Role of Fibroblasts and Myofibroblasts on the Pathogenesis and Treatment of Pelvic Organ Prolapse

Pelvic organ prolapse (POP) is a multifactorial connective tissue disorder caused by damage to the supportive structures of the pelvic floor, leading to the descent of pelvic organs in the vagina. In women with POP, fibroblast function is disturbed or altered, which causes impaired collagen metabolism that affects the mechanical properties of the tissue. Ideal surgical repair, either native tissue repair or POP surgery using an implant, aims to create a functional pelvic floor that is load-bearing, activating fibroblasts to regulate collagen metabolism without creating fibrotic tissue. Fibroblast function plays a crucial role in the pathophysiology of POP by directly affecting the connective tissue quality. On the other hand, fibroblasts determine the success of the POP treatment, as the fibroblast-to-(myo)fibroblast transition is the key event during wound healing and tissue repair. In this review, we aim to resolve the question of “cause and result” for the fibroblasts in the development and treatment of POP. This review may contribute to preventing the development and progress of anatomical abnormalities involved in POP and to optimizing surgical outcomes.

Impact of Sprouting under Potassium Nitrate Priming on Nitrogen Assimilation and Bioactivity of Three Medicago Species

Edible sprouts are rich in flavonoids and other polyphenols, as well as proteins, minerals, and vitamins. Increasing sprout consumption necessitates improving their quality, palatability, and bioactivity. The purpose of this study was to test how KNO3 priming affects the sprouting process species on three Medicago species (Medicago indicus, Medicago interexta, and Medicago polymorpha) and their nutritional values. Targeted species of Medicago were primed with KNO3, and the levels of different primary and secondary metabolites were determined. KNO3 induced biomass accumulation in the sprouts of the three species, accompanied by an increased content of total mineral nutrients, pigments, vitamins, and essential amino acids. Besides, our results showed that KNO3 enhanced the activity of nitrate reductase (NR), glutamate dehydrogenase (GDH), and glutamine synthetase (GS) enzymes, which are involved in the nitrogen metabolism and GOGAT cycle, which, in turn, increase the nitrogen and protein production. KNO3 treatment improved the bioactive compound activities of Medicago sprouts by increasing total phenolic and flavonoid contents and enhancing the antioxidant and antidiabetic activities. Furthermore, species-specific responses toward KNO3 priming were noticeable, where Medicago interexta showed the highest antioxidant and antidiabetic activities, followed by Medicago polymorpha. Overall, this study sheds the light on the physiological and biochemical bases of growth, metabolism, and tissue quality improvement impact of KNO3 on Medicago sprouts.

Methods to Study the Myenteric Plexus of Rat Small Intestine

The close interaction between the enteric nervous system, microbiome, and brain in vertebrates is an emerging topic of recent studies. Different species such as rat, mouse, and human are currently being used for this purpose, among others. The transferability of protocols for tissue isolation and sample collection is not always straightforward. Thus, the present work presents a new protocol for isolation and sample collection of rat myenteric plexus cells for in vivo as well as in vitro studies. With the methods and chemicals described in detail, a wide variety of investigations can be performed with regard to normal physiological as well as pathological processes in the postnatal developing enteric nervous system. The fast and efficient preparation of the intestine as the first step is particularly important. We have developed and described a LIENS chamber to obtain optimal tissue quality during intestinal freezing. Cryosections of the flat, snap-frozen intestine can then be prepared for histological examination of the various wall layers of the intestine, e.g. by immunohistochemistry. In addition, these cryosections are suitable for the preparation of defined regions, as shown here using the ganglia of the mesenteric plexus. This specific tissue was obtained by laser microdissection, making the presented methodology also suitable for subsequent analyses that require high quality (specificity) of the samples. Furthermore, we present here a fully modernized protocol for the cultivation of myenteric neurons from the rat intestine, which is suitable for a variety of in vitro studies.

Harnessing Endophytic Fungi for Enhancing Growth, Tolerance and Quality of Rose-Scented Geranium (Pelargonium graveolens (L’Hér) Thunb.) Plants under Cadmium Stress: A Biochemical Study

Heavy metal contamination in soil is increasing rapidly due to increasing anthropogenic activities. Despite the importance of rose-scented geranium as a medicinal plant, little attention was paid to enhancing its productivity in heavy metal-polluted soil. In this regard, endophytes improve plant resistance to heavy metal toxicity and enhance its tissue quality. Here, the impact of the three endophytic fungi Talaromyces versatilis (E6651), Emericella nidulans (E6658), and Aspergillus niger (E6657) on geranium growth, tolerance, and tissue quality under cadmium (Cd) stress was investigated. In contrast to E. nidulans, T. versatilis and A. niger enhanced geranium growth and the stimulatory effect was more pronounced under Cd-stress. The three endophytes significantly alleviated Cd accumulation and increased mineral content in geranium leaves. In addition, endophytic fungi successfully alleviated Cd-induced membrane damage and reinforced the antioxidant defenses in geranium leaves. Inoculation with endophytes stimulated all the antioxidant enzymes under Cd-stress, and the response was more obvious in the case of T. versatilis and A. niger. To reduce the toxicity of tissue-Cd levels, T. versatilis and A. niger upregulated the detoxification mechanisms; glutathione-S-transferase, phytochelatin, and metallothionein levels. Moreover, endophytic fungi improved the medicinal value and quality of geranium by increasing total antioxidant capacity (TAC), phenolic compound biosynthesis (phenylalanine ammonia-lyase), and vitamin content as well as the quantity and quality of essential oil, particularly under Cd-stress conditions. The variation in the mechanisms modulated by the different endophytic fungi was supported by Principal Component Analysis (PCA). Overall, this study provided fundamental insights into endophytes’ impact as a feasible strategy to mitigate the phytotoxicity hazards of Cd-stress in geranium and enhance its quality, based on the growth and biochemical investigations.

ACL Size, but Not Signal Intensity, Is Influenced by Sex, Body Size, and Knee Anatomy

Background: Little is known about sex-based differences in anterior cruciate ligament (ACL) tissue quality in vivo or the association of ACL size (ie, volume) and tissue quality (ie, normalized signal intensity on magnetic resonance imaging [MRI]) with knee anatomy. Hypothesis: We hypothesized that (1) women have smaller ACLs and greater ACL normalized signal intensity compared with men, and (2) ACL size and normalized signal intensity are associated with age, activity levels, body mass index (BMI), bicondylar width, intercondylar notch width, and posterior slope of the lateral tibial plateau. Study Design: Cross-sectional study; Level of evidence, 3. Methods: Knee MRI scans of 108 unique ACL-intact knees (19.7 ± 5.5 years, 62 women) were used to quantify the ACL signal intensity (normalized to cortical bone), ligament volume, mean cross-sectional area, and length. Independent t tests were used to compare the MRI-based ACL parameters between sexes. Univariate and multivariate linear regression analyses were used to investigate the associations between normalized signal intensity and size with age, activity levels, BMI, bicondylar width, notch width, and posterior slope of the lateral tibial plateau. Results: Compared with men, women had significantly smaller mean ACL volume (men vs women: 2028 ± 472 vs 1591 ± 405 mm3), cross-sectional area (49.4 ± 9.6 vs 41.5 ± 8.6 mm2), and length (40.8 ± 2.8 vs 38.1 ± 3.1 mm) ( P < .001 for all), even after adjusting for BMI and bicondylar width. There was no difference in MRI signal intensity between men and women (1.15 ± 0.24 vs 1.12 ± 0.24, respectively; P = .555). BMI, bicondylar width, and intercondylar notch width were independently associated with a larger ACL ( R 2 > 0.16, P < .001). Younger age and steeper lateral tibial slope were independently associated with shorter ACL length ( R 2 > 0.03, P < .04). The combination of BMI and bicondylar width was predictive of ACL volume and mean cross-sectional area ( R 2 < 0.3). The combination of BMI, bicondylar width, and lateral tibial slope was predictive of ACL length ( R 2 = 0.39). Neither quantified patient characteristics nor anatomic variables were associated with signal intensity. Conclusion: Men had larger ACLs compared with women even after adjusting for BMI and knee size (bicondylar width). No sex difference was observed in signal intensity, suggesting no difference in tissue quality. The association of the intercondylar notch width and lateral tibial slope with ACL size suggests that the influence of these anatomic features on ACL injury risk may be partially explained by their effect on ACL size. Registration: NCT02292004 and NCT02664545 ( ClinicalTrials.gov identifier).

Role of the facial artery musculomucosal flap in the reconstruction of palatal defects

The facial artery musculomucosal flap (FAMM flap) is a convenient option for covering complicated palatal defects, as it is a local flap inside the oral cavity with good tissue quality and minimal drawbacks. The present prospective study included 17 patients, 7 males and 10 females. Most of the patients had palatal fistulae, after cleft palate surgical repair; only one had a post-traumatic palatal defect. Superiorly based FAMM flaps were used for eight patients, and inferiorly based FAMM flaps were used for nine patients. A speech specialist assessed all patients, postoperatively. The range of follow-up time was from two to four years. In the majority of cases, reconstruction was successful and uneventful. Patients were satisfied regarding oral function. Complications were minor, which include incomplete coverage, bulkiness, temporary hardness and dimpling. The FAMM flap is a very useful, versatile, and technically easy flap for covering difficult palatal defects.

7-Tesla MRI Evaluation of the Knee, 25 Years after Cartilage Repair Surgery: The Influence of Intralesional Osteophytes on Biochemical Quality of Cartilage

Objective To evaluate the morphological and biochemical quality of cartilage transplants and surrounding articular cartilage of patients 25 years after perichondrium transplantation (PT) and autologous chondrocyte transplantation (ACT) as measured by ultra-high-field 7-Tesla (7T) magnetic resonance imaging (MRI) and to present these findings next to clinical outcome. Design Seven PT patients and 5 ACT patients who underwent surgery on the femoral condyle between 1986 and 1996 were included. Patient-reported outcome measures (PROMs) were assessed by the clinical questionnaires: Knee injury and Osteoarthritis Outcome Score (KOOS), International Knee Documentation Committee (IKDC), and Visual Analogue Scale (VAS) for knee pain. The morphological (MOCART score) and biochemical quality (glycosaminoglycans [GAGs] content and collagen integrity) of cartilage transplants and surrounding articular cartilage were analyzed by 7T MRI. The results of the PT and ACT patients were compared. Finally, a detailed morphological analysis of the grafts alone was performed. Results No statistically significant difference was found for the PROMs and MOCART scores of PT and ACT patients. Evaluation of the graft alone showed poor repair tissue quality and high prevalence of intralesional osteophyte formation in both the PT and ACT patients. Penetration of the graft surface by the intralesional osteophyte was related to biochemically damaged opposing tibial cartilage; GAG content was significantly lower in patients with an osteophyte penetrating the graft surface. Conclusions Both PT and ACT patients have a high incidence of intralesional osteophyte formation 25 years after surgery. The resulting biochemical damage to the opposing tibial cartilage might be dependent on osteophyte morphology.

Revision Quadriceps Tendon Repair With Bone-Achilles Allograft Augmentation

Background: Surgical repair of chronic quadriceps tendon ruptures can be daunting, especially after failure of a prior repair. In this setting, tissue quality is usually poor, necessitating graft augmentation. In this video, we describe our technique for Achilles tendon allograft augmentation for revision quadriceps tendon repair. Indications: Failed quadriceps tendon repair defined as ongoing extensor mechanism deficit including patella baja, functional deficit, or palpable quadriceps defect with confirmed retear on advanced imaging. Technique Description: Patient is placed in the supine position, and a midline incision is extended to the tibial tubercle. Full thickness medial and lateral flaps are raised, nonviable scar tissue is excised, and suprapatellar adhesions are released to ensure full mobilization of the viable remnant quadriceps. A plane is then developed deep to the patellar tendon paratenon from proximal to distal. A reamer is used to prepare a socket just medial to the tibial tubercle. The calcaneal bone block of the Achilles allograft is fashioned to match the recipient site on the tibia with a sagittal saw. The graft is shuttled deep to the paratenon, and the bone plug is fixed to the tibia with an interference screw. Suture from the patellar anchors is then used to place 2 running Krackow stitches spanning the remnant quadriceps tendon proximally. The remnant tissue is subsequently reduced and tied with an anchor pull-through technique. The soft tissue component of the Achilles graft is laid over the repair and oversewn with free nonabsorbable suture. Patient is placed in a brace locked in extension for 6 weeks and allowed to be weight bearing as tolerated. Results: Long-term patient-reported outcomes of Achilles allograft reconstruction for revision extensor mechanism repairs are limited. Two studies of 17 reconstructions each reported this to be a reliable and durable option at a mean follow-up of 65 and 52 months, respectively. Discussion/Conclusion: Revision surgery for extensor mechanism deficits can be a challenging procedure. Our preference is to perform augmentation with Achilles allograft with bone plug fixation on the tibial side. This allows for augmentation of the entire extensor mechanism, as well as bone-to-bone healing on the tibial side.

Anatomic Measurement and Variability Analysis of the Anterior Talofibular Ligament and Calcaneofibular Ligament of the Ankle

Background: The anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL) contribute greatly to the overall stability of the ankle joint; however, ATFL and combined ATFL-CFL sprains are common. Anatomic reconstruction of the lateral collateral ligament with grafts has been proposed for patients with poor tissue quality or inadequate local tissue. Anatomic reconstruction of the lateral ankle ligaments requires a good understanding of their anatomic location. Purpose: To describe the anatomy of the ATFL and CFL ligaments quantitatively and qualitatively and explore the relationship of some morphological parameters. Study Design: Descriptive laboratory study. Methods: A total of 66 adult ankle specimens were analyzed for ATFL band type, origin, length, width, thickness, and angle between the ATFL and CFL, and 73 adult ankle specimens were used for measuring the origin of the CFL. The coefficient of variation was used to describe and compare the respective variability of angle, length, width, and thickness. The origin of the ATFL was labeled as point A, and the leading edge of the CFL intersection with the articular surface of the calcaneus was considered point B. Results: The ATFL had a variable number of bands. A high degree of variability (coefficient of variation >0.2) was seen for most morphological measurements of the ATFL. In addition, the length of distance AB also varied. The CFL originated at the tip of the fibula in only 9% of specimens. It was found more commonly at the anterior border of the lateral malleolus (4.94 ± 1.70 mm from the tip). The angle between the ATFL and CFL was consistent at 100° to 105º. Conclusion: A fair amount of variability of ATFL length, width, and thickness were found in our study, with less variability in the ATFL-CFL angle. Most CFLs attached anterior to the tip of the fibula. Clinical Relevance: Providing relevant anatomic data of ATFL and CFL is important in ensuring proper surgical treatment of ankle joint injuries.