Role of Scaffolds,Subchondral, Intra-Articular Injections of Fresh Autologous Bone Marrow Concentrate Regenerative Cells in Treating Human Knee Cartilage Lesions: Different Approaches and Different Results

The value of bone marrow aspirate concentrates for treatment of human knee cartilage lesions is unclear. Most of the studies were performed with intra-articular injections. However, subchondral bone plays an important role in the progression of osteoarthritis. We investigated by a literature review whether joint, subchondral bone, or/and scaffolds implantation of fresh autologous bone marrow aspirate concentrated (BMAC) containing mesenchymal stem cells (MSCs) would improve osteoarthritis (OA). There is in vivo evidence that suggests that all these different approaches (intra-articular injections, subchondral implantation, scaffolds loaded with BMAC) can improve the patient. This review analyzes the evidence for each different approach to treat OA. We found that the use of intra-articular injections resulted in a significant relief of pain symptoms in the short term and was maintained in 12 months. However, the clinical trials indicate that the application of autologous bone marrow concentrates in combination with scaffolds or in injection in the subchondral bone was superior to intra-articular injection for long-term results. The tendency of MSCs to differentiate into fibrocartilage affecting the outcome was a common issue faced by all the studies when biopsies were performed, except for scaffolds implantation in which some hyaline cartilage was found. The review suggests also that both implantation of subchondral BMAC and scaffolds loaded with BMAC could reduce the need for further surgery.     

Longitudinal and temporal water temperature patterns in the Yangtze River and its influence on spawning of the Chinese sturgeon (Acipenser sinensis Gray 1835)

In this study, water temperature longitudinal and temporal patterns were investigated in four locations of the Yangtze River to identify their influence on spawning of the Chinese sturgeon (Acipenser sinensis Gray 1835). The results showed that, historically (1960–1980), the four locations, Pingshan (PS, 2,687 river kilometres [rkm]), Yichang (YC, 1,640 rkm), Chenglingji (CLJ, 1,265 rkm), and Hukou (HK, 805 rkm), all had similar average annual water temperatures (around 18.0 °C) but very different amplitudes: The upper two locations, PS (11.4–23.2 °C) and YC (9.1–25.9 °C), were relatively stable and had smaller amplitude than the lower two ones, CLJ (6.0–30.0 °C) and HK (5.9–30.8 °C). On the basis of the physiological effects for Chinese sturgeon, we defined three water temperature ranges: stressing (>23 °C), holding (23–20 °C), and spawning (20–16 °C). We found that the upper two locations with successful spawning of the Chinese sturgeon (PS and YC) had short or no stressing period but long holding and spawning periods. Currently (2013–2016), the impoundment of the Three Gorges Reservoir increases the time for water temperature decreasing to 20 °C at the current spawning ground (YC) in autumn, which increases the stressing and holding periods, and has significant negative impact on the spawning of Chinese sturgeon. Nonetheless, considering the very high stressing temperature and the short duration of the holding and spawning periods at CLJ and HK, we propose that YC is still the most suitable spawning location for the Chinese sturgeon.     

葛洲坝电厂调度对中华鲟产卵场水流条件的影响

建立中华鲟产卵场三维水流数学模型,模拟葛洲坝电厂不同调度情况下产卵场水流条件;分析了葛洲坝下游近坝段适宜中华鲟产卵的水深、近底层流速和两者叠加的区域范围;研究了葛洲坝电厂调度对中华鲟产卵场水流条件影响,并初步提出了中华鲟产卵场水流条件改善措施。研究结果表明:葛洲坝下泄流量在15 000～6 000 m3/s范围变化时,适宜中华鲟产卵的水流区域范围随下泄流量减小相应减小;在下泄流量为6 000 m3/s情况下,若下泄流量全由大江电厂泄流时有利于增大大江尾水区适宜产卵的水流区域范围。     

硫酸软骨素生产新工艺的研究

制备硫酸软骨素新工艺利用双酶酶解法提取。设计了正交试验，得到最优化反应条件。最优反应条件为温度50℃，pH8．6-8．9，加酶量0．03g／d1，收率达17．84％。     

Registration of confocal scanning laser microscopy and quantitative backscattered electron images for the temporospatial quantification of mineralization density in 18-month old thoroughbred racehorse articular calcified cartilage

Combined backscattered electron scanning electron microscopy (BSE SEM) and confocal scanning laser microscopy (CSLM) have been used to put tissue mineralization data into the context of soft tissue histology and fluorescent label information. Mineralization density (Dm) and linear accretion rate (LAR) are quantifiable parameters associated with mineralizing fronts within calcified tissues. Quantitative BSE (qBSE) may be used to determine Dm, while CSLM may be used to detect label fluorescence from which LAR is calculated. Eighteen-month old Thoroughbred horses received single calcein injections 19 and 8 days prior to euthanasia, labeling sites of active mineralization with fluorescent bands. Confocal scanning laser microscopy images of articular calcified cartilage (ACC) from distal third metacarpal condyles were registered to qBSE images of the same sites using an in-house program. ImageJ and Sync Windows enabled the simultaneous collection of LAR and Dm data. The repeatability of the registration and measurement protocols was determined. Dm profiles between calcein labels were explored for an association with time. Dm was 119.7 +/- 24.5 (mean +/- standard deviation) gray levels (where 0 = backscattering from monobrominated and 255 from monoiodinated dimethacrylate standards, respectively), while modal and maximum LAR were 0.45 and 3.45 microm/day, respectively. Coefficients of variation (CV) for Dm were 0.70 and 0.77% with and without repeat registration, respectively; CVs for LAR were 1.90 and 2.26% with and without repeat registration, respectively. No relationship was identified between Dm and time in the 11-day interlabel interval. Registration of CSLM to qBSE images is sufficiently repeatable for quantitative studies of equine ACC.     

Some experiences in the use of a polymeric cryoprotectant in the freezing of plant tissue

Recently it has been suggested that polymeric cryoprotectants might be usefully employed for reducing ice crystal size during ultrastructural and analytical studies of frozen biological tissues. Furthermore, it was reported that they have little physiological effect and cause negligible structural changes in the tissue. Our experiences with one such polymer, polyvinyl pyrrolidone (PVP), in the cryopreservation of mature plant roots prepared for electron microscopy, have led us to conclude that preservation deep into the structure of this tissue is not improved. Even short periods of exposure of tissue to polymer cause rapid withdrawal of water from vacuolated cells of plant roots, resulting in shrinkage and collapse. Low temperature techniques have confirmed that little if any improvement in the reduction of ice crystal size results if the root is first treated with PVP.     

Hydrogel‐mediated formation of living cartilage template for endochondral initiation

The use of hydrogel in cartilage tissue engineering is especially popular due to its high hydrophilic property which is similar to native cartilage matrix. Alginate hydrogel was used as a transient scaffold material to facilitate chondrocyte proliferation into a three‐dimensional scaffold‐free living hyaline cartilaginous graft (LhCG). As LhCG is purely cell‐based and has a marked resemblance to native hyaline cartilage, it served as an excellent in vitro platform for studying the endochondral ossification pathway. Due to the complexity of events involved throughout endochondral ossification, this study only focuses on early stages of the process where it involves chondrocyte hypertrophy and blood vessel invasion. Human umbilical vein endothelial cells (HUVECs) were selected as the target cells for possible endothelialization in the LhCG template. They were seeded onto the LhCG construct and subjected to vascular endothelial growth factor (VEGF) treatment. Results suggested that VEGF is indeed a potent driving force for initiation of the endochondral pathway. It alone is sufficient to induce hypertrophy in chondrocytes and the corresponding expression of osteogenic genes with or without the presence of HUVECs in the LhCG template. On the other hand, the effect of HUVECs in the LhCG system was less evident. It is hypothesized that this is attributed to the preservation of anti‐angiogenic properties in primary chondrocytes from the LhCG construct, inhibiting HUVECs from endothelialization in the LhCG+HUVEC construct. Based on the outcome from this study, it is recommended that hypertrophy in chondrocytes should be induced prior to endothelial cell introduction so that the microenvironment will be altered to favor angiogenesis within the cartilaginous template. © 2013 Society of Chemical Industry     

Injectable polyethylene glycol-laponite composite hydrogels as articular cartilage scaffolds with superior mechanical and rheological properties

In this study, injectable PEG-based hydrogels containing Laponite particles with mechanical and structural properties close to the natural articular cartilage are introduced. The nanocomposites are fabricated by imide ring opening reactions utilizing synthesized copolymers containing PEG blocks and nanoclay through a two-step thermal poly-(amic acid) process. Butane diamine is used as nucleophilic reagent and hydrogels with interconnected pores with sizes in the range of 100–250?µm are prepared. Improved viscoelastic properties compared with the conventional PEG hydrogels are shown. Evaluation of cell viability utilizing human mesenchymal stem cells determines cytocompatibility of the nanocomposite hydrogels.     

Mechanical and tribological properties of poly(hydroxyethyl methacrylate) hydrogels as articular cartilage substitutes

Poly(hydroxyethyl methacrylate) (p(HEMA)) hydrogels have been proposed as promising biomaterials to replace damaged articular cartilage. A major obstacle to their use as replacement bearing tissue is their poor mechanical properties in comparison with healthy articular cartilage. The purpose of this study was to obtain p(HEMA) hydrogels with physicochemical and mechanical properties close to healthy articular cartilage, by introducing a hydrophilic monomer, namely acrylic acid (AA). Formulations of hydrogels with different amounts of hydrophilic monomer (acrylic acid, AA) were synthesized and tested: p(HEMA), p(HEMA-co-5%AA), p(HEMA-co-25%AA). The macro-mechanical tests were reproduced at nanoscale in order to verify if the superficial properties of the hydrogels are similar to the bulk ones.