Human muscle-derived stem cells (hMDSCs) have been shown to regenerate bone efficiently when they were transduced with Lenti-viral bone morphogenetic protein 2 (LBMP2). However, whether the age of ...hMDSCs and the animal host affect the bone regeneration capacity of hMDSCs and mechanism are unknown which prompted the current study.
We isolated three gender-matched young and old populations of skeletal muscle stem cells, and tested the influence of cells' age on in vitro osteogenic differentiation using pellet culture before and after Lenti-BMP2/green fluorescent protein (GFP) transduction. We further investigated effects of the age of hMDSCs and animal host on hMDSC-mediated bone regeneration in a critical-size calvarial bone defect model in vivo. Micro-computer tomography (CT), histology, and immunohistochemistry were used to evaluate osteogenic differentiation and mineralization in vitro and bone regeneration in vivo. Western blot, quantitative polymerase chain reaction (PCR), and oxidative stress assay were performed to detect the effects of age of hMDSCs on cell survival and osteogenic-related genes. Serum insulin-like growth factor 1 (IGF1) and receptor activator of nuclear factor-kappa B ligand (RANKL) were measured with an enzyme-linked immunosorbent assay (ELISA).
We found LBMP2/GFP transduction significantly enhanced osteogenic differentiation of hMDSCs in vitro, regardless of donor age. We also found old were as efficient as young LBMP2/GFP-transduced hMDSCs for regenerating functional bone in young and old mice. These findings correlated with lower phosphorylated p38MAPK expression and similar expression levels of cell survival genes and osteogenic-related genes in old hMDSCs relative to young hMDSCs. Old cells exhibited equivalent resistance to oxidative stress. However, both young and old donor cells regenerated less bone in old than young hosts. Impaired bone regeneration in older hosts was associated with high bone remodeling due to higher serum levels of RANKL and lower level of IGF-1.
hMDSC-mediated bone regeneration was not impaired by donor age when hMDSCs were transduced with LBMP2/GFP, but the age of the host adversely affected hMDSC-mediated bone regeneration. Regardless of donor and host age, hMDSCs formed functional bone, suggesting a promising cell resource for bone regeneration.
Background:
Microfracture or bone marrow stimulation (BMS) is often the first choice for clinical treatment of cartilage injuries; however, fibrocartilage, not pure hyaline cartilage, has been ...reported because of the development of fibrosis in the repair tissue. Transforming growth factor β1 (TGF-β1), which can promote fibrosis, can be inhibited by losartan and potentially be used to reduce fibrocartilage.
Hypothesis:
Blocking TGF-β1 would improve cartilage healing in a rabbit knee BMS model via decreasing the amount of fibrocartilage and increasing hyaline-like cartilage formation.
Study Design:
Controlled laboratory study.
Methods:
An osteochondral defect was made in the patellar groove of 48 New Zealand White rabbits. The rabbits were divided into 3 groups: a defect group (defect only), a BMS group (osteochondral defect + BMS), and a BMS + losartan group (osteochondral defect + BMS + losartan). For the rabbits in the BMS + losartan group, losartan was administrated orally from the day after surgery through the day of euthanasia. Rabbits were sacrificed 6 or 12 weeks postoperatively. Macroscopic appearance, microcomputed tomography, histological assessment, and TGF-β1 signaling pathway were evaluated at 6 and 12 weeks postoperatively.
Results:
The macroscopic assessment of the repair revealed that the BMS + losartan group was superior to the other groups tested. Microcomputed tomography showed superior healing of the bony defect in the BMS + losartan group in comparison with the other groups. Histologically, fibrosis in the repair tissue of the BMS + losartan group was significantly reduced when compared with the other groups. Results obtained with the modified O’Driscoll International Cartilage Repair Society grading system yielded significantly superior scores in the BMS + losartan group as compared with both the defect group and the BMS group (F value: 15.8, P < .001, P = .012, respectively). TGF-β1 signaling and TGF-β-activated kinase 1 of the BMS + losartan group were significantly suppressed in the synovial tissues.
Conclusion:
By blocking TGF-β1 with losartan, the repair cartilage tissue after BMS was superior to the other groups and consisted primarily of hyaline cartilage. These results should be easily translated to the clinic because losartan is a Food and Drug Administration–approved drug and it can be combined with the BMS technique for optimal repair of chondral defects.
Clinical Relevance:
Biologically regulated marrow stimulation by blocking TGF-β1 (oral intake of losartan) provides superior repair via decreasing fibrocartilage formation and resulting in hyaline-like cartilage as compared with outcomes from BMS only.
The purpose of this study was to evaluate biomechanical and histopathological results of a retrieved acellular human dermal allograft (AHDA) after superior capsule reconstruction (SCR). A 67-year-old ...man with pseudoparalysis was treated with SCR for an irreparable posterosuperior rotator cuff tear. The patient failed clinically 4.5 months postoperatively and elected to undergo reverse total shoulder arthroplasty (RTSA). At the time of RTSA, the AHDA was harvested. Biomechanical and histopathologic analyses were performed and compared to native grafts. Failure loads for the explanted graft and native grafts 1 and 2 were 158, 790, and 749 N, respectively. The stiffness values were 20.2, 73, and 100.5 N/mm. The displacement at failure for each graft was 10.1, 27.9, and 17.0 mm. Hematoxylin and eosin and Masson's trichrome staining revealed the presence of cells in all portions of the AHDA. The medial portion presented extensive cellular infiltration, the middle portion moderate, and the lateral portion the least infiltration. Although the only identifiable cells in the lateral portions were found in pockets on the interior of the graft, cells were mainly localized on the exterior. Postoperative cell incorporation could be found in acellular dermal allograft after SCR. However, biomechanical properties in the early postoperative phase were inferior compared with unimplanted allografts.
Background:
A previous publication demonstrated that the oral intake of losartan promoted microfracture-mediated hyaline-like cartilage repair in osteochondral defects of a rabbit knee model. ...However, an intra-articular (IA) injection of losartan may have direct beneficial effects on cartilage repair and has not been studied.
Purpose:
To determine the dosage and beneficial effects of an IA injection of losartan on microfracture-mediated cartilage repair and normal cartilage homeostasis.
Study Design:
Controlled laboratory study.
Methods:
Rabbits were divided into 5 groups (n = 6 each): a microfracture group (MFX group) and 4 different losartan treatment groups that received varying doses of IA losartan (0.1, 1, 10, and 100 mg per knee). An osteochondral defect (5 mm) was created in the trochlear groove cartilage of 1 limb in each rabbit, and 5 microfracture perforations were made in the osteochondral defect. Both the injured and the contralateral knee joints were injected with IA losartan immediately after microfracture and at 2 and 4 weeks after surgery. Rabbits were sacrificed at 6 weeks after surgery for analysis including gross observation, micro–computed tomography, histology, and reverse transcription quantitative polymerase chain reaction.
Results:
Micro–computed tomography and gross observation demonstrated comparable subchondral bone healing and hyaline-like cartilage morphology in the 0.1-, 1-, and 10-mg losartan groups relative to the MFX group. Conversely, the 100-mg losartan group showed neither bony defect healing nor cartilage repair. Histology revealed higher O’Driscoll scores and hyaline-like cartilage regeneration in the 1-mg losartan group compared with the MFX group. In contrast, the 100-mg losartan group showed the lowest histology score and no cartilage repair. An IA injection of losartan at the doses of 0.1, 1, and 10 mg did not cause adverse effects on uninjured cartilage, while the 100-mg dose induced cartilage damage. Quantitative polymerase chain reaction results showed downregulation of the transforming growth factor β (TGF-β) signaling pathway after IA losartan injection.
Conclusion:
An IA injection of losartan at the dose of 1 mg was most effective for the enhancement of microfracture-mediated cartilage repair without adversely affecting uninjured cartilage. Conversely, a high dose (100 mg) IA injection of losartan inhibited cartilage repair in the osteochondral defect and was chondrotoxic to normal articular cartilage.
Clinical Relevance:
An IA injection of losartan at an optimal dosage represents a novel microfracture enhancement therapy and warrants a clinical trial for future clinical applications.
In semiconductor and Micro-Electro-Mechanical Systems production, thick resist films are indispensable for circuit formation, chemical mechanical planarization, or wafer-level packaging applications. ...We investigate multiple metrology protocols that aim at meeting the needs for fast, accurate, full-wafer determination of the thickness of up to 100μm thick resist layers deposited on silicon or glass wafers. We found very effective to use a two-step strategy consisting of: i/ an accurate determination of the optical characteristics of the photoresist based on prism-coupling technique, and ii/ the fast, full-wafer measurement of the wafers of interest, using high-resolution spectroscopic reflectometry (SR). The robustness of this strategy increased when analyzing the SR data with a Fourier transform (FT) based algorithm which combines FT-based filtering around the thickness of interest with spectral matching to improve measurement performance.
•Reliable protocols for the in-fab metrology of ultra-thick photoresist are required.•High-resolution spectroscopic reflectometry features ultra-thick resist capability.•Refractive index of thick resist is accurately determined by prism-coupling technique.•Analysis of reflectometry data is based on prism-coupling deduced optical constants.•The precision of reflectometry analysis is improved by Fourier transform algorithm.
Single-mode waveguides for GRAVITY Perraut, K.; Jocou, L.; Berger, J. P. ...
Astronomy and astrophysics (Berlin),
06/2018, Volume:
614
Journal Article
Peer reviewed
Open access
Context.
Within the framework of the second-generation instrumentation of the Very Large Telescope Interferometer of the European Southern Observatory we have developed the four-telescope beam ...combiner in integrated optics.
Aims.
We optimized the performance of such beam combiners, for the first time in the near-infrared
K
band, for the GRAVITY instrument dedicated to the study of the close environment of the galactic centre black hole by precision narrow-angle astrometry and interferometric imaging.
Methods.
We optimized the design of the integrated optics chip and the manufacturing technology as well, to fulfil the very demanding throughput specification. We also designed an integrated optics assembly able to operate at 200 K in the GRAVITY cryostat to reduce thermal emission.
Results.
We manufactured about 50 beam combiners by silica-on-silicon etching technology. We glued the best combiners to single-mode fluoride fibre arrays that inject the VLTI light into the integrated optics beam combiners. The final integrated optics assemblies have been fully characterized in the laboratory and through on-site calibrations: their global throughput over the
K
band is higher than 55% and the instrumental contrast reaches more than 95% in polarized light, which is well within the GRAVITY specifications.
Conclusions.
While integrated optics technology is known to be mature enough to provide efficient and reliable beam combiners for astronomical interferometry in the
H
band, we managed to successfully extend it to the longest wavelengths of the
K
band and to manufacture the most complex integrated optics beam combiner in this specific spectral band.
Analgesia for labor pain: a cost model Macario, A; Scibetta, W C; Navarro, J ...
Anesthesiology (Philadelphia),
03/2000, Volume:
92, Issue:
3
Journal Article
Peer reviewed
Epidural analgesia and intravenous analgesia with opioids are two techniques for the relief of labor pain. The goal of this study was to develop a cost-identification model to quantify the costs ...(from society's perspective) of epidural analgesia compared with intravenous analgesia for labor pain. Because there is no valid method to assign a dollar value to differing levels of analgesia, the cost of each technique can be compared with the analgesic benefit (patient pain scores) of each technique.
The authors created a cost model for epidural and intravenous analgesia by reviewing the literature to determine the rates of associated clinical outcomes (benefit of each technique to produce analgesia) and complications (e.g., postdural puncture headache). The authors then analyzed data from their institution's cost-accounting system to determine the hospital cost for parturients admitted for delivery, estimated the cost of each complication, and performed a sensitivity analysis to evaluate the cost impact of changing key variables. A secondary analysis was performed assuming that the cost of nursing was fixed (did not change depending on the number of nursing interventions).
If the cesarean section rate equals 20% for both intravenous and epidural analgesia, the additional expected cost per patient to society of epidural analgesia of labor pain ranges from $259 (assuming nursing costs in the labor and delivery suite do not vary with the number of nursing interventions) to $338 (assuming nursing costs do increase as the number of interventions increases) relative to the expected cost of intravenous analgesia for labor pain. This cost difference results from increased professional costs and complication costs associated with epidural analgesia.
Epidural analgesia is more costly than intravenous analgesia. How the cost of the anesthesiologist and nursing care is calculated affects how much more costly epidural analgesia is relative to intravenous analgesia. Published studies have determined that epidural analgesia provides relief of labor pain superior to intravenous analgesia, quantified in one study as 40 mm better on a 100-mm scale during the first stage of labor and 29 mm better during the second stage of labor. Patients, physicians, and society need to weigh the value of improved pain relief from epidural analgesia versus the increased cost of epidural analgesia.
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