicability of ET in pigs is now possible. There are, nevertheless, many factors that influence Ns DU- ET effectiveness that need to be addressed. The aim of this study was to evaluate the effects of the weaned recipients' parity on fertility and prolificacy following Ns DU- ET. The recipients (n = 120) were selected based on their reproductive history and body condition and grouped into three categories according to their parity: primiparous sows, sows of parity 2 and sows of parities from 3 to 5. Thirty fresh embryos (morulae and unhatched blastocysts) were non-surgically transferred into one uterine horn of each recipient. It was possible to insert the Ns DU- ET catheter through the cervix along a uterine horn in 98.3% of the recipients. The parity had no influence on the difficulty grade of the insertions or on the percentage of correct insertions. The cervix and uterine wall were not perforated during the insertions, and vaginal discharge was not observed after transfer in any of the recipients. There were no differences in the pregnancy rates (74.8%), farrowing rates (71.2%) or litter sizes (9.6 ± 3.3) between groups. Also, there were no differences between groups regarding to the piglets' birthweights or piglet production efficiency. In conclusion, these results demonstrate that weaned sows from parity 1 to 5 are appropriate to be used as recipients in Ns DU- ET programs, which increase the possibilities for the utilization of ET in the recipient farms. [ABSTRACT FROM AUTHOR]

Contents The objective of this study was to optimize protocols for the cryopreservation of sex-sorted boar spermatozoa. In the experiment 1, we evaluated the effects of a standard boar sperm cryopreservation procedure (3% final glycerol concentration) on the in vitro characteristics of sex-sorted sperm frozen at low sperm concentrations (20 × 106 sperm/ml; S20 group). Non-sorted spermatozoa frozen at 1000 × 106 (C1000 group) and 20 × 106 (C20 group) sperm/ml were used as the freezing control groups. In experiment 2, the effects of different final glycerol concentrations (0.16%, 0.5%, 1.0%, 2.0% and 3.0%) on post-thaw quality of the S20 and C20 groups were evaluated. In both experiments, the samples were evaluated prior to freezing (5°C) and at 30, 90 and 150 min after thawing. Experiment 1 indicated that freezing sperm at low concentrations decreased (p < 0.05) the total motility (TM) and progressive motility (PM) at 90 and 150 min after thawing regardless of whether the sperm were sorted or not. However, the sperm membrane integrity was not affected at any evaluation step. Inexperiment 2, significant effects on the TM and PM because of increased glycerol concentrations in the S20 and C20 groups were observed only at 90 and 150 min after thawing. The samples frozen in 3% glycerol showed lower (p < 0.05) TM and PM values when compared to those frozen in the presence of 0.5% and 1% glycerol. In both experiments, non-sorted control samples displayed higher percentages of spermatozoa with damaged DNA than sorted spermatozoa. In conclusion, the optimization of cryopreservation conditions by decreasing the glycerol concentrations can improve post-thaw motility of sex-sorted spermatozoa frozen at low concentrations. [ABSTRACT FROM AUTHOR]

Content Complement component 3 (C3) has well-established roles within immune system, but its roles outside of immune system are less characterized. The extensive presence of C3 throughout the female reproductive tract, and its temporal, and gamete-specific regulation of expression suggest a potential role for C3 in reproduction. In the present investigation, the effects of C3, C3b and iC3b on porcine oocyte maturation, fertilization and embryonic development were examined. We identified the ability of iC3b to positively influence oocyte maturation. No effects on fertilization efficiency, penetration rates, polyspermy and blastocyst formation were observed. However, C3, C3b and iC3b presence in embryo culture medium resulted in fewer total cells in test blastocysts compared to control blastocysts. The results of this study indicate a potential function for iC3b in oocyte maturation. Furthermore, it was demonstrated that the presence of either C3, C3b or iC3b has a negative influence on early embryonic development in the porcine species. [ABSTRACT FROM AUTHOR]

Contents The current cervical artificial insemination (CAI) procedure, involving deposition of excessive sperm numbers, is uneconomical for pig industry. The most obvious alternative requires uterine deposition in combination with fixed-time AI, which would reduce the number of sperm required per pregnant sow, thus allowing the best use of valuable boars and, ultimately, the commercial integration of frozen-thawed and sexed sperm. This review depicts possible best ways to implement an efficient use of liquid-stored, frozen-thawed and sexed sperm by the pig industry. [ABSTRACT FROM AUTHOR]

Contents Recent advances in new technologies to produce cloned and genetically modified pigs involve manipulating oocytes and/or embryos in vitro. Although a great deal of progress has been made, the current IVM–IVF systems still result in major problems: a high rate of polyspermy; and a low development rate and low quality of blastocysts for in vitro compared with the in vivo–produced embryos. This study summarizes recent advancements in IVM–IVF–IVC porcine systems. Recent methods to select monospermic embryos are also discussed. Finally, achievements in vitrification and in somatic cell nuclear transfer are discussed. [ABSTRACT FROM AUTHOR]

Contents The present study was designed to evaluate three different in vitro fertilization (IVF) systems: a straw-IVF system with 10 min of coincubation, a straw-IVF system with 6-h coincubation and the microdrop-IVF system with 6-h coincubation (the traditional IVF system used routinely in most of IVF laboratories) in an attempt to reduce polyspermic penetration ( Experiment 1 ). When the straw-IVF system was tested in combination with two coincubation times, the use of 10 min of coincubation significantly increased (p < 0.001) the penetration rate and the efficiency of fertilization (67.7 ± 6.4% vs 31.9 ± 6.5% and 41.5 ± 2.5% vs 17.6 ± 2.5% for 10 min and 6 h, respectively), while there were no significant differences in the incidence of monospermy between both systems (64.3 ± 5.1% and 67.7 ± 3.4%, for 10 min and 6 h, respectively). The penetration rate in the 6-h microdrop-IVF system was higher (93.8 ± 3.6%; p < 0.001) compared with the 10-min straw-IVF system (67.7 ± 6.4%), however, monospermy was severely reduced (25.0 ± 4.3% vs 67.7 ± 3.4%, for the 6-h microdrop-IVF system and 10-min straw-IVF system, respectively). The efficiency of the IVF showed similar values between microdrop and 6-h straw-IVF systems, but efficiency was significantly improved (p < 0.05) when the 10-min straw-IVF system was used. Experiment 2 was designed to compare porcine in vitro embryo production in two IVF systems, the 6-h microdrop-IVF system (1000 sperm per oocyte) and 10-min straw-IVF system (30 000 sperm per oocyte). The blastocyst formation rates tended (p = 0.06) to be higher when the 10-min straw-IVF system was used compared with the 6-h microdrop-IVF system. In addition, the number of total cells per blastocyst increased significantly (p < 0.05) in the 10-min straw-IVF system. These results showed that the 10-min straw-IVF system is an effective way to decrease polyspermic penetration, and improve the efficiency of fertilization and the quality of blastocysts in terms of cell number per embryo. [ABSTRACT FROM AUTHOR]

Contents The sorting of X- and Y-chromosome-bearing spermatozoa by flow cytometry is nowadays one of the most apt assisted-reproduction technologies in livestock production. Potential economic and biological benefits, as well as those related to easier management of herds, have been reported arising out of the application of this technique, especially in cattle. Yet, the sex-sorting procedure induces damage to spermatozoa, affecting their function and fertilizing ability. Different species present varying degrees of susceptibility to damage from the sorting process and each has its own requirements for sex-sorted insemination procedures. Thus, several new protocols and strategies have been designed for the handling of sorted spermatozoa, with the main objective of optimizing their fertilizing ability and the consequent application of flow-cytometric sex-sorting technology. This article reviews current advances in this technology, pointing out the components to be improved before this technology may be widely applied in different domestic species. [ABSTRACT FROM AUTHOR]

Contents Low-dose AI procedures are required by the pig industry to efficiently utilize emerging sperm technologies, such as cryopreservation and sex-sorting. Currently, several different procedures for inseminating with a low or very low number of spermatozoa have been described. Deep intrauterine insemination allows the deposition of the spermatozoa in the depth of the uterine horn, allowing a significant reduction in the number of spermatozoa inseminated with maintenance of optimal reproductive performance. Intra-oviductal laparoscopic insemination has been recently applied in pigs. This technique has proved to be applicable with diluted and sex-sorted spermatozoa. This review discusses several problems encountered during the development of deep intrauterine insemination and intra-oviductal laparoscopic insemination of pigs and provides potential solutions for the practical application of both the technologies. [ABSTRACT FROM AUTHOR]

Contents Semen extended in a liquid state, together with conventional artificial insemination (AI), is the sole sperm technology used by the pig industry. Sperm technologies known for many years, such as cryopreservation, and others developed during recent years, such as sperm sexing, have not yet been integrated into commercial use. Moreover, there has recently been an explosion of new technologies, such as sperm mediated gene transfer or encapsulated spermatozoa which need additional supportive techniques before they can be economically applied to pig breeding. The speed with which the aforementioned sperm technologies are accepted and utilized by the pig industry depends on the availability of efficient insemination procedures. Therefore, AI is entering a new dimension where it will be converted into a tool for the efficient application of current and new sperm technologies. Some new insemination procedures have been recently developed. This review examines the suitability of available insemination procedures for the efficient application of current, emergent and future sperm technology to the pig industry. [ABSTRACT FROM AUTHOR]

Contents A new procedure for non-surgical deep intrauterine insemination (DUI) in unrestrained sows hormonally induced to ovulate, has been reported. In comparison with standard artificial insemination (AI), with this procedure, the sperm numbers inseminated can be reduced 20-fold without reducing the reproductive performance of these hormonally treated sows. The present study evaluated, using two experiments, the reproductive performance applying 20-fold different sperm numbers per AI dose using DUI or standard AI in spontaneously ovulating sows, under field conditions. In experiment 1, AI was applied to crossbred sows at 12, 24 and 36 h after onset of spontaneous oestrus using one of the following two regimes: (i) DUI (treatment) with 0.15 × 109 fresh boar spermatozoa in 5 ml of Beltsville thawing solution (BTS) extender (n = 95), and (ii) standard cervical AI (control) with 2.85 × 109 fresh spermatozoa in 95 ml of BTS extender (n = 95). The farrowing rates of the two groups of sows were statistically similar (NS). However, a decrease (p < 0.002) in litter size and the total number of pigs born alive was observed in sows inseminated with the DUI procedure. In experiment 2, 42 post-weaned oestrus sows were inseminated following the same design described for experiment 1 during spontaneous oestrus. On day 6 after onset of oestrus, the proximal segment of the uterine horns of the sows were flushed under surgery to retrieve eventual embryos and evaluate the success of fertilization per cornua (e.g. occurrence of effective uni- vs bilateral sperm transport rendering uni- or bilateral, complete or partial fertilization). Retrieved embryos were assessed for cleavage and number of accessory spermatozoa. Although identical overall pregnancy rates were achieved in both insemination groups, the percentage of sows with partial bilateral fertilization and unilateral fertilization was markedly higher (p < 0.05) in the DUI group (35%) compared with the control (standard AI) group (5%), with a consequent lower (p < 0.001) percentage of viable early embryos after DUI. The number of accessory spermatozoa in the zona pellucida of the embryos was highly variable, but higher (p < 0.001) in control animals than in DUI-AI. No accessory spermatozoa were found in oocytes retrieved from sows depicting unilateral fertilization. In conclusion, DUI in spontaneously ovulating sows with 0.15 × 109 spermatozoa renders similar farrowing rates but a lower litter size compared with use of standard AI with a 20-fold higher sperm dose. The lower litter size ought to be related to a decreased distribution of spermatozoa after DUI leading to a higher incidence of partial bilateral and unilateral fertilization. [ABSTRACT FROM AUTHOR]

Over the past decade, there has been an increase in the development and/or in the improvement of emerging reproductive technologies in pigs. Among emerging reproductive technologies with potential short-term application in pig production are: artificial insemination with low number of spermatozoa, cryopreservation of spermatozoa and embryos, sperm sexing, and non-surgical embryo transfer. The following review will give emphasis to recent advancements in these reproductive technologies that are starting to show possibilities of serious applications under field conditions. [ABSTRACT FROM AUTHOR]

Sperm handling, associated to artificial reproduction technologies (ART) such asin vitrofertilization (IVF) or the use of flow cytometry for cell analysis or sorting imposes volumetric extension of the sperm suspension and decreases sperm viability, presumably because of the removal of seminal plasma (SP) components. This study evaluated whether a 10% v/v of autologous SP (retrieved from the same donor boar) or homologous SP (e.g. from any of the four fertile boars included, other than the one providing the spermatozoa) would differently affect the viability of boar spermatozoa subjected to large extension in a simple saline medium [phosphate-buffered saline and 0.1% ethylenediaminetetraacetic acid (EDTA), PBSm] to a concentration of 0.3 × 106 spermatozoa/ml and incubated for 2 h at 30°C. Sperm viability was monitored as membrane integrity [using the fluorophore carboxyfluorescein diacetate (C-FDA) and propidium iodide (PI)], mitochondrial function (using the fluorophore R-123) and motility characteristics [using Computer Assisted Sperm Analysis (CASA)]. Substraction of the SP and extension followed by incubation in PBSm significantly (p < 0.05) decreased sperm viability, which could be restored by addition of autologous SP. Furthermore, exposure of the extended spermatozoa to homologous SP (from any other individual boar) significantly (p < 0.05) varied with the source of the sire; some boars exerting beneficial effects (even surpassing the effects of the autologous SP; p < 0.05) while at least one boar negatively (p < 0.05) influencing the viability of the incubated spermatozoa. It is concluded that SP should be present when incubating highly extended spermatozoa. As a result of the obvious differences among boars, it would be advantageous to examine the ability of SP to maintain sperm viability prior to the use of SP pools during sperm handlingin vitro. [ABSTRACT FROM AUTHOR]

Contents Unilateral enlargement of left testicle and scrotum was detected in an 8-year-old West Highland White Terrier. The histopathological diagnosis after surgery was a seminoma (SEM) tumour, and a diagnosis of metastatic foci was also detected in vaginal tunic and scrotum. Two months later, new metastatic SEM foci in the skin were diagnosed. Twenty-two months after the initial orchiectomy new multiple cutaneous nodules and a swelling of periesophageal structures were observed. Finally, the necropsy revealed multiple malignant metastatic SEM focus. To the author's knowledge, this is the first description of a canine SEM with unusual widespread metastasis on the base of tongue, soft palate, trachea and pericardium. [ABSTRACT FROM AUTHOR]

This study was conducted to evaluate the combined effect of a short period of gamete coincubation and a new in vitro fertilization (IVF) system in straw (Li et al., 2003; Biol. Reprod. 69: 1580) to attempt to reduce polyspermic penetration. A total of 723 in vitro matured oocytes were inseminated with thawed spermatozoa. Various sperm concentrations (20 000, 30 000, 40 000, and 50 000 sperm/oocyte) and two coincubation times (6 h and 10 min) were used for straw-IVF and compared to control group (1000 sperm/ovocito, in microdrop). The oocytes from 10 min group were washed in mTBM medium to remove spermatozoa not bound to the zona and transferred to the same medium (containing no sperm) for 6 h. After 6 h, oocytes from each group were cultured in NCSU-23 medium for 12–15 h to assess fertilization parameters. The sperm concentration did not affect the efficiency of the IVF in both periods of coincubation. The short coincubation time significantly (p < 0.001) increased penetration rate and efficiency of the IVF (67.7 ± 6.4% vs 31.19 ± 6.5% and 41.5 ± 2.5% vs 17.6 ± 2.5% for 10 min and 6 h, respectively), while there were no significant differences in the incidence of monospermy between 10 min (64.3 ± 5.1) and 6 h (67.7 ± 3.4%) coincubation times. The penetration rate in the control group was higher (95.5 ± 5.6%; p < 0.001) than in the straw groups, but monospermy was severely reduced (25.0 ± 4.3%). These results show that short coincubation time together with the straw IVF system improves the efficiency of the porcine IVF. (Supported by SENECA). [ABSTRACT FROM AUTHOR]

Background: Currently, high polyspermy remains a significant obstacle to achieving optimal efficiency in in vitro fertilization (IVF) and in vitro embryo production (IVP) systems in pigs. Developing strategies that would prevent polyspermy is essential in overcoming this challenge and maximizing the potential of this reproductive biotechnology. Previous results have demonstrated that using boar spermatozoa subjected to a high-extension and reconcentration procedure and then cryopreserved resulted in significant improvements in IVF/IVP systems with high rates of monospermy and penetration., Objective: The aim of the present study was to unveil the molecular mechanisms that may underlie the changes in fertilization patterns exhibited by highly extended and cryopreserved boar spermatozoa., Materials and Methods: To achieve this goal, we used quantitative proteomic analysis (LC‒ESI‒MS/MS SWATH) to identify differentially abundant proteins (DAPs) between highly extended (HE) and conventionally (control; CT) cryopreserved boar spermatozoa. Prior to the analysis, we evaluated the in vitro post-thawing fertilizing ability of the sperm samples. The results demonstrated a remarkable improvement in monospermy and IVF efficiency when using HE spermatozoa in IVF compared with CT spermatozoa., Results: At the proteomic level, the combination of high-extension and cryopreservation had a significant impact on the frozen-thawed sperm proteome. A total of 45 proteins (24 downregulated and 21 upregulated) were identified as DAPs (FC > 1 or ≤1; p < 0.05) when compared with CT spermatozoa. Some of these proteins were primarily linked to metabolic processes and the structural composition of sperm cells. The dysregulation of these proteins may have a direct or indirect effect on essential sperm functions and significantly affect spermatozoa-oocyte interaction and, therefore, the sperm fertilization profile under in vitro conditions. While these findings are promising, further research is necessary to comprehend how the disturbance of specific proteins affects sperm fertilization ability., (© 2023 The Authors. Andrology published by Wiley Periodicals LLC on behalf of American Society of Andrology and European Academy of Andrology.)

Aim: To evaluate the efficacy and harms of a short (7-10 days) compared with a standard (10-14 days) duration of antibiotics in culture-proven neonatal sepsis for reducing all-cause mortality, treatment failure and duration of hospitalisation., Methods: Medline, EMBASE and Cochrane CENTRAL were searched for randomised trials., Results: We included five studies, all conducted in India (447 infants with a gestational age greater than 32 weeks). Except for one study, all studies were at high risk of bias. All-cause mortality was reported in three studies with only one death reported in the standard duration regimen arm (243 patients, very low certainty). A meta-analysis showed no evidence of the effect on treatment failure (RR of 1.47 [95% CI 0.48-4.50], 440 patients, five studies, very low certainty) of short-term antibiotics. Short-term antibiotic regimen shortened the duration of hospitalisation by 4 days (mean difference of -4.04 days [95% CI -5.47 to -2.61]; 4 studies; 371 patients; very low certainty)., Conclusion: Among studies focused on infants born with a gestational age greater than 32 weeks, short-term administration of antibiotics may shorten the duration of hospitalisation, but the evidence is very uncertain. The evidence on other predefined outcomes is very uncertain to draw definite conclusions., (© 2023 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.)

Background: The wide use of topical and systemic antifungal agents as the conventional treatment for oral candidiasis has caused Candida albicans to develop resistance to these agents. The aims of this study were to evaluate the effects of photodynamic therapy (PDT) and chitosan on buccal candidiasis and study the possible enhancive effect of chitosan on the photosensitizer methylene blue., Methods: Thirty-five DBA/2 immunosuppressed mice were orally inoculated with a suspension of Candida albicans. The animals were randomized into seven groups (n = 5 per group): Group 1 (control); Group 2 (nystatin); Group 3 (PDT); Group 4 (chitosan 1.5 mg/ml); Group 5 (chitosan 3 mg/ml); Group 6 (PDT + chitosan 1.5 mg/ml); and Group 7 (PDT + chitosan 3 mg/ml). The Candida albicans count was evaluated on days 3, 5, 7, and 11 after inoculation. At last, macroscopic and microscopic analyses of the tongue dorsa were performed., Results: On day 7 after inoculation, the control group showed a greater number of Candida albicans (5.25 ± 0.41 log10 CFU/ml), with significant differences compared to all other groups (P ≤ 0.05). On day 11 after inoculation, animals treated with PDT showed lower CFU/ml count. Groups 2, 3, 4, 5, and 6 showed fewer microscopic candidiasis lesions than Groups 1 and 7., Conclusions: PDT has an antifungal effect, even greater than nystatin. Chitosan has a powerful fungicide effect but did not possess any enhancive effect on methylene blue., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: To investigate the effects of intraarticular glucocorticoid treatment on macrophage infiltration, the expression of the chemokines monocyte chemoattractant protein 1 (MCP-1) and macrophage inflammatory protein 1alpha (MIP-1alpha), and the expression of matrix metalloproteinases 1 and 3 (MMPs 1 and 3) and their inhibitors, the tissue inhibitors of metalloproteinases 1 and 2 (TIMPs 1 and 2), in osteoarthritis (OA) synovial membranes., Methods: Forty patients underwent arthroscopic biopsy before and 1 month after intraarticular injection of glucocorticoids. Twenty-one patients received 120 mg of methylprednisolone acetate (Depo-Medrol; Upjohn, Kalamazoo, MI), and 20 patients received placebo (1 patient received placebo in 1 knee and methylprednisolone acetate in the other). Immunoperoxidase staining for the expression of CD68, MCP-1, MIP-1alpha, MMP-1, MMP-3, TIMP-1, and TIMP-2 was performed, and the immunostaining was quantified by color video image analysis., Results: CD68, MCP-1, MIP-1alpha, MMP-1, MMP-3, TIMP-1, and TIMP-2 immunostaining was observed in all synovial membranes. Intraarticular glucocorticoid treatment was associated with a small (30%) but statistically significant (P = 0.048) reduction in CD68+ macrophage staining in the synovial lining layer, but there was no change in the CD68 expression in the synovial sublining layer. No significant differences were observed for MCP-1, MIP-1alpha, MMP-1, MMP-3, TIMP-1, and TIMP-2 immunostaining in the synovial lining or sublining layers., Conclusion: Intraarticular glucocorticoids may reduce CD68+ macrophage infiltration into the synovial lining layer, but not the expression of MCP-1, MIP-1alpha, MMP-1, MMP-3, TIMP-1, and TIMP-2 in the synovial membrane, in patients with OA.