The effect of biofeedback pelvic floor training with ACTICORE1 on fecal incontinence A prospective multicentric cohort pilot study.

INTRODUCTION: Fecal incontinence refers to the inability to pass stool in a localized and timely manner resulting in the involuntary loss of intestinal contents such as air, intestinal mucus or stool. The prevalence of fecal incontinence in the general population is approximately 2-21%. Women are more frequently affected than men. Physiotherapeutically guided pelvic floor training, otherwise known as Kegel exercise, is the mainstay of treatment for fecal incontinence. The objective of this study was to evaluate the feasibility and potential benefits of a new biofeedback training, which uses a non-insertable pelvic floor sensor with digital interface, called ACTICORE1. METHODS: From January 2020 to April 2021, we conducted a prospective non-randomized multicentric clinical pilot study at the Alexianer St. Hedwig Hospital Berlin (Germany), private clinic Strack (Germany) and the University Hospital Magdeburg (Germany). Patients with fecal incontinence, defined as a Wexner score >2, were recruited and asked to either perform biofeedback training with ACTICORE1 (6 min daily for 16 weeks) or daily Kegel exercise (Physiotherapeutic guidance weekly for the first 6 weeks; biweekly for the remaining 10 weeks). The primary outcome was severity of fecal incontinence after 16 weeks of training assessed using the Wexner score. Secondary outcomes were severity of fecal incontinence after 12 weeks and patients' quality of life assessed using the EQ-5D-3L questionnaire after 16 weeks of training. The two-one-sided t-tests (TOST) procedure was used to determine if training with ACTICORE1 has equivalent or noninferior efficacies compared to Kegel exercise. RESULTS: A total of 40 individuals were included. Dropout occurred in 4 cases. The final sample included 19 patients who performed the ACTICORE1 training (ACTICORE-group) and 17 patients who performed guideline-based physiotherapy (PHYSIO-group). Univariate analysis of biometric parameters showed no statistically significant differences. Individuals in the ACTICORE-group were younger (M=46,6 (SD=18,9) years vs. M=57,1 (SD=17,3) years, p=0.093). In terms of endpoint evaluation, a non-inferiority of ACTICORE1 compared to the therapy standard (Kegel exercise) was detected. Both groups showed a statistically significant intraindividual improvement in fecal incontinence as measured by Wexner scoring after 16 weeks. The TOST detected a non-inferiority of ACTICORE1 training (98% confidence interval with equivalence bounds 5 for low and high; Results: 1.36, upper 6.75). CONCLUSION: Pelvic floor training with ACTICORE1 may enable sufficient pelvic floor training as a digital health application. The study at hand revealed a non-inferiority of ACTICORE1 training compared to Kegel exercise.

Impact of Retropubic vs. Transobturator Slings for Urinary Incontinence on Myofascial Structures of the Pelvic Floor, Adductor and Abdominal Muscles.

Suburethral tension-free slings (tapes or bands) are an essential component in the operative treatment of urinary incontinence. In the present contribution the influence of the type of suburethral sling (retropubic vs. transobturator) on the myofascial structures of the abdominal, adductor and pelvic floor muscles is examined. For this purpose, 70 patients were prospectively observed clinically and physiotherapeutically. Significant differences were seen in the improvement of the pelvic floor musculature (strength, endurance, speed) after placement of a suburethral sling, irrespective of whether it was of the retropubic or the transobturator type. Thus, after surgical treatment patients should be encouraged to undertake further pelvic floor exercising or this should be prescribed for them. There were no significant changes in the abdominal and adductor muscles but there were slight increases with regard to pain level, pain on palpation, and trigger points after placement of both types of sling; thus this is not a criterion in the decision as to which type of sling to use.

Efficient non-viral transfection of primary human adult chondrocytes in a high-throughput format.

OBJECTIVE: The development of a reliable high-throughput transfection protocol for primary human articular chondrocytes. METHODS: Primary human chondrocytes were isolated from adult knee cartilage by an optimized enzymatic digestion protocol and cultivated in high-density monolayer culture for 3-5 days. Isolated chondrocytes were transfected with a green fluorescent protein (GFP)-expressing reporter construct using amaxa's Nucleofector 96-well Shuttle System. Transfection efficiencies were measured by fluorescence activated cell sorting and cell viability was determined by an adenosine-5'-triphosphate (ATP) assay. siRNA oligonucleotides (against glyceraldehyde-3-phosphate dehydrogenase (GAPDH)) were transfected into the cells using the optimized nucleofection protocol and mRNA knockdown values were determined by a branched-DNA assay. RESULTS: Transfection efficiencies of more than 70% of surviving cells were achieved routinely with the nucleofection protocol presented in this article. Cell viability 24h post transfection was around 80%. The cell number used per transfection was reduced to 2x10(5) per sample. In addition, the protocol proved to be well suited for the transfer of siRNA molecules into primary human chondrocytes with suppression rates on the mRNA level of more than 95% (for GAPDH). CONCLUSIONS: We present the successful use of nucleofection on primary human chondrocytes using a microtiter plate compatible format that for the first time allows the efficient transfection of up to 96 samples in parallel. The optimized nucleofection protocol is offering maximum substrate flexibility by allowing transfer of DNA and siRNA oligonucleotides with the same set of parameters. Moreover, the transfection procedure requires substantially lower cell numbers than single cuvette protocols and is therefore perfectly suited for applications requiring multiple experimental replicates.

IL-1beta and BMPs--interactive players of cartilage matrix degradation and regeneration.

Intact human adult articular cartilage is central for the functioning of the articulating joints. This largely depends on the integrity of its extracellular matrix, given the high loading forces during movements in particular in the weight-bearing joints. Unlike the first impression of a more or less static tissue, articular cartilage shows - albeit in the adult organism a slow--tissue turnover. Thus, one of the most important questions in osteoarthritis research is to understand the balance of catabolic and anabolic factors in articular cartilage as this is the key to understand the biology of cartilage maintenance and degeneration. Anabolic and catabolic pathways are very much intermingled in articular cartilage. The balance between anabolism and catabolism is titrated on numerous levels, starting from the mediator-synthesizing cells which express either catabolic or anabolic factors. Also, on the level of the effector cells (i.e. chondrocytes) anabolic and catabolic gene expression compete for a balance of matrix homeostasis, namely the synthesis of matrix components and the expression and activation of matrix-degrading proteases. Also, there are multiple layers of intracellular cross-talks in between the anabolic and catabolic signalling pathways. Maybe the most important lesson from this overview is the notion that the anabolic-catabolic balance as such counts and not so much sufficient net anabolism or limited catabolism alone. Thus, it might be neither the aim of osteoarthritis therapy to foster anabolism nor to knock down catabolism, but the balance of anabolic-catabolic activities as a total might need proper titration and balancing.

MMP-2/gelatinase A is a gene product of human adult articular chondrocytes and is increased in osteoarthritic cartilage.

OBJECTIVE: Collagen fibril degeneration involves initially the cleavage within the triple helix by the collagenases (1 and 3), but then mainly involves also the gelatinases, of which gelatinase A (MMP-2) appears to play a central role in many tissues. The objective of this study was to determine the quantitative expression levels as well as the distribution in normal and osteoarthritic cartilage of gelatinase A and in cultured articular chondrocytes with and without stimulation by Il-1beta. METHODS: Conventional and online PCR technology and immunohistochemistry were used to determine MMP-2 expression levels on the mRNA and protein level. RESULTS: Conventional PCR analysis could demonstrate the presence of MMP-2 mRNA in normal and osteoarthritic chondrocytes. Online quantitative PCR confirmed the presence of MMP-2 mRNA expression in normal articular chondrocytes in vivo (and in vitro). An increase of 5x (p < 0.001) was observed in osteoarthritic cartilage in vivo. Of note, no significant up-regulation of gelatinase A was observed by Il-1beta in vitro. Immunostaining for gelatinase A confirmed the presence of MMP-2 with mono- and polyclonal antibodies in normal and osteoarthritic chondrocytes with somewhat higher levels observed in the latter. CONCLUSIONS: The presented results confirm the increased expression of gelatinase A by osteoarthritic articular chondrocytes as previously described. Of note, also normal adult articular chondrocytes expressed significant amounts of gelatinase A in vivo and in vitro suggesting gelatinase A as being also involved in physiological collagen turnover in human adult articular cartilage.