Infection after Anterior Cruciate Ligament Reconstruction: A Narrative Review of the Literature.

Infection is an uncommon side effect of arthroscopic surgery, and this percentage is higher in anterior cruciate ligament reconstruction (ACLR) surgery, where graft and fixation devices are used. Infections can not only lead to high re-admission rates and poor functional recovery of the knee but can also have a significant negative impact on the patient's psychological and economic health, especially in athletes, as it can affect their sports career. It is important to be aware of the many risk factors, especially the manifestation of symptoms. These may sometimes be non-specific to the infectious pathology and common to other situations, such as the presence of a significant intra-articular hematoma. Septic arthritis after ACLR can occur at any time after surgery but typically presents acutely, while late manifestation is relatively rare. Diagnosis of infection is based on patient history, physical examination, laboratory parameters, and analysis of synovial fluid after joint aspiration, which is the gold standard for diagnosing post-operative infection. Once symptoms appear and the diagnosis seems certain, it is necessary to intervene quickly with arthroscopic debridement and long-term antibiotic treatment to try to save the graft and resolve the infectious situation to avoid graft failure and arthrofibrotic sequelae. The aim of this paper is to provide an overview of the epidemiology, pathogenesis, risk factors, clinical presentation, diagnostic evaluation, and current treatment guidelines of septic arthritis after ACLR surgery by analyzing recent literature, in particular meta-analyses and systematic reviews.

Anterior Cruciate Ligament Reconstruction with Quadrupled Semitendinosus Graft or Synthetic Ligament: Knee Stability and Clinical Outcomes at Three Years Follow-Up.

The incidence of anterior cruciate ligament reconstruction (ACLR) surgeries is increasing and graft choice is important for a rapid return to activity, especially in patients older than 30 years. The aim of this study is to compare in term of quality of life and knee stability of patients who undergone ACLR using quadrupled semitendinosus (ST4) graft against patients who undergone ACLR with synthetic ligaments. Thirty-nine patients undergoing ACLR were enrolled in the study and were divided into two groups: ACLR with synthetic ligaments-LARS (group A) and ACLR with quadrupled semitendinosus graft ST4 (group B). They underwent surgery at Policlinico di Bari Orthopedic Unit between January 2017 and January 2020. Group A was composed by nineteen patients (36.16 ± 4.41 mean age-years, 22.47 ± 2.63 mean BMI-kg/m2, 39.37 ± 10.05 mean time evaluation after surgery-months) and group B was composed by twenty patients (34.95 ± 3.59 mean age-years, 21.1 ± 2.88 mean BMI-kg/m2, 36.75 ± 8.69 mean time evaluation after surgery-months). For each patient, the following data were recorded: age; side of injury, BMI, date of surgery, anterior knee laxity with the arthrometer, and Lysholm knee scoring scale. Mean value of anterior tibial translation (ATT) in group A was 3.09 mm ± 0.65 and in group B was 2.66 mm ± 1.61 (pvalue of 0.1139). Mann--Whitney U test used to compare the Lysholm means values between groups showed a pvalue of 0.9307. LARS has comparable clinical and functional outcomes compared with hamstring autografts at short-term of 3 years follow-up. Level of Evidence: IV.

Surgical strategies in pediatric supracondylar humeral fractures: our experience.

BACKGROUND AND AIM: Supracondylar humeral fractures are the most common skeletal injury of childhood elbow. Treatment option for Gartland type II-III-IV fractures is based on closed reduction and percutaneous pinning (CRPP) fixation using Kirshner wires. Seldom open reduction is needed. Literature described different method of CRPP. The aim of the study is to report our experience in the surgical management of supracondylar humeral fractures comparing it with the literature, in order to identify useful information for a correct and better approach to reduce complications and improve clinical outcomes. METHODS: 148 patients with a mean age of 5.72 ± 2.52 years and with Gartland type II-III-IV humeral supracondylar fractures were treated with CRPP at our Orthopedic Pediatric Unit. They were divided into three groups according to surgical technique. Group A was represented by patients treated with cross pinning (1 medial and 1 lateral pin), Group B represented by 2 lateral pins while Group C represented by 2 lateral and one medial pin. Evaluation criteria are based on Mayo Elbow Performance Index (MEPI); Bauman's and Carrying Angle and Flynn's criteria. Data were recorded at the following times: T0 (before surgical procedure); T1 (one-month post-surgery); T2 (six months post-surgery). RESULTS: The three surgical techniques showed comparable results according to MEPI, Bauman's angle, Carrying's angle and Flynn's criteria from T0 to T1. There is an improvement for all Groups. Group C reported the best MEPI outcome at T2. However, 2 patients in this group did not show excellent results according to Flynn's criteria. CONCLUSIONS: There is no single and superior treatment for displaced humeral supracondylar fractures and that each fracture has its own personality.

Graft Intra-Articular Remodeling and Bone Incorporation in ACL Reconstruction: The State of the Art and Clinical Implications.

The knee is one of the most frequently affected joints in sports trauma, and anterior cruciate ligament (ACL) injury and meniscal tears are the most common lesions. ACL reconstruction (ACLR) remains the treatment of choice for patients willing to return to their previous activity. There are different surgical techniques and different possible usable grafts. The graft used for ACLR surgery undergoes a bone incorporation process and an intra-articular remodelling named ligamentization until it reaches characteristics similar to the native ligament. After the first incorporation stage, the remodelling process is divided into an early stage that could last 4 weeks, a proliferative stage that lasts 4 to 12 weeks, and a final stage of ligamentization that could last over 1 year. The period of return to sport (RTS) after ACLR, which is becoming shorter and shorter, can be a high-risk period for athletes due to the risk of graft failure. This systematic review aims to define the phases of the ligamentization process considering graft type and fixation techniques, as well as the graft's anatomopathological and biomechanical characteristics, to evaluate a criterion-based rehab progression and maximize patient outcomes for an RTS respecting graft biology. The rehabilitative program has to promote and optimize the graft remodelling and incorporation processes; moreover, it has to accommodate physiological graft healing and avoid overloading. An early RTS and noncompliance with the biological characteristics of the graft in the various phases are associated with a high incidence of re-injury.

Quality of Life and Clinical Evaluation of Calcaneoplasty with a Balloon System for Calcaneal Fracture at 5 Years of Follow-Up.

Calcaneal fractures are a challenging clinical problem. Management of this type of injury remains controversial, especially in the context of intra-articular fractures. Surgical treatment with open reduction and internal synthesis (ORIF) is considered the standard treatment for CF, but it is associated with many complications. Several minimally invasive techniques such as balloon-assisted reduction, pin fixation, and tricalcium phosphate augmentation have been proposed to avoid the frequent and recurrent postoperative problems related to these fractures. We retrospectively examined 20 patients (mean age was 54.5), all undergoing minimally invasive calcaneoplasty surgery at our Department of Orthopaedics and Traumatology between 2012 and 2016. X-ray and CT scan were performed preoperatively and at 5 years of follow-up (57.9 ± 6 months). The American Orthopaedic Foot and Ankle Society (AOFAS) score was used for clinical examination, and the Short-Form (36) Health Survey (SF-36) score and Visual Analogue Scale (VAS) were used to assess the Health-Related Quality of Life (HRQoL). All 20 patients were available at the final follow-up. The mean AOFAS score was 82.25/100. The VAS results attest an overall average of 2.7/10 (0-9). The average of the parameters "Physical Health" and "Mental Health" was, respectively, 81.25 and 83.55. In terms of postoperative complications, we observed no cases of superficial or deep infections. Clinical response after balloon-assisted reduction, pin fixation, and tricalcium phosphate augmentation has shown a comparable or better outcome according to the AOFAS and VAS score. Quality-of-life scores, obtained according to the SF-36 questionnaire, are considered high. From both a clinical and quality-of-life point of view, our study highlights that there is not gender distinction. Further comparative studies with a higher number of patients are needed which assess the quality of life in the various techniques used to treat calcaneal fractures.

High-affinity sigma-1 (σ1) receptor ligands based on the σ1 antagonist PB212.

Aim: The σ1 receptor is a druggable target involved in many physiological processes and diseases. To clarify its physiology and derive therapeutic benefit, nine analogs based on the σ1 antagonist PB212 were synthesized replacing the 4-methylpiperidine with basic moieties of varying size and degree of conformational freedom. Results & methodology: 3-Phenylpyrrolidine, 4-phenylpiperidine or granatane derivatives displayed the highest affinity (Ki.#x00A0;= 0.12, 0.31 or 1.03 nM). Calcium flux assays in MCF7σ1 cells indicated that the highest σ1 receptor affinity are σ1 antagonists. Molecular models provided a structural basis for understanding the σ1 affinity and functional activity of the analogs and incorporated Glennon's σ1 pharmacophore model. Conclusion: Herein, we identify new compounds exploitable as therapeutic drug leads or as tools to study σ1 receptor physiology.

The σ1 receptor agonist (+)-pentazocine increases store-operated Ca2+ entry in MCF7σ1 and SK-N-SH cell lines.

BACKGROUND: The intracellular [Ca2+] is modulated by σ receptors. An important component of the cellular machinery governing the intracellular [Ca2+] is Store-Operated Calcium Entry (SOCE). Here we want to investigate whether ligands of σ receptors affect SOCE. METHODS: The intracellular [Ca2+] was monitored, with the fluorescent Ca2+-sensitive probe Fura-2, in four cell lines with a different expression of σ receptors, namely MCF7 (expressing σ1 receptors with a low density and overexpressing σ2 receptors), MCF7σ1 (overexpressing σ1 receptors), SK-N-SH, and HT-29. RESULTS: When thapsigargin was used to deplete intracellular Ca2+ stores, in a Ca2+-free incubation medium, the Ca2+ influx (following Ca2+ re-addition) was significantly increased by 1µM (+)-pentazocine (σ1 receptor agonist) in MCF7σ1 (by 22.5%) and SK-N-SH (by 45.6%), but not in HT-29 and MCF7 cells. We have used, as a second approach, the "Mn2+ quenching" protocol. In MCF7σ1 cells, after thapsigargin treatment, the fluorescence quenching induced by Mn2+ influx (evidence of Ca2+ influx) was significantly increased (by 25.8%) by 1µM (+)-pentazocine, significantly decreased (by 18.0%) by BD1063 (σ1 receptor antagonist), and not affected by the presence of both ligands. These effects were not observed in MCF7 cells. Finally, in MCF7 cells, 1µM PB28 (σ2 receptor agonist), did not affect both the Ca2+ response after Ca2+ re-addition and the fluorescence quenching induced by Mn2+ influx. CONCLUSIONS: We propose that the σ1 receptor agonist (+)-pentazocine increases SOCE in MCF7σ1 and SK-N-SH cell lines. The σ2 receptor agonist PB28 does not affect SOCE in MCF7 cells.

Investigation of σ receptors agonist/antagonist activity through N-(6-methoxytetralin-1-yl)- and N-(6-methoxynaphthalen-1-yl)alkyl derivatives of polymethylpiperidines.

A series of polymethyl-substituted piperidines linked to either a 6-methoxy-1,2,3,4-tetrahydronaphthalen-1-yl or a 6-methoxynaphthalen-1-yl moiety was generated with the aim of verifying a previously generated hypothesis: tetralin and naphthalene nuclei confer opposite activity at the σ1 receptor. Compounds 6, 9 and 10 displayed appreciable affinity at both σ subtypes, but none of the novel compounds displayed significant antiproliferative activity in MCF7wt and MCF7σ1 cell lines. The effect on bradikynin-triggered Ca(2+) mobilization was studied as a methodology to suggest σ receptors mediated activity.

The sigma-1 receptor antagonist PB212 reduces the Ca²âº-release through the inositol (1, 4, 5)-trisphosphate receptor in SK-N-SH cells.

Sigma-1 receptors are specifically located at the endoplasmic reticulum-mitochondrion interface, but upon stimulation by ligands or under prolonged cellular stress, they translocate to other areas of the cell. Sigma-1 receptors are involved in the regulation of intracellular [Ca(2+)] by affecting the Ca(2+)-influx or the release from intracellular stores. In SK-N-SH cells, we measured the affinity of 4-methyl-1-[4-(6-methoxynaphthalen-1-yl)butyl]piperidine (PB212) at sigma-1 receptor by using a competition binding assay with specific radioligand; we obtained a K(i) value=316 ± 19 nM. PB212 also showed an antiproliferative effect in SK-N-SH cells (EC(50)=32 ± 4 µM) but had no effect in MCF7 cells, which only express sigma-2 receptor; these findings suggest that PB212 behaves as a sigma-1 receptor antagonist. We have studied the effect of PB212 on Ca(2+) homeostasis of the SK-N-SH cell line with the fluorescent probe Fura-2. 100 µM PB212 induced a Ca(2+)-efflux from the endoplasmic reticulum through the inositol (1, 4, 5)-trisphosphate (IP(3)) receptor. Moreover, [PB212] ranging from 1 to 100µM reduced the Ca(2+)-response, triggered by carbachol or bradykinin that engage the phospholipase C/IP(3) pathway; such a response is generally increased by sigma-1 receptor agonists. On the other hand, PB212 did not reduce the Ca(2+)-response mediated by IP(3) in LoVo cells, which do not express neither sigma-1 nor sigma-2 receptors, and in MCF7 cells. The fact that the activity of the sigma-1 receptor can be experimentally modulated by agonists and antagonists supports the intriguing hypothesis that some endogenous molecules, unknown at the moment, modulate the sigma-1 receptor and its cellular targets.

MC70 potentiates doxorubicin efficacy in colon and breast cancer in vitro treatment.

A major limitation of cancer treatment is the ability of cancer cells to develop resistance to chemotherapeutic drugs, by the establishment of multidrug resistance. Here, we characterize MC70 as ABC transporters inhibitor and anticancer agent, alone or with chemotherapy. MC70 was analyzed for its interaction with ABCB1, ABCG2 and ABCC1 by specific transport assays. In breast and colon cancer cell lines, cell growth and apoptosis were measured by MTT assay and DNA laddering Elisa kit, respectively. Cell cycle perturbation and cellular targets modulation were analyzed by Flow-cytometry and Western blotting, respectively. MC70 interacted with ABC transporters. In breast cancer cells, MC70 slightly inhibited cell proliferation strongly enhancing doxorubicin effectiveness. By contrast, MC70 was found to inhibit cell growth in colon cancer cells without affecting doxorubicin efficacy and in combination with topoisomerase I inhibitors it could be a promising therapeutic approach. What is more, it was also observed that MC70 induced apoptosis, canceled in favor of necrosis when given in combination with high doses of doxorubicin. MC70 inhibited cell migration probably through its interaction with sigma-1 receptor. Modulations of i) cell cycle, ii) pAkt and the phosphorylation of the three MAPKs were highlighted, while any activity was excluded at transcription level, thus accounting for the phenotypic effects observed. MC70 might be considered as a new potential anticancer agent capable to i) enhance chemotherapy effectiveness and ii) to play a contributory role in the treatment of chemotherapy resistant tumors.

F281, synthetic agonist of the sigma-2 receptor, induces Ca2+ efflux from the endoplasmic reticulum and mitochondria in SK-N-SH cells.

We demonstrate that F281, a synthetic agonist of the sigma-2 receptor (s2R), induces a non transient increase in intracellular [Ca(2+)] ([Ca(2+)](i)) and cell death in SK-N-SH cells. Sigma receptors are classified into two subtypes, with different molecular weight and tissue distribution. While the sigma-1 receptor has been cloned, the s2r is less characterized and its physiological ligand and role need further investigation. In tumour cell lines, synthetic agonists of the s2R trigger apoptosis and modulate [Ca(2+)](i). In particular, CB-64D induces a Ca(2+) response while PB28 supresses Ca(2+) signalling. We have recently synthesized F281, by replacing the 5-methoxytetraline moiety of PB28 with a carbazole nucleus. Although this bioisosteric substitution should not affect the ligand affinity at the receptor, F281 (after 24h incubation) was more cytotoxic than PB28 (EC(50) values 65.4nM and 8.13 microM, respectively) in SK-N-SH cells. We used the fluorescent probes fura-2, rhod-2 and JC-1. F281 mobilizes Ca(2+) from mitochondria and from the endoplasmic reticulum, by opening its inositol 1,4,5-trisphosphate receptor; Ca(2+)-entry through the channels activated by store depletion was also observed. After the increase in [Ca(2+)](i) and within 10 min, we observed a sudden drop in metabolic activity and intracellular [ATP] leading to cell death.

The adrenergic receptor subtypes present in frog (Rana esculenta) skin.

Frog skin transports ions and water under hormonal control. In spite of the fundamental role played by adrenergic stimulation in maintaining the water balance of the organism, the receptor subtype(s) present in the skin have not been identified yet. We measured the increase in short-circuit current (ISC, an estimate of ion transport) induced by cirazoline, clonidine, xamoterol, formoterol, or BRL 37344, in order to verify the presence of alpha1, alpha2, beta1, beta2, or beta3 receptor subtypes, respectively. Only after treatment with formoterol, BRL 37344 and, to a lesser extent, cirazoline was measured a significant increase in ISC (57%, 33.2%, and 4.7%, respectively). The formoterol and BRL 37344 concentrations producing half-maximal effect (EC50) were 1.12 and 70.1 nM, respectively. Moreover, the formoterol effect was inhibited by treatment with ICI 118551 (antagonist of beta2 receptors) while SR 59230A (antagonist of beta3 receptors) had no effect; opposite findings were obtained when the BRL 37344 stimulation was investigated. Finally, by measuring the transepithelial fluxes of 22Na+ and 36Cl-, we demonstrated that Na+ absorption is increased by activation of beta2 and beta3 and is cAMP-sensitive, whereas the Cl- secretion is only increased by activation of beta2 receptors and is cAMP- and calmodulin-sensitive.

The sigma-2 receptor agonist PB28 inhibits calcium release from the endoplasmic reticulum of SK-N-SH neuroblastoma cells.

In this paper we demonstrate that PB28 abolishes the Ca2+ release through the inositol 1,4,5-trisphosphate (InsP3) receptors and ryanodine receptors in SK-N-SH cells. Sigma receptors are divided into the subtypes sigma-1 and sigma-2, which are expressed in tumor cell lines and characterized by distinct pharmacological profiles. The sigma-1 receptor has been recently cloned, whereas the sigma-2 receptor is less well characterized. The endogenous ligand(s) of both subtypes remain unclear. In isolated guinea pig ileum, PB28 inhibits the contraction induced by carbachol dose dependently and in a non-competitive manner. In SK-N-SH cells PB28 challenge does not affect the intracellular Ca2+ concentration but incubation with PB28 for 45 min abolishes the cytosolic Ca2+ increases evoked by carbachol or histamine. This effect, not sensitive to cycloheximide, is caused by direct inhibition of the InsP3 receptors, since PB28 abolishes the response elicited by InsP3 administration in permeabilized SK-N-SH cells. Finally, incubation for 45 min with PB28 also abolishes the cytosolic Ca2+ increase evoked by caffeine.

Inositol 1,4,5-trisphosphate and ryanodine receptors mobilize calcium from a common functional pool in human U373 MG cells.

This investigation concentrates on the change in Ca(2+) concentration ([Ca(2+)]) caused by ryanodine in U373 MG cells. This cell type from a human astrocytoma is a unique cellular model because it only expresses the type 3 ryanodine receptor (RyR3), which is generally the least abundant isoform. In the presence of physiological [Ca(2+)] in the extracellular medium, U373 MG cells are caffeine-insensitive, even after forskolin treatment, and ryanodine-sensitive only when an unusually high concentration (30 microM) is applied. Xestospongin C behaves like thapsigargin and therefore cannot be used as a selective antagonist of inositol 1,4,5-trisphosphate receptors (InsP(3)Rs). After ryanodine challenge, addition of an analog of Substance P (SP), which should deplete InsP(3)-sensitive stores, has no effect on [Ca(2+)](i). After thapsigargin treatment, which unmasks the calcium leak from intracellular stores, neither ryanodine nor SP change [Ca(2+)](i), suggesting that thapsigargin completely depletes the ryanodine-sensitive and the InsP(3)-sensitive stores of U373 MG cells. Finally, in experiments monitoring the [Ca(2+)] in intracellular stores, InsP(3) stimulation of permeabilized cells causes a decrease in [Ca(2+)] that is not affected by subsequent ryanodine treatment. Our results support the conclusion that U373 MG cells express both InsP(3)Rs and RyRs that can individually or in combination mobilize only one functional Ca(2+) pool.

Calcium response after stimulation by substance P of U373 MG cells: inhibition of store-operated calcium entry by protein kinase C.

In this paper we investigate the Ca2+ response after Substance P (SP) stimulation of U373 MG cells. SP is a tachykinin and physiologically acts as a neurotransmitter and neuromodulator in the nervous system, but pathologically triggers malignant glial cells, such as U373 MG, to release cytokines and increase proliferation rate. In this paper we show that SP increases the proliferation rate of U373 MG cells and the intracellular Ca2+ concentration by mobilizing Ca2+ only from thapsigargin-sensitive stores. In fact, Ca2+ entry through store-operated calcium entry (SOCE) channels, which was observed after thapsigargin treatment, was not detected after stimulation by SP. The inhibition of SOCE after SP stimulation must be mediated by protein kinase C (PKC), because it was not observed in the presence of calphostin C (an inhibitor of PKC). Moreover, stimulation by SP-induced membrane potential hyperpolarization. Our results are consistent with the following sequence of events: (i) SP interacts with NK(1) receptors; (ii) fast homologous receptor desensitization occurs; (iii) reuptake by endoplasmic reticulum Ca(2+)-ATPase quantitatively overwhelms the extrusion by plasma membrane Ca2+-ATPase. These results have two important consequences. In U373 MG cells the SOCE does not contribute to the Ca2+ response after SP, and is not necessarily involved in promoting cell proliferation.