Autologous platelet-rich plasma effects on Staphylococcus aureus–induced chondrocyte death in an in vitro bovine septic arthritis model

Andrew J.T. Muir ◽  
Andrew J. Niehaus ◽  
Joseph W. Lozier ◽  
Sara L. Cole ◽  
Zarah A. Belacic ◽  

Abstract OBJECTIVE To investigate the chondroprotective effects of autologous platelet-rich plasma (PRP), ampicillin-sulbactam (AmpS), or PRP combined with AmpS (PRP+AmpS) in an in vitro chondrocyte explant model of bovine Staphylococcus aureus–induced septic arthritis. SAMPLE Autologous PRP and cartilage explants obtained from 6 healthy, adult, nonlactating Jersey-crossbred cows. ProcedureS Autologous PRP was prepared prior to euthanasia using an optimized double centrifugation protocol. Cartilage explants collected from grossly normal stifle joints were incubated in synovial fluid (SF) alone, S aureus–inoculated SF (SA), or SA supplemented with PRP (25% culture medium volume), AmpS (2 mg/mL), or both PRP (25% culture medium volume) and AmpS (2 mg/mL; PRP+AmpS) for 24 hours. The metabolic activity, percentage of dead cells, and glycosaminoglycan content of cartilage explants were measured with a resazurin-based assay, live-dead cell staining, and dimethylmethylene blue assay, respectively. Treatment effects were assessed relative to the findings for cartilage explants incubated in SF alone. RESULTS Application of PRP, AmpS, and PRP+AmpS treatments significantly reduced S aureus–induced chondrocyte death (ie, increased metabolic activity and cell viability staining) in cartilage explants, compared with untreated controls. There were no significant differences in chondrocyte death among explants treated with PRP, AmpS, or PRP+AmpS. CLINICAL RELEVANCE In this in vitro explant model of S aureus–induced septic arthritis, PRP, AmpS, and PRP+AmpS treatments mitigated chondrocyte death. Additional work to confirm the efficacy of PRP with bacteria commonly associated with clinical septic arthritis in cattle as well as in vivo evaluation is warranted.

2018 ◽  
Vol 8 (1) ◽  
pp. 1 ◽  
Konstantinos Sfakianoudis ◽  
Mara Simopoulou ◽  
Nikolaos Nitsos ◽  
Anna Rapani ◽  
Athanasios Pappas ◽  

This report presents the case of a woman aged 40 who has experienced premature menopause from the age of 35. Having rejected oocyte donation, she opted for intraovarian injection of autologous platelet-rich plasma with the aim to rejuvenate the ovarian tissue and enable the employment of her own gametes through in-vitro fertilization. Six weeks following the autologous platelet-rich plasma treatment, a significant reduction in the patient’s follicle-stimulating hormone (FSH) levels were noted. A natural in-vitro fertilization cycle led to a biochemical pregnancy, resulting in a spontaneous abortion at the 5th week of pregnancy. This is the first report of a successful autologous platelet-rich plasma application leading to pregnancy in menopause. This report uniquely contributes to the medical knowledge and challenges current practice in the context of infertility. The efficiency and safety of this treatment with regard to the reproductive system merits further investigation.

Andrology ◽  
2019 ◽  
Vol 8 (1) ◽  
pp. 191-200 ◽  
R. Bader ◽  
J. N. Ibrahim ◽  
M. Moussa ◽  
A. Mourad ◽  
J. Azoury ◽  

2018 ◽  
Vol 7 (7) ◽  
pp. 457-467 ◽  
I. D. M. Smith ◽  
K. M. Milto ◽  
C. J. Doherty ◽  
S. G. B. Amyes ◽  
A. H. R. W. Simpson ◽  

ObjectivesStaphylococcus aureus (S. aureus) is the most commonly implicated organism in septic arthritis, a condition that may be highly destructive to articular cartilage. Previous studies investigating laboratory and clinical strains of S. aureus have demonstrated that potent toxins induced significant chondrocyte death, although the precise toxin or toxins that were involved was unknown. In this study, we used isogenic S. aureus mutants to assess the influence of alpha (Hla)-, beta (Hlb)-, and gamma (Hlg)-haemolysins, toxins considered important for the destruction of host tissue, on in situ bovine chondrocyte viability.MethodsBovine cartilage explants were cultured with isogenic S. aureus mutants and/or their culture supernatants. Chondrocyte viability was then assessed within defined regions of interest in the axial and coronal plane following live- and dead-cell imaging using the fluorescent probes 5-chloromethylfluorescein diacetate and propidium iodide, respectively, and confocal laser-scanning microscopy.ResultsHla-producing mutants caused substantial chondrocyte death compared with the toxin-deficient control (Hla-Hlb-Hlg-), whilst mutants producing Hlb and Hlg in the absence of Hla induced minimal chondrocyte death. Coronal studies established that Hla-induced chondrocyte death started in the superficial zone of cartilage and spread to deeper layers, whereas Hlb and Hlg toxins were without significant effect.ConclusionThis study identified Hla as a highly potent S. aureus toxin that caused rapid chondrocyte death in bovine cartilage, with other toxins or metabolic products produced by the bacteria playing a minor role. The identification of Hla in mediating chondrocyte death may assist in the development of therapeutic strategies aimed at reducing the extent of cartilage damage during and after an episode of septic arthritis. Cite this article: I. D. M. Smith, K. M. Milto, C. J. Doherty, S. G. B. Amyes, A. H. R. W. Simpson, A. C. Hall. A potential key role for alpha-haemolysin of Staphylococcus aureus in mediating chondrocyte death in septic arthritis. Bone Joint Res 2018;7:457–467. DOI: 10.1302/2046-3758.77.BJR-2017-0165.R1.

PLoS ONE ◽  
2014 ◽  
Vol 9 (9) ◽  
pp. e107813 ◽  
Lorenzo Drago ◽  
Monica Bortolin ◽  
Christian Vassena ◽  
Carlo L. Romanò ◽  
Silvio Taschieri ◽  

E. Scott Sills ◽  
E. Scott Sills ◽  
J. L. Petersen ◽  
N. S. Rickers ◽  
Samuel H. Wood ◽  

This registered, prospective clinical trial assessed serum anti-Mullerian hormone (AMH) patterns after treatment with activated platelet rich plasma (PRP). Patients with low ovarian reserve and/or at least 1 prior failed in vitro fertilization (IVF) cycle (n=182) received PRP injected into ovarian tissue under ultrasound guidance. Pretreatment AMH, BMI and platelet (PLT) concentration were recorded and serum AMH, follicle stimulating hormone, and estradiol were then measured at 2-week intervals for up to three months. Mean±SD patient age was 45.4±6.1yrs. Improved serum AMH was observed in 51 patients (28%) with median increase of 167% [95%CI 91; 280] after treatment; mean interval to maximum AMH increase was 4 weeks (range 2-10 weeks). Improved post-treatment AMH was not limited to younger patients; when stratified by age (<42 vs. ≥42yrs), significant AMH improvements were seen in both groups after treatment (p=0.03 and 0.009, respectively). Among responders, mean basal PLT count was higher (274K) vs. non-responders (250K); p<0.001. This is the first clinical trial to describe an intraovarian PRP technique for low reserve and finds the treatment safe and associated with significant increases in serum AMH for some patients, usually within four weeks. The substantially different pre-treatment PLT concentrations measured across PRP response groups warrants further investigation. Additional research can characterize ovarian response better, optimize PRP protocols, and collect outcomes data from those who subsequently undergo IVF with autologous oocytes.

2021 ◽  
Aleksandar Ljubić ◽  
Tatjana Božanović ◽  
Andrea Pirkovic-Cabarkapa ◽  
Andjela Perovic ◽  
Dušica Ljubić ◽  

Abstract Background: Patients with premature ovarian failure (POF) exhibit a diminished ovarian reserve and hormonal dysfunction. Case presentation: We aimed to restore normal hormonal function and folliculogenesis in a 31-year-old patient with POF, who had been amenorrheic for two years. We designed and performed three different ovarian regeneration procedures for three consecutive years, from 2015 to 2017: 1) intraovarian injection of activated autologous platelet-rich plasma (PRP); 2) activated autologous PRP and bone marrow-derived stem cell injection into the ovaries (SEGO); 3) ovarian cortical tissue resection and fragmentation; and in vitro ovarian tissue activation with autologous PRP and bone marrow stem cell retransplantation into the ovaries (the SEGOVA). The patient exhibited no improvement following the PRP treatment. The patient regained regular menstrual cycles after the SEGO procedure, although no follicular growth was observed yet. One month after the SEGOVA procedure, follicular growth was detected, and the patient underwent several stimulation protocols without obtaining oocytes. Eight months after the SEGOVA, the patient underwent in vitro fertilization (IVF) in a spontaneous cycle, when an oocyte of good quality was retrieved. Following intracytoplasmic sperm injection (ICSI), the oocyte failed to be fertilized. Eleven months after the SEGOVA, the patient reported a spontaneous pregnancy via natural conception. Pregnancy resulted in birth at term by uncomplicated vaginal delivery. After three different ovarian rejuvenation procedures, normal hormonal function and follicular growth were restored in the patient with POF, and the patient had a successful natural pregnancy following the last SEGOVA procedure. Although no ovarian function was detected after the first two procedures, they may have contributed to the outcomes from the SEGOVA procedure, a treatment that showed promising results in recovering ovarian function in patients with POF. Conclusions: It cannot be ruled out that ovarian rejuvenation with bone marrow‑derived stem cells and autologous growth factors, together with ovarian tissue fragmentation, took time to exhibit its effects and contributed to the final result – a successful natural conception and pregnancy.

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