The gelling effect of platelet-rich fibrin matrix when exposed to human tenocytes from the rotator cuff in small-diameter culture wells and the design of a co-culture device to overcome this phenomenon.

OBJECTIVES: Platelet-rich fibrin matrix (PRFM) has been proved to enhance tenocyte proliferation but has mixed results when used during rotator cuff repair. The optimal PRFM preparation protocol should be determined before clinical application. To screen the best PRFM to each individual's tenocytes effectively, small-diameter culture wells should be used to increase variables. The gelling effect of PRFM will occur when small-diameter culture wells are used. A co-culture device should be designed to avoid this effect. METHODS: Tenocytes harvested during rotator cuff repair and blood from a healthy volunteer were used. Tenocytes were seeded in 96-, 24-, 12-, and six-well plates and co-culture devices. Appropriate volumes of PRFM, according to the surface area of each culture well, were treated with tenocytes for seven days. The co-culture device was designed to avoid the gelling effect that occurred in the small-diameter culture well. Cell proliferation was analyzed by water soluble tetrazolium-1 (WST-1) bioassay. RESULTS: The relative quantification (condition/control) of WST-1 assay on day seven revealed a significant decrease in tenocyte proliferation in small-diameter culture wells (96 and 24 wells) due to the gelling effect. PRFM in large-diameter culture wells (12 and six wells) and co-culture systems induced a significant increase in tenocyte proliferation compared with the control group. The gelling effect of PRFM was avoided by the co-culture device. CONCLUSION: When PRFM and tenocytes are cultured in small-diameter culture wells, the gelling effect will occur and make screening of personalized best-fit PRFM difficult. This effect can be avoided with the co-culture device.Cite this article: C-H. Chiu, P. Chen, W-L. Yeh, A. C-Y. Chen, Y-S. Chan, K-Y. Hsu, K-F. Lei. The gelling effect of platelet-rich fibrin matrix when exposed to human tenocytes from the rotator cuff in small-diameter culture wells and the design of a co-culture device to overcome this phenomenon. Bone Joint Res 2019;8:216-223. DOI: 10.1302/2046-3758.85.BJR-2018-0258.R1.

Autofluorescence reveals menstrual phase in the endometrium.

OBJECTIVE: Tissue contains fluorophores that autofluoresce without additional dye or photosensitizer with the appropriate light excitation. This technique has been widely applied for discrimination between normal and precancerous tissue. The aim of this study was to explore the capability and reliability of autofluorescence phase determination in samples of human endometrium. METHODS: A total of 70 measurement sites from 31 endometrial tissue samples from hysterectomy were enrolled. Xenon light (330 nm) was directed at the endometrial tissue and the resultant autofluorescence intensity recorded. Spectra were then grouped according to the proliferative and secretory phase, with multivariant analysis, partial least square (PLS) and analysis of variance (ANOVA) used for evaluation of the statistical significance of phase determination. RESULTS: Both proliferative and secretory autofluorescence spectra showed a similar characteristic triple-peak curve shape pattern, however, each of the intensities at the three peaks between the two phases varied markedly (p < 0.01). PLS analysis confirmed that collagen, NADH and FAD autofluorescence were the principle determinants of endometrial spectrum; the sensitivity and specificity of phase determination by autofluorescent was 100% and 97%, respectively. CONCLUSION: Autofluorescence measurement provides real-time information on endometrial phase status and, based on our results, it appears reasonable to suggest that it may be promising as a clinical tool for prompt phase interpretation.

Effects of hyperbaric oxygen and platelet derived growth factor on medial collateral ligament fibroblasts.

PURPOSE: This study investigated hyperbaric oxygen (HBO2) and platelet-derived growth factor-BB (PDGF-BB) to determine their combined effects on fibroblasts from rabbit medial collateral ligament (MCL). METHOD: Cells were divided into four groups: (I) Control, (II) HBO2 treatment, (III) PDGF-BB treatment and (IV) HBO2 combined with PDGF-BB treatment. All hyperoxic cells were exposed to 100% O2 at 2.5 atmospheres absolute (ATA) in a hyperbaric chamber for 120 minutes per 48 hours. Measurement of cell growth was based on increase in cell number. Cell cycle modulations were analyzed by fluorescence-activated cell sorter (FACS). Quantity of Type I and Type III collagen was determined by western blotting and image analyzer. RESULTS: Treatment doses of HBO2 alone or PDGF-bb alone dependently increased cell growth. A combination of HBO2 treatment plus PDGF-bb treatment had an additive effect on cell growth in comparison with HBO2 treatment alone or PDGF-bb treatment alone. FACS analysis revealed that HBO2 alone, PDGF-bb alone and PDGF-bb plus HBO2 treatment increase the percentage of cells accumulated in S-phase. Western blotting analysis revealed that Type III collagen content was decreased significantly after HBO2 treatment alone or HBO2 plus PDGF-bb treatment but not in PDGF-bb treatment alone. In contrast, although Type I collagen content was increased after HBO2 treatment, the increase in Type I collagen (increase /original) was not statistically significant. CONCLUSION: HBO2 or HBO2 plus PDGF-bb treatment decreases the Type III collagen/Type I collagen content, which could result in mechanically stronger collagen fibrils. We propose HBO2 therapy as a potentially effective treatment for MCL healing.

The assessment of stability and reliability of a virtual reality-based laparoscopic gynecology simulation system.

PURPOSE OF INVESTIGATION: Surgeons require practiced skills in laparoscopic surgery. A virtual reality-based simulator system was developed for extensive training. The purpose of this study was to assess the feasibility of a virtual reality-based laparoscopic gynecology simulation system. METHODS: Laparoscopic tools and three-dimensional virtual environments were included in the simulation system. Ten healthy, non-disabled volunteers were recruited. The surgical procedure is a process of tubal sterilization by cauterization. Volunteers followed the training procedure, 15 trials in the first test and retest, respectively. RESULTS: Stable performances were obtained after about seven trials for all subjects. The intraclass correlation coefficients were 0.935 and 0.425 for task time and error frequency, respectively. CONCLUSION: The results of this study indicate that the system is stable and has a fair high test-retest reliability. Therefore, the VR-based laparoscopic gynecology system is feasible.

Correlation of three-dimensional tumor volumetry with cervical cancer prognostic parameters.

OBJECTIVE: Estimation of tumor size by a single diameter is used in the current Federal International Gynecology Oncology (FIGO) cervical cancer staging system. However, the role of three-dimensional volumetry is rarely mentioned. In this study, three kinds of tumor volume estimations were performed and the relationship of these volume estimations among associated cervical cancer prognostic parameters were evaluated. METHODS: Thirty patients with newly diagnosed cervical cancer at Taipei Veterans General Hospital were enrolled in the study. Final pathology and associated parametric information was obtained by chart review. The original magnetic resonance (MR) image was reappraised by an experienced radiologist and the tumor volume was estimated using either three axial measurements or the longest axial measurement. The corresponding three-dimensional (3-D) volume was calculated by the integration of lesion areas in different images using volumetric software developed by engineers at National Central University, Taiwan. All data were analyzed separately for correlation with clinical staging, histological differentiation, pelvic lymph node metastasis and survival status. RESULTS: A statistically significant difference in tumor volume estimates was found between 3-D-integration and one-dimensional estimates among tumors with lymph nodes metastasis, cell differentiation and parametrial involvement. CONCLUSION: This study found that the parameters of lymph node metastasis, parametrial involvement and tumor differentiation are volume-dependent. 3D-tumor volumetry showed superior discrimination of these parameters compared to current single diameter evaluation suggesting its potential as a rapid method for initial prediction of prognostic factors in cervical cancer.