Phosphatidylserine controls calcium phosphate nucleation and growth on lipid monolayers: A physicochemical understanding of matrix vesicle-driven biomineralization.

Bone biomineralization is an exquisite process by which a hierarchically organized mineral matrix is formed. Growing evidence has uncovered the involvement of one class of extracellular vesicles, named matrix vesicles (MVs), in the formation and delivery of the first mineral nuclei to direct collagen mineralization. MVs are released by mineralization-competent cells equipped with a specific biochemical machinery to initiate mineral formation. However, little is known about the mechanisms by which MVs can trigger this process. Here, we present a combination of in situ investigations and ex vivo analysis of MVs extracted from growing-femurs of chicken embryos to investigate the role played by phosphatidylserine (PS) in the formation of mineral nuclei. By using self-assembled Langmuir monolayers, we reconstructed the nucleation core - a PS-enriched motif thought to trigger mineral formation in the lumen of MVs. In situ infrared spectroscopy of Langmuir monolayers and ex situ analysis by transmission electron microscopy evidenced that mineralization was achieved in supersaturated solutions only when PS was present. PS nucleated amorphous calcium phosphate that converted into biomimetic apatite. By using monolayers containing lipids extracted from native MVs, mineral formation was also evidenced in a manner that resembles the artificial PS-enriched monolayers. PS-enrichment in lipid monolayers creates nanodomains for local increase of supersaturation, leading to the nucleation of ACP at the interface through a multistep process. We posited that PS-mediated nucleation could be a predominant mechanism to produce the very first mineral nuclei during MV-driven bone/cartilage biomineralization.

Agreement between direct, oscillometric and Doppler ultrasound blood pressures using three different cuff positions in anesthetized dogs.

OBJECTIVE: To evaluate the agreement between invasive blood pressure (IBP) and Doppler ultrasound blood pressure (DUBP) using three cuff positions and oscillometric blood pressure (OBP) in anesthetized dogs. STUDY DESIGN: Prospective study. ANIMALS: Nine adult dogs weighing 14.5-29.5 kg. METHODS: The cuff was placed above and below the tarsus, and above the carpus with the DUBP and above the carpus with the OBP monitor. Based on IBP recorded via a dorsal pedal artery catheter, conditions of low, normal, and high systolic arterial pressures [SAP (mmHg) <90, between 90 and 140, and >140, respectively] were induced by changes in isoflurane concentrations and/or dopamine administration. Mean biases ± 2 SD (limits of agreement) were determined. RESULTS: At high blood pressures, regardless of cuff position, SAP determinations with the DUBP underestimated invasive SAP values by more than 20 mmHg in most instances. With the DUBP, cuff placement above the tarsus yielded better agreement with invasive SAP during low blood pressures (0.2 ± 16 mmHg). The OBP underestimated SAP during high blood pressures (-42 ± 42 mmHg) and yielded better agreement with IBP for mean (MAP) and diastolic (DAP) arterial pressure measurements [overall bias: 2 ± 15 mmHg (MAP) and 0.2 ± 16 mmHg (DAP)]. CONCLUSIONS: Agreement of SAP determinations with the DUBP is poor at SAP > 140 mmHg, regardless of cuff placement. Measurement error of the DUBP with the cuff placed above the tarsus is clinically acceptable during low blood pressures. Agreement of MAP and DAP measurements with this OBP monitor compared with IBP was clinically acceptable over a wide pressure range. CLINICAL RELEVANCE: With the DUBP device, placing the cuff above the tarsus allows reasonable agreement with IBP obtained via dorsal pedal artery catheterization. Only MAP and DAP provide reasonable estimates of direct blood pressure with the OBP monitor evaluated.

Wettability and pre-osteoblastic behavior evaluations of a dense bovine hydroxyapatite ceramics.

In this study, the wettability, cell viability, and roughness of an experimental dense bovine hydroxyapatite [Ca10(PO4)6(OH)2] ceramic block were evaluated so that, in the future, it could be used as a base material for dental implants. The results to commercial zirconia and a commercially pure titanium (Ti) alloy were compared. The surface roughness and contact angles were measured. An in vitro evaluation was conducted by means of tests in which pre-osteoblastic MC3T3-E1 cells were placed in indirect and direct contact with these materials. For cell viability, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and crystal violet test were conducted. A qualitative analysis was conducted using variable pressure scanning electron microscopy (SEM). No statistically significant differences were observed in wettability and roughness tests among the groups. In both the MTT assay and crystal violet test, all groups demonstrated satisfactory results without cytotoxicity. SEM showed cell adhesion and cell proliferation results on the material surfaces after 24 h and 48 h. In conclusion, this dense Ca10 (PO4)6(OH)2 ceramic can be considered as a potential biocompatible material.